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Housing and homeless resources
For questions regarding eligibility including how to apply, click here.
For general questions and inquiries please email firstname.lastname@example.com or contact 507-431-5780
Emergency assistance
MNPrairie offers emergency assistance if you are experiencing a crisis that may result in homelessness or loss of utilities. Please refer to our emergency assistance page for more information
If you don't qualify for emergency assistance, you may be able to get similar help from another homeless prevention program. Eligibility for these programs is determined by your residential address, income and specific housing circumstances. Before you visit one of these organizations in person, call ahead to get information on when you can apply and whether funds are still available for the month.
Secure and safe housing for all ages and people is a basic need. Dodge, Steele and Waseca take part in the Coordinated Entry system to support people who may be at risk of becoming homeless or who are currently homeless. To access homeless support services, follow the steps below:
- If you are currently working with a MNPrairie case manager, start by contacting your case manager.
- If you do not currently work with a MNPrairie case manager, contact one of these agencies for a homeless assessment for services.
Coordinated entry
Is a process developed to ensure that all people experiencing a housing crisis have fair and equal access and are quickly identified, assessed for, referred, and connected to housing and assistance based on their strengths and needs. It is a priority list for homeless services and programs.
For more information about Coordinated Entry and continuum of care, visit Three Rivers Continuum of Care.
Agency: Semcac
Entry point: Individuals and families with children
Phone number(s): Steele: 507.451.7134 - Dodge: 507.634.4350
Agency: Partners for Affordable Housing
Entry point: Families with children and homeless adult females. This agency operates a shelter.
Phone number: Waseca: 507.387.8189
Agency: Minnesota Valley Action Council
Entry point: Everyone
Phone number: Waseca: 507.835.8240
Agency: Lutheran Social Services
Entry point: Youth who are 24 years old and younger
Phone number(s): Dodge & Steele: 507.316.8273 -
Waseca: 507.381.6670
Agency: MN Assistance Council for Veterans
Entry point: Veterans
Phone number: Dodge, Steele & Waseca: 507.345.8258
Agency: Committee Against Domestic Abuse
Entry point: Victims of domestic and sexual violence
Phone number: Waseca: 507.835.7828
Agency: Women's Shelter Inc.
Entry point: Victims of domestic and sexual violence
Phone number: Dodge: 507.634.6070 ext. 160
Agency: MNPrairie
Entry point: Any population
Contact information: firstname.lastname@example.com for a referral
Housing resources
Housing Crisis - Who to Call
- Minnesota landlord and tenant rights
- Legal assistance for renters and homeowners
- Other Housing programs offered by MNPrairie:
For general questions pertaining to housing resources within the MNPrairie region, you can reach the housing team.


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Kim Lewis, an entrepreneur, is now in the rare club of the 1% of black-owned women wine companies.
Lewis, an avid traveler, often enjoyed tasting different wines from various regions during her travels. She often joked with friends that she should start a wine company. Those jokes became a reality when Lewis decided to create Ole' Orleans Wines.
"Being a black woman with a wine company makes me feel on top of the world," Lewis told Travel Noire.
Initially, Lewis reached out to major labels hoping they could help her start a wine company, but the response was lacking. She then later learned another option would be to hire a consultant for around $20,000 to help her navigate the complicated process.
"When I learned how much money it cost, I said let me see if I can complete this paperwork on my own," Lewis said. "I was able to do it, submit it, and get approved for my license on the first try."
The next step in the process was to find a vineyard to partner with and procure the necessary grapes. Lewis' first choice was to look in Napa Valley, California, but that proved to not be cost-effective. She also looked at a vineyard in Texas, before her ultimate choice in Louisiana.
From there, Ole' Orleans Wines was born. According to the website: "Ole' Orleans Wines, a tribute to New Orleans and it's historic legacy. Richly flavored, New Orleans very own handcrafted wines, created in Louisiana. La Nouvelle-Orléans, located along the Mississippi River is well known for its Creole and Cajun roots. Displaying its French and Spanish heritage through an elegant decay."
Two white blends are currently available, Ole Carollton and Tchoupitoulas, and can be found in the local New Orleans market and on the Tasting Room website. Lewis plans to release 10 additional blends in the future.

subset: commoncrawl


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To safely remove moles (cancerous, non-cancerous, or inconclusive), a procedure must be performed by a professional, and at Atlas Dermatology, Dr. Trent Massengale and Dr. Joshua Pitre are prepared to assist you in the process of removing any unwanted or precarious moles. Visit one of our two convenient locations in Baton Rouge or Prairieville, LA, to begin discussing your options to remove your moles.
Keeping An Eye On Your Moles
Pigmented growth clusters on the skin are just moles, but it's important to know if it's cancerous or not. By monitoring your moles, you can notice any unusual changes in the size, shape, color, or outline. These are good indicators to have your mole(s) checked to catch potential signs of cancer early. Following the ABCDE guidance from the American Academy of Dermatology Association is recommended to properly identify a cancerous or non-cancerous mole.
(A)symmetry—Is one half of your mole dissimilar to the other?
(B)order—Is the outline of your mole abnormal or irregular?
(C)olor—Does your mole show a multitude of tones?
(D)iameter—Cancerous moles are often compared to the size of a pencil eraser. So when observing moles on your skin, be sure to consider if your mole is akin to the size of a pencil eraser.
(E)volving—Is your mole changing in shape compared to your other moles?
Diagnosing Non-Cancerous and Cancerous Moles
Recognizing any abnormalities that develop should promptly be examined by your Baton Rouge and Prairieville, LA, Atlas Dermatology professional. The first step in checking any moles of concern is to have a biopsy done to properly identify the mole, and if it needs to be surgically removed.
Dr. Massengale and Dr. Pitre can help discern the seriousness of your mole(s) by presenting you options of treatment or removal. For peace of mind, book an appointment in Baton Rouge and Prairieville, LA, with Atlas Dermatology by calling (225) 313-4560.


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Due to their exceptional quality, performance and style, Rolex is one of the most iconic, respected and coveted brands in the world. Unfortunately, their popularity also means that there are endless fake Rolex watches on the market, some of which can be very convincing to the average buyer. The good news is that Rolex utilizes many anti-counterfeit measures in their timepieces and once you can identify those, you'll know how to spot a real Rolex among fakes. While it's easy to tell that a $25 dollar "Rolex" is fake, we'll show you how to spot even the most high end of fake Rolex's.
How to Spot a Real Rolex
While all Rolex collections have their own signature features, below are general characteristics that will help you spot a real Rolex. See our experts' top tips for spotting a fake Rolex:
- Everything on the dial will be perfectly printed, even when under extreme magnification. Any uneven print or misspellings are a big red flag.
- Rolex's signature five-point crown will be at 12 o'clock.
- Current Rolexes have their serial number printed on the inner bezel ring at 6 o'clock with the word Rolex printed multiple times in the remaining space.
- If the watch features a date window, the date will automatically change at exactly midnight. A fake Rolex will have a slow date change.
- Also, if the watch has a date window, it will be covered by a cyclops lens which works to magnify the date by 2.5 its normal size. Fake Rolexes either have a flat cyclops, weak magnification or are offset from the date.
- Rolexes made after 2002 will have a small five-point crown laser etched within the crystal at 6 o'clock. Typically, you will need very good lighting and a magnifier to see the etching.
- Real rolexes are heavier than their fake counterparts because genuine Rolexes are made with high quality materials including Oystersteel, 18kt gold and platinum.
- Rolex's signature five-point crown will be on the watch's winding mechanism, known as the crown. The exact number of dots below the crown will vary depending on the specific model.
- Genuine Rolex watches have smooth case backs that are free from engravings and decorations, expect for the Deepsea Dweller.
- All Rolex watches have waterproof Oyster Cases except for the Cellini which is a dress watch.
- Rolex's do not have transparent case backs, except for the Prince Cellini collection.
- Older Rolex's will have the serial numbers printed between the lugs at 6 o'clock. The watch's bracelet needs to be removed in order to see these numbers and should only be removed by professional watchmakers.
- The watch's model number will be printed between the lugs at 12 o'clock and the bracelet will also need to be removed to see it. Fake Rolexes will be missing these numbers, have numbers that do not reflect their actual model or be faintly printed.
- Rolex bracelets are only made in Oystersteel, 18kt gold, Oystersteel and 18kt gold or platinum.
- All Rolex bracelets have their signature five-point crown on the clasp and the straps have the five-point crown on the buckle.
- The bracelets will have the Rolex's arabesque hallmark printed inside the folding clasp.
- Rolex movements are always mechanical with the exception of the Oysterquartz Rolexes made from 1970's into the 1990's and certain Cellini models.
The best way to buy a real, pre-owned Rolex is to buy from reputable sellers. With myGemma, you never have to worry about fake Rolex watches as our team of experts authenticate every one of our products to guarantee their authenticity and exceptional quality. We are proud of our A+ Better Business Bureau rating and committed to selling expertly authenticated, pre-owned luxury watches and jewelry.
Shop a wide array of pre-owned luxury watches and designer jewelry from your favorite brands at fantastic prices. Affording your perfect Rolex is easier than ever with myGemma - enjoy up to 80% off retail prices, easy monthly payments with our partner Affirm and free US shipping.


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An apical brush border is a characteristic of many mature epithelia. This dynamic structure consists of dense microvilli supported by F-actin bundles that protrude into the apical cytoplasm, where they are crosslinked by spectrin and myosin II to form the terminal web. Little is known about the terminal web, through which vesicles transit to and from the apical membrane. Analysis of mutations in βHeavy-spectrin, the Drosophila brush border spectrin, reveals that this protein is necessary for the maintenance of Rab5 endosomes in the midgut. As a consequence, an apical H+ V-ATPase that is probably responsible for lumenal acidification is lost both from the brush border and Rab5 endosomes. Epistasis tests indicate that βHeavy-spectrin is required during endocytosis after Dynamin and before Rab5-mediated endosome activities. These data are consistent with the location of spectrin in the terminal web, and suggest that this molecule is required for correct sorting decisions at the early endosome.
