text:
- Research
- Open Access
Immunomodulator expression in trophoblasts from the feline immunodeficiency virus (FIV)-infected cat
© Scott et al; licensee BioMed Central Ltd. 2011
- Received: 24 June 2011
- Accepted: 5 July 2011
- Published: 5 July 2011
FIV infection frequently compromises pregnancy under experimental conditions and is accompanied by aberrant expression of some placental cytokines. Trophoblasts produce numerous immunomodulators that play a role in placental development and pregnancy maintenance. We hypothesized that FIV infection may cause dysregulation of trophoblast immunomodulator expression, and aberrant expression of these molecules may potentiate inflammation and compromise pregnancy. The purpose of this project was to evaluate the expression of representative pro-(TNF-α, IFN-γ, IL-1β, IL-2, IL-6, IL-12p35, IL-12p40, IL-18, and GM-CSF) and anti-inflammatory cytokines (IL-4, IL-5, and IL-10); CD134, a secondary co-stimulatory molecule expressed on activated T cells (FIV primary receptor); the chemokine receptor CXCR4 (FIV co-receptor); SDF-1α, the chemokine ligand to CXCR4; and FIV gag in trophoblasts from early-and late-term pregnancy.
We used an anti-cytokeratin antibody in immunohistochemistry to identify trophoblasts selectively, collected these cells using laser capture microdissection, and extracted total RNA from the captured cell populations. Real time, reverse transcription-PCR was used to quantify gene expression.
We detected IL-4, IL-5, IL-6, IL-1β, IL-12p35, IL-12p40, and CXCR4 in trophoblasts from early-and late-term pregnancy. Expression of cytokines increased from early to late pregnancy in normal tissues. A clear, pro-inflammatory microenvironment was not evident in trophoblasts from FIV-infected queens at either stage of pregnancy. Reproductive failure was accompanied by down-regulation of both pro-and anti-inflammatory cytokines. CD134 was not detected in trophoblasts, and FIV gag was detected in only one of ten trophoblast specimens collected from FIV-infected queens.
Feline trophoblasts express an array of pro-and anti-inflammatory immunomodulators whose expression increases from early to late pregnancy in normal tissues. Non-viable pregnancies were associated with decreased expression of immunomodulators which regulate trophoblast invasion in other species. The detection of FIV RNA in trophoblasts was rare, suggesting that the high rate of reproductive failure in FIV-infected queens was not a direct result of viral replication in trophoblasts. The influence of placental immune cells on trophoblast function and pregnancy maintenance in the FIV-infected cat requires additional study.
- Placental Tissue
- Laser Capture Microdissection
- Feline Immunodeficiency Virus
- Reproductive Failure
- Late Gestation
Precisely-regulated placental and decidual immunomodulatory molecules such as cytokines and chemokines are necessary to establish and maintain pregnancy [1–6]. While many of the immunomodulators expressed at the maternal-fetal interface are products of decidual leukocytes such as T cells, NK cells, dendritic cells, and macrophages [2, 5, 7], leukocytes are not exclusive producers of these molecules. Trophoblasts, including both cytotrophoblasts and syncytiotrophoblasts, are fetal-derived cells of epithelial origin that form the placental chorionic villi. These cells contact and attach to the maternal endometrium and serve as a barrier between maternal and fetal blood supplies in human pregnancy [8]. Trophoblasts are a source of many cytokines and other immunomodulatory molecules that support both placental development and parturition [9–11].
Mother-to-child transmission of HIV remains an important source of pediatric infection worldwide [12]. The role that HIV-infected trophoblasts play in vertical transmission and pregnancy outcome is incompletely understood. In in vitro experiments using trophoblastic cell lines, cells were efficiently infected with cell associated virus [13], but not cell free virus [14] due to differences in entry pathways into the cell. In the latter case, virus production was marginally increased by treatment with TNF-α and IL-1β. HIV infections can dysregulate cytokine expression in trophoblasts [15, 16]. It has been speculated that aberrant expression of cytokines results in placental inflammation, facilitating transfer of the virus across inflamed membranes to infect the fetus [16].
Using the FIV-infected cat model for lentivirus vertical transmission, we demonstrated frequent reproductive failure in litters delivered at early-or late-gestation by cesarean section. Placental and fetal infection occurred in nearly all pregnancies in FIV-infected queens [17, 18]. We recently reported that FIV-infection during early-and late-term gestation affects immunomodulator expression, producing a pro-inflammatory microenvironment at early, but not late pregnancy [19]. However, the specific cell populations expressing the targeted immunomodulators were not identified. In the present study, we hypothesized that FIV infection may cause dysregulation of immunomodulator expression in trophoblasts and contribute to reproductive failure. Our objectives were to determine whether feline trophoblasts microdissected from early-and late-term placentas of infected queens were infected with FIV, to quantify trophoblast expression of representative pro-and anti-inflammatory cytokines and other immunomodulators known to be important in pregnancy, and to assess whether gene expression was related to pregnancy outcome.
Animals and virus
All procedures utilizing cats (Felis domesticus) were performed with approval of the Institutional Animal Care and Use Committee of Mississippi State University. As previously reported [17, 18], cats were reproductively mature, specific-pathogen-free (SPF) animals of less than 12 months of age when obtained from a commercial cattery. Ten cats were inoculated intravenously with 1 ml of a feline plasma pool containing approximately 1.3 × 104 copies/ml of FIV-B-2542 (provided by Dr. Edward Hoover) [20]. Ten cats were uninoculated and served as normal controls. Infection was confirmed within 6 weeks p.i. by detection of FIV provirus by PCR and for seroconversion using the PetChek® FIV Antibody Test Kit (IDEXX Laboratories, Inc., Westbrook, ME), then queens were allowed to breed naturally with SPF toms. Toms used to breed uninfected females were never exposed to infected females and vice versa. Breeding was observed and pregnancy was confirmed by palpation and ultrasonography.
Placental Tissue Evaluated in Study
Queen number and placenta
Pregnancy term
Queen FIV status
Fetal viability
Fetus FIV
Trophoblast FIV
Status/Ct value
2779 Placenta B
2779 Placenta D
2779 Placenta E
Non-viable; resorption
0326 Placenta A
8035 Placenta B
Non-viable; arrested
8035 Placenta C
Non-viable; arrested
0866 Placenta A
0866 Placenta B
Non-viable; resorption
9581 Placenta A
9581 Placenta B
9746 Placenta C
9730 Placenta B
9730 Placenta R1
Non-viable; resorption
9730 Placenta R3
Non-viable; resorption
+/ > 37
13226 Placenta C
Non-viable; arrested
13226 Placenta D
Non-viable; arrested
Immunohistochemistry (IHC) to detect trophoblasts
All IHC procedures were performed using RNase-free, sterile conditions according to established protocol using polyclonal antibody to cytokeratin to label trophoblasts [21]. To enhance the capture of trophoblasts by laser capture microdissection (LCM), sections were dehydrated in 75%, 95%, and 100% ethanol. Sections were then placed in undiluted xylene until LCM was performed.
Laser capture microdissection to capture trophoblasts
Following immunohistochemical detection in placental specimens, trophoblasts were microdissected using the Veritas™ Microdissection Instrument (Arcturus Bioscience, Inc., MountainView, CA). Microdissection was performed using the Capture IR laser (Power: 70 mW; Pulse: 2500 μsec; and Intensity: 200 mV) and UV Cutting laser (Laser spot size: ~8.0 μm and Laser power: 7.0 mW). Similar-sized fields of ~2.00 mm2 were collected onto CapSure Macro LCM Caps (Arcturus). All sections and capture areas were photographed using the dissection instrument.
