text:
Ironies In The Story Of An Hour
Essay, Research Paper
Kate Chopin employs the tool of irony in "The Story of an Hour" to illustrate the problem relative to marital relationships in which one individual imposes his "private will" upon the other. She presents, through the story of Mrs. Mallard, an issue not socially accepted at the end of the 19th century.
This is the story of Mrs. Mallard, a woman with a heart condition who finds out her husband has died in a train accident. She reacts with sadness at first, but after seeking solitude, realizes that she is free. She is ready to begin her new life when her husband, who was not involved in the train accident, comes home alive. The woman dies from heart failure on the spot.
The purpose of irony in Kate Chopin's "The Story of an Hour" is to convey a message without saying it explicitly. In the context that the story was written, at the end of the 19th century, woman were often not allowed many rights. Their role in society was trifling compared to what men had. Chopin, a feminist ahead of her time, uses irony in this particular story to show the unequal role women had in relationships in the late 1800's. Mrs. Mallard's discovery of her long lost freedom and desire to live for herself only comes after her husband's death. The ironic tone in the story is employed by Chopin to present a socially unaccepted concept in a more acceptable format.
In "The Story of an Hour", Chopin makes use of different types of irony. The first type of irony encountered is situational irony, where there is a contrast between what is expected to happen in a particular situation and what actually happens. After grieving with "wild abandonment" the death of her husband, Mrs. Mallard seeks solitude in her room. Now the reader starts to see the world through her eyes, a world full of new and pure life. As she looks out of the window, she sees spring and all the new life it brings. The descriptions used now are far away from death. Mrs. Mallard stares out the "open window" at "the new spring life". As for the weather, instead of being gloomy and dark to symbolize death, she sees "patches of blue sky showing here and there through the clouds." She also mentions that birds are singing and there is a "delicious breath of rain" in the air, all images not usually associated with death. She is expected to mourn her husband's death, but in contrast, she is thinking about new life. At the end of the story, Chopin uses dramatic irony, where there is a contrast between what the audience knows and what the characters think is happening. "When the doctors came they said she had died of heart disease – of joy that kills." The other characters are still unsuspecting of her actual joy in death. They believe her joy corresponds with the love she had for her husband. In contrast, the reader knows that the love she had for her husband pales in comparison to the joy she feels upon the discovery of her newfound freedom.
Mrs. Mallard begins to fantasize about living her life for herself. "Free, free, free!" are the words Mrs. Mallard whispers in her room. Coming from a woman who just lost her husband, one can wonder how was their relationship. However, Mrs. Mallard clarifies that their relationship is one of love. Brently Mallard "had never looked save with love upon her." And likewise, "she knew that she would weep again when she saw the kind, tender hands folded in death." However, love does not seem to be the problem here. Regardless of the love she has for her husband Brently Mallard, the problem she sees is the unequal relationship in which one individual exercises their "powerful will" on the other. Even though at times she had loved him, she is now regaining her freedom. Another sign that informs the reader of her new liberation is the revelation of her first name. Her name is Louise, she is no longer Mrs. Mallard, she is Louise, she has her own identity because she is free. It is ironic to see that it took Brently Mallard's death for Louise to realize that she was not "Body and soul free!" It seems as if she finds personal strength in her husband's death, ready to face the world as a whole person. Once Louise Mallard recognizes her desire to live for herself, desire that her marriage will not grant her, her heart will not allow her to turn back.
"The Story of an Hour" is a story of great irony. One that carries a message of hope and freedom. The title itself is self explanatory: The story refers to that of Louise Mallard's life. A woman that lived in the true sense of the word, with the will and freedom to live for only one hour.
ДОБАВИТЬ КОММЕНТАРИЙ [можно без регистрации]
перед публикацией все комментарии рассматриваются модератором сайта - спам опубликован не будет
Хотите опубликовать свою статью или создать цикл из статей и лекций?
Это очень просто – нужна только регистрация на сайте.


text:
We have some guests with us in this episode. We asked the guys from Saving the Game to come on the podcast and talk with us a bit about the wide and wonderful world of tabletop role playing games. Grant and Peter share some games we've never even heard of before and explain the value of playing rpgs as a whole.
If you were curious about role playing games but wasn't into the sword and sworcery style of D&D – definitely give this a listen.
Some great resources mentioned in this episode:
–The Big Friendly List of Rules from the Fear the Boot forums
–Everyone is John
–The Trouble with Rose
And, naturally, be sure to check out the guys on their podcast site or find them on the InnRoads Ministries podcast page.
Podcast: Play in new window | Download
Subscribe: Apple Podcasts | Android | RSS

subset: commoncrawl


text:
eBay Inc. (EBAY - Free Report) is scheduled to announce its third quarter fiscal 2012 results on October 17, and we notice limited movement in analyst estimates.
Prior-Quarter Synopsis
eBay's second quarter 2012 earnings per share (EPS) of 51 cents (excluding one-time items and stock-based compensation) exceeded the Zacks Consensus Estimate by 3 cents on strong top-line growth, the strengthening Marketplaces segment and solid payments business.
The company reported gross revenue of $3.40 billion, up 3.7% sequentially, exceeding consensus expectations of $3.36 billion, aided by improved customer experience. Over 85% of eBay's total revenue was transactions-based, while the remaining 15% came from marketing services.
Pro forma gross margin was 71.5%, up 87 bps sequentially and down 92 bps year over year. The sequential increase was attributable to higher volumes and lower transaction expense.
Third Quarter Guidance
eBay expects revenue of $3.3–3.4 billion (down 1.4% sequentially and up 13.0% year over year at the mid-point), GAAP EPS in a range of 42 cents – 44 cents and non GAAP EPS in the range of 53 cents – 55 cents.The Zacks Consensus Estimate is pegged at 46 cents, down a penny since the company reported results but still over the range guided by management.
For 2012, management expects revenue of $13.8–$14.1 billion, GAAP EPS of $1.91 to $1.96 and non GAAP EPS of $2.30 to $2.35. The Zacks Consensus Estimate is pegged at $2.03.
Detailed earnings results can be viewed in the blog titled: eBay Delivers Another Good Quarter
Agreement of Analysts
Over the last 30 days, 1 out of the 17 analysts providing estimates made an upward revision, while 2 analysts moved their estimates in the opposite direction both for the third quarter as well as fiscal 2012.
The majority of analysts expect a strong third quarter with revenue and earnings per share coming in at the higher end of the management guidance. They see continued traction in eBay's U.S. Marketplaces business and continued online and offline share gains in the PayPal segment.
The analysts believe that PayPal is expanding its market opportunity by aggressively enabling mobile payments and expanding Point-of-Sale (POS) offerings. The analysts expect the Payments business to deliver strong total payment value (TPV) and revenue growth, benefiting from mobile payments, the overall strength in eCommerce, as well as positive momentum in Marketplaces. According to management, PayPal expects to process $10 billion in mobile payments this year.
However, a few analysts believe that macro weakness in Europeand weak currency trends could limit upside potential. Hence, they expect management to provide a conservative fourth quarter guidance, in-line or below the consensus estimate of $3.94 billion and $0.68 for revenues and EPS, respectively.
Magnitude of Estimate Revisions
In the past 30 days, the Zacks Consensus Estimate decreased by a penny to 46 cents for the third quarter but remain unchanged for fiscal 2012.
Over the 90-day period, the Zacks Consensus Estimate was down by a penny for the third quarter and 4 cents for fiscal 2012.
Aided by growth in its PayPal business and mobile commerce, we expect eBay to report a decent third quarter.
We remain impressed with eBay's strategy and executionand believe that eBay is well positioned for sustained growth, with both Payments and the Marketplaces business showing signs of improvement.
PayPal, which accounts for almost half of eBay's annual revenue is expanding beyond its online payments service and finding new ways to get a foothold in the offline world. Last year, the company expanded its offline partnerships and announced that PayPal point of sale was gaining traction as few retailers, including Home Depot Inc and Office Depot Inc (ODP) had agreed to accept PayPal as a payment mode.
Recently, eBay inked a deal with Discover Financial Service, which will extend PayPal's services to 7 million merchants next year. We believe that eBay is expanding its addressable market and taking all the necessary measures for continued growth in mobile payments in the near future.
PayPal has also reinforced its mobile payments segment through the PayPal Mobile Express Checkout system and the Zong acquisition, which are expected to boost mobile payment volumes.
However, we are concerned about the increasing competition from major online retailers, such as Amazon.com (AMZN), as well as many other smaller players. Additionally, Google Inc. (GOOG) has been eyeing the online payment space, thus threatening eBay's position.
While eBay's payments business shows great promise, here too competition may be expected to pick up as banks and other companies announce their own payment systems and Google's digital wallet gathers steam.
Currently, eBay has a Zacks #3 Rank, implying a short-term Hold rating.


text:
A purpose-filled life.
(By Janifer Wheeler, Certified Life Coach)
I've been thinking about this topic, and the children's magazine Highlights keeps running through my mind. Remember "Fun with a Purpose™"? Of course, the purpose of Highlights is to provide kids with interesting articles, pictures, games, etc. that, while reading, teach a valuable lesson or reinforce academic skills — all neatly hidden in an entertaining format. But doesn't that also describe life in general… a series of events, interactions, and experiences from which we learn and grow, sometimes without even realizing it?
What does it really mean to live a fun life with purpose? The dictionary defines fun as "enjoyment," and purpose as "a reason, goal, plan or intention." So, if you combine the two, you're having fun intentionally, while also achieving your goals. If it's this easy, though, why are so many people unhappy and dissatisfied with their lives?
How can you regain that childhood notion of fun, but also live with purpose?
(One of our favorite muses rewarding herself for losing 30 lbs.! Congrats Diane!)
Start by weeding out the activities, relationships or tasks that are stealing your joy. Misery, even within the grieving process, should always be a temporary state. If you can barely drag yourself out of bed every day because you hate your job, or if you're sending every call from that certain someone straight to voicemail, then it's probably time to make some changes. (Example: I recently got a new smartphone and as I transferred the contacts, I deleted every person who did not truly enhance or bring a smile to my world.)
Removing the dead weight can be hard and make you feel guilty, but it's a necessary step if you truly want a life that is enjoyable and lived to its fullest.
Your next step is to evaluate why you do the things you do. Last year, I decided not to return to teaching after eight years in the classroom. I realized I was getting burned out and actually made a written list of all the reasons why I was a teacher: it kept me on the same schedule as my child… I enjoyed the content… had summers off… it was in my degree field… I was in a good school system… I had a steady paycheck. All of those things were great, but nowhere on this list was changing lives, making a difference, or molding the next generation.
Those were the reasons I'd started teaching, but they had faded as the years went by – overshadowed by parents' unrealistic demands, scorn and blame for society's ills, and a very unbalanced work/family existence. Leaving the security of a guaranteed income was a bit stressful, but then I started my own Life Coaching business, Wheeler for Life, and found a new purpose. Now, I'm able to help others employ the tools and strategies they need to live their lives on their own terms – finding both the fun AND the purpose.
Sometimes, the word "purpose" can be daunting. Not everyone's a planner or has a grand scheme for his/her life — I certainly don't! My idea of a five-year-plan is to still be alive; who knows what I'll be doing or where I'll be? Flexibility, creativity, and spontaneity DO reside within goals, plans and intentions. In fact, I've found that if you intentionally look for the fun, the wonder, and the beauty in your life and the people traveling the path with you, your purpose will present itself.
A Purpose Filled Life
Janifer Wheeler, Certified Life Coach


