Cloning and characterization of the proximal promoter region of rainbow trout (Oncorhynchus mykiss) interleukin-6 gene.

Interleukin-6 (IL6) is a pleiotropic cytokine with important immunoregulatory functions. Its expression is inducible in immune cells and tissues of several fish species. We also found that IL6 mRNA abundance was significantly increased in spleen, liver, and gill of rainbow trout after experimental infection with Aeromonas salmonicida. Genomic DNA sequences of IL6 orthologs from three salmonid species revealed a conserved exon/intron structure and a high overall nucleotide identity of >88%. To uncover key mechanisms regulating IL6 expression in salmonid fish, we amplified a fragment of the proximal IL6 promoter from rainbow trout and identified in-silico conserved binding sites for NF-κB and CEBP. The activity of this IL6 promoter fragment was analyzed in the established human embryonic kidney line HEK-293. Luciferase- and GFP-based reporter systems revealed that the proximal IL6 promoter is activated by Escherichia coli. Essentially, both reporter systems proved that NF-κB p50, but not NF-κB p65 or CEBP, activates the IL6 promoter fragment. Truncation of this fragment caused a significant decrease in IL6 promoter activation. This characterization of the proximal promoter of the IL6-encoding gene provides basic knowledge about the IL6 gene expression in rainbow trout.

Secondhand tobacco smoke exposure is associated with increased risk of failed implantation and reduced IVF success.

BACKGROUND: Infertility and early pregnancy loss are prevalent as is exposure to secondhand tobacco smoke (STS). Previous research has suggested a relationship between STS exposure and early pregnancy loss, but studies have been limited by small study sizes and/or imprecise methods for exposure estimation. IVF allows for the collection of follicular fluid (FF), the fluid surrounding the pre-ovulatory oocyte, which may be a more biologically relevant sample media than urine or serum in studies of early reproduction. METHODS: In a retrospective analysis of a prospective cohort study, we measured cotinine in FF collected during 3270 IVF treatment cycles from 1909 non-smoking women between 1994 and 2003 to examine the relationship between STS exposure and implantation failure. RESULTS: In adjusted models, we found a significant increase in the risk of implantation failure among women exposed to STS compared with those unexposed [odds ratio (OR) = 1.52; 95% confidence interval (CI) = 1.20-1.92; risk ratio (RR) = 1.17; 95% CI = 1.10-1.25]. We also found a significant decrease in the odds for a live birth among STS-exposed women (OR = 0.75; 95% CI = 0.57-0.99; RR = 0.81; 95% CI = 0.66-0.99). CONCLUSIONS: Female STS exposure, estimated through the measurement of cotinine in FF, is associated with an increased risk of implantation failure and reduced odds of a live birth.

Acidities of arsenic (III) and arsenic (V) thio- and oxyacids in aqueous solution using the CBS-QB3/CPCM method.

A number of calculations of acidic As(III) and As(V) species formed with sulfur and oxygen (H(3)AsS(3), H(2)AsS(3)(-), HAsS(3)(2-), AsS(3)(3-), H(3)AsS(4), H(2)AsS(4)(-), HAsS(4)(2-), AsS(4)(3-), H(3)AsO(3), H(2)AsO(3)(-), HAsO(3)(2-), AsO(3)(3-), H(3)AsO(4), H(2)AsO(4)(-), HAsO(4)(2-), and AsO(4)(3-)) are presented. pK(a)s for successive deprotonations in both the gas phase and aqueous solution (using both explicit water molecules and a self-consistent reactive field conductive polarizable continuum model (SCRF CPCM) for solvation) are fitted to known experimental values for the H(3)PO(4) and H(3)AsO(4) series' of deprotonations with a linear extrapolation showing r(2) values of 0.97 for the CBS-QB3 method with a single explicit water molecule in the CPCM. Though the unfitted pK(a)s of H(3)AsO(4) (pK(a1) = 4.6, pK(a2) = 17.7, and pK(a3) = 28.6) compare unfavorably with experimental values of pK(a1) = 2.3, pK(a2) = 7, pK(a3) = 13, by the linear fit: 0.42907 x pK(a) - 0.23, the predicted pK(a)s become pK(a1) = 1.7, pK(a2) = 7.3, and pK(a3) = 12.1, which correspond well. The H(3)PO(4) calculation shows a similar trend; pK(a1) = 4.9, pK(a2) = 19.9, and pK(a3) = 29.8 becomes pK(a1) = 1.9, pK(a2) = 8.3, pK(a3) = 12.6. Experimentally, H(3)PO(4) has pK(a1) = 2.2, pK(a2) = 7.1, and pK(a3) = 12.3. With the same extrapolation, we predict H(2)SO(3) to have pK(a1) = 1.1 and pK(a2) = 8.1, which compare favorably with the experimental values of pK(a1) = 1.9 and pK(a2) = 7.2.