RNA purification and quantification from microdissected tissues
RNA was extracted from microdissected trophoblasts of FIV-B-2542-infected and uninfected queens at early gestation (n = 5 and n = 3, respectively) and late gestation (n = 5 and n = 3, respectively), using the PicoPure RNA Isolation Kit (Arcturus). All RNA samples were treated with DNase to remove contaminating genomic DNA. RNA was quantified using the NanoDrop™ 3300 Fluorospectrometer (Thermo Fisher Scientific, Inc. Waltham, MA). All RNA was stored at -80°C until used.
Reverse transcription-PCR to generate cDNA
Using the following cycling program: 25°C, 10 min; 37°C, 120 min; 85°C, 5 sec; 4°C, hold, cDNA was synthesized using the High Capacity cDNA Reverse Transcription Kit (Applied Biosystems, Inc., Foster City, CA) according to the manufacturer's instructions. Briefly, each reverse transcription reaction was performed in a final reaction volume of 20 μl containing 2 μl of the 10X RT buffer, 2 μl RT random primers, 0.8 μl of 25X dNTP, 1 μl of Multiscribe Reverse Transcriptase, 1 μl RNase inhibitor, 3.2 μl of RNase-free water, and 10 μl of 30-800 pg RNA template. All cDNA was stored at -20°C until used.
Primer design of feline immunomodulators and internal control
Primers and Probes used to Evaluate Immunomodulator and Virus Expression
Primer Sequence
(5' to 3')
Probe Sequence (5' to 3')
Accession # or Reference
β actin
Ref 23
Ref 23
Ref 23
Ref 25
Ref 26
Ref 24
Ref 26
Ref 17
Evaluation of expression of immunomodulators using TaqMan real-time PCR
The real-time PCR (qPCR) used an iCycler (Bio-Rad) and the following cycling program: 50°C, 2 min; 95°C, 10 min; 60 X (95°C, 15 s; 60°C, 1 min). Each reaction contained 10 μl of the 2X TaqMan® Universal PCR Master Mix (Applied Biosystems, Life Technologies Corp., Carlsbad, CA), 1 μl of forward and reverse primers (10 pmol/μl), 1 μl of the respective probe (100 fmol/μl), and ~2-4 μg cDNA. For every placental cDNA sample, parallel reactions were performed in triplicate simplex reactions on separate plates for each gene. Due to the low concentration of RNA isolated from extracted cells, and consequently, the limited amount of cDNA available for use in qPCR, standard curves for internal control and target gene amplicons were generated using pooled, serially-diluted whole placental tissue RNA from representative uninfected cats. Each titration simplex reaction contained 6.25 μl of the 2X Thermoscript™ reaction mix, 0.25 μl of Thermoscript™ RT/Platinum®Taq Mix (Invitrogen), 1 μl of forward and reverse primers (10 pmol/μl), 1 μl of the respective probe (100 fmol/μl), and five-fold diluted 2 μg RNA as starting titration template. The standard curves were used to normalize for differences in PCR efficiency between the internal control and target genes and between sample plate runs. Differences in the amount of template cDNA in each reaction was corrected by the cycle threshold (Ct) value for β-actin. Normalized samples were divided by the calibration, generating the relative expression levels.
Statistical analysis
Statistical evaluation of fetal viability from early-and late-term gestation was previously reported [17, 18]. Statistical analyses of immunomodulator expression for early-and late-term control and FIV-B-2542-infected trophoblasts were done using single-factor ANOVA and the two independent sample Wilcoxon rank sum test (SOCR Analysis, University of California, Los Angeles, CA).
Microdissection of trophoblasts from placental tissues
Expression of FIV gag in trophoblasts
FIV gag was detected in only a single, late-term trophoblast specimen, collected from a placenta associated with a fetal resorption (Table 1). The high Ct value (greater than 37) indicated low level infection of this trophoblast specimen. FIV gag was previously detected in RNA from whole placental tissues using the same protocol [27].
Expression of immunomodulators in trophoblasts
Messenger RNA for IL-4, IL-5, IL-6, IL-1β, IL-12p35, IL-12p40, and CXCR4 was expressed in all early-and late-term trophoblasts. Messenger RNA for TNF-α, IFN-γ, IL-2, IL-18, IL-10, GM-CSF, CD134, and SDF-1α was not detected in trophoblasts from either stage of pregnancy. All immunomodulators were amplified from RNA extracted from whole placental tissues used for generating standard curves (data not shown), indicating that failure to detect some of them in trophoblasts was not a result of inadequate qPCR conditions.
To determine the effect of FIV infection on expression of these immunomodulators, trophoblasts from FIV-infected and control queens at both early-and late-term gestation were compared. At early gestation, the marginally-reduced expression of nearly all immunomodulators in FIV-infected cats did not reach the level of significance. The expression of the anti-inflammatory cytokine IL-5 alone was decreased to a significant level (p = 0.038) (Figure 2B). At late gestation, FIV-infection resulted in decreased expression of the anti-inflammatory cytokine IL-4 (p = 0.021) and the pro-inflammatory cytokine IL-12p35 (p = 0.021); decreased expression of the pro-inflammatory cytokine IL-12p40 (p = 0.057) approached, but did not reach significance. There was no significant effect on the expression of the remaining immunomodulators (Figure 2C).
Using the FIV-infected, pregnant cat to evaluate lentivirus-induced placental immunopathology and pregnancy outcome, we and others reported vertical transfer of FIV-B-2542 at early-and late-term pregnancy [17, 18, 28], with more than 50% of offspring infected. We found fetal non-viability to be significantly higher in FIV-infected queens at both stages of pregnancy [17, 18]. While the mechanism of FIV-induced reproductive failure is unresolved, placental inflammation appears to accompany FIV infection, especially at early pregnancy. In a previous report by our laboratory which evaluated immunomodulator expression in whole placental specimens [19], an FIV-induced, pro-inflammatory microenvironment at early, but not late pregnancy, was evident, including increased expression of IL-6 and IL-12, decreased expression of IL-10, and IL-12:IL-10 ratios that favored the pro-inflammatory cytokine in infected cats at early pregnancy.
In the present study, the effect of FIV infection on immunomodulator expression in a targeted cell population, trophoblasts, was evaluated at early and late gestation. Immunomodulators produced by trophoblasts play important roles in the progression of pregnancy. Thus, understanding how infections or perturbed pregnancy correspond to altered gene expression specifically in trophoblasts, in situ and in vitro, has been the focus of many investigations [29–32]. We were able to detect expression of IL-4, IL-5, IL-6, IL-1β, IL-12p35, IL-12p40, and CXCR4 in feline placental trophoblasts at both time points during feline gestation. The relative expression of all immunomodulators increased in normal specimens as gestation progressed, indicating increased immunomodulatory function of feline trophoblasts at late pregnancy. At early gestation, FIV infection was associated with slightly decreased expression of all cytokines, although only the expression of IL-5 (an anti-inflammatory cytokine) decreased to significant levels. At late gestation one anti-inflammatory cytokine (IL-4) and one pro-inflammatory cytokine (IL-12p35) were decreased in FIV-infected queens. The data suggest that FIV infection of the queens caused a minor effect on trophoblast function, and an FIV-induced pro-inflammatory effect in trophoblasts was not clearly evident.