text:
We studied the ability of typical unmyelinated cortical axons to conduct action potentials at fever‐like temperatures because fever often gives CNS symptoms. We investigated such axons in cerebellar and hippocampal slices from 10 to 25 days old rats at temperatures between 30 and 43°C. By recording with two electrodes along axonal pathways, we confirmed that the axons were able to initiate action potentials, but at temperatures >39°C, the propagation of the action potentials to a more distal recording site was reduced. This temperature‐sensitive conduction may be specific for the very thin unmyelinated axons because similar recordings from myelinated CNS axons did not show conduction failures. We found that the conduction fidelity improved with 1 mmol/L TEA in the bath, probably due to block of voltage‐sensitive potassium channels responsible for the fast repolarization of action potentials. Furthermore, by recording electrically activated antidromic action potentials from the soma of cerebellar granule cells, we showed that the axons failed less if they were triggered 10–30 msec after another action potential. This was because individual action potentials were followed by a depolarizing after‐potential, of constant amplitude and shape, which facilitated conduction of the following action potentials. The temperature‐sensitive conduction failures above, but not below, normal body temperature, and the failure‐reducing effect of the spike's depolarizing after‐potential, are two intrinsic mechanisms in normal gray matter axons that may help us understand how the hyperthermic brain functions.
Most central and peripheral mammalian axons conduct action potentials (spikes) with great fidelity at normal body temperature and down to 5–15°C (Franz and Iggo 1968), but above physiological temperature, peripheral axons sometimes fail. Temperature‐induced conduction failures have been studied for more than 100 years (Bernstein 1902). They are known to occur in several nonmammalian preparations (Swadlow et al. 1980), and also in humans with demyelinating disease (Rasminsky 1973; Howells et al. 2013), but whether conduction failures occur in normal axons in the mammalian brain at elevated temperatures as seen during fever is not known.
Theoretically, conduction failures could contribute to the drowsiness we experience during fever, and to clinical syndromes like "febrile seizures", which is the most common pathological brain activity in young children (Dube et al. 2007). The identification of temperature‐sensitive conduction failures in mammalian brain may therefore contribute to a more mechanistic understanding of such symptoms and pathology.
The very thin unmyelinated axons in cortex and many other gray matter areas may face greater challenges than other thicker or myelinated axons at elevated temperatures because of their very high axial electrical resistance, very large surface‐to‐volume ratio, extensive branching, and variable diameter along their paths. Particularly, variations in diameter and branching (Swadlow et al. 1980) reduce conduction reliability by creating regions of "impedance mismatch", for example, in squid (Westerfield et al. 1978), leech (Yau 1976), and vertebrate sensory ganglia (Dun 1955). High temperature in combination with such impedance mismatch is known to elicit failures in some branching axons, for example, in the squid (Westerfield et al. 1978) and vertebrate sensory axons (Barron and Matthews 1935).
The mechanism by which high temperature elicits conduction failures is probably by reducing the width and amplitude of the spike (Hodgkin and Katz 1949). Conversely, factors that increase spike amplitude and/or duration, for example, pharmacological block of spike repolarizing K+ currents, are expected to reduce conduction failures (Bostock et al. 1978; Westerfield et al. 1978). In the cerebellar parallel fibers, spike broadening has been recorded with voltage‐sensitive dyes in response to 1 mmol/L TEA (Sabatini and Regehr 1997). We will utilize this relatively well‐established pharmacology of the parallel fiber spike's repolarization to test if conduction failures, as expected, are reduced by spike broadening.
Furthermore, if failures are due to geometric factors combined with a temperature‐induced reduction of spike amplitude and/or width, one may expect moderate depolarization of the axonal membrane to reduce failures, at least in a temperature range where the spikes are not severely attenuated. Interestingly, typical cortical thin axons and several peripheral thin axons have an intrinsic mechanism, the depolarizing after‐potential (DAP), that increases axonal excitability for 50–100 msec after a spike (Gardner‐Medwin 1972; Wigstrom and Gustafsson 1981; Palani et al. 2012), and may therefore reduce conduction failures.
To identify conduction failures, the spikes need to be recorded at least at two positions along the axons, one close to the stimulation site to confirm that spikes are initiated, and the other more distal to check if spikes arrive further out in the axon. The reason to distinguish initiation failures and conduction failures is that those two types of failures probably have very different causes and consequences (Swadlow et al. 1980). Initiation of spikes, which can be temperature sensitive (Takeya et al. 2002), usually takes place in the axon's initial segment and is a regulated, homeostatic process (Adachi et al. 2015). Conduction failures on the other hand are probably not under the same control and may be caused by a wide range of normal or pathological factors, including axonal structure and nonuniform distribution of membrane currents. Also, the network effect will be different if the whole axon fails, compared to if it fails at a specific point far from the soma.
Although two recording sites on individual typical unmyelinated, cortical axons have not been achievable, it is possible to record from bundles of them at different distances from the stimulation site. If higher temperature gives conduction failures between the proximal and distal electrode, it may be detected as a preferential drop in the population response at the distal recording site. For example, the cerebellar parallel fibers showed such distal‐recording drop at high frequencies when spikes had to pass the branch point, but not when spikes were recorded at two sites after the branch point (Baginskas et al. 2009).
Despite their structural predispositions for conduction failures, typical unmyelinated cortical axons, like cerebellar parallel fibers and axons in hippocampal stratum radiatum, conduct spikes reliably at normal body temperature (Eccles et al. 1966; Frenguelli and Malinow 1996; Mackenzie et al. 1996; Cox et al. 2000; Forti et al. 2000; Koester and Sakmann 2000; Raastad and Shepherd 2003; Brenowitz and Regehr 2007). An important remaining question is therefore if temperatures above normal body temperature could elicit conduction failures in such axons.
Materials and Methods
Experiments were conducted according to the National Institutes of Health Guidelines for Animal Research. All animals were treated in accordance with the regulations of the Institutional Animal Care and Use Committee at Emory University, GA. We obtained 177 successful recordings from 95 rats.
Slice preparation
Wistar and Sprague–Dawley rats of either sex (10–25 days old) were used. Slices of cerebellum and hippocampus were prepared as described previously (Pekala et al. 2014). Briefly, the animal was exposed to isoflurane and decapitated after loss of consciousness and pain reflexes.
The brain was quickly removed and submerged in oxygenated (5% carbogen), ice‐cold artificial cerebrospinal fluid (ACSF, in mmol/L): glycerol 250, KCl 1.25, KH2PO4 1.25, MgCl2 7, NaHCO3 25, CaCl2 0.5, glucose 16. The cerebellum was dissected free and 300 μm slices were cut along the folia, perpendicular to their surface, to obtain long uncut parallel fibers. Hippocampal 300 μm slices were cut transverse to hippocampus' longitudinal axis. For recording from alveus fibers, hippocampus was split longitudinally approximately along the fissure separating dentate gyrus from the CA1 area. The slices were cut with a VF‐200 (Precisionary Instruments Inc., San Jose, CA) or, in a few cases, with a Leica VT1200 (Leica Microsystems GmbH, Wetzlar, Germany) and were stored in an oxygenated chamber for at least 0.5–1 h at room temperature before recording. The ionic composition of ACSF used for storage and recording was (in mmol/L): NaCl 125, KCl 1.25, KH2PO4 1.25, MgCl2 1, NaHCO3 25, CaCl2 2 and glucose 16 (pH = 7.4). Fast synaptic potentials were always blocked, in all experiments, by adding DNQX (30 μmol/L), APV (50 μmol/L), and picrotoxin (25 μmol/L) to the ACSF.
During recording, the slices were kept submerged in a 3 mL tissue chamber with oxygenated ACSF flowing at 4–7.5 mL/min. The temperature was controlled by a temperature controller (MTC‐20/2SD, npi electronic GmbH, Tamn, Germany).
6,7‐dinitroquinoxaline‐2,3‐dione (DNQX) and (DL)‐amino‐5‐phosphonovaleric acid (APV), were purchased from Tocris Bioscience, (Bristol, UK). Picrotoxin, tetraethylammonium (TEA), and salts were purchased from Sigma–Aldrich (Milwaukee, WI).
Compound action potentials recordings
Compound action potentials (cAPs) were recorded in cerebellum from parallel fibers, and in hippocampus from stratum radiatum and alveus fibers. The recording electrodes were pulled to a tip size of ~30–60 μm in diameter, from borosilicate glass capillaries (BioMedical Instruments, Germany). Outer and inner diameters were 1.5 and 0.86 mm, respectively. A proximal recording electrode was positioned, 0.3–0.8 mm from the stimulating electrode, and the distance between the proximal and distal recording electrodes was 0.5–2.1 mm (Fig. 1A).
Axons were activated by short electrical pulses (50–150 μsec) from an ISO‐stim 01MD (npi electronic GmbH, Tamm, Germany), delivered through a monopolar glass pipette with a tip diameter of 30–70 μm. Trains of stimuli with identical intervals of 10, 20, or 30 msec were repeated every 10–20 sec, although some experiments used single stimuli repeated every 10–20 sec. When amplitude and latency were stable at a temperature around 34–36°C, evaluated >5 min, temperature was increased, approximately 0.5–1.6°C per minute until it reached 43–44°C, or alternatively until cAP amplitudes were undetectable. The temperature was then reduced to 34–36°C.
Grease‐gap recordings
To monitor changes in the axonal action potential with its depolarizing after‐potential (DAP) during fever‐like temperatures, we used the grease‐gap technique as previously described (Palani et al. 2012; Pekala et al. 2014). Briefly, a grease‐gap pipette was made from pipette tips, usually 20 μL (Molecular BioProducts, 3521) and the tip cut to make an opening of ~1.5 mm. An arch, ~0.5 mm wide and 0.3 mm high, was carved out at the bottom. The cut surface of the pipette along with the arch was covered with grease (petroleum jelly, Vaseline, Chesebrough‐Pond's Inc.). The bottom of the plastic recording chamber was covered with Sylgard 184 (Dow Corning, Auburn, MI). By positioning the arched window over a bundle of axons, we could record a monophasic signal with distinct fast and slow, positive components, corresponding to a compound Na+‐dependent spike and its DAP, respectively.
For ordinary extracellular and grease‐gap recordings, we used a DPA‐2FL amplifier (npi electronic GmbH, Tamm, Germany) with a 10× at the head‐stage gain. Signals were amplified 100× at the amplifier (with the head stage gain that is 1000×), low pass filtered at 1 kHz, digitized at 10 kHz with National Instruments NI USB 6343, and stored on computer hard disk.
Whole‐cell patch‐clamp recordings
To record from cerebellar granule cells, we used the blind approach (Blanton et al. 1989) as described previously (Palani et al. 2012; Pekala et al. 2014). Pipettes were pulled from borosilicate glass (OD 1.5 mm ID 0.86 mm, BioMedical Instruments, Germany) and had tip resistance of 5–8 MΩ. The intracellular solution contained (in mmol/L): K‐gluconate 120, KCl 20, MgCl2 2, Na2ATP 2, HEPES 10, pH 7.2. Intracellular patch‐clamp recordings were amplified by SEC‐10LX (npi electronic GmbH, Tamm, Germany), voltage clamped in linear mode, current clamped in switching mode. The electrode's access resistance, between 6 and 20 MΩ, was not compensated. Axons were electrically activated by a 20–50 μm wide monopolar glass electrode.
cAP magnitude.
To measure the cAP, we used the standard deviation (SD) of the trace in a time window bracketing the action potential volley appearing at all temperatures during the experiment (gray area in Fig. 1B and G). The window is necessary because the latency shifts with temperature. Using the maximal peak‐to‐peak amplitude within a time window has the disadvantage of picking up the maximal peak‐to‐peak noise when cAP amplitudes are small. In experiments with relatively large cAP amplitudes, the peak‐to‐peak measure gave results similar to SD (Fig. 1D and E). However, because the amplitude often dropped considerably, we used SD as the magnitude of the cAP.
For the analysis of temperature effects on the cAP amplitudes during the four‐stimuli trains, we used measurement from the distal electrode because they were always well separated from the stimulus artifact, and would theoretically, be most influenced by conduction failures due to their longer conduction distance.
Noise subtraction and cutoff.
Baseline noise was measured at the beginning of the sweep, separately at proximal and distal electrode, in a time window similar to the one used to measure the cAP. This noise (SDnoise) was subtracted from cAP (SDcAP), according to the equation: sqrt(SDcAP2 − SDnoise2). The term "cAP magnitude" is used throughout this article for the magnitude of cAP, measured as SD, with subtracted noise. We included only recordings with detectable signals at both proximal and distal electrode, leaving data with a high signal‐to‐noise ratio of the noise‐subtracted cAP at both electrodes, signal‐to‐noise ratio = 26.5 ± 2.8 and 3.4 ± 0.4 on average at the proximal and distal electrodes, respectively, in the 40–42°C range. The strategy to exclude recordings with undetectable responses resulted in exclusion of many recordings that had undetectable responses only at the distal electrode at high temperature, and many of these probably were due to conduction failures. Therefore, this strategy most likely reduced, but could not have increased, the opportunity to detect conduction failures.
cAP latency.
Latency was measured for each response in the train of four stimuli as the time from the beginning of the stimulus artifact to the time of the minimum of the biphasic cAP.
Temperature binning.
All noise‐subtracted cAPs and their latencies were ordered with respect to the temperature at which they were taken, collected in 1°C bins, and the median value of each temperature bin was used for further calculations. The temperature of a temperature bin is given as its lower limit + 0.5°C, for example, the bin of 37–38°C is called "37.5°C".
Grease‐gap recorded cAP and DAP.
The amplitude of the cAP was measured as a maximum value of cAP relative to the baseline. The DAP was measured 3 msec before the artifact of the following stimulus, to obtain DAP amplitude immediately before the next cAP.
Data are presented as mean ± SEM unless otherwise specified. To compare log‐ratios, like in Figure 3B, we used Student's t‐test, two‐tailed, assuming unequal variance. Significance of changes in parameters between three or more groups of means (like in Figs. 5, 7) were tested with one‐way ANOVA, followed by Fisher's least significant difference (LSD) multiple comparisons test. To determine if log‐ratios were different from 0, as in Figures 2D, E, and F, 3C, and 4E, we used one‐sample z‐test. Statistical analyses were performed in Excel or IBM SPSS Statistics 22. P < 0.05 were considered significant.
Conduction failures at moderately elevated temperatures in cerebellum and hippocampus