A brush border is characteristic of many mature transporting epithelia, and is conserved across vertebrate and invertebrate species (Bement and Mooseker, 1996; Bonfati et al., 1992). It can be divided into three subdomains: the microvilli, where active transport proteins reside; the exocytic/endocytic zone at the base of the microvilli, where protein delivery and recovery occur; and the terminal web in the apical cytoplasm, where the F-actin crosslinking proteins spectrin and non-muscle myosin II form a dense meshwork between the F-actin bundles protruding from each microvillus (Bement and Mooseker, 1996). Remarkably, whereas brush border components are rapidly turned over, this structure remains ordered and stable (Stidwill et al., 1984).
The development, structure and composition of the microvillar domain have been well characterized, and the delivery and operation of individual transporters and some cytoskeletal components are well understood (Bement and Mooseker, 1996). By contrast, very little is known about the function of the terminal web. This meshwork is an extremely dense and potentially formidable barrier to vesicle transport (Hirokawa et al., 1983b), and probably utilizes unique variations on exocytic and endocytic processes. For example, the close juxtaposition of a spectrin-based membrane skeleton (SBMS) to a membrane has long been thought to antagonize the endocytic machinery passively by excluding other coats, and proteolytic removal (Kamal et al., 1998) or network disassembly makes way for vesicle formation. However, the SBMS in the terminal web is not so closely juxtaposed to the membrane at the endocytic zone, suggesting that here it might not require removal to permit endocytosis. In addition, the high degree of order within the brush border cytoskeleton (Hirokawa et al., 1983a) makes is hard to envision how F-actin polymerization could assist in the initial departure of endocytic vesicles (Merrifield, 2004). Nonetheless, the dynamic nature of the brush border requires that vesicular transport across the terminal web must be vigorous, and many vesicles are seen in this region (Hirokawa et al., 1983a; Hirokawa et al., 1983b). It therefore seems likely that novel mechanisms of vesicle processing might exist in the terminal web. Vesicles are seen to associate with spectrin during their traverse (Hirokawa et al., 1983a), suggesting a role for spectrin in transport at the brush border.
Spectrins are rope-like cytoskeletal proteins largely comprising spectrin repeats of ∼106 amino acids that form heterotetramers of α and β subunits; the Drosophila genome encodes a single α and two β subunits (β-spectrin and βHeavy-spectrin; βH). (αβ)2- and (αβH)2-spectrin are basolaterally and apically polarized, respectively. Several modular protein-protein and protein-lipid interaction motifs within each spectrin isoform confer specific functionalities upon these molecular scaffolds. Spectrins form two-dimensional networks, in conjunction with F-actin, that are associated with various membranes and have a wide array of cellular functions (Bennett and Baines, 2001; De Matteis and Morrow, 2000). Mutations in all non-erythroid spectrin genes examined to date cause substantial or complete lethality and pleiotropic phenotypes, demonstrating their essential role in cell structure (Bennett and Baines, 2001). However, the precise roles of most non-erythroid SBMS have yet to be fully defined.
Recently, we demonstrated that over-expression of the C-terminal domain of βH in Drosophila salivary glands leads to the sequestration of dynamin and the inhibition of endocytosis (Williams et al., 2004). These data suggest that the apical SBMS is interacting directly or indirectly with endocytic components and that the SBMS has an active role in protein turnover. We therefore sought to test the hypothesis that brush border spectrin is involved in endocytosis (Hirokawa et al., 1983a) by examining the role of fly βH in protein turnover specifically in the terminal web of fly middle midgut. Several lines of evidence support the existence of a typical brush border organization in insects. In Manduca sexta, both biochemical and ultrastructural evidence support this notion (Bonfati et al., 1992). Similarly, in fly cells that have a well-developed brush border (Filshie et al., 1971), it is subtended by the apical SBMS (Thomas et al., 1998). Moreover, fly brush borders contain both unconventional myosin 1 isoforms typical of this structure: myosin IA in the terminal web, and myosin 1B in the microvilli (Morgan et al., 1995).
We now report that acid secretion and nuclear positioning in Drosophila cuprophilic cells (CCs) are dependent on the apical SBMS. Furthermore, we demonstrate that these cells contain defects in their early endosomes, and that loss of acid secretion is correlated with the dispersal of an apical H+ V-type ATPase and Rab5 from both the apical membrane and the early endosome system. Genetic interaction experiments suggest that the SBMS is required for correct sorting decisions at the early endosome. These data demonstrate that apical spectrin in the terminal web has a role in establishing the level of brush border proteins through the regulation of the early endosome recycling pathway.
karst mutant CCs are polarized and exhibit a subset of α-spectrin mutant phenotypes
The Drosophila middle midgut CCs possess a deeply invaginated apical domain, with a prominent brush border (Fig. 1A). A pore connects the apical invagination to the gut lumen. (Fig. S1, supplementary material, contains a more detailed guide to the complex geometry of these cells that may aid the reader in interpreting the micrographs in this paper.) CCs are arranged along the middle midgut in a checkerboard pattern interspersed with the complementarily shaped interstitial cells. The CCs mark a region of intense gut acidification that disappears if the fate of the middle midgut cells is changed (Dubreuil et al., 2001). Since the midgut is a secondary epithelium, these cells have no zonula adherens and rely exclusively on smooth septate junctions (SJs) for lateral adhesion and as a barrier to permeability (see Filshie et al., 1971; Tepass, 1997).
As in other epithelia, βH and α-spectrin co-localize at the apical domain of CC and interstitial cells, whereas β- and α-spectrin co-localize basolaterally (Fig. 1B-D) (Dubreuil et al., 2001; Dubreuil et al., 2000; Lee et al., 1993; Thomas et al., 1998). Additionally, α-spectrin is enriched in the SJ at the pore of the CC apical invagination (Fig. 1B-D, inset), where it co-localizes with the SJ marker Coracle (Fig. 1G-I, and insets). βH abuts but does not overlap with Coracle (Fig. 1J-L, and inset), identifying the apical margin of the SJ as the apical-lateral boundary in this cell type.
Previous analyses have shown that mutants for the common α-spectrin subunit have CCs with an enlarged pore and mispositioned nuclei, and exhibit an acidification defect in the middle midgut (Dubreuil et al., 1998; Lee et al., 1993). By contrast, mutants for basolateral β-spectrin exhibit only the enlarged pore phenotype (Dubreuil et al., 2000). Together, these results imply a role for the apical SBMS in nuclear positioning and gut acidification. We therefore examined karst mutant guts that lack wild-type βH. Staining for α-spectrin in karst mutant guts confirms that α-spectrin is selectively lost from the apical membrane (Fig. 1E-F). This indicates that the karst mutation disrupts only the apical SBMS in this cell type, and supports earlier findings that (αβ)2 tetramers do not substitute at the apical domain in the absence of βH (see Zarnescu and Thomas, 1999). As predicted, we find that nuclei are often mispositioned in karst mutant CCs. Instead of their central wild-type location (Fig. 1E), 13-71% of the nuclei in each gut are found at the outer edges of the cell, causing a bulge near the pore (Fig. 1F). The variable expressivity of this phenotype suggests that (αβH)2-spectrin has only an accessory role in nuclear positioning, or normally prevents stochastic shifts in position during gut peristalis and/or locomotion. We also find that gut acidification is defective in karst mutants (see below) but that CC pore sizes are normal. Together, these results show that the structural and functional defects caused by loss of the common α-spectrin are thus the sum of the disruption of the individual apical and basolateral SBMS.
Acid secretion by CCs requires the apical SBMS
Using Bromophenol Blue (BPB) as a pH indicator, it was shown that α-spectrin, but not α-spectrin, mutants lack the ability to acidify their guts (Dubreuil et al., 1998). Third-instar karst larvae were similarly fed BPB and a statistically significant decrease in overall acidification was apparent (Fig. S2, supplementary material). However, we found this assay to be susceptible to misinterpretation near the color transitions for BPB and ineffective at cellular resolution. Acridine Orange (AO), which undergoes a pH-dependent shift in emitted fluorescence, both provides a more consistent assay and is amenable to high-resolution microscopy. AO has been widely used as a marker of cellular compartments with a negative charge, such as lysosomes. Near neutral pH, light is primarily emitted at green wavelengths, but at an acidic pH the fluorescence is red-shifted.
In third-instar wild-type guts, red-shifted AO is found in the lumen and in the CC apical invaginations (Fig. 2A, arrows, and E), with the apical invaginations often exhibiting a greater red shift than the lumen. In individual CCs, we frequently see plumes of red-shifted fluorescence streaming out of the apical invagination into the gut lumen passing through a ring-shaped area of green fluorescence in the intersitial cells (Fig. 2A, arrowheads, B-D). The origin of this ring is unknown, but appears to be an internal feature near the boundary of the intersititial cell and CC. Since the CC apical invaginations are the most acidic regions we observe, these data support the notion that they, and not interstitial cells, are the source of midgut acid.
AO staining in third-instar karst mutants reveals a consistent loss of acidification. The typical karst mutant gut lumen demonstrates very little, if any, red shift, indicating that the pH is significantly higher than in wild type (Fig. 2, compare A with G and E with F). The apical invaginations are typically devoid of fluorescence and acid plumes are no longer observed (Fig. 2G, arrows). These data indicate that the loss of acidification is associated with defects in the apical SBMS, and reinforce the hypothesis that the CC apical invagination is the source of acid for the midgut. In addition, there does not appear to be a ring of green fluorescence at the pore connecting the apical invagination and lumen, suggesting that either karst mutant interstitial cells are unable to produce it, or that this cell type only does so in response to acid secretion by adjacent CCs.
Staining with AO reveals an additional phenotype in karst mutant CCs. Faint acidic vesicles that are occasionally seen in wild-type cells are prominent and numerous in karst guts (Fig. 2G). This is suggestive of a defect in protein trafficking in karst mutants that leads to an increase in the number of lysosomes.
The H+ V-type ATPase is mislocalized in karst mutant CCs
The proton pump responsible for midgut acidification has not yet been identified. However, the preeminent candidate is a H+ V-type ATPase (V-ATPase), a multi-subunit complex thought to energize all insect epithelia (Wieczorek et al., 2000). In Drosophila, just one candidate V-ATPase exists in the genome, and has been localized to the apical membranes of Malpighian tubules, where it drives fluid secretion (Dow et al., 1994). We used the C23 antibody raised against Manduca V-ATPase subunit B (97% identical to Drosophila subunit B) (Dow, 1999) to localize the homologous fly protein. We confirmed that this antibody detects a single band of the predicted size in fly extracts (57 kDa, Fig. S3, supplementary material).