Non-viable pregnancies were accompanied by decreased trophoblast expression of both pro-and anti-inflammatory immunomodulators at early-(IL-4, IL-6, IL-12p35, and CXCR4) and late-term (IL-4, IL-12p35, and IL-12p40). One would predict reduced placental function in dying pregnancies. However, decreased gene expression was not significant for all immunomodulators, suggesting that reduction in immunomodulator function was not solely attributable to deterioriation of the placenta in non-viable pregnancies. The decreased expression of IL-4 and IL-6 in early placental trophoblasts from non-viable pregnancies parallels the IL-4 and IL-6 expression patterns found in the endometrial tissue and deciduas of women with frequent implantation failures[33] and recurrent abortions [34]. IL-4 is a Th2 cytokine that is mainly involved in the peri-implantation period of placental development [35]. Along with transforming growth factor-β (TGF-β), IL-4 mediates the process of trophoblast invasion during human placentation [36–39]. IL-4 and IL-6 contribute to the angiogenesis of trophoblastic villi [40]. Silencing of IL-6 in JEG-3 cells (choriocarcinoma cells maintaining trophoblast-like characteristics) using siRNA resulted in significantly reduced proliferation of those cells [41]. IL-1β activates matrix metalloprotease 3 (MMP-3), a protease that digests extracellular matrix proteins and allows trophoblast penetration of the maternal endometrium [42]. In the present study, the decrease in expression of IL-1β approached significance (p = 0.052) at early pregnancy. In the human placenta, CXCR4, expressed by trophoblasts, is also important in trophoblast invasion via interaction with its ligand, SDF-1α, expressed on decidual stromal cells [43]. In addition, this interaction protects trophoblasts from apoptosis via stimulation of anti-apoptotic pathways [44]. IL-12, a heterodimeric cytokine consisting of 35 and 40 kd protein subunits, is also an important regulator of trophoblast invasion. It inhibits trophoblast invasion by causing an IFN-γ-dependent reduction in expression of MMP-2, MMP-9, urokinase-type plasminogen activator, and other proteases that digest extracellular matrix proteins. IL-12 also causes an increase in expression of inhibitors of these proteases [45]. The decreased expression of IL-4, IL-6, IL-12p35, and CXCR4 in trophoblasts from early-term non-viable pregnancies shows dysregulation of putative mediators of feline trophoblast invasion during early feline gestation, an outcome that could result in nutrient and oxygen deprivation of the developing fetus and account for the increased reproductive failure that occurred in infected animals.
In contrast, cultured placental trophoblasts from HIV-infected women expressed significantly higher amounts of IL-1β, IL-6, and TNF-α than placentas from uninfected women [16]. These data indicate that HIV-infection during pregnancy results in increased cytokine expression and that a pro-inflammatory microenvironment (an environment potentially conducive to vertical transfer) exists in HIV-infected placentas. We did not see this trend in trophoblasts collected from the cat model. However, the study of human tissues used cultured trophoblasts, whereas in the present study, RNA was extracted from trophoblasts microdissected from frozen placentas. Whether culturing the cells affected gene expression is unknown.
While we previously reported the detection of mRNA for the primary FIV receptor CD134 in whole feline placental tissues [46], we did not detect CD134 in any trophoblast sample from early-or late-term pregnancy. Correspondingly, FIV was detectable at a low level (Ct > 37) in only one trophoblast specimen, a placenta from a late-term fetal resorption, indicating that trophoblasts may not be highly susceptible or permissive to FIV infection. This result was surprising, given the high frequency of fetal infection that was documented in these same cats [17, 18], and suggests that infected trophoblasts may not be the source of fetal infection. We speculate that immune cells in the surrounding microenvironment, such as regulatory T cells (Tregs) or Th17 cells, the former of which are known to support the replication of FIV [47, 48], may influence trophoblast function and serve as the source of fetal infection. Our laboratory is currently investigating feline Tregs and Th17 cells in lentivirus-induced placental immunopathology and pregnancy failure.
A limitation of this study was the small number of tissues evaluated by LCM, particularly in terms of separating viable versus non-viable tissues. LCM is a procedure which is laborious, time-intensive, and expensive to perform, so limiting the samples collected was a necessity. We attempted to minimize inter-cat variability by evaluating multiple placentas from the same queen where possible. The differences that we have historically observed between placentas from the same queen assured us that we can consider placentas from the same queens as independent specimens [17, 19, 27]. Yet, the inclusion of additional placental specimens may have strengthened the statistical power and allowed us to identify additional viral effects. In addition, it is impossible to eliminate all cell contamination from the microenvironment surrounding the trophoblasts due to limitations in the precision of focusing the laser. However, our samples were highly enriched for trophoblasts, as is evident by the well-defined tissue lifts that we were able to achieve (Figure 1). LCM followed by qPCR has been used as a means to collect trophoblasts from whole placental specimens and measure gene expression in these cells by others [49]. Regardless of the limitations, this study provides novel information. Our previous reports clearly reveal that FIV infection results in significantly higher rates of reproductive failure, probably by inducing placental immunopathology [17–19]. The present data shed additional light on this anomaly, suggesting that aberrant expression of trophoblast cytokines that may regulate trophoblast invasion accompanies fetal demise.
Feline trophoblasts expressed an array of pro-and anti-inflammatory immunomodulators whose expression increased from early to late pregnancy in normal tissues, while some immunomodulators that were expressed in whole placental tissues were not detected. A virus-induced, pro-inflammatory environment was not detected in trophoblasts. Non-viable pregnancies were associated with decreased expression of immunomodulators which regulate trophoblast invasion in other species, a finding which may explain the high rate of reproductive failure in FIV-infected cats. The detection of FIV RNA in trophoblasts was rare. This result suggests that reproductive failure and altered trophoblast gene expression in FIV-infected queens was not a direct result of viral infection of trophoblasts, but perhaps was influenced by cells in the microenvironment adjacent to trophoblasts. The effects of placental immune cells on trophoblast function and pregnancy maintenance in the FIV-infected cat requires additional study.
We thank Dr. Shane C. Burgess, College of Veterinary Medicine, Mississippi State University, for the use of laboratory facilities and advice in quantitative PCR. We are grateful to Dr. Edward A. Hoover, Colorado State University, for providing the FIV-B-2542 inoculum. We thank Aimee Bell for assistance with LCM and the veterinary staff at the College of Veterinary Medicine, Mississippi State University for assistance with surgeries and animal care. This project was supported by the National Institutes of Health (2R15AI048419-02A1) and (3R15AI048419-02A1S1).