To detect conduction failures, we recorded compound action potentials (cAPs) from cerebellar parallel fibers with two recording electrodes positioned at different distances along the axonal path (Fig. 1A). Fast synaptic signals were blocked in all experiments presented in this article (see Methods). Elevation of the bath temperature from 33 to 42°C reduced the cAP amplitudes at both electrodes (Fig. 1B). The cAPs increased again as temperature was reduced to 33°C, showing that the amplitude drop was not caused by irreversible damage (Fig. 1C).
The observation that the cAP almost disappeared at the highest temperatures is likely due to failure to activate or failure to propagate the spikes. To distinguish those two possibilities, we compared the reduction of the proximal and distal cAP (cAP‐p and cAP‐d, respectively). The cAPs were measured as the SD of the response in a time window that bracketed the cAP at all temperatures (gray area in Fig. 1B and G), and noise was subtracted, detailed in Methods. Because cAP‐p was usually larger than cAP‐d, we normalized them to their average values <37°C. We could then see that cAP‐d dropped more than cAP‐p at temperatures >37°C (displayed as logarithmic values from a typical experiment in Fig. 1D). The measurement of cAP as peak‐to‐peak amplitude (Fig. 1E) within the gray window gave similar results as SD at high amplitudes, but because of better performance at low amplitudes, as explained in Methods, we used SD for the remaining experiments. The bigger drop in amplitude of cAP‐d, relative to cAP‐p, at high temperatures was also illustrated by normalizing both cAPs to cAP‐p (so that cAP‐p has the peak‐to‐peak value 1.0) at 35, 37, and 40°C (Fig. 1F). Myelinated fibers in alveus (Fig. 1G) did not show a selective drop in cAP‐d, when compared with cAP‐p (Fig. 1H). This suggests that the temperature conduction failures were specific for thin, unmyelinated axons.
The temperature‐sensitive drop of cAP‐d found in unmyelinated fibers of cerebellum was confirmed by comparing cAP‐p and cAP‐d in 78 experiments from rats 10–25 days old (average 17.9 days) at different temperatures. Increasing temperature reduced all cAPs to very small values (Fig. 2A), displayed in Figure 2B as logarithmic values. Similar to the example in Figure 1D, we normalized cAP‐p and cAP‐d to their average temperatures <37°C (Fig. 2C). Visual inspection of cAP‐p and cAP‐d from all experiments suggested that increasing temperature reduced cAP‐d more than cAP‐p (Fig. 2C).
To evaluate if cAP‐d dropped more than cAP‐p, we calculated the ratio cAP‐d/cAP‐p in a range of temperatures (Fig. 2D). The average of these ratios showed that cAP‐d dropped more than cAP‐p at temperatures >39°C (first temperature bin significantly < 0, P = 0.013, was the bin 39–40°C). This preferential distal‐recording (cAP‐d) drop contrasted with the proportional drop of cAP‐p and cAP‐d at temperatures <39°C (values around zero in the logarithmic plot in Fig. 2D) and strongly suggested that some spikes failed to propagate between the two recording electrodes at temperatures >39°C.
The effect was not isolated to the youngest animals because the groups 10–14 and 17–25 days old, both showed significant drops in cAP‐d/cAP‐p ratio at high temperatures (Fig. 2D, right), although the youngest group was slightly more temperature sensitive because their cAP‐d/cAP‐p ratio dropped significantly >39°C while the oldest group dropped significantly >40°C.
The temperature‐sensitive conduction failures may be typical for gray matter axons because such axons in hippocampus also showed preferential drop at the distal recording electrode, at temperatures >38°C (Fig. 2E, average age 20.5 days, n = 25), although of smaller magnitude than in the parallel fibers in cerebellum. Furthermore, thin, unmyelinated CNS axons may be uniquely temperature sensitive because myelinated CNS axons (hippocampal alveus fibers) did not show signs of failures at comparable temperatures (Fig. 2F, average age 21.4 days, n = 16).
Block of voltage‐sensitive K+ channels reduced temperature‐induced failures
One of the hallmarks of conduction failures is that spike broadening, either by reducing temperature or by drugs, help overcome some of the failures (Bostock et al. 1978; Westerfield et al. 1978; Swadlow et al. 1980). We tested this idea using 1 mmol/L TEA in the extracellular solution, shown to efficiently block fast Kv3 channels (Rudy and McBain 2001) and also known to broaden the spike in cerebellar parallel fibers (Sabatini and Regehr 1997; Matsukawa et al. 2003; Pekala et al. 2014). Figure 3A presents two typical experiments with approximately 2°C higher threshold for conduction failures when exposed to TEA.
On average, there were fewer conduction failures at 39.5 and 40.5°C with TEA than without (Fig. 3B). The average may underestimate the TEA effect because the drop in cAP‐d/cAP‐p ratio occurred at slightly different temperatures in different experiments (like the two illustrated in Fig. 3A). We therefore calculated the difference between log (cAP‐d/cAP‐p) with and without TEA in the individual experiments. The average of these pairwise differences confirmed that TEA significantly increased the threshold for conduction failures at 39.5 and 40.5°C (Fig. 3C). At higher temperature (41.5°C), the variability prevented clear significance (P = 0.053), which may be due to the fact that cAP magnitude was very small, and also that conditions for spike conduction deteriorated beyond rescue.
The finding that 1 mmol/L TEA reduced the preferential distal‐electrode drop supports the hypothesis that voltage‐sensitive potassium channels (Kv) contributed to the conduction failures observed at high temperatures, although TEA may have reduced failures caused by other mechanisms too.
Do spikes improve the conduction fidelity of the following spikes?
Many thin, unmyelinated CNS axons have a depolarizing after‐potential (DAP) that offers an explanation of the hyper‐excitable period following individual spikes in such axons (Gardner‐Medwin 1972; Wigstrom and Gustafsson 1981; Palani et al. 2012). We therefore tested if the DAP could help spikes propagate at high temperatures.
We repeated the parallel fiber stimulus four times with 20 msec intervals, which is within the duration of the DAP (Palani et al. 2012; Pekala et al. 2014) and within the range of granule cell firing frequencies in vivo (Eccles et al. 1966; Chadderton et al. 2004; Jorntell and Ekerot 2006). With increasing temperature, all four cAPs declined. The first response at the proximal and distal electrodes were analyzed previously (Figs. 1 and 2) and showed that cAP‐d fell more than cAP‐p at temperatures >38.5°C, interpreted as conduction failures. Interestingly, during the train of stimuli, we observed that the last cAP (cAP4) in the train declined less than the first cAP at elevated temperatures (cAP1, shown at 36 and 40°C in Fig. 4A from a typical experiment).
The reduced drop of cAP4 relative to cAP1 was quantified similarly to the analysis of cAP‐p and cAP‐d (details in Methods). First, we normalized cAP1 and cAP4 to their average values <37°C, showing that cAP1 fell more than cAP4 at temperatures >38°C (Fig. 4B, same experiment as in A).
Example traces from 37, 38, and 40°C, normalized to the cAP1 peak‐to‐peak values (Fig. 4C) show that relative to cAP1, cAP4 was much larger at 38 and 40°C, meaning that cAP4 fell less than cAP1 at those temperatures. Similarly, the cAP4/cAP1 ratio showed that cAP4 declined less than cAP1 at temperatures >37°C (Fig. 4D). On average, the cAP4/cAP1 ratios increased significantly at temperatures from 37.5 to 41.5°C (Fig. 4E). A possible explanation for this is that there were fewer conduction failures at cAP4 compared to cAP1 at the high temperatures because this was in the temperature range in which conduction failures were detected by the cAP‐d/cAP‐p ratio (Fig. 2). If so, larger values of cAP4/cAP1 would be expected to be associated with the smaller values of cAP‐d/cAP‐p, and more so with increasing temperature, exactly as seen in Figure 4F (n = 78).
Theoretically, excitability may have increased during the four‐stimulus train due to accumulation of extracellular K+. If more K+ accumulated at high than low temperature, it may offer an explanation for the temperature‐sensitive correlation shown in Figure 4F. However, based on the results described in the following section, we find that unlikely.
Latency changes suggested an excitability increase of constant time course and magnitude following each spike
A spike with a DAP would be expected to speed up conduction of the spike that follows at its tail. This was exactly what we observed, the latencies of the second (Lat2), third (Lat3), and fourth (Lat4) cAPs were shorter than the latency of first (Lat1) cAP (interval = 20 msec, at 37°C, Fig. 5A and B). The reduced latency already at Lat2 confirmed that the axons were more excitable after being activated once (Gardner‐Medwin 1972; Wigstrom and Gustafsson 1981; Soleng et al. 2004). This effect did not accumulate during the train, observed as overlap of the three last responses (Fig. 5A, right) and was confirmed by similar reduction of Lat2‐4 in 20 experiments (at 37°C, Lat2‐4 were: 85 ± 2%, 84 ± 1%, and 84 ± 1% of Lat1, respectively, Fig. 5B).
Although the latency to each cAP was shorter at higher temperature, the relative reductions in Lat2‐4 were similar (Fig. 5C). At 40°C, Lat2‐4 were 89 ± 3%, 91 ± 3%, and 89 ± 3% of Lat1, respectively (all P < 0.003). Lat2‐4 were not significantly different from each other (all P > 0.4, n = 20, data not shown).
Such a very stable reduction of Lat2‐4 is unlikely to be due to factors that accumulate with repeated spikes, such as extracellular K+, unless the maximal velocity was already reached at Lat2 (a "ceiling effect"). To test for a ceiling effect, we used different stimulus intervals (10, 20, and 30 msec). We found that the maximal velocity was not reached because the different intervals resulted in different latency changes. Average values of Lat2‐4 were different, 82 ± 1%, 84 ± 1%, and 89 ± 2% of Lat1, for 10, 20, and 30 msec stimulus intervals, respectively (Fig. 5D).
At the longest intervals (30 msec), the latency reduction was not only the smallest but also had the lowest variability between Lat2‐4 (Fig. 5E, coefficient of variation 3.2 ± 0.8%, 2.0 ± 0.7%, 0.8 ± 0.2% at 10, 20, and 30 msec intervals, respectively). This is opposite of what would be expected by a ceiling effect, which would occur at the shortest latency when the axon could not conduct faster. We therefore conclude that the reduced latency was due to an excitability‐increasing process of constant time course and magnitude, relatively activity independent.
The DAP in parallel fibers helps propagate spikes
Our hypothesis was that the axonal DAP increased excitability after individual spikes and helped conduction in the temperature range where some axons failed. To test this, we electrically activated the parallel fibers while recording antidromic spikes from the granule cell soma. The recorded cells were assumed to be granule cells based on their location in stratum granulosum, activation from stratum moleculare with a latency, and their high resistance to somatic current injections (Fig. 6A, mean = 2.10 GΩ, SD = 0.72 GΩ, n = 14, similar to e.g., Diwakar et al. 2009). The average latency between stimulus and the peak of the spike was 2.00 msec (SD = 1.39 msec, n = 14, Fig. 6B, green traces).
At moderately hyperpolarized potentials (−72 mV, Fig. 6B, red traces) compared to rest (−66 mV, Fig. 6B, green traces), the fast component of the spike disappeared, probably because spike conduction failed at some distance from the soma and the fast component was attenuated by the axonal cable (Sheffield et al. 2011). At even more hyperpolarized potentials, also the slow component disappeared (−75 mV, Fig. 6B, gray traces). We adjusted the somatic potential to give ~50% failures. The responses to identical stimuli showed spike‐like all‐or‐none behavior (red and gray traces).
With ~50% failures at a constant stimulus strength, we repeated the stimulus four times and observed that if the axon conducted one spike, the following stimulus always resulted in a successfully conducted spike (Fig. 6C and D). By counting failures and spikes (including attenuated spikes, like the red traces in B), we observed 1216 failures immediately after a failure, but only two failures immediately after 1784 successfully propagated spikes (16 experiments). This confirms that a successful spike (full or attenuated) increased the chance of successful conduction for the following spikes.
Unless a successful conduction occurred, the success rate did not increase during the train (red bars Fig. 6E). This means that extracellular K+ or other excitability‐increasing effects from adjacent axons or other cells had minor effects on the excitability compared to the strong effect of a successfully propagated spike which abolished almost all failures at the following stimulus.
A likely explanation for the failure‐reducing effect of a single spike is that residual depolarization (the DAP) left from the preceding spike helped overcome regions of low excitability. The DAP may therefore offer an explanation for the larger cAP amplitudes at the fourth stimulus compared to the first in the temperature range that induced failures (Fig. 4).
Constant amplitude and waveform of the DAP
For the DAP to be an acceptable explanation for the very similar latency reduction at all three cAPs (Lat2‐4, Fig. 5), the DAP would need to have a relatively constant amplitude at a given interval after each spike, which we will address in this section. The intracellular DAPs of the antidromic spikes, measured on the decay of responses 1–4 immediately before the following stimulus (arrows in Fig. 7A) were very similar during the train of four stimuli (Fig. 7B, −57.0 ± 2.6, −56.7 ± 1.9, −54.7 ± 2.5, and 56.6 ± 2.7 mV). The SD of the membrane potential of those four DAPs in the individual experiments was only 0.75 ± 0.11 mV on average for 14 experiments. That low variability in the membrane potential may explain the similar latency reductions at cAP2–4 (Fig. 5).
Because the initial segment of the axon has other membrane currents, axial resistance, and input resistance than more distal parts (Kole and Stuart 2012) which theoretically could influence our somatically recorded DAPs, we also recorded the waveform of the axonal spikes far away from the soma using a miniaturized grease‐gap method (Palani et al. 2012).
Grease‐gap recordings give information about the temporal changes of the transmembrane voltage, but not the true voltage. Therefore, to compare intracellular and grease‐gap recordings, we used a measure we could obtain from both recording techniques, namely the difference between DAP amplitude and baseline measured before the first stimulus (arrows in Fig. 7A and C). To compare the DAP variability recorded by different techniques on a similar scale, we normalized the amplitudes to the value of the first DAP amplitude, both in intracellular and grease‐gap recordings (Fig. 7D).
The DAP variability between responses 1–4, measured as coefficient of variation (SD/mean) was 0.095 ± 0.019 (n = 14) and 0.090 ± 0.011 (n = 23, P = 0.84, data not shown) for the intracellular and grease‐gap measurements, respectively. This similarity between the variation of the DAPs measured intrasomatically and by the grease‐gap technique supports the hypothesis that both those techniques detect features of the axonal DAP. The fact that the variability was similar also suggests that the axons may have a DAP variability of approximately 0.75 mV, as recorded at the soma, also further away from the soma. Additionally, these data show that the axonal DAP was relatively resistant to moderate activity (the repetition of four spikes).
Influence of temperature on action potential and membrane potential
When Hodgkin and Katz (1949) described the temperature effects on the propagating spike in the squid giant axon, they noted that increasing temperature caused a small depolarization of the resting membrane potential, but a large amplitude‐ and width‐reducing effect on the spike. This led them to conclude that the spike failure was not simply due to loss of membrane potential, but depended on spike‐specific mechanisms.