Influence of curli expression by Escherichia coli 0157:H7 on the cell's overall hydrophobicity, charge, and ability to attach to lettuce.

Curli fibers are produced by some Escherichia coli cells in response to environmental stimuli. These extracellular proteins enhance the cell's ability to form biofilms on various abiotic surfaces. E. coli 0157:H7 cells readily attach to a variety of fruit and vegetable surfaces. It is not known whether the expression of curli influences the cell's ability to attach to produce surfaces. In this study, the effect of curli expression on the cell's overall hydrophobicity, charge, and ability to attach to cut and whole iceberg lettuce surfaces was examined. All strains, regardless of curli expression, attached preferentially to the cut edges of lettuce (P < 0.05). The curli-producing cells of E. coli 0157:H7 strain E0018 attached in significantly greater numbers to both cut and whole lettuce pieces than did the non-curli-producing E0018 cells (P < 0.05); however, no significant attachment differences were observed between the curli-producing and non-curli-producing cells of E. coli 0157:H7 strains 43894 and 43895. All curli-producing E. coli 0157:H7 strains were significantly more hydrophobic (P < 0.01); however, no association between the cells' hydrophobic characteristics and lettuce attachment was observed. Overall surface charge of the cells did not differ among strains or curli phenotypes. Results indicate that overall hydrophobicity and cell charge in E. coli 0157:H7 strains do not influence attachment to iceberg lettuce surfaces. The presence of curli may not have any influence on attachment of E. coli 0157:H7 cells to produce items. Additional factorsmay influence the attachment of E. coli 0157:H7 to plant surfaces and should be further examined.

Effects of modified atmosphere packaging on toxin production by Clostridium botulinum in raw aquacultured summer flounder fillets (Paralichthys dentatus).

Packaging fishery products under vacuum atmosphere packaging (VAC) and modified atmosphere packaging (MAP) conditions can significantly extend the shelf life of raw, refrigerated fish products. There is considerable commercial interest in marketing VAC and MAP refrigerated (never frozen) raw fish fillets. The objective of this study was to determine if Clostridium botulinum toxin development precedes microbiological spoilage in raw, refrigerated flounder fillets. Aquacultured flounder (Paralichthys dentatus) individual fish fillets either were packed with a film having an oxygen transmission rate (OTR) of 3000 cm3 m(-2) 24 h(-1) at 22.8 degrees C or were vacuum packaged or packaged under 100% CO2 with a film having an OTR of 7.8 cm3 m(-2) 24 h(-1) at 21.1 degrees C and were stored at 4 and 10 degrees C. Samples were analyzed by aerobic plate count (APC) for spoilage and qualitatively for botulinum toxin with a mouse bioassay. The results demonstrate that flounder fillets (4 degrees C) packaged with a film having an OTR of 3,000 were microbiologically spoiled (APC, > 10(7) CFU/g) on day 15, but there was no toxin formation, even after 35 days of storage. However, at 10 degrees C, toxin production occurred (day 8), but it was after microbial spoilage and absolute sensory rejection (day 5). Vacuum-packaged fillets and 100% CO2 fillets (4 degrees C) packaged with a film having an OTR of 7.8 were toxic on days 20 and 25, respectively, with microbial spoilage (APC, >10(7) CFU/g) not occurring during the tested storage period (i.e., >35 days). At 10 degrees C, in vacuum-packaged flounder, toxin formation coincided with microbiological spoilage (days 8 to 9). In the 100% CO2-packaged fillets, toxin formation occurred on day 9, with microbial spoilage occurring on day 15. This study indicates that films with an OTR of 3,000 can be used for refrigerated fish fillets and still maintain the safety of the product.