Authors' Affiliations
- Bennett WA, Lagoo-Deenadayalan S, Stopple JA, Barber WH, Hale E, Brackin MN, Cowan BD: Cytokine expression by first-trimester human chorionic villi. Am J Reprod Immunol 1998, 40: 309-318.View ArticlePubMedGoogle Scholar
- Challis JR, Lockwood CJ, Myatt L, Norman JE, Strauss JF, Petraglia F: Inflammation and pregnancy. Reprod Sci 2009, 16: 206-215. 10.1177/1933719108329095View ArticlePubMedGoogle Scholar
- Robertson SA, Seamark RF, Guilbert LJ, Wegmann TG: The role of cytokines in gestation. Crit Rev Immunol 1994, 14: 239-292.View ArticlePubMedGoogle Scholar
- Saito S: Cytokine cross-talk between mother and the embryo/placenta. J Reprod Immunol 2001, 52: 15-33. 10.1016/S0165-0378(01)00112-7View ArticlePubMedGoogle Scholar
- Munoz-Suano A, Hamilton AB, Betz AG: Gimme shelter: the immune system during pregnancy. Immunological Reviews 2011, 241: 20-38. 10.1111/j.1600-065X.2011.01002.xView ArticlePubMedGoogle Scholar
- Tokmadz̆IĆ VS, Tsuji Y, BogoviĆ T, LaŠKarin G, Cupurdija K, Š Trbo N, Koyama K, Okamura H, Podack ER, Rukavina D: IL-18 is Present at the Maternal-Fetal Interface and Enhances Cytotoxic Activity of Decidual Lymphocytes. American Journal of Reproductive Immunology 2002, 48: 191-200. 10.1034/j.1600-0897.2002.01132.xView ArticlePubMedGoogle Scholar
- Beer AE, Kwak JYH: Immunology of normal pregnancy. Immunol Allergy Clin N Am 1998, 18: 249-270. 10.1016/S0889-8561(05)70004-7View ArticleGoogle Scholar
- Senger PL: Chapter 14. Placentation, Gestation, and Parturition. In Pathways to Pregnancy and Parturition. 1st edition. Pullman, WA: Current Conceptions, Inc; 1997:233-247.Google Scholar
- Hauguel-de Mouzon S, Guerre-Millo M: The placenta cytokine network and inflammatory signals. Placenta 2006, 27: 794-798. 10.1016/j.placenta.2005.08.009View ArticlePubMedGoogle Scholar
- Weetman AP: The immunology of pregnancy. Thyroid 1999, 9: 643-646. 10.1089/thy.1999.9.643View ArticlePubMedGoogle Scholar
- Bowen JM, Chamley L, Keelan JA, Mitchell MD: Cytokines of the Placenta and Extra-placental Membranes: Roles and Regulation During Human Pregnancy and Parturition. Placenta 2002, 23: 257-273. 10.1053/plac.2001.0782View ArticlePubMedGoogle Scholar
- Key facts by region-2008 Report on the global AIDS epidemic[http://data.unaids.org/pub/GlobalReport/2008/20080715_fs_regions_en.pdf]
- Lagaye S, Derrien M, Menu E, Coito C, Tresoldi E, Mauclere P, Scarlatti G, Chaouat G, Barre-Sinoussi F, Bomsel M: Cell-to-cell contact results in a selective translocation of maternal human immunodeficiency virus type 1 quasispecies across a trophoblastic barrier by both transcytosis and infection. J Virol 2001, 75: 4780-4791. 10.1128/JVI.75.10.4780-4791.2001PubMed CentralView ArticlePubMedGoogle Scholar
- Vidricaire G, Tardif MR, Tremblay MJ: The low viral production in trophoblastic cells is due to a high endocytic internalization of the human immunodeficiency virus type 1 and can be overcome by the pro-inflammatory cytokines tumor necrosis factor-alpha and interleukin-1. J Biol Chem 2003, 278: 15832-15841. 10.1074/jbc.M210470200View ArticlePubMedGoogle Scholar
- Lee BN, Ordonez N, Popek EJ, Lu JG, Helfgott A, Eriksen N, Hammill H, Kozinetz C, Doyle M, Kline M, et al.: Inflammatory cytokine expression is correlated with the level of human immunodeficiency virus (HIV) transcripts in HIV-infected placental trophoblastic cells. J Virol 1997, 71: 3628-3635.PubMed CentralPubMedGoogle Scholar
- Shearer WT, Reuben J, Lee BN, Popek EJ, Lewis DE, Hammill HH, Hanson IC, Kline MW, Langston C: Role of placental cytokines and inflammation in vertical transmission of HIV infection. Acta Paediatr Suppl 1997, 421: 33-38.View ArticlePubMedGoogle Scholar
- Weaver CC, Burgess SC, Nelson PD, Wilkinson M, Ryan PL, Nail CA, Kelly-Quagliana KA, May ML, Reeves RK, Boyle CR, Coats KS: Placental immunopathology and pregnancy failure in the FIV-infected cat. Placenta 2005, 26: 138-147. 10.1016/j.placenta.2004.04.013View ArticlePubMedGoogle Scholar
- Boudreaux CE, Lockett NN, Chemerys DN, Clay BT, Scott VL, Willeford B, Brown T, Coats KS: Maternal hematological and virological characteristics during early feline immunodeficiency virus (FIV) infection of cats as predictors of fetal infection and reproductive outcome at early gestation. Vet Immunol Immunopathol 2009, 131: 290-297. 10.1016/j.vetimm.2009.04.011PubMed CentralView ArticlePubMedGoogle Scholar
- Scott VL, Boudreaux CE, Lockett NN, Clay BT, Coats KS: Cytokine Dysregulation in Early-and Late-Term Placentas from Feline Immunodeficiency Virus (FIV)-Infected Cats. American Journal of Reproductive Immunology 2011, 65: 480-491. 10.1111/j.1600-0897.2010.00919.xPubMed CentralView ArticlePubMedGoogle Scholar
- Rogers AB, Hoover EA: Maternal-fetal feline immunodeficiency virus transmission: timing and tissue tropisms. J Infect Dis 1998, 178: 960-967. 10.1086/515692View ArticlePubMedGoogle Scholar
- Scott VL, Wallace K, Mays S, Ryan P, Coats KS: An immunohistochemical assay to detect trophoblasts in frozen feline placenta. J Vet Diagn Invest 2011, 23: 275-281. 10.1177/104063871102300212View ArticlePubMedGoogle Scholar
- Scott VL, Burgess SC, Shack LA, Lockett NN, Coats KS: Expression of CD134 and CXCR4 mRNA in term placentas from FIV-infected and control cats. Vet Immunol Immunopathol 2008, 123: 90-96. 10.1016/j.vetimm.2008.01.014PubMed CentralView ArticlePubMedGoogle Scholar
- Taglinger K, Van Nguyen N, Helps CR, Day MJ, Foster AP: Quantitative real-time RT-PCR measurement of cytokine mRNA expression in the skin of normal cats and cats with allergic skin disease. Veterinary Immunology and Immunopathology 2008, 122: 216-230. 10.1016/j.vetimm.2007.11.014View ArticlePubMedGoogle Scholar
- Leutenegger CM, Mislin CN, Sigrist B, Ehrengruber MU, Hofmann-Lehmann R, Lutz H: Quantitative real-time PCR for the measurement of feline cytokine mRNA. Veterinary Immunology and Immunopathology 1999, 71: 291-305. 10.1016/S0165-2427(99)00100-2View ArticlePubMedGoogle Scholar
- Foley JE, Rand C, Leutenegger C: Inflammation and changes in cytokine levels in neurological feline infectious peritonitis. J Feline Med Surg 2003, 5: 313-322. 10.1016/S1098-612X(03)00048-2View ArticlePubMedGoogle Scholar
- Kipar A, Meli ML, Failing K, Euler T, Gomes-Keller MA, Schwartz D, Lutz H, Reinacher M: Natural feline coronavirus infection: differences in cytokine patterns in association with the outcome of infection. Vet Immunol Immunopathol 2006, 112: 141-155. 10.1016/j.vetimm.2006.02.004View ArticlePubMedGoogle Scholar
- Lockett NN, Scott VL, Boudreaux CE, Clay BT, Pruett SB, Ryan PL, Coats KS: Expression of regulatory T cell (Treg) activation markers in endometrial tissues from early and late pregnancy in the feline immunodeficiency virus (FIV)-infected cat. Placenta 2010, 31: 796-802. 10.1016/j.placenta.2010.06.019PubMed CentralView ArticlePubMedGoogle Scholar