Since it has not yet been possible to measure membrane potential directly in thin unmyelinated cortical axons, we measured steady‐state membrane potential at the soma using tight‐seal recordings. However, fast voltage changes in the axon, like the spike, are probably not well represented at the soma. For example, Kv10.1 is located almost exclusively to the axon, and most likely influences spike repolarization, but no effects of this could be detected by somatic recordings (Mortensen et al. 2015). We therefore measured cAP amplitude from the parallel fibers using a grease‐gap method (Palani et al. 2012). Somatic membrane potential and grease‐gap recorded cAP will be influenced by axonal membrane potential and spike amplitude, respectively, although further qualifications will be given in the Discussion.
The results were qualitatively very similar to Hodgkin and Katz (1949) (Fig. 8A). Membrane potential depolarized by 3% (0.97 ± 0.02, n = 8) at 40°C compared to its value at 37°C (average membrane potential before normalization at 37°C = −66.0 ± 3.4 mV). In contrast, grease‐gap peak cAP decreased by 37% (0.63 ± 0.06, n = 13) at 40°C compared to its value at 37°C.
If the DAP helps to reduce failures of spike propagation during elevated temperatures, as we propose, it should be present also at the temperatures at which axons start to fail. This was indeed the case, as shown by the average shape of potentials from nine grease‐gap recordings at 36 and 40°C, temporally aligned at their peak amplitude (Fig. 8B).
We have identified two intrinsic mechanisms that influence spike propagation in typical gray matter axons: a temperature‐sensitive mechanism that gives conduction failures at temperatures above, but not below normal body temperature, and a temperature and activity‐resistant mechanism that improves conduction fidelity at temperatures immediately above normal body temperature.
It is important to distinguish spike conduction and initiation failures because the mechanisms for those failure types are often different. One difference is that conduction failures occur in axons that have an essential function intact, namely the ability to initiate spikes. In general, two recording sites along the axonal path is necessary to identify conduction failures to confirm that axons can initiate spikes, at a proximal site, and to check if the spikes can travel to a more distal recording site.
The two recording sites also gave the opportunity to better interpret the extracellular population recordings, which is still the only practical method to study the electrical behavior of typical mammalian, small‐bouton, unmyelinated cortical axons. For example, increasing temperature reduced the population response (cAP). This may have many causes, like smaller amplitude and width of the individual spikes, and reduced number of axons activated by the electrical stimulus, but such factors would be expected to contribute equally at both proximal and distal recording sites. This assumption was supported by the observation that the cAPs changed proportionally at those two sites at temperatures <38°C for the unmyelinated cerebellar and hippocampal axons. Furthermore, the myelinated alveus axons, measured in exactly the same way as the unmyelinated axons, showed proportional changes at the two electrodes over the full temperature range.
Therefore, when the distal cAP dropped more than the proximal one only in the unmyelinated axons, and only at temperatures >39°C, we concluded that axons failed between the two recording sites. This conclusion assumes that the axons giving rise to the cAPs at the two electrodes had similar properties, but does not require them to be exactly the same axons.
What caused the failures?
A definite answer requires details about electrical properties locally along the axons, which is not available for any axon, but the question has been explored comprehensively with calculations and computer models based on accurate morphological measurements (Westerfield et al. 1978). In general, conduction failures occur because a segment of the axon has low excitability, or because the current from one segment is insufficient to bring the membrane of adjacent segments to threshold. The latter may happen with abrupt changes in diameter, capacitance, or branching, making regions of "impedance mismatch" (Van Essen 1973; Goldstein and Rall 1974; Yau 1976; Luscher et al. 1994).
In combination with such impedance, mismatch increases in temperature and/or activity can cause conduction failures (Rasminsky 1973; Westerfield et al. 1978). We will comment only on the effects of temperature because we deliberately used few stimuli (four) repeated every 20 sec, which is below the activity reported to give activity‐induced failures (Smith 1980; Baginskas et al. 2009).
An important mechanism that contributes to temperature‐induced conduction failures is that increasing temperature reduces the duration and amplitude of the spike (Hodgkin and Katz 1949; Thompson et al. 1985; Yu et al. 2012). These effects are due to faster rate constants of inactivation and activation of the Na+ and K+ currents, respectively. They become 2.6–3.0 times faster if temperature increases by 10°C (Hodgkin et al. 1952; Schauf 1973; Westerfield et al. 1978). Current amplitude, membrane resistance, excitation threshold, rate of rise of the spike, and cytoplasmic resistivity seem to have less effect (Westerfield et al. 1978; Yu et al. 2012). Such a temperature‐induced reduction in amplitude and/or width is expected to influence the compound action potential, both in the regular extracellular recordings and the grease‐gap recordings, which is compatible with our observation (Fig. 8).
There is, however, a theoretical possibility that temperature causes conduction failures by changing the membrane potential. Depolarization will inactivate Na+ channels, and hyperpolarization will increase threshold for spike. From somatic recordings, there are examples of both hyperpolarization and depolarization with increasing temperature, although from low temperatures increasing temperature tend to hyperpolarize (6–33°C in Volgushev et al. 2000), but closer to 36°C, there seems not to be any systematic effect of temperature changes (Thompson et al. 1985; Trevelyan and Jack 2002). Above 36°C, we found a small depolarization (3%, or about 2 mV), similar to Yu et al. (2012), Kim and Connors (2012), and qualitatively similar to Hodgkin and Katz (1949) in the squid axon. Thus, it may be a general tendency to depolarize when temperature deviates from normal body temperature. It is unlikely that the small change in membrane potential we observed would change the spike amplitude enough to cause failures because previously we found that an increase in extracellular K+ from 2.5 to 6 mmol/L clearly increased the conduction velocity, meaning that a moderate depolarization gave increased excitability, but the amplitude of the grease‐gap recorded cAP did not change (Pekala et al. 2014).
Our finding that the unbranching myelinated alveus axons did not have conduction failures may, of course, have many causes because electrical properties of myelinated axons are very different from the typical gray matter axons. It is interesting, though, that we have not been able to find any report of conduction failures in normal myelinated, unbranching axons. They do fail to function at high temperatures (Bernstein 1902), but without pathology‐like demyelination or local compression, they seem not to fail at specific locations while the spike is being propagated. This may indicate that branching is usually necessary to stop an already propagating spike.
What prevented the failures?
Drugs that broadens the spike often prevent failures (Bostock et al. 1978; Westerfield et al. 1978; Swadlow et al. 1980). Our results show that this is the case also in thin, unmyelinated axon in the mammalian cortex.
We used TEA because it is known to broaden the parallel fiber spike (Sabatini and Regehr 1997). However, cerebellar granule cells express more than 26 K+ channel α‐subunits, including six delayed rectifier types (Mathie et al. 2003), and TEA reduces several of them, but usually at higher concentration than what is needed to block the Kv3 family of channels (Rudy and McBain 2001). Based on pharmacological studies of the grease‐gap recorded parallel fiber cAP (Pekala et al. 2014), we find it likely that the Kv3 family dominates the fast repolarization. In that article, the Kv1‐blocking toxins changed the cAP much less than 1 mmol/L TEA. Only when TEA‐sensitive channels were blocked, Kv1‐blocking increased the cAP significantly. Quinine, that blocks not only Kv2 channels but also several other channels, did not have any detectable effect on the fast cAP, unless both Kv3 and Kv1 channels were blocked. Together, those results suggest that Kv1 channels participated less than Kv3 as long as Kv3 channels were operational, and that the spike broadening was the main mechanism by which 1 mmol/L TEA reduced conduction failures.
Since Kv3‐family channels are among the fastest spike repolarizing channels, it is tempting to suggest that these channels may be limiting for the spike amplitude and width, and therefore cause failures when they become even faster at high temperature. However, the rate of Na+ channel inactivation and K+ activation will both increase with temperature, and it seems difficult to determine which of those changes are most important for reduction of spike amplitude and width (Yu et al. 2012) without having voltage‐clamp control of the membrane.
The DAP reduced failures
The DAP has been studied in several preparations and so have conduction failures, but the influence of a DAP on conduction failures has to the best of our knowledge not been studied. The DAP has never been recorded directly, as a membrane potential, in unmyelinated small‐bouton CNS axons, but several lines of evidence, some illustrated by experiments in the present article, strongly support its existence in cerebellar parallel fibers and hippocampal stratum radiatum axons (Soleng et al. 2004; Palani et al. 2012; Pekala et al. 2014). For example, with threshold axonal activation and somatic membrane potentials that allowed antidromic somatic spike invasion, the fast spike and the DAP always occurred or failed concurrently, meaning that the DAP was a part of the spike. At somatic potentials that blocked invasion of the fast spike component, the DAP could still be detected, meaning that the DAP was created in the axon and not by the soma.
The hypothesis that the DAP reduced the failure rate is supported by results from the following experiments:
In intrasomatic recordings, the DAP eliminated somatic invasion failures. Since the axon‐to‐soma transition is the largest transition from small to large diameter found in most neurons, introducing a particularly large impedance mismatch, the finding that the DAP eliminated these somatic invasion failures makes it likely that the DAP can reduce failures also further out in the axon.
In trains of four stimuli, increasing temperatures reduced the first more than the last cAP. Importantly, this tendency was strongest at the temperature where the two‐electrode recordings showed signs of conduction failures. A likely interpretation is therefore that an excitability‐increasing effect of the preceding spikes made it easier for the following spikes to propagate successfully. The DAP offers an explanation for such spike‐dependent excitability‐increasing effect.
The cAP latency was reduced from the first to the second response, but almost identical for response 2, 3, and 4. This is compatible with an excitability‐increasing process of equal magnitude after each spike in the four‐stimulus train. When we showed that the DAP, measured either on antidromic action potentials, or far out in the axon with grease‐gap recordings, have very similar amplitude in such four‐stimulus trains, it is likely that the DAP was the process that facilitated propagation, and helped the axons overcome temperature‐induced conduction failures.
Unfortunately, antidromic spikes detected in somatic recordings were not a suitable approach for demonstrating that the DAP could reduce temperature‐induced failures (we used somatic hyperpolarization to induce invasion failures). Firstly, the intracellular recordings were often not stable at temperatures >38°C, which added to the already difficult task of activating the axon belonging to the cell recorded from. Secondly, there is a theoretical but likely problem that only a narrow temperature range allows both successful activation and conduction failures. Below that temperature range, the axons never fail to propagate action potential, and above that range, they may not be able to initiate and action potential. Therefore, to investigate if the DAP influenced temperature‐induced failures further away from the soma, we had to use population recordings.
Could higher temperature increase stimulus‐dependent K+ efflux, and thereby contribute to larger cAPs at the end compared to the beginning of the four‐stimulus trains? That is unlikely because such K+‐mediated excitability increase would accumulate during the train, conflicting with the finding that excitability, measured as latency reduction jumped from the first to the second stimulus, but thereafter was almost constant for stimuli 2–4. Furthermore, potassium accumulation is unlikely to be the major excitability‐increasing factor because the failure rate for antidromic invasion at the soma actually increased with number of stimuli for stimuli that were not preceded by a successfully propagated spike. This strongly suggests that it was the propagation of a spike, and not the stimulus by itself, that increased the excitability.
The most direct proof that the DAP reduces failure rates would be to compare failure rates with and without the DAP present. Unfortunately, this is currently not possible because there is no known method to reduce the DAP (although it can be enhanced by Kv blockers as described in Pekala et al. 2014).
DAP is robust
The remarkably constant excitability increase, measured in terms of latency reduction of the responses following the initial one in trains of four stimuli, suggests that the DAP is relatively insensitive to activity. This was supported by somatic recordings of antidromic spikes, and grease‐gap recordings of axonal spike populations. The relatively activity‐independent DAP amplitude is similar to recordings using voltage‐sensitive dye presynaptically in crayfish neuromuscular junction (Lin 2012), but has to the best of our knowledge not been studied before in mammals. Furthermore, the DAP was still present at fever‐like temperatures in axons far away from the soma.
In conclusion, we have found that typical unmyelinated, small‐bouton CNS axons can fail even after a spike has been successfully initiated. The theoretical but very likely possibility that regions of impedance mismatch, sensitive to geometrical factors, play a role in these failures is important because axonal geometry may change during development and in disease. For example, large axonal swellings occur on both myelinated and unmyelinated axons in neurodegenerative‐relevant mouse models (Sanchez‐Varo et al. 2012).
The relatively moderate temperature increase giving conduction failures makes them potentially important for understanding confusion, fatigue, and seizures occurring at high body temperatures. The tendency of the young animals to fail at a lower temperature than the older ones (Fig. 2D) may contribute to an understanding of the higher seizure susceptibility of young rats and perhaps humans (Dube et al. 2007). However, evolution could obviously have pushed the failure temperature higher, so since these mammalian axons have this failure mechanism at temperatures just above normal body temperature, it may serve a protective function, for example, by limiting the axons' energy expenditure at high temperatures.
Conflict of Interest
None declared.
Funding Information
This work was supported by the National Institutes of Health, Bethesda, MD, USA (grant no. R21NS082680).
- Manuscript Received: August 24, 2016.
- Manuscript Revised: August 29, 2016.
- Manuscript Accepted: August 31, 2016.
- © 2016 The Authors. Physiological Reports published by Wiley Periodicals, Inc. on behalf of the American Physiological Society and The Physiological Society.
This is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.