Effect of gonadorelin, lecirelin, and buserelin on LH surge, ovulation, and progesterone in cattle.

Analogs of gonadoliberin (GnRH) are widely used in cattle to synchronize estrus and to induce ovulation, as well as for the treatment of ovarian cysts. The aim of this study was to compare the plasma profiles of LH and progesterone and the follicular dynamics in response to the administration of gonadorelin, lecirelin, or buserelin at the dose recommended to induce ovulation. In addition, the biological response to a half dose of lecirelin was assessed. Twelve healthy Holstein female cows were divided into four sequence groups, according to a Latin square design and received the four treatments during the four periods of the study. Before each period, the estrous cycle was synchronized, and on Day 6 or 7 of the ensuing cycle, the time at which it was most likely to have a dominant follicle, 100 µg of gonadorelin, 25 µg of lecirelin, 50 µg of lecirelin, or 10 µg of buserelin was administered to the cows. Blood samples were regularly collected for up to 4 days after the GnRH administrations. The plasma LH response was evaluated for up to 6 hours after administration, and the plasma progesterone response and ovarian follicular dynamics were evaluated for up to 4 days. There was a significantly lower LH release after gonadorelin treatment compared to lecirelin at the doses of 25 or 50 µg and the buserelin treatment. The mean maximal LH concentration after gonadorelin treatment was 2.5 lower than after lecirelin or buserelin treatment and was reached 1 hour earlier. Four days after the GnRH administration (i.e., at Days 10-11 of the estrous cycle), the overall mean increase in plasma progesterone concentration was 70% and did not differ between the treatment groups. The percentage of disappearance of the dominant follicle (84.8% of ovulation and 4.3% of luteinization) after GnRH treatment was high (73%, 82%, 100%, and 100%, for gonadorelin, lecirelin at the doses of 25 and 50 µg, and buserelin, respectively) and did not differ between the GnRH treatments. The follicle disappearance was followed by the emergence of a synchronous follicle wave within 2 days in almost all the heifers. Altogether, our data show that the three GnRH analogs, at the doses indicated for the induction of ovulation or at a half dose for lecirelin, are almost equally effective to induce the disappearance of the dominant follicle at Day 6 to 7 of the estrous cycle.

Efficacy of detergents in removing Salmonella and Shigella spp. from the surface of fresh produce.

Fresh produce has been implicated in several foodborne disease outbreaks. Produce surfaces can be primary sites of contamination during production and handling. One approach to reduce contamination is to treat fresh produce with rinsing agents. In this study, different detergent agents were used at 22 and 40 degrees C to determine their efficacy in removing Salmonella and Shigella spp. from the surfaces of strawberries, tomatoes, and green-leaf lettuce. Produce was inoculated at 22 degrees C with a cocktail of nalidixic acid-resistant organisms (6 to 6.5 log CFU/ml). After air drying for 1 h, samples were rinsed with either 0.1% Tween 80, 0.1% sodium lauryl sulfate (SLS), or water (control) at 22 or 40 degrees C. Rinse solutions were spiral plated onto tryptic soy agar supplemented with 50 mg of nalidixic acid per liter. In trials involving strawberries and lettuce, Salmonella and Shigella were removed at levels of 4 and 3 log CFU/ml, respectively, except from Salmonella-inoculated strawberries rinsed with SLS, for which minimal removal rates were 1.5 log CFU/ml at 22 degrees C and < 1 log CFU/ml at 40 degrees C. When whole strawberries were analyzed after rinsing with SLS, few organisms were recovered. This result suggests that SLS may have a lethal or sublethal effect on Salmonella, especially when a 40 degrees C solution is used. Salmonella and Shigella removal rates for tomatoes were 1 and 1.5 log CFU/ml lower, respectively, than those for strawberries or lettuce. Overall, detergents were no more effective in removing organisms from produce than water was. The detergents examined would not constitute effective overall produce rinse treatments.

Effect of organic acids and hydrogen peroxide on Cryptosporidium parvum viability in fruit juices.