- O'Neil LL, Burkhard MJ, Hoover EA: Frequent perinatal transmission of feline immunodeficiency virus by chronically infected cats. J Virol 1996, 70: 2894-2901.PubMed CentralPubMedGoogle Scholar
- Burrough ER, DiVerde KD, Sahin O, Plummer PJ, Zhang Q, Yaeger MJ: Expression of Toll-like Receptors 2 and 4 in Subplacental Trophoblasts From Guinea Pigs (Cavia porcellus) Following Infection With Campylobacter jejuni. Veterinary Pathology Online 2011, 48: 381-388.View ArticleGoogle Scholar
- Riewe SD, Mans JJ, Hirano T, Katz J, Shiverick KT, Brown TA, Lamont RJ: Human trophoblast responses to Porphyromonas gingivalis infection. Molecular Oral Microbiology 2010, 25: 252-259. 10.1111/j.2041-1014.2010.00573.xPubMed CentralView ArticlePubMedGoogle Scholar
- Aldo PB, Mulla MJ, Romero R, Mor G, Abrahams VM: Viral ssRNA Induces First Trimester Trophoblast Apoptosis through an Inflammatory Mechanism. American Journal of Reproductive Immunology 2010, 64: 27-37.PubMed CentralPubMedGoogle Scholar
- Tayade C, Black GP, Fang Y, Croy BA: Differential Gene Expression in Endometrium, Endometrial Lymphocytes, and Trophoblasts during Successful and Abortive Embryo Implantation. The Journal of Immunology 2006, 176: 148-156.View ArticlePubMedGoogle Scholar
- Stewart-Akers AM, Krasnow JS, Brekosky J, DeLoia JA: Endometrial leukocytes are altered numerically and functionally in women with implantation defects. Am J Reprod Immunol 1998, 39: 1-11. 10.1016/S0165-0378(98)00025-4View ArticlePubMedGoogle Scholar
- Piccinni MP, Romagnani S: Regulation of fetal allograft survival by a hormone-controlled Th1-and Th2-type cytokines. Immunol Res 1996, 15: 141-150. 10.1007/BF02918503View ArticlePubMedGoogle Scholar
- Wilczynski JR: Th1/Th2 cytokines balance--yin and yang of reproductive immunology. Eur J Obstet Gynecol Reprod Biol 2005, 122: 136-143. 10.1016/j.ejogrb.2005.03.008View ArticlePubMedGoogle Scholar
- Irving JA, Lala PK: Functional role of cell surface integrins on human trophoblast cell migration: regulation by TGF-beta, IGF-II, and IGFBP-1. Exp Cell Res 1995, 217: 419-427. 10.1006/excr.1995.1105View ArticlePubMedGoogle Scholar
- Klentzeris LD: The role of endometrium in implantation. Hum Reprod 1997, 12: 170-175.PubMedGoogle Scholar
- Piccinni MP, Maggi E, Romagnani S: Role of hormone-controlled T-cell cytokines in the maintenance of pregnancy. Biochem Soc Trans 2000, 28: 212-215.View ArticlePubMedGoogle Scholar
- Piccinni MP, Scaletti C, Maggi E, Romagnani S: Role of hormone-controlled Th1-and Th2-type cytokines in successful pregnancy. J Neuroimmunol 2000, 109: 30-33. 10.1016/S0165-5728(00)00299-XView ArticlePubMedGoogle Scholar
- Jauniaux E, Gulbis B, Schandene L, Collette J, Hustin J: Distribution of interleukin-6 in maternal and embryonic tissues during the first trimester. Mol Hum Reprod 1996, 2: 239-243. 10.1093/molehr/2.4.239View ArticlePubMedGoogle Scholar
- Dubinsky V, Poehlmann TG, Suman P, Gentile T, Markert UR, Gutierrez G: Role of regulatory and angiogenic cytokines in invasion of trophoblastic cells. Am J Reprod Immunol 2010, 63: 193-199.View ArticlePubMedGoogle Scholar
- Husslein H, Haider S, Meinhardt G, Prast J, Sonderegger S, Knöfler M: Expression, Regulation and Functional Characterization of Matrix Metalloproteinase-3 of Human Trophoblast. Placenta 2009, 30: 284-291. 10.1016/j.placenta.2008.12.002PubMed CentralView ArticlePubMedGoogle Scholar
- Zhou WH, Du MR, Dong L, Yu J, Li DJ: Chemokine CXCL12 promotes the cross-talk between trophoblasts and decidual stromal cells in human first-trimester pregnancy. Hum Reprod 2008, 23: 2669-2679. 10.1093/humrep/den308View ArticlePubMedGoogle Scholar
- Jaleel MA, Tsai AC, Sarkar S, Freedman PV, Rubin LP: Stromal cell-derived factor-1 (SDF-1) signalling regulates human placental trophoblast cell survival. Mol Hum Reprod 2004, 10: 901-909. 10.1093/molehr/gah118View ArticlePubMedGoogle Scholar
- Karmakar S, Dhar R, Das C: Inhibition of Cytotrophoblastic (JEG-3) Cell Invasion by Interleukin 12 Involves an Interferon γ-mediated Pathway. Journal of Biological Chemistry 2004, 279: 55297-55307. 10.1074/jbc.M407013200View ArticlePubMedGoogle Scholar
- Scott VL, Burgess SC, Shack LA, Lockett NN, Coats KS: Expression of CD134 and CXCR4 mRNA in term placentas from FIV-infected and control cats. Vet Immunol Immunopathol 2008, 123: 90-96. 10.1016/j.vetimm.2008.01.014PubMed CentralView ArticlePubMedGoogle Scholar
- Joshi A, Garg H, Tompkins MB, Tompkins WA: Preferential feline immunodeficiency virus (FIV) infection of CD4+ CD25+ T-regulatory cells correlates both with surface expression of CXCR4 and activation of FIV long terminal repeat binding cellular transcriptional factors. J Virol 2005, 79: 4965-4976. 10.1128/JVI.79.8.4965-4976.2005PubMed CentralView ArticlePubMedGoogle Scholar
- Joshi A, Vahlenkamp TW, Garg H, Tompkins WA, Tompkins MB: Preferential replication of FIV in activated CD4(+)CD25(+)T cells independent of cellular proliferation. Virology 2004, 321: 307-322. 10.1016/j.virol.2004.01.014View ArticlePubMedGoogle Scholar
- Chan S, Murray PG, Franklyn JA, McCabe CJ, Kilby MD: The use of laser capture microdissection (LCM) and quantitative polymerase chain reaction to define thyroid hormone receptor expression in human 'term' placenta. Placenta 2004, 25: 758-762. 10.1016/j.placenta.2004.02.007View ArticlePubMedGoogle Scholar
This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.


text:
"STRONG. The Magazine for Girls."
STRONG. The Magazine for Girls.
"STRONG aims to empower girls to be the best version of themselves they can possibly be by presenting them with strong female role models and encouraging their interests and passions, wherever they may lie."
Hi, my name is Sarah Beach and I'm the founder of "STRONG. The Magazine for Girls". Since I started this endeavor I have received an overwhelming amount of support. So many people believe in this project and have offered their time, services and expertise free of charge to help bring the first issue to life. I'm now looking to the Kickstarter community to give us the push we need to really get this magazine off the ground! You can click on a link and read the first issue of STRONG at the bottom of the page.
What is STRONG?
STRONG is a new magazine aimed at tween and teenaged girls that seeks to break the mold. While many other publications on offer for this age group focus on celebrities, STRONG focuses on real girls with diverse interests. While other publications focus on body image and fashion, STRONG focuses on keeping a healthy body and mind. STRONG also presents great role models - girls who have overcome adversity and thrived, and women who are breaking glass ceilings in areas previously dominated by men.