text:
I am reading everything and watching every video I can about this phone. It sounds great. I am going tomorrow to handle it again and make sure I'm ok with the size. I just have a couple questions. They seem simple or stupid but I want to be sure I'm making the right choice for me.
I have use both android and iOS in the past. I'm typing this on an iPhone 4. I had the galaxy s3 and loved it except for the GPS. It was awful. It took forever to connect. This was a huge problem. So my first question is, how is the GPS on the Note 2? Does it connect quickly?
Also I think I would use the S pen to cut and paste a lot. When sending to other devices does it work well? In other words does what I paste show up clean in text messages or emails regardless of the phone used by the recipient?
How does the phone handle exchange email? I have used Touchdown in the past but would like to keep my email together like I so on iOS. Does it work well or do people use another email app?
My whole family is on iOS devices so its easy for us to share pictures and things. Is anybody else in a similar situation? How do you handle sharing? I was thinking that Google + would be the way to go. Any thoughts?
I appreciate any feedback. I really want the bigger screen. I hate this small screen and I just don't see the iPhone 5 being any better really. Thank you.
Sent from my iPhone using Tapatalk


text:
Richard and June Ross Genealogy Fund
Promoting, Encouraging, and Sustaining Genealogy
The purpose of the Richard and June Ross Genealogy Fund is to promote, encourage and sustain the research, documentation and preservation of genealogy and history of the Keweenaw Area of Michigan's Upper Peninsula.
The Houghton-Keweenaw County Genealogical Society exists to educate and support our members in their research and the preservation of family histories. Our goal is to foster interest in local area history and genealogy, provide instruction in research techniques, and preserve information for use by future generations.
Richard Albert Ross was born on August 30, 1920, in Racine, Wisconsin. He graduated from Hyde Park High School in Chicago, Illinois, in 1938. Richard, who was known as "Dick" to his friends, served as a Warrant Officer in the US Army during World War II.
June Jennings Ross grew up in Houghton, Michigan, and graduated from Houghton High School in 1938. Richard and June met in California, where Richard started working for the California State Department of Employment in 1941. They were married in 1947. Richard retired in 1975. In 1977, Richard and June began the genealogical research that became their life's work and their lasting legacy.