Immunofluorescent staining of wild-type third-instar Drosophila midguts with the V-ATPase antibody reveals apically concentrated staining at the base of the microvilli (Fig. 3C, arrows in I) and in large cytoplasmic concentrations that are found in both CC and interstitial cells (Fig. 3C, arrowheads in I). Staining is not seen in the microvilli because of the high density of pumps in the membrane, which excludes antibodies without antigen retrieval procedures (Wieczorek et al., 2000). In our hands, sufficient staining is seen for us to follow the fate of the pump at the apical membane. Apical membrane accumulation of this protein is not evident in interstitial cells, again consistent with the CCs being the source of acid. In karst mutants, the V-ATPase is undetectable at the apical surfaces of CCs, and is no longer found in a cytoplasmic compartment. Instead, only a diffuse cytoplasmic staining is seen (Fig. 3D-F, J-L). This perturbation indicates that the stability and/or subcellular location of this pump is dependent on an intact apical SBMS, and is consistent with this V-ATPase being the primary proton pump for gut acidification.
The early endosome is disrupted in karst mutants
In order to determine the nature of the V-ATPase-containing compartments, we performed immunofluorescent co-localizations for V-ATPase and markers of various intracellular compartments. Neither Lava Lamp, a Golgi marker (Sisson et al., 2000), nor Rab11, a marker of the recycling endosome (see Jankovics et al., 2001, and references therein), co-localizes with the V-ATPase-labeled cytoplasmic compartment (Fig. 4A-C, D-F, respectively), although Rab11 does exhibit a partial overlap with the V-ATPase in the terminal web region. However, Rab5, an early endosome marker (Wucherpfennig et al., 2003), co-labels the V-ATPase-positive compartment (Fig. 4G-I). Rab5 is also found at the apical surface and in several smaller vesicular structures, consistent with its known roles in endocytosis and recycling.
To refine further the identity of the Rab5-positive endosomal compartments, we performed an additional immunolabeling experiment using larvae expressing the UAS-2xFYVE transgene using the mex-GAL4 driver, which expresses GAL4 in first- and second-instar middle midgut. The 2xFYVE protein is a widely used marker that specifically labels phosphatidylinositol 3-phosphate [PtdIns(3)P]-positive early endosomes, reflecting the activation of phosphoinositide 3-kinase (PI 3-kinase) by Rab5 (Wucherpfennig et al., 2003). This experiment further subdivides the CC early endosomes into three populations of Rab5-positive compartments: (1) those with Rab5 alone found near the apical membrane; (2) those co-labeling with Rab5 and 2xFYVE found in the cytoplasm; and (3) those with Rab5 and the V-ATPase found near the basal membrane at the pore (Fig. 4J-M). No triple labeling of any single compartment was observed. These data indicate that a major pool of the V-ATPase resides in an early endosomal compartment that is PtdIns(3)P negative. An additional observation is the labeling of the entire basolateral membrane by the 2xFYVE construct whereas it is not detected at the apical domain. Such an absolute segregation of a membrane lipid along the apicobasal axis has, to our knowledge, not been documented before.
The presence of V-ATPase in a Rab5-positive endosome suggests that this represents a pool of the pump that is recycling from the apical membrane. To test this hypothesis, we examined the distribution of the V-ATPase in a temperature-sensitive Shibire2Ts (Shi2Ts) background where dynamin can be inactivated by raising the temperature of the larvae. After 1 hour at the non-permissive temperature, we found that both Rab5 and V-ATPase strongly accumulated at the apical membrane (see Fig. 6, compare A,B with E,G). This demonstrates that the pump is actively internalizing and that dynamin is not required for its delivery to the apical membrane. In addition, the intensity of Rab5 staining at the V-ATPase-positive endosomes is noticeably decreased (Fig. 6E-G, insets), suggesting that the inactivation of dynamin decreases the movement of Rab5 to this compartment, consistent with the notion that the pump is recycling from the apical membrane to this location.
In karst mutant third-instar larvae, the endosomal structures that stain for V-ATPase are lacking. To determine if this compartment is still present, Rab5 antibodies were used to stain karst mutant third-instar midguts. This revealed that no structures corresponding to Rab5-positive endosomes are detectable in any mutant cells. Instead, only low-level diffuse cytoplasmic staining of Rab5 is seen, resembling the mis-distributed V-ATPase (Fig. 4N-P). By contrast, Lva and Rab11 distributions appear normal in karst guts (Fig. 4Q-S).
To examine the etiology of this phenotype, we examined earlier larval stages. Staining karst mutant first-instar larvae with Rab5 and V-ATPase antibodies reveals an intact early endosome (Fig. 5, compare A-C with D-F). Moreover, AO feeding experiments indicate that the lumen is acidified in both wild-type and karst mutant guts at this stage (M.D.P. and C.M.T., unpublished data). In the second instar, the Rab5 compartment has disappeared in some of the karst mutant guts, although the majority still display a near wild-type appearance (Fig. S4, supplementary material, compare A-C with D-F). Together, these data indicate that karst midgut cells contain a selective defect in their recycling pathway that leads to the loss of early endosomes around the transition from second to third instar.
The loss of the early endosome in karst mutant cells is reminiscent of the phenotype induced in neurons by expressing a dominant-negative Rab5 (Rab5S43N), which is constitutively bound to GDP (Wucherpfennig et al., 2003). Using the mex-GAL4 driver line, we expressed Rab5S43N in the midgut. This induced a marked loss of Rab5 from V-ATPase-labeled endosomes in both CC and interstitial cells in the first instar (Fig. 5G-I). By the second instar, the V-ATPase and Rab5 no longer co-localize (Fig. 5J-L). However, V-ATPase-labeled structures remain prominent both in CCs and in interstitial cells, whereas the Rab5 antibody now labels a distinct compartment near the CC pore. This continued presence of a Rab5-labeled compartment upon expressing Rab5S43N in the first-instar larval midgut differs from its effects in the fly nervous system, where a total loss of particulate Rab5 staining was observed, suggesting the disintegration of the early endosome (Wucherpfennig et al., 2003). This might result either from differing levels of expression from the different drivers or from tissue-specific responses. Although Rab5 is thought to have roles in both the formation of endocytic vesicles and their fusion at the early endosome (Pfeffer, 2003), we do not observe an accumulation of V-ATPase at the apical membrane when Rab5S43N is expressed in wild-type cells. Thus, our results suggest that Rab5-mediated events at the early endosome are most sensitive to inhibition in this tissue.
In order to test whether βH and Rab5 operate in the same pathway, we expressed Rab5S43N in a karst mutant background and examined early larval stages where karst-induced loss of Rab5 has not yet occurred. Strikingly, the loss of βH prevents Rab5S43N-induced dissociation of Rab5 from the V-ATPase-labeled compartment in both first and second-instar larvae (Fig. 5M-O and Fig. S4J-L, supplementary material, respectively). This epistatic relationship strongly suggests that Rab5 and βH operate in a common pathway that is required for endosomal integrity but that βH is upstream of Rab5 and that the karst mutation limits the loss of endosomal identity induced by Rab5S43N.
To determine more precisely the stage at which βH-spectrin functions in the recycling pathway, the Shi2Ts allele was employed to inhibit endocytosis. In third-instar Shi2Ts/Y larvae at the permissive temperature, the distribution of Rab5 and V-ATPase appears normal (Fig. 6A-B); whereas Rab5 puncta and a broad band of V-ATPase signal became concentrated at the apical membrane at the restrictive temperature (Fig. 6E-G). In addition, Rab5 staining at the V-ATPase-labeled endosome is reduced (compare insets in Fig. 6E,G with Fig. 4G,I), indicating it is not being replenished.
The Rab5 and V-ATPase staining patterns in third-instar Shi2Ts/karst double-mutant larvae (Shi2Ts/Y; kst2/Df(3L)1226) at the permissive temperature are identical to karst mutants - weak and diffusely cytoplasmic with no endosomal concentration (Fig. 6C-D; compare with Fig. 3 and Fig. 4N-P). At the restrictive temperature, Rab5 reappears in numerous cytoplasmic puncta with a strong accumulation near the apical membrane, particularly in the interstitial cells (Fig. 6J,M). Also at the restrictive temperature, the V-ATPase staining sometimes becomes concentrated at the apical domain, but remains diffuse in most cells (Fig. 6H). A lower level of apical pump accumulation is to be expected in the karst mutant background, since the endosomal pool is gone and its delivery is presumably dependent on de novo synthesis of protein. These data indicate that we are achieving a significant inhibition of endocytosis. In cells that have accumulated apical V-ATPase, it does not co-localize with any Rab5-positive vesicles, and no staining typical of the large V-ATPase-positive endosome is evident (Fig. 6H-J). Thus, Shi2Ts is epistatic to karst at the plasma membrane. Together with the previous experiment combining karst and Rab5S43N, our results are fully consistent with a role for βH after dynamin pinches off endocytic vesicles but before Rab5-mediated endosome fusion occurs.
Here, we demonstrate that βH-spectrin in the terminal web has a role in Rab5-mediated endosomal recycling. Specifically, we report that karst mutant cells of the Drosophila midgut specifically lose all Rab5-positive endosomes at around the transition from second to third instar, whereas other endomembrane compartments remain intact. This defect results in the loss of functional H+ V-ATPase from the apical membrane and a failure to acidify the larval gut lumen. This phenotype correlates with an increase in the number of Acridine Orange (AO)-accumulating acidic vesicles, suggesting that late endosomes/lysosomes become more numerous, and that recycling/degradation decision-making is disrupted in the absence of spectrin. This is the first function ascribed to spectrin in the brush border, and represents a novel role for the F-actin cytoskeleton during endocytosis that might be a functional specialization of brush border organization.
Polarization of the membrane skeleton reflects specialized roles
In primary epithelia (αβ)2- and (αβH)2-spectrin are basolateral and apical, respectively, and this characteristic distribution is also evident in the midgut (Fig. 1B-D) (Dubreuil et al., 2000). Here, detailed examination of loss-of-function mutations was performed in these cells for all three spectrin loci and we conclude that the apical and basolateral SBMS have distinct and separable functions. Thus, mutation of the common α-spectrin chain causes defects in nuclear positioning, acid secretion and pore size (Dubreuil et al., 1998; Lee et al., 1993). Defects in pore size and Na+K+ ATPase localization have been ascribed to basolateral β-spectrin (Dubreuil et al., 2000), whereas defects in nuclear localization and acid secretion arise from loss of apical βH-spectrin (shown here). In agreement with previous studies in primary epithelia (Zarnescu and Thomas, 1999), apical spectrin is not required to establish the apicobasal axis and no evidence of redundancy between the two SBMS is evident.