Cryptosporidium parvum has historically been associated with waterborne outbreaks of diarrheal illness. Foodborne cryptosporidiosis has been associated with unpasteurized apple cider. Infectious oocysts are shed in the feces of common ruminants like cattle and deer in and near orchards. In this study, the ability of organic acids and hydrogen peroxide (H2O2) added to fruit juice to inhibit the survival of C. parvum was analyzed. Oocyst viability was analyzed by a cell culture infectivity assay with the use of a human ileocecal cell line (HCT-8) whose infectivity pattern is similar to that for human oral infectivity. Cell monolayers were infected with 10(6) treated oocysts or a series of 10-fold dilutions. Parasitic life stages were visualized through immunohistochemistry with 100 microscope fields per monolayer being counted. In vitro excystation assays were also used to evaluate these treatments. Organic acids and H2O2 were added to apple cider, orange juice, and grape juices on a weight/volume basis. Malic, citric, and tartaric acids at concentrations of 1 to 5% inhibited C. parvum's infectivity of HCT-8 cells by up to 88%. Concentrations ranging from 0.025 to 3% H2O2 were evaluated. The addition of 0.025% H2O2 to each juice resulted in a >5-log reduction of C. parvum infectivity as determined with a most-probable-number-based cell culture infectivity assay. As observed with differential interference contrast and scanning electron microscopy, reduced infectivity may be mediated through effects on the oocyst wall that are caused by the action of H2O2 or related oxygen radicals. The addition of low concentrations of H2O2 can represent a valuable alternative to pasteurization.

Survivability of Salmonella and Shigella spp. in sodium lauryl sulfate and tween 80 at 22 and 40 degrees C.

Fresh produce has been implicated in several foodborne disease outbreaks. A primary site of contamination during production and handling is the surface of produce. One approach to reducing contamination is to treat fresh produce with rinsing agents. Studies have examined the efficacies of detergents and other rinses in recovering pathogens from produce surfaces. The determination of how these detergents affect bacterial cells may aid in understanding the mechanisms behind their removal. This study examines the survivability of Salmonella and Shigella in two detergents. A 0.1% sodium lauryl sulfate (SLS) solution, a 0.1% Tween 80 solution, and water were inoculated with a cocktail of stationary-phase organisms (3 log CFU/ml) and incubated for up to 32 h at 22 degrees C and 40 degrees C. Samples were taken over time and plated on tryptic soy agar supplemented with 50 ppm of nalidixic acid. Salmonella survived in all solutions and exhibited significant growth in water (0.8 log CFU/ml at 22 degrees C and 1.9 log CFU/ml at 40 degrees C) and Tween 80 (1.0 log CFU/ml at 40 degrees C). Shigella survived in all solutions at 22 degrees C and exhibited a growth level of 2.0 log CFU/ml in SLS. Shigella also survived in all solutions at 40 degrees C, although its populations decreased significantly in Tween 80 over time. Elevated temperatures may allow Tween 80 to kill Shigella spp. over time. Overall, the detergents tested were not detrimental to the cells; therefore, if these solutions were to be used as produce rinse agents, they would aid in removal of organisms from surfaces rather than kill the cells.

Examination of attachment and survival of Toxoplasma gondii oocysts on raspberries and blueberries.

The consumption of Toxoplasma gondii oocysts on fresh produce may be a means of its transmission to humans. Cats shed T. gondii oocysts, which contaminate produce directly or contaminate water sources for agricultural irrigation and pesticide and fertilizer applications. Cyclospora cayetanensis is a related coccidial parasite, and outbreaks of diarrhea caused by C. cayetanensis have been associated with the ingestion of contaminated raspberries. The oocysts of these coccidians are similar in size and shape, indicating that they may attach to and be retained on produce in a similar manner. In the present study the attachment and survival of T. gondii oocysts on 2 structurally different types of berries were examined. Raspberries and blueberries were inoculated individually with 1.0 x 10(1) to 2.0 x 10(4) oocysts of sporulated T. gondii. Berries inoculated with 2.0 x 10(4) oocysts were stored at 4 C for up to 8 wk. Oocyst viability and recovery were analyzed by feeding processed material to mice. Mice fed T. gondii-inoculated berries stored at 4 C for 8 wk developed acute infections. In other experiments mice fed raspberries inoculated with > or = 1.0 x 10(1) oocysts became infected, whereas only mice fed blueberries inoculated with > or = 1.0 x 10(3) oocysts became infected. This study demonstrates that T. gondii oocysts can adhere to berries and can be recovered by bioassays in mice and that raspberries retain more inoculated oocysts than do blueberries. The results suggest that T. gondii may serve as a model for C. cayetanensis in food safety studies.