text:
Biography: I began drawing at a very early age. While I was in high school I was offered a scholarship to attend the Rhode Island School of Art and Design. Unfortunately, I declined this offer and have since regretted it. I currently attend the University of Advancing Technology, majoring in Game Design with emphasis on Modeling and Animation. I primarily use Maya and Photoshop, and also have experience with Illustrator and 3D Studio Max. The majority of my early self-taught studies focused on illustration. I did a lot of comic book drawings as well as portraits when I was younger. I have always been fascinated with painting but have not done much with it as I find it far too messy!
Country: United States
E-mail: email@example.com
Development by Magic Web Solutions.


text:
Have you ever wondered if you should be using prebiotics or probiotics in your horse feed? The info below helps explain the benefits of both!
Click the image to be able to zoom in.
Ever wonder how your horse's digestive system works? What goes on in there? Why are they so sensitive? And why should I divide the feed ration into 2 or 3 feedings per day? Let's take a closer look to better understand.
Mouth & Teeth: Teeth are the beginning of the entire process. Designed to grind foodstuffs into smaller pieces, the act of chewing also stimulates three glands in the mouth to produce saliva. These glands can produce up to 10 gallons per day of saliva. The saliva contains bicarbonate (a natural acid buffer) and amylase (assists with carbohydrate digestion). Teeth are an important component to digestion and should be checked annually to insure proper function. A horse that is unable to effectively chew long stem forage, such as a senior horse, is at higher risk of impaction colic. If you have a horse like this, be sure to consult with your veterinarian for a comprehensive care and feeding program.
Esophagus: The purpose of the esophagus is to funnel food from the mouth to the stomach. Approximately sixty inches in length, this is a one-way passage. Unlike humans, horses cannot vomit. This is why horses who 'bolt' their feed (eat too fast and don't chew adequately) can get into trouble and feeding practices need to be adjusted to reduce the risk of bolting feed.
Stomach: Small in size compared to the rest of the horse's body, food will only spend about 15 minutes in the stomach before it moves on. The stomach is designed to function best when it is ¾ full; therefore, care takers are encouraged to provide horses with a steady supply of forage throughout the day. Because of the small size, a horse should not be fed more than 0.5% of body weight in one meal. Meals of grain are best divided into 2 or 3 portions per day.
Small Intestine: After leaving the stomach, food will spend anywhere from 30 to 60 minutes here; a good thing with the many nutrients that are absorbed. Nutrients such as proteins (amino acids), vitamins A, D, E and K, calcium, phosphorus, and other minerals along with starches and sugars. Cereal grains such as oats, barley, and corn that are high in carbohydrates (starch and sugar) are easily digested here. The horse doesn't have a gall bladder, so bile from the liver flows directly into the small intestine to aid in the digestion of fat.
Large Intestine: is comprised of the cecum, large colon, small colon and rectum.
With this short tour and explanation, we hope you have a better understanding of how your horse digests and absorbs nutrients, and that this also sheds light on why good feeding management and regular dentistry care are important aspect for the digestive health and overall well-being of your horse.
While scanning information about various horse feeds, you may have come across the phrase 'contains prebiotics and probiotics'….Hmmm, sounds impressive, but what are they, what do they do for your horse, and why are they important? Pre- and probiotics are considered "functional ingredients" that are added to horse feed to provide benefits to your horse. Here is some information about them and what they can mean for the digestive tract and overall wellbeing of your horse.
It starts with the gut. The environment of the intestine (a.k.a. gut) contains naturally occurring beneficial microorganisms commonly called 'bugs'. Gut bugs are found in all species, including humans, and are essential to the digestion process. For the horse, gut bugs work to break down components of forage and feed as they pass through the digestive tract. The bugs deconstruct complex molecules within feedstuffs, which releases nutrients and allows the synthesis of energy substrates and important vitamins. Those nutrients are then absorbed through the intestines into the bloodstream, where they become available to cells in the body to support basic maintenance, growth and activity.
Feed that is broken down and digested more completely allows for more nutrients to be readily available for absorption. This is essentially what probiotics do. Adding probiotics to a horse feed means adding more beneficial bugs to the existing population in the horse's gut. Probiotics such as yeast culture, work with the naturally occurring bug population to enhance the digestive process, further breaking down complex protein and fiber fractions in the gut and making them more available for absorption into the blood stream. As a probiotic, yeast culture has also been shown to balance and stabilize the digestive microbial ecosystem in the cecum of the horse, as well as help prevent the colonization of bad bacteria in the gut. A stable microbial ecosystem is beneficial to the horse beyond improvements in digestive and absorption efficiency, it also reduces the risk of digestive upset, such as gas colic, that a horse might experience with changes in feed or hay, or while under stress from transportation, shows, changes in weather or other.
Prebiotics can be thought of as an energy jolt for the gut bugs. Prebiotics are a rich nutrient source for the gut bugs (e.g. lactobacilli, bifidobacterial) which in turn stimulates their growth and activity, making them more effective at their job. Research has shown that prebiotics help stabilize the population of gut bugs even through sudden changes in the diet, which helps to reduce incidents of digestive upset. For performance horses who require energy dense diets that include higher levels of starches and sugars, prebiotics can help reduce the incidence of digestive disorders and support optimal performance. Prebiotics such as inulin and oligofructose are selectively fermented by the gut bugs, stimulating their growth and activity, which benefits the horse by enhancing the absorption and retention of certain minerals, which in turn can support the immune system, skeletal tissue, and more.
The population of bugs in the gut are sensitive to changes in their environment brought on by stress, illness or the ingestion of undesirable materials .The addition of pre- and probiotics to a diet has been shown to be beneficial in reducing the incidence of digestive upset, namely diarrhea. For senior horses, the use of pre- and probiotics in feeds has been shown to improve the digestibility and absorption of nutrients, which can translate into an enhanced quality of life. In summary, pre- and probiotics work with the naturally occurring gut bugs to support optimal gut health, aid in the digestion process, as well as provide a buffer against negative bacteria.
There is a lot of discussion these days about carbohydrates, the good and the bad. In reality, they are all related. Carbohydrates contain carbon, hydrogen and oxygen in varied chemical relationships.
Starches and sugars are produced by plants as a means to store energy, so they contain energy that is more readily available than the energy in fiber.
Because horses produce less amylase, the enzyme used to break down starch, than some other animals, say perhaps pigs, they are more limited in their ability to digest large quantities of starch. However, they digest smaller quantities very efficiently and very effectively!
Starches and sugars are a fairly concentrated source of Calories to be used as a source of energy when extra energy is required to maintain body condition and do more work. In the wild, horses would consume seed heads (grain) when available in the fall to help them gain weight (added Calories) to be ready for winter, so starches and sugars are a part of the diet in horses in the wild. Today's working horse requires Calories, and a combination of fiber, starch, sugar and fat can be the best way to support the horse to maintain the balance of optimal health and optimal performance.
A high-performing horse can have up to twice the calorie requirement as the same horse in a maintenance stage. Owners and trainers of performance horses often give more feed to meet that calorie need. Because horses can use fat as a calorie source efficiently, and fat contains more than double the calories of starch, high-fat horse feeds make perfect sense to increase the energy intake without greatly increasing the quantity of feed needed.
Feeding higher-fat, controlled starch level feeds can play a role in lowering the chance of colic and laminitis by reducing the amount of starch (carbohydrates) in the ration. Here's why:
Keep in mind that starch is still a crucial part of a horse's diet, and is required for proper muscle function. As horse owners, it is best to work towards an optimal balance of all nutrients in the diet, not the use of one to replace another. A proper balance of controlled starch levels, along with increased fat levels in the horse feed, will help deliver a horse that is ready to go and has the fuel in the tank to keep on going.
Lately there has been tremendous interest in the horse world about fat. In regards to human nutrition, "fat" is often considered a bad word, and low-fat diets are popular. But we should remember that in people, some fats are necessary and healthy. This is equally true for horses: fats play a very important role in horse feeds and nutrition.
There are many reasons to feed horses added fat. The best reason for using added fat is for an energy (calorie) source. The primary purpose for grain feeding is to provide energy for maintenance, performance, growth and reproduction. Because fat contains more than twice the calorie content of starch, and horses digest and utilize fat well, higher fat horse feeds offer an excellent opportunity to increase calorie intake without greatly increasing the quantity of feed.
Other reasons for adding fat to a feed ration include improved endurance, heat tolerance, hair coat and attitude:
It is important to keep in mind that indiscriminate fat supplementation can create deficiencies of other nutrients. This is known as empty calories; where energy levels are adequate, but protein, lysine and mineral levels are not. Developmental bone problems can result which may precipitate injuries in young horses.
There also is a period of adjustment of about three to four weeks for horses to receive benefits from added fat. Any change in diet should be done gradually over seven to10 days to avoid the possibility of digestive upsets.
A balanced diet, tailored to the use and age of the horse, is the most important consideration. A trained nutritional consultant can make recommendations that will best fit your horse and the activity involved.
In our previous post, we learned what fiber is and how a horse digests it,and we also learned that a horse consuming 1-1.5% of it's bodyweight in quality roughage will satisfy its daily fiber requirements.
When it comes to any grain sources that may be added to your horses diet, fiber plays a much smaller role since the amount of fiber that is added by grains is relatively little, but the effect and digestive process is similar.
When feeding the grain portion of the diet, ensure that your horse is not receiving high quantities of grain meals all at once – typically no more than 5-6 pounds of grain per meal at most. Because grains tend to be higher in starch than roughage, feeding too much at once can overwhelm the small size and quick rate of passage of food through the stomach and small intestine, and allow starches to pass undigested to the hindgut. Digestion of starches in the hindgut releases lactic acids that are toxic to the fiber-digesting microorganisms, which can result in a gas colic episode or laminitis.
Generally speaking, when you look at a the tag from a basic equine ration, the higher the crude fiber level listed, the lower the energy content of the feed. Of course, there are other factors that must be looked at, such as the fat level, and also possibly the sources of fiber.
Beet pulp, for example, is often referred to as a "super-fiber" due to the high level of fiber it provides while also providing roughly the same energy level as oats. While soy hulls and dehydrated alfalfa are common ingredients used to increase fiber levels, a performance horse ration with a higher fiber level may make use of beet pulp to achieve both increased energy and increased fiber levels.
Last week a horse owner contacted me about changing her horses diet. She stated that they are ¾ of the way through show season and he is just "off his game". It seems that the horse was showing a lack of appetite and not finishing his grain. In addition, his disposition seemed to have changed, being rather grumpy and his performance level was suffering. A few times he had shown signs of mild colic over the past two months.
I suggested the owner contact her veterinarian, as it sounded like the horse may have an ulcer. I explained that the percentage of horses with ulcers continues to increase, and that higher intensity levels of training are correlated with an increase in ulcer incidence. The ulcers often occur in the upper third of the stomach, which does not have a mucus layer and does not secrete bicarbonate that helps to buffer stomach acid. It is also interesting to note that ulcers have not been founded on pastured horses. This is likely due to the fact that as a horse grazes, it produces large amounts of saliva, which contain the bicarbonate and amylase needed to provide a buffer for the stomach lining.
The owner was not pleased with my answer, but agreed to call the vet. Within the week she contacted me and said the horse had been diagnosed with a gastric ulcer. He was now on medication, but we needed to make dietary changes as well. I suggested the following "back to basic" steps to help manage her horses condition:
In essence, to help keep them from suffering the ill effects of ulcers, we need to let our horses just be horses.
Feeding bran mashes to horses is a common tradition dating back a long time, and is often thought to be a help in preventing colic through a laxative effect. Bran is believed to be a laxative in people, but to get that effect in horses, you would actually have to feed it in quantities bigger than your horse could eat. Some horses do produce softer stools the day after eating bran, but this probably reflects bran's tendency to irritate the lining of equine intestines. If fed daily over a long period of time, bran may actually contribute to the formation of enteroliths.
The bigger danger in feeding bran to horses is the calcium:phosphorus ratio of bran. Calcium and phosphorus work together to build sound bones and assist muscle function. To do so, they must be absorbed in appropriate proportions (preferably 1.2 or more parts calcium to 1 part phosphorus). If, over a long period of time, phosphorus exceeds calcium in the diet, the horse's body will pull extra calcium from the horses bones to meet it's needs, and eventually weaken the skeleton.
Wheat bran and rice bran contain approximately 10 times more phosphorus than calcium. Therefore, an occasional bran mash won't harm the horse, and he will likely relish the treat. However, daily bran regimens in large quantity should be avoided, unless calcium is supplemented in sufficient quantities elsewhere in the diet.
Colic is one of the leading health problems facing horse owners. According to the USDA's National Health Monitoring System (NAHMS) Equine Study 1998, about 4% of the horse population experiences colic each year. Colic rated second only to old age as the cause of death in equines. The same study indicated that horse owners most commonly identified "unknown" causes for colic, followed by gas colic and feed related.
Feeding management and non-feeding-related management practices can all have an impact on reducing the risk of colic.
The following management practices can aid in reducing the risk of colic:
Colic prevention—rather than colic treatment—is clearly much better for both the horse and the horse owner.