βH-spectrin is required for acid secretion
Drosophila CCs define a region of intense gut acidification. Several lines of evidence indicate that it is this cell type that secretes protons. First, our observations that AO accumulates in the lumen of the CCs and is most protonated in the pores exiting these cells (Fig. 2A-D) provides direct evidence that this cell type is responsible acidification of the lumen. Second, proton-secreting cells in insect epithelia are energized by a H+ V-type ATPase (V-ATPase) located at their apical surface (Wieczorek et al., 2000), and we have localized the Drosophila V-ATPase to the apical surface of CCs (Fig. 3C,I), but not the interstitial cells. Furthermore, in karst CCs, where the gut lumen is no longer acidified (Fig. 2F,G, Fig. S2, supplementary material), this V-ATPase is no longer detected at the apical membrane (Fig. 3F,L) and AO no longer accumulates in their apical invaginations (Fig. 2F-G), correlating the presence of this pump with proton secretion. These data are supported by reports demonstrating that gut acidification is dependent upon the differentiation of the CC and interstitial cell region (Dubreuil et al., 2001), and that the development of a deeply invaginated apical domain in CCs is characteristic of oxyntic cells from other animal systems (Dow, 1992; Forte and Yao, 1996).
An additional reservoir of V-ATPase is seen in a subset of the Rab5-positive endosomes that do not contain PtdIns(3)P (Fig. 4J-M). The apical enrichment of V-ATPase when dynamin is inhibited (Fig. 6E-F) indicates that this pump is actively recycling. This situation is reminiscent of pumps found in some vertebrate cells (e.g. Forte and Yao, 1996; Pastor-Soler et al., 2003), where V-ATPases also recycle between internal compartments and the apical membrane. Definitive evidence that this is indeed the V-ATPase responsible for acid secretion must await future analyses. Nonetheless, this pump serves as a useful maker to assess the effects of karst mutants on the endomembrane system.
βH-spectrin regulates protein recycling through the early endosome
karst mutant midgut cells accumulate numerous and prominent AO-labeled compartments (Fig. 2G). These vesicles are often red-shifted, which is indicative of a low pH and thus the presence of more-numerous late endosomes or lysosomes. This suggests that loss of (αβH)2-spectrin has consequences for membrane trafficking outside of the terminal web region. This is further corroborated by the observation that karst mutant cells in third-instar larvae not only lack V-ATPase at the apical membrane but also the endosomal pool of this enzyme and any detectable Rab5-positive cytoplasmic compartments (Fig. 4N-P). This extensive perturbation of Rab5 endosomes is highly selective since markers for recycling endosomes (Rab11) and the Golgi apparatus (Lava Lamp) are still present in a wild-type distribution (Fig. 4Q-S). The loss of Rab5-positive compartments appears to occur around the transition from the second to third instar, since Rab5 and V-ATPase exhibit a normal distribution in first-instar larvae (Fig. 5, compare A-C with D-F), are sometimes absent in (presumably late) second-instar larvae and are uniformly absent by the third instar (Fig. 4N-P). This timing correlates with the onset of a major growth phase during which the third-instar larva will more than quadruple its mass (Ashburner, 1989). The cells of the gut also grow to accommodate this increase in size and we speculate that the run down of the recycling system in karst mutants at this time results from an underlying weakness in endosome dynamics coupled with this rapid growth, rather than a complete block in one step. This effect cannot be the result of maternal perdurance, because the interaction between the karst mutation and dominant-negative Rab5 activity reveals this defect during the first instar (see below).
The selective perturbation of one intracellular trafficking pathway arising from the loss of spectrin in the terminal web is not an obvious prediction from its location (Hirokawa et al., 1983a). However, the results of the epistasis experiments are consistent with this position between the apical membrane and the endosome (Fig. 7). In karst mutant third-instar larvae, the apical accumulation of Rab5 and sometimes V-ATPase when dynamin is inhibited (Fig. 6H-J) demonstrates that βH is not required for the delivery of V-ATPase or the recruitment of Rab5 during endocytosis, and places spectrin function downstream of initial internalization events. This result also indicates that Rab5 is still present in the cells in the absence of βH. However, in first- and second-instar karst mutant larvae, Rab5S43N disruption of Rab5 localization to endosomes is abrogated. This places βH function upstream of vesicle fusion and/or other events at the early endosome. Although it remains possible that βH resides undetected at the early endosome, it is striking that our data reflect the location of spectrin in the terminal web in relation to the exit of newly internalized vesicles - between the microvilli where dynamin pinches off vesicles, but above the apical cytoplasm where Rab5 must be active for endosome fusion. Immunolocalization of spectrin in the mouse terminal web in quick-freeze deep-etch specimens reveals numerous examples of contact with newly endocytosed vesicles (Hirokawa et al., 1983a), an observation that is also consistent with this hypothesis.
The early endosomal system is a dynamic network of sorting organelles that is maintained by localized activation of specific Rab GTPases and requires cooperative interactions within and between its constituent domains (Seabra and Wasmeier, 2004). Thus, a wholesale redirection of the sorting process would have severe consequences for endosomal maintenance. The epistatic relationship of βH, dynamin and Rab5 mutations, together with the superabundance of acidic vesicles in karst mutant cells that indicates increased lysosomal trafficking, suggest the hypothesis that brush border spectrin `processes' newly internalized proteins to make them competent to enter the recycling pathway at the early endosome (Fig. 7). Thus, in the absence of spectrin, the diversion of large quantities of protein and membrane to the lysosomal pathway, coupled with the interlinked nature of the endosomal system, would result in the run down of the entire Rab5 network. An attractive hypothesis in this scenario that would satisfy the observed location of spectrin in the terminal web, yet the effects of its loss are seen at the endosome, is that spectrin might be important for polarizing activities that modulate the ubiquitylation state of freshly internalized proteins (Raiborg et al., 2003).
Materials and Methods
Fly stocks
karst alleles have been previously described (Thomas et al., 1998; Zarnescu and Thomas, 1999). Wild-type controls utilized either Oregon-R or the karst progenitor line mwh ve red. The midgut UAS driver P{w+mC GAL4::Hsp70Bbmex.P5′2.1 = mexGAL4} was created as follows: a 2.1 kb promoter fragment from the mex1 locus (a gift from S. Newfeld, Arizona State University, Tempe, AZ) was excised with EcoR1 and Kpn1 and inserted into pGaTB upstream of the GAL4 gene. The mex-GAL4 cassette was then subcloned into pP{CaSpeR-4} using Kpn1 and Not1, the sequence was verified and used to transform yw; Sb Δ2-3/TM6, Ubx according to standard methods (Rubin and Spradling, 1982). This driver expresses GAL4 from late embryogenesis through the second instar in the middle midgut, reflecting regulation of the mex1 gene (Newfeld et al., 1996). P{Rab5S43N.Scer\UAS=UAS-Rab5.S43N} (dominant-negative Rab5) was a gift from M. Gonzalez-Gaitan (Max-Planck-Institut, Dresden, Germany). The temperature-sensitive Shibire2Ts (Dynamin) line was acquired from the Drosophila stock center (Bloomington, IN). Recombinant kst2 UAS-DRab5S43N/TM6B and Shi2Ts; Df(3L)1226/TM6 lines were generated using standard genetic methods.
Dye feeding experiments
Bromophenol Blue (BPB; 0.2% w/v) was added to freshly made yeast paste (2.78 g instant yeast, 8 ml dH2O). Up to 40 third-instar larvae were fed this yeast paste for ≥2 hours before observation. The recessive marker red on the karst chromosomes was used to identify mutant larvae. Larvae that had filled their guts with dye were dissected in 1 × PBS containing 5 mM EGTA to paralyze continuing peristaltic motions of the intestine, facilitating rapid observation and preventing the mixing of gut contents and rupture. Observations on each live gut were made immediately during each dissection. Our titration experiments for BPB are: yellow/green/blue at pH 1.8/1.8-3.0/3.0, respectively.
Acridine Orange (AO; 100 μM) was fed to larvae in a solution of 10% sucrose 30 to 60 minutes before dissection as above, and immediately observed. AO imaging was carried out using a Zeiss LSM 510 META confocal microscope (Carl Zeiss) with a 488 nm laser line. Unshifted and acid-shifted fluorescence was detected using 500-530 nm and 565-615 nm filters, respectively. To facilitate comparison, both wild-type and karst mutant larvae were always examined in each session using the same 40× water immersion lens and identical microscope settings.
All antibodies were raised against Drosophila proteins unless otherwise stated, and were used at the following concentrations: mouse monoclonal anti-human-Myc (Ab-1, clone 9E10; Oncogene Research Products) and rabbit anti-βH (#243) (Thomas and Kiehart, 1994) were used at 1:300. Mouse anti-Coracle (from R. Fehon, University of Chicago, Chicago, IL) was used at 1:50. Mouse anti-α-spectrin (ascites #N3; from D. Branton, Harvard University, Cambridge, MA) was used at 1:600. Guinea-pig anti-Manduca V-ATPase subunit B (#C23; from M. Huss, University of Osnabruck, Osnabruck, Germany) was used at 1:150 for immunofluorescence and 1:1500 on immunoblots. Rabbit anti-Lava Lamp (from J. Sisson, University of Texas, Austin, TX) was used at 1:5000, rat anti-Rab11 (from R. Cohen, University of Kansas, Lawrence, KS) was used at 1:1000, and rabbit anti-Rab5 (MP1; from M. Gonzales-Gaitan, Max-Planck-Institut, Dresden, Germany) was used at 1:75. For secondary antibodies: Alexa Fluor 546 goat anti-mouse and Alexa Fluor 488 goat anti-rabbit antibodies (Molecular Probes) were used at 1:250. Cy5 goat anti-guinea-pig, Cy3 goat anti-rat, and FITC goat anti-mouse antibodies (Jackson ImmunoResearch) were used at 1:250. HRP-conjugated anti-guinea-pig antibodies (Sigma-Aldrich) were used at 1:2000.