[Elements of shoulder mechanics]. / Eléments de mécanique de l'épaule.

An understanding of shoulder movement is difficult. The author has devised a schematic diagram which gives a better definition of the basic movements. It allows a simpler analysis of limitation of movement and of the relative importance of the scapulo-humeral, sterno-clavicular and acromio-clavicular joints. The role of the peri-articular muscles is stressed as motors and stabilizers. This analysis accounts for difficulties encountered in prosthetic surgery.

[How to perform oblique-plane osteotomy]. / Comment réaliser une ostéotomie plane oblique.

Oblique plane osteotomy allows correction of associated angular and rotational deformity by a single bony section and also allows modification of the existing angle at the cervico-diaphyseal level in the femur. Exact assessment of the direction of the osteotomy and of extent of rotation are essential to success. This paper proposes a simple method by use of a table and performance is facilitated by the of Kirschner wires.

[Resection-arthrodesis of the knee for bone tumours (author's transl)]. / La résection-arthrodèse du genou pour tumeur.

The authors describe the technique, devised by Juvara and modified by Merle d'Aubigné of resection-arthrodesis of the knee with fixation by medullary nailing and using massive bone grafts. They report 38 cases operated on for various conditions (20 giant-cell tumours, 7 chondro-sarcoma, 6 juxta-cortical sarcoma, 3 other types of sarcoma, one desmoid fibroma, one chondroma). The best results were obtained in cases in which surgical excision had been sufficiently wide and when very thick nails were used. The use of homografts increased the rate of infection. Resections of the lower and of the femur resulted in more complications than tibial resections. In femoral lesions, the part of the tibia that is transplanted should not be turned upside-down but should be mobilised upwards to provide better bone contact. The indications are discussed and compared with those of total replacement. The authors believe that, despite the number of complications, this technique is more practicable than total knee replacement.

[A peculiar tumor. Adamantinoma of the long bones. Apropos of 7 consecutive cases followed up after resection]. / Une tumeur singulière. L'adamantinome des os longs. A propos de sept cas consécutifs suivis après résection.

Seven cases of tibial adamantinoma have been reviewed. All were treated by en bloc resection and followed for one to twenty years after operation. Of five patients followed for more than three years, three have had a good result after 3, 12 and 20 years of follow-up. One patient suffered a recurrence in the soft tissues seven years after resection and remained well after thirteen years follow-up. One patient required amputation two years after resection and died from a mediastinal tumour of undetermined nature sixteen years after amputation. The two failures occurred in patients seen secondarily following recurrence after initial curettage. The tibial shaft was reconstructed in three cases by massive allografts. One of these failed because of infection but was reconstructed using autografts; one had a late fracture which united spontaneously. The third, after sound union, had to be amputated for recurrence. Two smaller resections were treated successfully by tibial and iliac autografts. Two cases were treated by free vascularised fibular transplants which united, though one suffered a late fracture which united spontaneously. Curettage, which is almost always followed by recurrence, should be rejected in favour of extensive resection. Resection is successful when done primarily but poses severe problems in reconstruction and requires a prolonged period of treatment. The small numbers in this series do not allow a judgement to be made as to the superiority of one technique over another.

Infertility, Pregnancy Loss and Adverse Birth Outcomes in Relation to Maternal Secondhand Tobacco Smoke Exposure.