text:
This post is syndicated (?) from WhiskyIntelligence.com
Ardbeg Embassy Launch Tasting at TWE Vinopolis – Scotch Whisky News
Ardbeg Embassy Launch Tasting at TWE Vinopolis
As mentioned a few posts ago we ran a tasting at the end of May to celebrate our shop's ascension to the rank of Ardbeg Embassy. We were joined by Distillery Manager Mickey Heads and an audience of peat fans eager to try…


text:
Europe, relegation and the Scudetto are in the balance in Livorno-Lazio, Torino-Udinese, Cagliari-Parma, Verona-Catania and Sassuolo-Juventus.
Click here for match previews of every single Serie A Week 35 fixture, plus reports from Inter-Napoli, Roma-Milan and Bologna-Fiorentina.
The action begins today at 11.30 UK time (10.30 GMT) with an intersection of the two races, as Verona chase Europe by hosting bottom of the table Catania.
This afternoon Parma visit Cagliari and Lazio go to Livorno, as yet more teams looking for continental qualification face off against sides in the battle to stay in Serie A.
Chievo also take on Sampdoria, while Torino hope to maintain their form against Udinese.
At 19.45 UK time (18.45 GMT), Atalanta and Genoa face off in a bid to end their disappointing runs.
The round concludes on Monday evening with leaders Juventus taking on relegation strugglers Sassuolo – could there be an upset with the Europa League semi-final on Bianconeri minds?


text:
US Army Sergeant and Wife Portrait in Dress UniformAdd to Cart Add to Lightbox Download
US Army sergeant in Class A uniform with beret and decorations with wife. Both model released. ..Image does not comply with USDOD regulations for advertising use without modification of insignia and decorations.
- Filename
- MSPEH1111-013.jpg
- Copyright
- ©Hans Halberstadt
- Image Size
- 5184x3456 / 4.1MB
2010 army ARMY USAF STOCK PHOTO 27 NOV 11 chris class CLASS A UNIFORM couple COUPLE WIFE HUSBAND WIFE FAMILY Desktop family hans Hans HP 2010 husband MSP MSP111127046 nov photo stock UNIFORM USAF users wife sergeant man men us army released portrait formal dress class a medals decorations rank beret together mary family couple wife husband
- Contained in galleries


text:
We know that the kids love watching television and playing on the computer, Wii, PlayStation and Xbox. We're parents too and we can categorically say that getting them outside for a bit of exercise can be excruciatingly painful at times.
But now, we have found the ultimate solution for all mums and dads! To be honest, we can't believe we didn't think of this before… Instead of persuading the kids to go outside and reap the benefits of playing in the garden, we're just going to use the climbing frame as a punishment!
We're sure they will rant at the rock wall and despise the slide and swinging will be absolutely loathsome! It will also give us parents the opportunity to enjoy a bit of peace and quiet without having to hear the squabbles over console controllers or the television remote. Bliss.
Another benefit of using this form of punishment comes when you've finally let them back through the back door. They'll traipse in with tummies rumbling because they've used up so much energy outside. There will be no whinging about their evening meal – and let's face it, that would be rather welcome (certainly at my dinner table!). Come bedtime, they'll be feeling incredibly drowsy and will sleep through the night without a peep. The next day, they'll feel brighter, bolder and more alert which is always a good mood to be sent to school in!
Come the evening when they get home from school and start squabbling over that TV remote again, repeat yesterday's solution!
Our wooden swing sets are all designed for children aged three to twelve years. All of the Selwood climbing frames we stock come with a 10-year guarantee on the wood, which means that this form of punishment will last throughout childhood!
N.B: In the unlikely event that you have to find an alternative form of punishment because your child ends up enjoying playing outside, Climbing Frames UK cannot be held responsible.