Immunofluorescence and immunoblots
Larvae were dissected in fresh 4% paraformaldehyde in 1 × PBS, and incubated for 20-45 minutes with rocking at room temperature. Hemizygous temperature-sensitive Shi2Ts third-instar larvae were submerged in vials in a 34°C water bath for 1-2 hours immediately prior to dissection. Third-instar middle midguts were dissected away from the rest of the intestine for better antibody penetration. For all intestines, fixation was followed by three washes in 1 × PBS and blocking for 10-30 minutes in incubation solution (1 × PBS, 5% normal goat serum, 1% Triton X-100). Primary antibodies were applied in incubation solution and kept at 4°C overnight with rocking. Following four washes in incubation solution, secondary antibodies were applied in incubation solution and kept at 4°C overnight with rocking. Following four additional washes in incubation solution, the guts were equilibrated in mounting medium (80% glycerol, 100 mM Tris pH 8.5). Intestines were imaged using either an Olympus Fluoview 300 (Olympus America) or a Zeiss LSM 510 META confocal microscope. Images were processed using Adobe PhotoShop v4.0 (Adobe Systems). To detect Drosophila V-ATPase subunit B on immunoblots using Manduca antibody C23, two third-instar Oregon-R larvae were ground in loading buffer and the protein separated on a 12% polyacrylamide gel, then blotted and probed according to standard protocols.
The authors gratefully acknowledge M. Huss, D. Branton, R. Fehon, J. Sisson and R. Cohen for gifts of antibodies, S. Newfeld for the mex promoter fragment, and M. Gonzalez-Gaitan for Rab5 antibodies and the Rab5S43N and myc-2xFYVE fly stocks. We also thank S. Gilroy and R. Cyr for use of their confocal microscope, C. Gay for suggesting the use of Acridine Orange, D. Hess for technical assistance, and D. Cavener for his support. Finally, we thank R. Patterson, E. Siegfried, A. LeBivic and members of the Thomas lab for their critical evaluation of this manuscript. This work was partially funded by NIH grant #GM52506.


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On Truth and Fake News
Pastor and writer John Hudson takes his cues from the prophet Jeremiah on how to preach God's truth to power in an era of "fake news."
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Teen Mom stars Kail Lowry and Briana DeJesus will once again be going on under the knife of the Snapchatting doctor, Dr. Miami.
The gals, who are both currently pregnant, will apparently be getting post-baby nip 'n' tuck sessions with the good doctor.
In an interview with Radar Online, Dr. Miami told us all about Kail's upcoming surgery.
"Kail is scheduled to have surgery with me after she has this baby," Dr. Miami said.
Of course, this is not Kail's first time on Dr. Miami's operating table. She already had a butt lift, tummy tuck and liposuction last year.
Dr. Miami didn't specifically say what surgery the mom-to-be was having this time but he hinted that it could be another tummy tuck.
"There is a myth that you can only have a tummy tuck once you are done having kids," he said. "But it's not really true. You can have a tummy tuck after your first kid and if you get pregnant again you can have another one. The body has an amazing elastic."
Briana DeJesus, the Teen Mom 3 star who will soon star alongside Kail on 'Teen Mom 2,' is also a former patient of Dr. Miami who will be coming back for more surgery after she gives birth.
Briana took to Twitter to brag about her upcoming surgery, tweeting a screenshot of a conversation she had with Dr. Miami.
"…I'm pregnant again so hopefully when I give birth. I would love to work with you again! Things are falling out of place," she texted the doctor.
He congratulated Briana and said he would be glad to work with her again.
"This is why I love him so much," Briana tweeted about Dr. Miami. "Got so many new ideas on what I wanna do! Stay tuned!"
On the same day as Kail's surgery last year, Briana and her sister Brittany went under the knife, getting a BBL (Brazilian Butt Lift) as well as liposuction and a labiaplasty, all of which was broadcast to Dr. Miami's thousands of Snapchat followers.
Both girls are currently pregnant due to have their babies this summer. This will be Kail's third child and Briana's second.
It's not yet known whether Kail and Briana will have their surgeries shown on Dr. Miami's new WEtv reality show, which is currently airing Fridays at 10 p.m.
(Photos: Instagram)


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Analysts raise Apple targets, see shares going as high as $156 after best-quarter ever
Led by the blockbuster debut of the iPhone 7 series, Apple exceeded Wall Street's expectations with its holiday quarter, sending shares of the company's stock higher and prompting analysts to revise their price targets upward.
In particular, Macquarie Securities, RBC Capital Markets, and Guggenheim all increased their estimates for AAPL following the company's December quarter results, which reached a record-setting $78.4 billion in revenue and $17.9 billion in net profit.
Analyst Ben Schachter of Macquarie, in particular, has the highest 12-month valuation for Apple at $156. In a note to investors on Wednesday, a copy of which was provided to AppleInsider, he said that the continues growth of Apple's "Services" business should command more attention going forward.
Driven by the success of the App Store, Apple's Services business drew $7.17 billion in revenue in the company's first quarter of fiscal 2017. With 30 percent growth year over year already, Apple has said it plans to double its Services revenue over the next four years.
Schachter has been bullish on Apple's Services business, but the ambition expressed by the company's management during its quarterly earnings conference call on Tuesday exceeded even the analyst's lofty expectations. As a result, he now expects that the Services business segment will generate 25 percent of Apple's gross profits in fiscal year 2017, despite accounting for only 13 percent of the company's revenue.
Amit Daryanani of RBC Capital Markets also increased his price target on AAPL from $125 to $140. Shares of the company exceeded $125 in intraday trading on Wednesday, overing above $127 as of publication.
Thanks to a number of factors — including an anticipated "super cycle" for the rumored redesigned "iPhone 8," as well as the possibility of tax breaks for overseas cash — Â Daryanani wondered if Apple could manage to exceed $10.00 in earnings per share.
Guggenheim analyst Robert Cihra also increased his target for Apple from $140 to $150, declaring the company's holiday quarter should be "more than enough to scare bears and wake up investors." He cited a number of upcoming catalysts, including the anticipated "iPhone 8," more than $231 billion in overseas cash that could be repatriated, and spending on research and development that has more than doubled over the last four years.
"We see investors being paid to own the stock," Cihra wrote.
Finally, Cowen and Company analyst Timothy Arcuri did not increase his price target from $135, but called that price now a "veritable layup." In his view, share prices as high as $150 would be possible if his bullish estimates hold.


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Sword Art Online: Fatal Bullet Original Costumes, Character Creation and More Revealed
Bandai Namco has released a slew of new details for the upcoming Playstation 4 game Sword Art Online: Fatal Bullet original costumes, character creation and more!
Here's the story for the game:
According to Bandai Namco, "YOU" are the protagonist in the story. Kirito and the rest of the SAO gang will be present but the main narrative lies in you. The game also asks "will you walk a different path from Kirito?"
New to the game and the series, particularly in this iteration of Gun Gale Online is the "ArFA-sys" support AI together with the "New Space Ship Area: SBC Fluegel".
Sword Art Online: Fatal Bullet Main Character
This new Sword Art Online game will let you use your own character who just so happens to be a party member / teammate for the series' protagonist Kirito. This means you get to design your own character from scratch. But for default purposes, here's what the male and female avatar would look like.
Sword Art Online: Fatal Bullet Character Development
In Fatal Bullet, you can customize not only you character's gender and hair style but also allows you to grow skills to become better suited as the story progresses. There are four main parameters you can develop in the game namely AGI, STR, VIT, INT, LUK and DEX.
Here are the breakdowns for each skill
- STR: Increases the max weight limit of the equipment you can carry. Increases close range attack strength. Some equipment requires a specific level of STR.
- VIT: Increases max HP. Increases the rate of damage reduction.
- INT: Increases the amount of time some skills and gadget effects last. Increases the effects of attacks that deal status ailments. Decreases the cool time for skills and gadgets.
- AGI: Increases movement speed. Increases the rate of damage reduction. Increases handling speed for reloading and such.
- DEX: Increases the effects of some gadgets. Increases critical hits and damage dealt during weak point attacks. Increases the accuracy of the Bullet Circle. Some equipment requires a specific level of DEX.
- LUK: Increases the item drop rate. Increases the rate of critical hits.
New Characters
Kureha (voiced by Ayane Sakura)
As per Bamco:
A girl who invites the protagonist to Gun Gale Online, creating his chance to log in. She is the protagonist's childhood friend, and a veteran Gun Gale Online player who has been playing since its early days. She is bright and cheerful, and has a meddlesome personality. She also considers the protagonist to be the object of her personal protection. She plays Gun Gale Online in order to realize her own strength, and spares no effort in order to discover that strength.
Zeliskam (voiced by Mamiko Noto)
A mysterious woman accompanied by an ArFA-sys that you meet during the story. She is calm-mannered, but is a talented solo player who has made a name for herself in Gun Gale Online. She tries to get a grasp of the protagonist, who obtained an ArFA-sys by luck. She will sometimes give you advice that is a sore to the ears. She dotes on her partner ArFA-sys, and enjoys wearing cute-looking clothes.
Its official name is the "Artificial Financial Adviser System." Its short name is "ArFA-sys." They were added to Gun Gale Online as a new feature in a major update rolled out by Zaskar, the company that operates the game.
ArFA-sys are support AI that learn, grown, and participate in battle with you. As its name of "Financial Adviser" implies, they can also manage in-game assets and items. They are reliable partners that assist the player in both battle and fortune.
ArFA-sys are divided into several types, and the one the protagonist obtained is the particularly high level and valuable Type-X. Its appearance and gender can also be customized by the player.
New Costumes for Classic Characters
It looks like all our favorite (or non-favorite, looking at you Kirito) will be getting new costumes here in Fatal Bullet. Check out the new designs and costumes they carry for this story. More than that there will be some really fresh ones from characters who did not appear in the Phantom Bullet Arc from the anime series. Kirito and Sinon are confirmed to have new costumes as well. These designs were done by Kotaro Yamada, the same artist who worked on Sword Art Online: Project Alicization manga which appeared in Dengeki Bunko Magazine.
A girl who, during the "Sword Art Online" period, was sub-leader of the game's leading guild, "Knights of the Blood." She is soft-mannered, but strong at her core. She met Kirito and decided to live with him. Her magnificent sword skills earned her the nickname "Flash" in Sword Art Online and "ALfheim Online." In Gun Gale Online, she uses a thin assault rifle that resembles the small swords she is accustomed to using.