To us, STRONG girls are girls who are confident to be themselves.
Why is STRONG necessary?
Women make up 50% of the workforce but hold just 26% of tech industry jobs. Only 11% of executives at Forbes 500 companies are female. We have never had a female President, Vice President, Head of the FBI, UN Secretary General or world chess champion. Things may be slowly improving, but the list of careers in which women are underrepresented is very, very long.
As our girls reach adolescence and start to really think about where they fit into the world, they are confronted by a lack of female role models. In the adventure books they read and the movies they watch, the protagonists are usually male. When they turn on the news they will see men both in the news and presenting the news (I once saw a split screen with a female anchor interviewing three other women and was so surprised to see no men on the screen that I actually snapped a picture on my phone and posted it to Facebook!). A picture of the G2O - the leaders and bank governors from 20 of the world's richest and most powerful nations - features just four women and 32 men.
In school their teachers are still predominately female, and they are far more likely to see a male doctor than a female one if they get sick because only 37% of physicians and surgeons are women.
Young girls I know are interested in all kind of different things; they love math and science, digging in the dirt, dressing up and dancing to music equally. But around the time they start to look to society for reassurance that they're OK, society instead sends them the message that all girls should be interested in romance and appearance; that wearing the latest fashion is crucial to their likability and that they should be concerned about what boys think of them. There is very little mention of women in industry, women in engineering, women firefighters or women in space. Society is still trying to put girls in a box.
What are STRONG's goals?
- We want STRONG to be a magazine that genuinely reflects girls today with their diverse lives and interests.
- We want girls to be inspired by reading about other girls like them who are pursuing their dreams.
- We want to help girls realize that there are women out there all over the world breaking glass ceilings and taking their place alongside men.
- We want to encourage girls to take responsibility for looking after their own bodies and minds.
- We want to teach girls fundamental life skills that every young adult should learn: from managing their money and their time, to doing their laundry, basic first aid and everything in between.
- We want to help girls understand the world they are growing up in by talking about current and historical events and by showing them other countries and cultures.
- We want to promote tolerance and kindness.
My Inspiration
My inspiration to create STRONG came from within my own family. I have three kids, 2 girls and a boy. It was a conversation I had with my daughters, Lucy and Daisy, that really convinced me that I had to take action. Despite being talented kids who are interested in math, science, and engineering, it transpired that they were getting the message that these weren't really things they could do long term, because "Girls don't really do that sort of thing.". I looked at the world through their eyes and saw school photo companies offer "magazine cover" photo options featuring video games and sports for boys and fashion for girls. I looked on their bookshelves and saw that all the action and adventure books they read had male protagonists. I thought about the movies we've watched and realized that that was mainly true for them too. I looked at the news and the messages that my kids had been getting there and I looked at our own family, where I gave up a successful career to raise our kids whilst my husband works in a male dominated field (he's a software engineer). I had to do something. I had to show my girls and girls like them that they can truly be whoever they want to be.
Issue 1 of STRONG.
We were fortunate to receive backing to print the first issue of STRONG and we are now making it available, in print and online, free of charge! We want as many people as possible to read STRONG so they can see for themselves how great this magazine is. We are going to place copies in libraries, schools and waiting rooms in order to further spread the word. Please click on the link at the bottom of the page to read STRONG. We hope you like what you see. It's not perfect, it's been a project that has evolved and I have been on a steep learning curve, but I feel we've made a good start!
What We Hope to Achieve from this Kickstarter
There are two main reasons for launching this Kickstarter. The first is to spread the word about STRONG. We want as many people as possible to read the first issue of STRONG so they can see what we are trying to achieve. In order to truly represent all girls, we need to hear from them! So we are encouraging our readers to click on the links within the magazine or to visit the "Just for Girls" part of our website to take part in polls, and submit reviews, photos and stories. If you are a STRONG girl with a story to tell, we want to hear from you!
The Just for Girls section of the website will grow over time to incorporate more articles and interactive content.
The second reason is that we need funding! STRONG is my baby. I am writing, editing, arranging the layout, marketing, constructing websites, writing business plans and more (like I said, I'm on a steep learning curve!). Having said that, many wonderful people have stepped in and volunteered their time to help me out because they truly believe that STRONG is an important project. I have had friends (and strangers!) offer to write, copy edit, help with graphic design, offer advice marketing, and take photographs. I am so confident that people want to see STRONG succeed that I have opened up online subscriptions and committed to producing the next 4 issues. However, in order for STRONG to grow and to be sustainable, we need a boost. Funding from this Kickstarter will help to pay for the printing and distribution of the second issue. It will also provide seed money for other fundraising events and for things such as print runs to look at different paper qualities as well as for professional help to take the magazine to the next level. I also plan to use the money to continue to market STRONG to build more subscriptions with the aid of a subscription agency.
I would really love your backing to help my dream become a reality and, in doing so, to help all of our daughters to do the same.
Thank you - and happy reading!
Sarah Beach
Founder, STRONG. The Magazine for Girls
Facebook: @strongmagazineforgirls
Twitter @strongforgirls
Instagram: @strongmagazineforgirls
Risks and challenges
Many of you visiting this page will know that this is actually a relaunch of the campaign for STRONG. The first time around we gained a LOT of supporters. Not only did 195 people pledge $11,500, but I also had countless people approach me offering to help and get involved, or simply offering encouragement. Unfortunately, our goal was $15,000, and as Kickstarters are all or nothing, not reaching our goal meant not getting funded. This time around I am reducing the goal amount to what is hopefully a more manageable $10,000 and encouraging all our previous backers to pledge early in the campaign to boost its visibility. I'm able to reduce the goal amount by cutting down on the number of issues we'll print, and offering online subscriptions which will offset some of the printing costs.
Putting STRONG out there again is a big risk! I'm hearing that lots of you think it's a great project, so I just hope we can get enough traction this time to achieve our goal.Learn about accountability on Kickstarter
Questions about this project? Check out the FAQ
Funding period
- (30 days)

subset: commoncrawl


text:
Cheeses, meats, pastries and a thousand other specialties make up the outstanding and authentic Vaudois terroir that will delight your senses.
Offering a charming landscape of fertile plains and wooded hills, the north of the Lake Geneva Region is quite different from the Alps and the shores of Lake Geneva. This region of vegetable crops and forests favoured the emergence of original terroir specialties. In the villages of the Jura and the plateau, thoroughness, the love of work well-done and ingenuity are at the root of the know-how used in the precision industry and in gastronomy. Although some cured meat types claim a millennial history, the Bonvillars truffles only became popular about ten years ago. They are, however, all offered in the region's restaurants and traditional inns (called "pintes").


text:
West Palm Installers of Hurricane Shutters in Miami, Florida provide your home or business with the best line of defense against severe winds and dangerous debris. We install Hurricane Shutters, Accordion Shutters, Bahama Shutters, Colonial Shutters, Storm Panels, Rolldown Shutters, Impact Doors and Impact Windows.
Our Hurricane Shutters Products & Services:
Our factory is equipped with the most advanced machinery and we have the largest inventory of products waiting for your order. Our office personnel are ready to answer your call and answer any and all questions that you may have about our products & services.
Why Choose West Palm Installers:
At West Palm Installers we manufacture our own hurricane protection systems and we are proud to say that we use only the best, strongest materials and components in the industry, as well as keeping a highly demanding quality control in both the manufacturing and installation.