text:
Hei all.........
ANother batch starting...........
This course is designed to enable the students to develop lighting techniques for glamour, beauty and fashion photography. The course syllabus covers lighting techniques for both indoor and outdoor location shoots. There will be sessions where models are hired for the shooting sessions. Participants are required to provide their own film. Model, makeup artist and stylist fees are not included in the course. Students may like to have their images taken during the shoot to be scanned into Kodak PhotoCD and have the chance to use the Societys large format Epson Stylus 7600 professional printer for printing. Upon completion of the course, the students are required to submit a portfolio of 6 pictures for assessment. A course certificate will be issued to students who clear the assessment.
Duration : 6 weeks, 2 hours per session
Course Fee : S$150 (member), S$250 (non-member)
Language : English/Mandarin
Pre-requisite : Basic Photography Course
Instructor : Goh Kim Hui
Course Syllabus
(Students of this course must have knowledge of the use of a 35mm camera)
1. Introduction to the equipment used in fashion photography. Lighting ratio and the effects of soft and hard light.
2. Lighting methods: high key, glamour shots, mood shots. styling and makeup for fashion and glamour photography.
3. Studio fashion shoots with model - face shots (glamour and beauty shots.)
4. Outdoor fashion shoot - shooting with natural light. methods used to modify lighting on location.
5. Studio fashion shoot with model - full-length shots.
6. Critique and course wrap-up.
Interested individuals, do drop us an e-mail to firstname.lastname@example.com
for more details............


text:
Author has written 3 stories for NCIS.
I hold absolutely no disillusions that I could ever do this for a living, but it's nice to send my thoughts out into the internetosphere and think that maybe somebody out there would enjoy reading them.
Reviews and constructive criticism are always appreciated, just remember: it's all in good fun.


text:
Maywood's First Recreational Pot Shop Opens TuesdayThe first recreational marijuana dispensary in the city of Maywood opened Tuesday to major fanfare.
Flames Erupt At Maywood Baseball FieldFirefighters were able to keep the blaze from spreading to a nearby building.
Man Fatally Stabbed By Mother Of His ChildrenAuthorities say a man was stabbed to death in Maywood while arguing with the mother of his children.
Out-Of-Control Car In Maywood Slams Into 2 Pedestrians, At Least 3 CarsThe driver reportedly took off running.
Police Arrest Man In Fresno On Suspicion Of Raping Woman In MaywoodInvestigators are trying to figure out if Martinez is linked to four other sexual assaults in Maywood last month.
Maywood Rape Suspect Arrested Near FresnoA man suspected of raping a woman last weekend in Maywood has been arrested near Fresno, authorities said Saturday.
Man Identified As Maywood Rape Suspect Is At LargeThe victim of the assault was an adult female who was home alone at the time of the attack.
Police In Maywood Searching For Suspect Who Breaks Into Homes To Grope Women, ChildrenThe latest incident authorities said happened about 4 a.m. Saturday morning.
Sexual Predator Allegedly Breaking Into Maywood Homes To Grope WomenPolice asked for the public's help in finding a sexual predator breaking into houses and groping female residents inside their own homes.
EPA Begins $3M Cleanup At Site Of Maywood Magnesium FireThe Environmental Protection Agency has begun a weeks-long cleanup at the site of an explosive industrial fire that forced about 300 people from their homes in Los Angeles County last spring.
State Audit Finds Debt, Poor Oversight In City Of MaywoodA state audit concludes the tiny Los Angeles County city of Maywood faces significant financial problems that threaten its ability to provide services to its residents.
Local Emergency Declared To Help Clean Up Maywood Warehouse FireThe Board of Supervisors declared a local emergency Tuesday in the wake of last week's explosive fire at a Maywood warehouse, saying hazardous levels of magnesium were found in the fire ash.


text:
Quality and craftsmanship at honest prices
Style is about self expression. Build a wardrobe of beautiful, flattering essentials that are both timeless and on trend. A Brisbane based label where everything is designed and made under one roof, using fabrics that are friendly to animals and the environment. They look better, feel better and wear longer. Simple, classic staple pieces that never go out of style. It is my hope that by creating and consuming mindfully, we will discourage the pervasive view of clothing as disposable.


text:
- Age / Gender:
- n/a, Unspecified
- Location:
- Location not disclosed
- Joined:
- 4/12/17
- All Stats >
User Statshot
- Community Stats
Level 5 Blank Slate
Normal Whistle
Ranked as Town Watch
Contact Info / Websites
Latest Art
Recent Game Medals
Kerrigan 5 Points from Kerrigan and the Frog, etc. Medal Stats.
Bitey 5 Points from the Brackenwood series Medal Stats.
Beebo 5 Points from BEEBO Medal Stats.
Zoe 5 Points from XOMBIE Medal Stats.
Tankgirl 5 Points from Newgrounds Medal Stats.
ZONE-tan 5 Points from ZTV News Medal Stats.
Chocolate Man & Cripple Boy 5 Points from Nameless Medal Stats.
Leo & Satan 5 Points from Leo & Satan Medal Stats.
Detective Nwar 5 Points from NWAR Medal Stats.
Edd Gould 5 Points from Eddsworld Medal Stats.
Total Medals Earned: 37 (From 3 different games.)


text:
|( 9 upvotes )
|See...to me, bleak IS funny. I'm just twisted that way...
|Preach on, brother space... preach on!
|Funny comes in different styles, phydeaux. I like this poster, but because it's bleak. Not because it's funny.
|Gives a whole new meaning to the word "Dropcloth" LMAOPMPAC!!
|Remember to lay out some plastic sheeting first Skeeter...no one likes a messy suicide.
|Wow that's depressing. I guess I'll just go kill myself.
|( 8 upvotes )
|( 17 upvotes )
|Ha! Great picture. That's funny. 5L
|Thanks, Sick, I'm glad you like it. Hopefully, you see the message behind it. Always recycle. lol
|Okay you made me LOL 5L's
|( 3 upvotes )
|( 9 upvotes )
|Modern Warfare 3 Bling Super Pro++
Misc info


text:
Nhl 96 [gameboy Game]
Imported 1 year ago
Share this item:
Strap on those helmets and hit the ice for realistic nhl battle. Choose from a complete list of real NHL teams and 1996 rosters, then get set to march toward the Stanley Cup. Compatible with the SNES by the use of the Super Game Boy adapter,


text:
Cut Paste Waterfall Background for Android has been downloaded by 100,000+ users and rated with an average rating of 3.90 from 1,085 reviewers. To download and install Cut Paste Waterfall Background.apk / com.shivashivam.photocutpastewaterfallbg.apk on your device, you can use any of the download methods provided below. If you like this free app, feel free to share it with your friends or leave a comment.
Warning Downloading Cut Paste Waterfall Background APK might result in charges if you are using mobile data to download this free app. To avoid any additional charges while downloading Cut Paste Waterfall Background or any other Andoid apps, always download via Wi-Fi network if possible.


text:
Jaipur, Rajasthan India (PressExposure) April 09, 2011 -- SEO Content Online, a brand in SEO by content is now coming up with yet another festive bonanza for the people of Jaipur, where its two offices are based other than in Colorado Springs, USA.
Since long time, it was felt by the owner of SEO Content Online that still people in India are not much aware of copywriting or content writing services and their role in search engine optimization. In order to promote the use of content in SEO and to help students from all disciplines (especially those from the stream of Master In Arts, Literature) is coming up with excellent job opportunities this year.
SEO content Online is conducting campus interviews with all the best collages of Jaipur to find out hidden talent of writers from the students. When asked from Mr. Swadesh Rohilla, the director of this company and Thoughtful Minds, how he wants to turn up this plan into action, he said.' Well we believe if we want to promote anything then first of all we need to pay emphasis on the ways by which people can be made to know about it. And as we are from content writing and SEO stream we know how to present our thoughts and how we can popularize it worldwide."
It is important to note that SEO Content Online that was started after the establishment of Thoughtful Minds has grown to a very big company and that is just because of the company's fair image and Google's appreciation for ethical ways of promotion that company strictly follows.
In one of the seminars in Jaipur, Mr. Swadesh stated, when we started our first venture Thoughtful Minds for web content and content writing services in Jaipur we were not much sure if we will be able to take this small firm to heights. As the problem was dual- it was difficult to find both clients as well as people to take up the writing job, it was looking like a wrong decision to start up this venture. However with our constant work and full dedication of few of our staff members today we are standing on a position where we are able to have two branch offices in Jaipur, India and one co office in Colorado Springs U.S.A. with over 142 members working for us.
Now in order to help students of Jaipur get benefitted with this high profile, white collar job in I.T. Sector of content writer, SEO content Online and Thoughtful Minds are inviting applications from graduates and post graduates to work on a full time basis. As it is one such stream where students of all streams can apply, (not excluding the students from Mass Communication), we are anticipating good job opportunity for the students of Jaipur.
When asked how the company initially is fulfilling the requirements of writers, Mr. Swadesh said other than students or pass outs from convent schools and collages we have doctors (M.B.B.S) and engineers working on a part basis for us. So thankfully we are able to serve our clients worldwide in a proper manner.
When asked why Mr. Swadesh selected this line of copywriting instead of natural stone business where he was also very much successful when he was working for Germany he said, well why to do things that are easy, it's better to go for difficult ones as they make you more dynamic.


text:
Hi Meltingpot Readers,
Sometimes I really love my job as a writer and cultural thinker. It sounds very nebulous to describe myself that way and granted, there's not a lot of money in the business of thinking and talking, but it does provide some wonderful opportunities. Like last night. I had the honor of playing 'hostess' at a literary dinner celebrating this wonderful new book called, Soul Food:The Surprising Story of an American Cuisine, One Plate at a Time. The menu for the meal was pulled from recipes in the book and was so delicious. Meanwhile, the author, Adrian Miller, entertained the audience with his fascinating story of the origins of Soul Food as well some of the myths and misunderstandings often associated with this under appreciated cuisine. The book is well-written and engaging as well as chock full of fascinating information. I highly recommend it for any foodie, soul food scholar and/or Black culture enthusiast. And last night's dinner was so good, I highly recommend the restaurant, Geechee Girl Rice Cafe, if you happen to be in Philly.
Here are some photos of our delicious meal. Try not to drool over dinner.
What do you think of when you hear soul food? I'm listening.