Japan gets first dibs with Sword Art Online: Fatal Bullet with a release date of February 2018 while North America and Europe gets it at a later time in 2018.


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I have had a VERY mixed experience with Anker durability, and was wondering what other people's experience was.
I've never had a negative experience with them. At some point everyone will get a bad product from any manufacture. So if I get a single product from Anker and it has issues. It doesn't surprise me. If it fails within their warranty period I know they will take care of it. I follow their policy and only buy from Anker.com, Amazon (Anker Direct,) and other authorized dealers.
There are a ton of happy customers out there. You can find happy customers on the various review sites, reviews on Amazon (Anker doesn't pay give away free products for reviews on Amazon), and on the community here.
Those that have issues with Anker, usually get a hold of a fake product, a product purchased from a 3rd party and has to deal directly with them, those buying from an unauthorized dealer, the product was damaged caused by the user, or the product is out of warranty. These types of issues aren't covered by warranty and customers get upset.
Anker for the most part will take care of their customers.
Thanks for the info!
I have 22 Anker items and have 0 issues. Oldest items are from 2011.
Damn, that's pretty good.
Out of over 30+ cables I have only had to have 1 replaced as it flat out stopped charging everything. No damage to the cable or connector as it stayed plugged in and stationary, but they replaced under warranty.
I have had 1 bluetooth headphone and 1 bluetooth speaker replaced, the headphones were manufacturing defect in right earbud that shorted and the speaker was the battery not holding charge. But aside from those 3 issues, I have had many Anker products without any problems and will continue to use and buy their products
Even my very old A 3143 speaker works fine
and has still a good battery after 3 years of intensive use.
You put that well!!!
Of the couple of dozen products I've had from Anker, over 4ish years, I've had 3 replaced. 1 car charger, an astro powercore and a pair of headphones.
I'm pretty thought with a lot of my equipment. They get thrown into bags regularly.tossed around in cars, traveling etc. So pretty good going I reckon. I have never had a cable fail, which surprises me as those are normally the point at which many things fails.
Never had an issue so far with durability through acceptable usage (what Anker have said can be done to the item)…out of the 30+ items I've had over the past 3 years, only had two replaced due to a hardware fault (battery related in most cases)…
I'm in the same boat… out of dozens of Anker purchases over the years, I've only needed warranty service on 2 of them.
That's not bad.
Woah. What? How does it still have a good battery?
I don't know.
I charge it with an ANKER charger and I use it only indoor (never outdoor or in a car were it might be exposed to the sun).
This are the only ideas I have.
The least reliable for me are the soundbuds (Slim, Life) as the thin cables fail usually at the connection into the bud. But I use these for hours/day so for a $20-$30 device to last daily for months which equates to hundreds of hours does not feel bad value. I haven't used warranty claims for the last failings as I have hammered the products so hard I felt it was my own use which made them fail rather than designed failure.
Powercore, Powerport, Powerline. Been very reliable.
If I were to combine these experiences, I would say the neck-weighted bud products need Powerline+ skills of better cables into the bud in the ear, so they last longer. So something like the Soundbuds Life with a thicker/stronger cable into the buds.
I have 10-15 Anker products…
Soundbud slims stopped working after 3-4 months, but the warranty replacement has had no issues.
Powerline+ lightning cable - rubber came apart and wires became exposed after using daily at work for 5-6 months. Anker sent me another cable, but I still used the damaged one; just taped it up with electrical tape, still using daily and it's going on almost 2 years old now.
Soundcore 2 survived a fall from the roof of my car going 15-20 mph. Granted it was inside an Anker case. I still use it daily at work 8-10 hours a day with no issues. It's a lil over 1 year old now.
Everything is working great with no complaints.
If I was to total up all the products including cables I own/bought I would say easily over 30. From these I have to say I've had to use the warranty on about 3 items.
As far as durability I've got a powerpack that is still working more than 6 years later - which says a lot about a product.
Don't forget though even "big" brands have product issues including Apple and Samsung etc
I think I had 1 issue I can recall with my Powercore 20100 and they handled it well. Took it back and gave me an updated version.
Sounds like the best recall from any Chinese company ever! Great job Anker!
Really bad. I purchased two 26000 units for long distance hiking in 2020 and because of COVID, only used them about 5 times, usually for only for 2 or 3 days at a time, and not even when hiking. On both units the USB A port stopped working. Anker is "surprised" but won't stand behind their products, so I am left with two paper weights. My recommendation is that you avoid Anker. There are a lot of other brands that have quality products.


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On the podcast is Inge Kristensen. Inge is the Chief Executive at the Danish Society for Patient Safety. She is an experienced leader in healthcare and social services, dedicated to quality improvement and patient safety. Inge has extensive experience in leading large-scale projects in hospitals and community care and creating cross-sectoral collaboration focusing on quality improvement. Inge is an experienced moderator, public speaker, and contributor to whitepapers, books, and journals on improvement in healthcare.
A few key takeaways:
On how to approach life
"I get up every morning and think we could do better today than we did yesterday."
On psychological safety and its importance
"You feel secure when you are in your role, in your function in healthcare. It has a lot to do with the ability to have an open and free discussion about errors, about improvement, at your workplace. "
Book recommendations from Inge:
Managing the Unexpected: Resilient Performance in an Age of Uncertainty
Download Psychological Safety resources (English / Danish)
Psychological Safety Toolbox
On the podcast is Inge Kristensen. Inge is the Chief Executive at the Danish Society for Patient Safety. She is an experienced leader in healthcare and social services, dedicated to quality improvement and patient safety. Inge has extensive experience in leading large-scale projects in hospitals and community care and creating cross-sectoral collaboration focusing on quality improvement. Inge is an experienced moderator, public speaker, and contributor to whitepapers, books, and journals on improvement in healthcare.
A few key takeaways:
On how to approach life
"I get up every morning and think we could do better today than we did yesterday."
On psychological safety and its importance
"You feel secure when you are in your role, in your function in healthcare. It has a lot to do with the ability to have an open and free discussion about errors, about improvement, at your workplace. "
Book recommendations from Inge:
Managing the Unexpected: Resilient Performance in an Age of Uncertainty
Download Psychological Safety resources (English / Danish)
Psychological Safety Toolbox
Mon, 7/19 12:59PM • 30:05
organization, psychological safety, patient safety, patients, healthcare workers, healthcare, healthcare sector, hospital, danish, health care, nursing homes, work, program, victims, speak, listeners, errors, impact, mid-Staffordshire, problems
Dr. Marco Bo Hansen, Intro, Inge Kristensen
Intro 00:00
You are listening to the hygiene and infection prevention network. We are dedicated to sharing clinical expertise and hand hygiene resources with healthcare communities worldwide.
Dr. Marco Bo Hansen 00:13
Hi, this is Dr. Marco Hansen, and I'm the medical director at sani nudge. I'm very excited to welcome a special guest to today's podcast. Inge Kristensen is the chief executive at the Danish society for patient safety, is an experienced leader in health care and social services dedicated to quality improvement and patient care. Inga has extensive experience in leading large-scale projects in hospitals and community care and creating cross-sectoral collaboration. Focusing on quality improvement is an experienced moderator, public speaker, and contributor to white papers, books, journals, and healthcare improvement. We are excited to have you in here. Welcome.
Inge Kristensen 00:57
Thank you very much, Marco; it's a pleasure to be here.
Dr. Marco Bo Hansen 01:00
Inge, we would like the listeners to know you a little more. How would you describe yourself?
Inge Kristensen 01:05
I'm not a healthcare professional by background, but I am deeply in love with healthcare and the healthcare sector. And I have been working in the healthcare sector for Well, 20 plus years, and I would describe myself as very curious, hopefully, also humble; I try to be at least but also very ambitious. I get up every morning and the things that we can do better today than we did yesterday.
Dr. Marco Bo Hansen 01:44
That's a great mindset. And also mentioned in the intro, You are the chief executive at the Danish society for patient safety. Can you explain to us what the Danish society for patient safety is?
Inge Kristensen 01:57
The Danish Society of patient safety is an independent organization working to improve quality and patient safety in the Danish healthcare system. We were founded in 2001 by all the leading stakeholders in Danish healthcare and had been working ever since. And work with improvement programs in healthcare and, hopefully, sometimes to set the agenda for patient safety in Denmark and work very internationally. We have collaborated with the IHI, were a strategic partner with the Institute for Healthcare Improvement IHI in Boston, and work with organizations in the UK and other European countries.
Dr. Marco Bo Hansen 02:55
And what is your main focus at the moment, for some
Inge Kristensen 02:57
time we have been working with diagnostic error. And we think that that's a somewhat overlooked phenomenon in healthcare. And also, we are working a lot with them. What you can say is, well, the old problems in patient safety, we don't seem to have the song. The classic problems are medication errors, infections, and other issues. So that's some of our main focus areas.
Dr. Marco Bo Hansen 03:40
How do you work with the patient safety part? Are you out in the healthcare sector and the hospitals and nursing homes? How do you collect the information and turn it into patient safety?
Inge Kristensen 03:52
Well, we try to harvest ideas. So, we go out internationally to find out what is also working what is working in a Danish setting, and then we try to translate it. As you say, we are working very closely with healthcare professionals and the patients and carers around improving. So, we enjoy our work most when we can be in the field and very close to the problems because our methods are also to create robust pathways and routines to get it right every time for everyone.
Dr. Marco Bo Hansen 04:49
Speaking of work, and also the work you have done in the Danish society for patient safety. When looking into your career track record, you have An impressive Korea already. What do you consider some of the highlights of your career so far?
Inge Kristensen 05:07
Well, some of the highlights have been working with a lot of stakeholders around infection control, creating a program for infection control in the capital region of Denmark and, well, nationally. But I'm also very proud of some of the cross-sectoral work I've been doing to find new solutions for some of the work between hospitals and the primary sector.
Dr. Marco Bo Hansen 05:43
That sounds very interesting. And another important topic that I know you're working with that I'm very interested to hear more about is psychological safety. Can you tell the listeners what psychological safety is and why it is so important in all the work you're doing? And in patient safety?