Contact West Palm Installers:
5141 NW 79th Avenue
Doral, FL 33166
Dade: (305) 406.3575
Toll Free: (866) 219.4710
Email: shutters @ westpalminstallers.com


text:
My friend John
(Sis: "Have you ever met him?"
Me: "No."
Sis: "I think it's funny that you call him your friend."
Me: shrug shoulders- "Whatever!")
over at thebestwaytohaveagoodideaistohavelots posted a great video that I want to share. It's a spoof both on news shows and on people who are over-concerned about what schools are teaching:
Don't click here to read more. Click here to read full entry.
Saturday, April 26, 2008
Spoof Videos, Whales, America's Schools
My friend John
Thursday, April 24, 2008
Fire Ants: Giving Regular Ants a Bad Name
"Sorry little ant," said me to the ant who froze as he saw me about to squish him on my kitchen floor, "but I just returned from fire ant territory. I have no mercy for you."
Before I went to Mississippi, the words "fire ants" conjured images of tiny red ants that you might run across in say, a crevice on one outside wall of your home. You know, somewhere you never go. El-wrongo.
The truth about fire ants: they are normal, small-ish ant size, and they are everywhere. Everywhere that there is grass-- in your yard, on the grass beside the road where you jog, in the graveyard-- you have ants. I think there is some poison you can put down, but that ain't good for kids and pets.
Fortunately, fire ants live in large mounds, so you can spot them. However, if you inadvertently step on one while wearing sandals, you are f#@%&! My nephew had 50 or so red pus-filled welts on his legs for a week after he stepped on an ant hill.
Ever since then, I'm on hyper alert in parks and yards. If grass is really really short, I'll walk on it in sandals, but I would NEVER walk in grass over 2 inches high in anything but sneakers, and even then, I would watch the ground constantly.
Taking a walk in the neighborhood? La la la, *GASP*- a 5 inch high mound that you know is filled with 1,000 biting ants! It's not like there's a specific danger ant area; it's everywhere.
Jogging in the park? *AUGH*!!! Leap sideways to avoid a food-wide ant estate.
That little quote in the intro? I actually said that out loud. Just me, home alone, talking to an ant. Click here to read full entry.
Friday, April 18, 2008
Harrison Ford, Nephews, Dreams
I just had the most insane dream. I was about to get in the car to go to work when Harrison Ford walked by. We chatted a bit, and I asked him if he was depressed, and he said, "All I can say is, bless any man who marries a younger woman." I was sitting on the ground, and he leaned down and kissed me three times on the lips, then walked away. I was a little taken aback, but I can roll with a lot of unexpected things. I considered calling after him, "Say hi to Calista!" but thought she might not know who I was, so I said, "I like those lips!" then got in my car.
Nephew 2: Jack, look. (Trying to put huge coin in his piggy bank.)
Nephew 1: I told you it was too big.
Nephew 2: See? It don't work.
Nephew 1, looking in pantry: No peanut butter crackers!?! Rrrrrrr! It can't be true.
I keep a journal, so if I have a dream I want to write in, I write my dream in it sideways. That way, the dream is in with the current entries, so when I go back and read, I can see what in my life inspired certain dreams. They often relate in interesting ways. I put them sideways so I can find them easily if I just want to go back and read my dreams. Click here to read full entry.
Thursday, April 17, 2008
Why Do People Need Friends Other Than Me?
I'm in Mississippi, and right now my sister is talking on the phone to her friend Jenn and I'm feeling really annoyed that she has other people she's friends with other than me. I think it's because I live far away, so the thought that anyone else gets to be with her when I can't bothers me.
It's the same thing when my friend in Portland OR sends me her pics on flickr, and I see pictures of her friends over at her house. I think, I hate those people. I say to myself, KOR, that's not rational, I'm sure they're very nice, and she needs to have friends in her town.
My friend in Ohio (yet another friend living far away!) agreed, in college, that we should all move to the same town. The friends I have from college are so great, and it stinks that they are far away. I don't think it's too late to implement that plan. Click here to read full entry.
Sunday, April 13, 2008
Stalking the Cast of Lost 3/20
On Tuesday, I start thinking about Thursday's upcoming Lost episode, and on Thursday, I start trolling Lost blogs. I found this among some cast interviews. It's a very funny interview- Dominic Monaghan poses as a German interview and fools his friend, and Lord of the Rings co-star, Elijah Woods.
I actually laughed out loud several times during this.
Click here to read full entry.
Trash Day: Antic. and Spying
6 am Thursday. There's a certain excitement in the air. My trash can is out front, and it's going to be picked up.
This is one of my favorite days of the week, which might sound odd. Part of the thrill is the happiness of knowing I'll have an empty trash can to put trash into, but part of it is the waiting. You might think I'm kidding when I say I run to the window when I hear the rumble of the truck. I crawl to the window and spy as the garbage men move like an efficient swat team, quickly drag my can over, lift it into the truck, shove it back into my driveway, leap on the truck, and speed away.
I'm not sure why this excites me. Maybe it's because we didn't have trash pick up where I grew up. I'm sure they had it available, but I don't remember any trash trucks in my neighborhood. I think we all just took our trash to the dump. It's a sort of small county (now it's growing because it's an hour south of DC) so the dump, along with the grocery store and post office, was one of the places you could feasibly bump into someone.
Since it's pitch dark at 6:15, I can't just look out the window; they'll see me. I have to be in a dark room, but near a window on the street side, so I'm sitting in my office, ready to switch off the light when I see them. I wanted to read and write in my sunroom (pitchblackroom) to watch the sunrise, but I had to come here for trash day. I can't have them know I'm watching them, although I did sit on my steps and wave one day. That was when I was on a detective mission for my city's new trash cans. You can read more about that on my other blog, Stamford Talk. The new trash cans were a big saga.
Hold on a sec. I think I hear something.
Click here to read full entry.
Saturday, April 12, 2008
I Destroyed Fancy Pancakes, Now I Have to Fix It
Yeah. I wanted to switch this blog over to a different google account, and rather than researching if there was an easy way to do it, I took off all my old posts and pasted them into Word documents with the idea of just reposting them. I knew it would take a while, but part of me likes creating problems to fix, especially when the problem involves reorganizing papers. I also wanted to clean Fancy Pancakes up a little bit, because I want the site to be less about piddly day to day stuff and more essay-ish like my friend Christopher's blog.
Turns out I could have just switched the blog over by sharing admin privileges and then deleting the old account. That would have taken about 4 minutes. Instead, it will take me around 4 hours to repost it all. Oh- and I also need to redo the "add more" function, my profile, and my blogroll.
So. I need to get back to work. Click here to read full entry.


text:
This "Mommy, Somebody Needs You" article has been making its rounds on my FB feed. It resonates so much with us moms it makes my heart ache a little when I read it.
That 'Mommy' is my duty, privilege and honor. I am ready to be there when somebody needs me, all day and all night. Mommy means I just put the baby back down after her 4am feeding when a 3-year-old has a nightmare. Mommy means I am surviving on coffee and toddler leftovers. Mommy means my husband and I haven't had a real conversation in months. Mommy means I neglect myself and put others before my needs, without a thought. Mommy means that my body is full of aches and my heart is full of love.
True stories.
The author couldn't have put it better. It is definitely a huge privilege to be a mommy. I actually love being very much needed by my babies. Since the arrival of our #2, I told myself that I need to cherish every bit of this since she could very well be our last baby.
Somebody needs me at bedtime.
To read books with. To revise Chinese words together. To hold to sleep with.
Despite the comfortable cot and cot bed we got them, Mommy's bed is always the best place to fall asleep on. Almost at every bed/nap time, I would have such conversations with the kids.