A substantial proportion of the etiology involved in female infertility and adverse pregnancy outcomes remains idiopathic. Recent scientific research has suggested a role for environmental factors in these conditions. Secondhand tobacco smoke (STS) contains a number of known or suspected reproductive toxins, and human exposure to STS is prevalent worldwide. Robust evidence exists for the toxic effects of active smoking on fertility and pregnancy, but studies of passive exposure are much more limited in number. While the association between maternal STS exposure and declined birth weight has been fairly well-documented, only recently have epidemiologic studies begun to provide suggestive evidence for delayed conception, altered menstrual cycling, early pregnancy loss (e.g. spontaneous abortion), preterm delivery, and congenital malformations in relation to STS exposure. There is also new evidence that developmental exposures to tobacco smoke may be associated with reproductive effects in adulthood. To date, most studies have estimated maternal STS exposure through self-report even though exposure biomarkers are less prone to error and recall bias. In addition to utilizing biomarkers of STS exposure, future studies should aim to identify vital windows of STS exposure, important environmental co-exposures, individual susceptibility factors, and specific STS constituents associated with female infertility and adverse pregnancy outcomes. The role of paternal exposures/factors should also be investigated.

Secondhand tobacco smoke exposure is associated with prolactin but not thyroid stimulating hormone among nonsmoking women seeking in vitro fertilization.

Prolactin (PRL) and thyroid stimulating hormone (TSH) serve important roles in the reproductive and other systems. Active smoking is associated with changes in PRL and TSH secretion, but the relationship between secondhand tobacco smoke (STS) exposure and these hormones is unclear. We measured PRL and TSH in serum as well as cotinine in follicular fluid (to estimate STS exposure) among 314 nonsmoking women undergoing in vitro fertilization treatment. We observed a significant increase in PRL concentrations (p=0.03) among STS-exposed nonsmokers compared to unexposed nonsmokers. There was no significant difference in TSH concentration (p>0.4) among those exposed to STS compared to those who were unexposed. STS exposure is associated with an increase in circulating PRL but not TSH levels. Future studies are needed to confirm our results, identify biological mechanisms involved, and better understand the potential clinical and public health implications.

Cotinine concentrations in follicular fluid as a measure of secondhand tobacco smoke exposure in women undergoing in vitro fertilization: inter-matrix comparisons with urine and temporal variability.

We examined the relationship between cotinine measures in follicular fluid (FF) and urine to inform our exposure assessment strategy for an ongoing epidemiological study of secondhand tobacco smoke (STS) exposure and early pregnancy loss. Among subjects undergoing in vitro fertilization (IVF), we compared cotinine levels in paired urine and FF samples from the same women and examined FF cotinine levels over time. We found a weak rank-order relationship (Spearman r<0.2) and poor agreement for classifying nonsmoking individuals as exposed to STS (sensitivity=0.29-0.71; specificity=0.35-0.72) between cotinine concentrations in FF and urine. We observed fair reliability (ICC=0.42-0.52) in FF cotinine concentrations from women undergoing multiple IVF cycles. If available, FF cotinine concentrations may be desired as a biomarker of low-level tobacco smoke exposure over urinary cotinine in studies of early reproduction. Collection of multiple FF samples for cotinine analysis may be needed to accurately represent long-term STS exposure.

The relation between depression and parkin genotype: the CORE-PD study.

BACKGROUND: Mutations in parkin are a known genetic risk factor for early onset Parkinson's disease (EOPD) but their role in non-motor manifestations is not well established. Genetic factors for depression are similarly not well characterized. We investigate the role of parkin mutations in depression among those with EOPD and their relatives. METHODS: We collected psychiatric information using the Patient Health Questionnaire and Beck Depression Inventory II on 328 genotyped individuals including 88 probands with early onset PD (41 with parkin mutations, 47 without) and 240 first and second-degree relatives without PD. RESULTS: Genotype was not associated with depression risk among probands. Among unaffected relatives of EOPD cases, only compound heterozygotes (n = 4), and not heterozygotes, had significantly increased risk of depressed mood (OR = 14.1; 95% CI 1.2-163.4), moderate to severe depression (OR = 17.8; 95% CI 1.0-332.0), depression (score ≥ 15) on the Beck Depression Inventory II (BDI-II) (OR = 51.9; 95% CI 4.1-657.4), and BDI-II total depression score (ß = 8.4; 95% CI 2.4-11.3) compared to those without parkin mutations. CONCLUSIONS: Relatives of EOPD cases with compound heterozygous mutations and without diagnosed PD may have a higher risk of depression compared to relatives without parkin mutations. These findings support evidence of a genetic contribution to depression and may extend the phenotypic spectrum of parkin mutations to include non-motor manifestations that precede the development of PD.