Inge Kristensen 06:02
Yeah, I wouldn't mention it because I knew you would ask about it. So but, but some of the work we're doing around psychological safety is very dear to me. And very important, I think for the healthcare sector in the future. Psychological safety is a paradigm you could say what it's developed. Mainly our has come on the agenda by the work of Professor Amy Edmondson, Harvard professor, what it is about is that you feel secure when you are in your role in your function in health care. So it's, it's really about the work environment. And also has a lot to do with the ability to have an open and free discussion about errors about improvement at your workplace.
Dr. Marco Bo Hansen 07:07
And what in your mind, what do you think happened? If an organization doesn't have the level of psychological safety needed? What are the consequences,
Inge Kristensen 07:18
More and more research shows that the lack of psychological safety can lead to patient safety incidents, where patients are harmed. And also, it has been very clear and obvious, I think, during the COVID era, that a while lacking psychological safety has led to that healthcare professionals go out of the work. They leave healthcare, get depressed, a burnout, all kinds of negative implications of psychological safety, and negative implications for the safety of the patients.
Dr. Marco Bo Hansen 08:12
And if you are listening to this podcast now and you are a healthcare worker or a manager leader, thinking, does my organization have the right level of psychological safety? How would you How would you know?
Inge Kristensen 08:29
Well, there are more and more tools you can use for the assessment of psychological safety. You can work systematically with the website, psychological safety, more and more. And do we have together with the organization called the Copenhagen Academy for medical simulation, developed some tools that we are using in the healthcare sector in Denmark. Still, I know there are more and more, for instance, Cincinnati Children's Hospital, I would say, well leading in some of this work, and it's amazing to see how they work with the speed of culture. And that the impact on the safety incidents has been very clear.
Dr. Marco Bo Hansen 09:29
Can you perhaps come up with an example of an organization where psychological safety was lacking and what the consequences of these were,
Inge Kristensen 09:40
I can propose or find an example by where it had a serious impact, and that's around. We had some cases around young boys with meningitis who died, and at least in one of these cases, we saw That a nurse tried to speak up. It was difficult for her, and she wasn't listening to in the concrete situation. And eventually, the young boy died. And of course, it was not that thing in itself that led to these avoidable deaths, but it certainly had an impact.
Dr. Marco Bo Hansen 10:29
Yeah, and these are, of course, tragic events, and we must do everything we can to prevent these events in the future and learn from the events. And some of the things that I've also seen in the healthcare culture are the fear of making mistakes, making errors, and these fears can be so intense that you are afraid to talk about errors in the first place. Is that something you're also working within in your organization?
Inge Kristensen 10:58
Yes, we work; we have quite a large program around mental health for healthcare professionals. And, it has evolved around the notion about a second victim. So the patient, of course, is the first victim of a serious adverse event. But on the back of that, the second victims and we all know that the healthcare professionals are in grave danger, for depressions, developing depressions, and even suicide. So. So it is very important that, that as a part of psychological safety, we have an environment where it is legitimate to talk about errors, and where there is a support system if errors occur and that no, not if when they occur, because they do whether there are people that there will be made mistakes. So So that's, that's obvious, and there should be a systematic approach to that. So we're developing that right now together.
Dr. Marco Bo Hansen 12:22
with a hospital. And these second victims, that's, that's one of the new, you can say, definitions that probably many of the listeners haven't heard before, have understood it correctly, to say that patients are first victims, healthcare workers, can be the second victims if we don't have a culture to embrace them and ensure that they don't get mentally distressed.
Inge Kristensen 12:48
Exactly. And actually, we also talk about third victims because the whole organization can be affected by a series of events. And, over time, their psychological safety and environment can suffer from these incidents. So if they, if they're not dealt with timely, it will show.
Dr. Marco Bo Hansen 13:16
overtime that is so interesting. I've learned something new first, second, and third victims; it's important to know these things and deal with it. Okay, so I want to shift focus a little bit too patient perspectives in nursing homes. And in 2020, your opinion was featured in a chronicle in a Danish news newspaper, and it is a respected Danish opinion piece for our listeners. So you shared that in nursing homes and healthcare organizations, the five most dangerous words are it could not having here. So can you please explain to us what does it mean? It could not happen here, and why is it so dangerous to say this sentence as an organization?
Inge Kristensen 14:05
Well, using those five words, I think they are? Well, intuitive. Sometimes. I think that when you hear that something dreadful is happening, you would say, well, it couldn't happen to us. It couldn't happen here. We have an organization with which is solid and where these events couldn't take place. But you have to pause, and you have to reflect. Because while most things can happen to anyone, and they and that's not well, that's not the problem is not to face it and find out how we can avoid these events? And how can we ensure that we are proactive in our ways to ensure that we have a safe system for our patients? So, for the citizens and also for the healthcare.
Dr. Marco Bo Hansen 15:22
workers, it is especially timely considering the tragedy that unfolded in nursing homes due to the COVID-19 Is it a particular phenomenon we see in nursing homes, the phrase it could not have been here, or do we also see it other places in society,
Inge Kristensen 15:40
the Chronicle built on the scandal, the Mid Staffordshire scandal in the UK. And I was trying to point out some of the learning issues, the learning point from that scandal, and that was the hospital where it looked from the outside as if they were, you could see in green. So you couldn't save from any of the control mechanisms so that there was anything wrong, but it was clear, when they looked into data in new ways, they could see that there were more or the mortality was sky-high. And the patient's stories and the carrier stories that surfaced were so grave that you can't find Mid Staffordshire on the map today because they shut that place down. So so it happens, it can happen everywhere. And in a way, I've had some cases in nursing homes these recent years. And when I look back and see media stories, we can find them from time to time. They surface so that it can happen in every sector. And usually, it happens, where there are a lot of patients, so on citizens who are witless, not able to, to take care of themselves.
Dr. Marco Bo Hansen 17:35
And how to work with this mindset as an organization. Any comments on that?
Inge Kristensen 17:42
It is a cultural phenomenon. And you have to work consistently with the culture. And you have to do that every day. You have to be proactive and have to set out some guidelines for how you want to work in our institution, workplace hospital, whatever unit you are in, and go for that relentlessly. And always have high standards. I also think that we need to talk more about ethics. There are in some hospitals and, and hospital organizations and also other types of welfare organizations. There are a wealth of ethical committers. But I don't think it's an I don't think it's it's common. You can find it, but it's not common. I believe we need to talk a lot more about these things regularly. Because if we adjust Well if you just adjust to things, they will deteriorate over time.
Dr. Marco Bo Hansen 19:15
Yeah. And that's a really important point that even though you might organize ethical or ethics bodies and talk about it in the management wards, it might not go down to all the others who work in an organization, others and they might not embrace it. So it's how you put it. You get it in the culture. That's the difficult part, I think.
Inge Kristensen 19:41
Yeah, and, and every journey begins with the first step. So so you have to be very specific. And if you pass by something you accepted, so it's, it's day-to-day work.
Dr. Marco Bo Hansen 20:00
Yeah. And that's, that's a really good point, I know that. The thing about if, if you pass something, you see something, you can speak up. I remember, not that long time ago, I was visiting my granddad in the hospital. And I could see, one of the doctors was wearing a watch on the wrist. And you are not allowed to do that for hygiene purposes. But I didn't dare to speak up because I was afraid that it would impact the treatment of my granddad. So why do you think you think that is? That we don't speak up? And don't they add to say something if we see something's wrong?
Inge Kristensen 20:49
Yes, but that that's about your psychological safety. I don't know about you, but what I have experienced is going into a hospital. I can feel or a nursing home, and I can feel immediately how things are; I can think that this is a good place. People say hello to me. Greet me, they, it seems there's an open culture there. There's, it's, it's nice, you can feel it. And sometimes you also go into places, and you say, I don't feel comfortable here. I'm not sure where things are. No one says hello to me. They just have this, you know, this eyesight where they are just not looking to the sides when they walk down the hallway. So whatever you experience, and then and that, of course, imposes. Subconsciously imposes a feeling in us and our ability to speak up. And I think that we should all be able to speak up because one of the most important sources of information for treatment and the well-being of patients is the patient itself or other relatives. So so you have to listen. And I think it's part of the well, the ethical conversation, and the psychological safety that you see some of these comments as a gift. And I see it in other industries. They use complaints very systematically and see them as a gift to the organization on how to improve, should try to impose on the health care workers said it's a gift.
Dr. Marco Bo Hansen 23:07
And I guess you can also start from today talking about psychological safety and just be aware of it wherever you go. Because in the supermarkets, when you see people not sanitizing hands, you most often don't say anything because you also think that you don't want to be rude to them, or you're afraid of them, seeing it as an accusation or something. So I guess society also likes a bit of psychological safety, you can say.
Inge Kristensen 23:40
Absolutely. And perhaps there is a little crack now from the COVID situation that we can take on that, that it's more legitimate to speak up also in Well, the supermarket or wherever you are.
Dr. Marco Bo Hansen 24:06
Exactly, exactly. And speaking of the corona pandemic, I really would like to talk about the impact of the pandemic on healthcare workers' mental health and patient safety. You briefly touched upon it, but what are your thoughts and insights into the impact of the corona pandemics on healthcare workers' mental health and impact on patient safety?
Inge Kristensen 24:31
I think that some of the issues that have come up, we knew that already but the corona period has been a pandemic has been like a giant light shining on health care to see all the cracks. You can see where some of the problems are, and I think For, for healthcare workers, psychological safety has become much more legitimate to talk about psychological safety. And I think the link between psychological safety and patient safety has become much more apparent. And we can use that forward. So that is one of the things, I also think that equity is for patients is one of the issues that the that we have got some, well, you could say negative insight on in Denmark, we have experienced that, that we have made a lot of systems and done it very quickly. But the well, the vaccination programs where you get an email or an electronic message, well, maybe 20% of the population cannot either receive it, read it, or respond adequately on it. So the system itself, is this making some of these errors, you could say, and we should learn from that because we see very clearly during the corona pandemic, but it happens all the time.
Dr. Marco Bo Hansen 26:34
I completely agree. And do you think we will? Will? Will we see more of it moving forward if we don't do anything about it? And how can we do something about it? What are you What's your take on this?
Inge Kristensen 26:47
Well, we have to go back to your very good example about the wristwatch and not say anything about it; we have to listen very carefully to what the patients tell us. And the relatives tell us because some of the insights we need to improve health care come from the relatives and the patients.