Me: Go to your bed and sleep, baby.
Cub: I sleep your bed first?
Me: I will roll you back to your bed after you fall asleep ok?
Cub: Ok! *rolls over to his bed to bring his bolster and panda*
Me: Can you go to sleep now? It's getting late.
Cub: Mommy, pat pat.
Me: *pat pat*
Cub: Mommy, hold hand.
Me: Do you want to hold my hand or want me to pat you to sleep?
Cub: *pauses* pat / hold hand (depending on preference for the night)
Cub: Mommy, blanket!
Me: Meimei, close your eyes and sleep!
Dragonlady: Okayyyyyy *closes her eyes and opens them again*
Me: *pats her bum*
Dragonlady: NOOOOO!!
Me: *cuddles her*
Dragonlady: NOOOOO!!!
Me: Go to sleep then!
Dragonlady: Blanket! Baby (bear)! Buls (Bolster)!!
Me: *covers her with the blanket and throw the bear and bolster at her*
Dragonlady: Theenk keww!
Somebody needs me at mealtimes.
Me: Meimei, do you want Mommy to feed you?
Dragonlady: NOOO! I feed.
Me: Good girl!
*turns to the cub*
Cub: Mommy, feed!
Me: Aren't you feeding yourself in school?
Cub: I want Mommy feed!
*Dragonlady joins in the fun*
Dragonlady: I want Mommy feed!
Somebody needs me when they are not well.
For medication. For nasal spray. For taking of temperatures. For oiling. For sponging. All day long and all through the night.
To clean them up when they throw up. To cuddle and tell them it's okay.
Somebody needs me even if I am unwell.
The kids have to be fed. The bottles have to be washed. The bags have to be packed. The bedtime routine has to go on. The Chinese revision playtime the kids love has to go on!
Mommy means ensuring you don't wake the kids up when you want to throw up in the middle of the night.
Mommy means you carry and cuddle your kids when they need you even when the masseuse thinks you walk funny cos of your back aches.
Somebody needs me during my shower.
For some reason, the dragonlady tends to stir in her sleep and wake up looking for me the moment I turn on the shower.
Mommy means you will have company in the shower probably until the kid(s) turns 3. (I HOPE!)
Somebody needs me all the time.
Mommy means letting the kids know you will always be there for them and always love them.
Cherish every single moment of being needed cos there will come a day when the children no longer need you as much as they did in the early years of their lives. And you can never turn back time again.


text:
Oh bacon, is there no end to the blessings you bestow? Stuffing raw bacon up your nose stops uncontrollable nosebleeds, according to research recently published in a respected medical journal. What other medical cure smells so delicious?
Four doctors at Detroit Medical Center in Michigan treated a child who had a rare hereditary disorder that causes prolonged nose bleeding by using what they called a "nasal tampon" made out of "cured salted pork," the Guardian reports. The so-called bacon tampon "stopped nasal hemorrhage promptly, effectively, and without sequelae [negative consequences]," they write in the January 2012 issue of the Annals of Otology, Rhinology and Laryngology. "To our knowledge, this represents the first description of nasal packing with strips of cured pork for treatment of life-threatening hemorrhage in a patient with Glanzmann thrombasthenia" (a platelet disorder).
The doctors used the method on the 4-year-old girl twice. "In both applications, the patient had complete cessation of nasal bleeding within 24 hours, and was discharged within 72 hours after treatment," they report.
This is an old folk cure that dates at least to the 1800s, but doctors stopped stuffing noses with bacon because "packing with salt pork was fraught with bacterial and parasitic complications," the physicians state in their article. They speculate that the meat may have certain tissue factors that help the body stop bleeding. According to the University of Minnesota Medical Center, it could be that the cold and salt work to constrict blood vessels and the fat stimulates the clotting system. "We suggest you leave the strip in place until it falls out," the university's website states. "Some people are uncomfortable putting salt pork up their or their child's nose. This is just a suggestion that has been successfully used by many patients."
If you like this story, consider signing up for our email newsletters.
You have successfully signed up for your selected newsletter(s) - please keep an eye on your mailbox, we're movin' in!
"Salt pork placed in the nose and allowed to remain there for about five days has been used, but the method is rather old-fashioned," Dr Henry Beinfield in Brooklyn, N.Y., said in a treatise called General Principles in Treatment of Nasal Haemorrhage published in 1953. In 1940, Dr. A.J. Cone of Washington University School of Medicine in St Louis praised the method in a paper called Use of Salt Pork in Cases of Haemorrhage.
"As newer synthetic hemostatic agents and surgical techniques evolved, the use of packing with salt pork diminished," doctors Ian Humphreys, Sonal Saraiya, Walter Belenky and James Dworkin write in their current Annals of Otology piece. However, modern methods of curing bacon using sodium nitrate or sodium nitrite preserve the pork and prevent bacteria from growing.
The mind boggles. What other medical miracles might this smoky, salty elixir accomplish? Treat erectile dysfunction? Cure cancer? Maybe Paula Deen should try it for her diabetes.
Follow Samantha Bonar @samanthabonar.


text:
Maybe even more important than the resume is your letter of application. It serves as a personal business card to your future employer. The letter of application allows you to explain who you are, what good qualities you have, what your ambitions are and why you should be invited for an interview. The letter gives you the opportunity to put forward your personality and mentality, where a resume mainly serves as an overview of your recent pursuits and activities. Even though there are many ways of making your letter of application successful, we particularly find the following points essential.
- Personal information
Always start your letter with personal information. Start with your name, address and residence. Then mention the address of the company where you're applying. Then follows the subject of the letter. Optional is to choose a standard subject: the job vacancy or function. To start more striking with 'stop the search!' or 'I am the candidate you are looking for' is also an option. After the subject, mention the date and write the salutation. Most of the time you don't know the contact person, so recommendable is to use formal greeting, such as Dear Ms./Mr. Smith. If you're unsure about the person's gender, write the full name as in Dear John Smith.
- Function
In the first paragraph, emphasize which function you are applying for and how you came across this function. There is nothing wrong with standard sentences as 'recently I saw your vacancy in' or 'recently one of your employees told me', but they are definitely not catchy. Try to come up with an original opening sentence, one that sticks with the reader. For example: 'every time I come across your company's commercials I regret not being part of your organisation, therefore I did not hesitate once when encountering the vacancy'
- Work experience
In the second paragraph, explain important and relevant work experiences/qualities mentioned in your resume. Focus on the work experience/qualities important for the company and function. Make it less abstract by mentioning examples from your recent job or past jobs. These examples will strengthen the argumentation why you are suitable for the function. If you are employed somewhere else, explain why you want to switch jobs. Maybe mention the possibility to make a promotion, or that you're eager for new challenges.
- Why that specific company?
Often, there are multiple employers involved in the same field. What makes this company so special that you want to work there? In the third paragraph, explain why you want to work for that company instead for one of their rivals. It might come in handy to check the company's website for useful information. Questions to think about are 'what is their mission and vision?', 'what feeling do I get with this employer?', 'where does this employer stand for?'. Try to show your genuine interest in the company. Also, define the additional value you will have for the employer and why you are suitable for the job.
- Closing
In the fourth paragraph, finish your letter by mentioning that you are willing to explain your letter/resume in an interview. Again, try to make this as original and catchy as possible. This way, the reader will remember you more easily. Even though it is important to be original, do not forget to stay formal and professional. The final sentence in a letter of application consists of a formal closing, such as Yours faithfully.
For a template of an application letter, click here.