Dr. Marco Bo Hansen 27:20
And I agree, we are not good at that. This mutual awareness and mutual communication between patients and relatives and healthcare workers lack many healthcare organizations. So that is, of course, an important focus area. A for all of the podcast guests, I'm always asking for expert insight, whether you have a book or an article, Chronicle or something that you have just stumbled upon, that you find very interesting and that you think others should perhaps read or listen to, is there anything that you can recommend to our listeners?
Inge Kristensen 28:04
Oh, I think there's so much interesting going on out there and beyond. Well, diving deeper into psychological safety, I would certainly recommend that you read if you haven't done that already. the book by Karl Weick and Catherine Sutcliffe, which is calling, managing the unexpected. Because it's a very comprehensive work on building a resilient organization, and how you can be proactive and endure some of these well, shortcomings or situations where there is a crisis, and even some of the day-to-day problems around staffing and things like that, so, so that's a very good book.
Dr. Marco Bo Hansen 29:08
Awesome. This is well noted. Thank you. So our audience includes hygiene preventionists, healthcare workers, researchers, and leaders across the healthcare space. Do you have a tip for those seeking to make a difference in health care?
Inge Kristensen 29:25
Ask yourself, what can I do by next Tuesday, or even by tomorrow? Because there's always something you can change and, and well, as I said, every journey begins with the first step. So I would recommend that certainly,
Dr. Marco Bo Hansen 29:47
that's great advice. And this is well notice, and Inga, thank you for your time and for sharing your thoughts and insights, and thank you to our listeners. Be safe,, and remember to clean your hands.


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Next week's events by Impactscool – 24th to 30th September
21 September 2018 | Written by La redazione
Come ogni settimana, il team di Impactscool sarà impegnato in diversi appuntamenti in tutta Italia e non solo. Scopri dove puoi trovarci nei prossimi giorni.
Training, innovation, workshops, labs: the last two dates with #Futura, the event designed by the Italian Education Minister to present the National Plan for the Digital School throughout Italy, have made Varese and Bergamo two gravitational centers for the future of education. In the program of events, in addition to hackathons and conferences, there was also the #woMEST format, organized by Impactscool: a real Vision Hack, created with the aim of bringing students closer to STEM subjects (science, technology, engineering, mathematics). The closing ceremony of the Varese event was also attended by the Italian Minister of Education, University and Research, Marco Bussetti. Impactscool managed to interview him exclusively: here is the opinion of the minister on school and technology.
The next date with #Futura will be in Rapallo (GE) from September 30th to October 2nd: here too Impactscool will propose a new #woMEST event.
And after the first workshop dedicated to the topic of emerging technologies, the cycle of workshops at the Tor Vergata University in Rome continues: Andrea Geremicca, CMO of Impactscool, will examine the theme of Artificial Intelligence with the University's students on September 25th. On September 28th, instead, a debate on the Blockchain is scheduled for the third meeting. The closing dates of the workshop cycle will be held on October 12th and 19th, and they will talk about the 3D printing and the future scenarios brought into play by the new technologies. Participation is reserved for students of the Tor Vergata University.
A real "Journey into the future", finally, will be what the Impactscool team will undertake on September 27th at the Tenuta Artimino at Villa Ferdinando in Artimino (Prato). In collaboration with the company Audionova, a debate will start at 18.00 on the impacts that the exponential technologies are having on our society: knowing its potential and learning how to use it can be an extraordinary resource to drive change, rather than having to undergo or try to resist. In addition to Cristina Pozzi, CEO of Impactscool, and Andrea Geremicca, CMO, two other guests will be among the speaker of the event: Massimo Chiriatti, a technologist, who also collaborates with universities for debates on the digital economy and is a member of Assob.it, a non-profit association that promotes the use of Blockchain technology, and Hady Ghassabian, Impactscool Ambassador and suborbital astronaut selected for a joint research project between the Embry-Riddle Aeronautical University and NASA. The event is reserved for members.


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Performance Contract Template 14 Download Free from music performance contract template , image source: www.sampletemplates.com
Every week brings task lists, emails, documents, and new projects. How much of this is totally different from the job you've done? Odds are, maybe not much. A number of our tasks are variations on something we have done hundreds of times before.
Don't reinvent the wheel each single time you start something fresh. Use templates–as starting point for new 17, standardized documents. As soon as you save a separate variant of the template, simply add, eliminate, or alter any info for that record, and you'll have the job.
Templates work anywhere: in word processors, spreadsheets, project management apps, survey platforms, and email. Here is the way to use templates in your favorite apps–and the way to create documents from a template–so it's possible to get your ordinary tasks faster.
Templates take time to construct, and it's easy to wonder whether they are worth the investment. The answer: absolutely. Editing a template takes far less time than formatting some thing. It's the distinction between copying and pasting some text, or retyping it.
That is not the only advantage: Using a template means you are not as inclined to leave out crucial info, too. By way of example, if you want to send freelance writers a contributor agreement, changing a standard contract template (rather than writing a new contract every time) ensures you won't depart out the crucial clause regarding possessing the material as soon as you've paid for this.
Templates also guarantee consistency. Maybe you send regular job updates to investors or clients. With a template, you know the upgrade will constantly have the same formatting, layout, and structure.
How to Produce Great Templates
Not many templates are created equal–and a few things don't need a template. Here are a couple of tips to follow.
First, templates must be comprehensive. So err on the side of including rather than too small, it's more easy to delete info than add it .
Imagine you are developing a template of your own resume. You would want to record details about your responsibilities and achievements, and that means you are going to have.
You always have the option to delete less-important notes later on, but you might forget it if it's not from the template.
Some tools will automatically fill in these variables for you (more on that in a little ). But should you have to fill in the information by yourself, include some text that is obvious and easy to look for so you can locate.


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Judo, literally 'the gentle way' is a martial art, a sport and a philosophy which originated in Japan. Judo is born from Jujutsu and founded in 1882 by Professor Jigoro Kano. He envisioned it as a way of becoming physically and mentally fit through disciplined training.
The principles of Judo are based on both maximum efficiency with minimum effort and mutual welfare and benefit. The end result is the accomplishment of a goal with the best use of energy. Success in this martial art depends upon the skill of using an opponent's own weight and strength against them.
Although Professor Kano died in 1938, Judo has since spread rapidly around the world. The first World Championships were held in 1953 and by 1964, it was included in the Olympic Games. Judo's sport popularity has led its physical and competitive aspects to dominate, but it should first and foremost remain a way of life.
Accessible to everyone, Judo affects all areas of a practitioner's life. Physically, it improves strength, flexibility, coordination, balance and reflexes. It teaches a self-defense method that requires skill rather than just size or strength. Mentally, it promotes concentration, self-confidence, awareness and leadership.
Rooted in strong moral and ethical principles, Judo fosters a sense of respect and discipline. Camal and Cruz teaches students to strive for physical, intellectual and moral perfection to improve their own life and contribute to the greater good. Judo encourages a balance of skill, physical ability, and mental strength. The longer one trains, the more benefits the person sees.


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How I left Amazon Prime and managed to survive | Economy | News
The real magic of Amazon and especially Prime is that they stay away from shopping.
New York times
You have given up on the Prime Minister. that's good.
This makes me weird Nearly 98 percent of Americans are members of the Amazon Shopping Club For at least two years they kept renewing their membership. Prime is one of America's toughest consumer products.
I was a Prime member for about three years until I stopped renewing my membership in 2019. My friends and colleagues are often surprised when I tell them this. But do not worry. I'll be fine.
This does not mean that I live without buying on Amazon. Let me tell you something: you are allowed to order from the website without being a prime member, although it does take a bit more planning for the free delivery.
Why do I disclose my buying habits to strangers? Each of our personal experiences is a way of thinking about whether we are holding on to love or out of habit. In my case, once I questioned the value I was getting from the subscription, it was very easy to cancel the subscription.
This is not a tip to get you off Prime. You do your thing. Also, my experience is not representative. I don't have children and live in New York, where there are many things I might need in a hurry – a roll of aluminum foil or a new phone charger – often within walking distance.
I know that many Americans live far from stores, have caring responsibilities, live with disabilities, or have other circumstances that don't give them the same amount of flexibility in shopping.
Over 300 readers wrote to On Tech about Prime and many said they felt they got more than their money for the convenience, Reliable shipping and other Prime benefits.
"I got my prime when my husband was sick and was caring for him," wrote a reader in Carol Stream, Illinois. "I couldn't shop and that was perfect for me."
You're too late for the prime minister. I signed up in 2015, I think because I've been moving and rating entertainment streaming services. I basically subscribed to Amazon Prime Video and thought the option to get faster shipping at no extra cost was a plus. (Hope you don't lose my memory. I can't quite remember what my mind was like in 2015).
In surveys, the majority of Prime members said that free shipping is the main reason to sign up and keep the service. Several On Tech readers, like me, have said that Prime Video helped make membership worthwhile.
I bought more from Amazon during the first few years I was a member. This is typical behavior of PM members. But over time, I discovered that I was asking for less.
It was time to renew and I shrugged my shoulders and stopped paying. It has changed my behavior. In 2020 and the first few months of 2021, when online shopping skyrocketed as many people tried to avoid going to stores, my order history showed that I didn't buy anything for myself on Amazon except for a few e-books for my Kindle reading device.
I often shop online at Walmart, Target, and other sites. I did my shopping locally, in part because I didn't want Amazon and other giant retailers to be the only ones left after the pandemic. I don't feel like I need to get value out of a Prime membership, Amazon was just an extra option rather than the only store I thought of.
For me, online shopping now requires more patience. I keep a list on my phone of things I might need and order multiple items at once to meet minimum order amounts in order to get free shipping from Amazon and other websites. (Amazon orders over $25 often qualify for free delivery.)
The list now includes a non-stick frying pan, light bulbs, a toilet brush, puzzles, and a pair of headphones. My life is very exciting, yes. I may not receive the parcel in a couple of days, but that's fine for me.
I also think I buy less stuff without being a prime member because it's not easy to get a buying impulse off my couch. Some On Tech readers said they had the same experience.
The real magic of Amazon and especially Prime is that they stay away from shopping. Prime members tend to go to Amazon without much thought, and this often works out well for them and the company. For me, the lack of Prime makes me pause for a minute before buying. This is perfect. (I)
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