Shigella infection in children under 5 years old in western French Guiana.

French Guiana, a tropical country, is characterised by a young and multi-ethnic population. Difficulties in accessing safe water sources lead to outbreaks of gastroenteritis. The objectives of this study were (1) to describe the microbiological profile of shigella strains isolated in western French Guiana, including antimicrobial susceptibility and the distribution of strains in terms of species and serotypes and (2) to estimate the incidence of shigellosis in children under 5 years old. A retrospective observational study was conducted of 213 cases of shigellosis diagnosed in the biology department of the hospital centre for western French Guiana between 2000 and 2012 in children under 5 years old. The serogroups (formerly known as species) that predominates in French Guiana was Shigella flexneri. No resistance was observed to fluoroquinolones or to third-generation cephalosporins. The average incidence of shigellosis in children under 5 years old in western French Guiana was estimated at 189.6 cases per 100 000 inhabitants per year. Shigellosis is a public health problem in western French Guiana. These infections suggest the difficulties in accessing safe water sources and the lack of public sanitation. A quadrivalent vaccine containing Shigella sonnei and three serotypes of S. flexneri (S. flexneri 2a, 3a and 6) could provide broad coverage against shigella infections.

Association between Zika virus and fetopathy: a prospective cohort study in French Guiana.

OBJECTIVE: To establish the incidence of fetal central nervous system (CNS) anomalies (including microcephaly), signs of congenital infection and fetal loss in pregnant women infected with Zika virus (ZIKV) and non-infected pregnant women in western French Guiana. METHODS: This prospective cohort study was conducted between 1 January and 15 July 2016. We evaluated and compared clinical and fetal ultrasound examinations of 301 pregnant women with biological confirmation of ZIKV infection and 399 pregnant women who were negative for ZIKV infection. RESULTS: Overall, the total number of fetuses with CNS involvement was higher in the infected than in the control group (9.0% vs 4.3%; relative risk, 2.11 (95% CI, 1.18-4.13)). Anomalies of the corpus callosum and presence of cerebral hyperechogenicities were significantly more common in the infected group. There was an increased risk of microcephaly in the infected compared with the control group (1.7% vs 0.3%; relative risk, 6.63 (95% CI, 0.78-57.83)), although this was not statistically significant. When the mother was infected during the first or second trimester, there was a greater risk of severe CNS involvement, more signs of infection and intrauterine fetal death than with infection in the third trimester. The rate of vertical transmission in the exposed group was 10.9%. CONCLUSION: ZIKV infection during pregnancy is associated with a significant risk of fetal CNS involvement and intrauterine fetal death, particularly when infection occurs during the first or second trimesters. Microcephaly was not present in every case of congenital ZIKV syndrome that we observed. Until more is known about this disease, it is paramount to evaluate suspected cases by detailed neurosonography on a monthly basis, paying particular attention to the corpus callosum and the presence of hyperechogenic foci. Copyright © 2017 ISUOG. Published by John Wiley & Sons Ltd.

HIV-associated disseminated histoplasmosis in western French Guiana, 2002-2012.

Disseminated histoplasmosis (DH) is the most current revelation mode of AIDS in French Guiana. We describe the clinical and paraclinical presentation of DH, diagnostic tools, evolution and factors associated with 1-year mortality in HIV-infected patients from western French Guiana. Microbiologically proven AIDS-related DH in Saint Laurent du Maroni's hospital between May 2002 and May 2012 were retrospectively included. Among the 82 patients included, 58 (71%) were male, 44 (53%) presented concurrent histoplasmosis and HIV diagnosis and 63 (80%) had a CD4 cell count under 50 cells µL(-1). Almost all patients had weight loss (97%) and fever (95%), while 84% had digestive symptoms (63% diarrhoea), 55% lymphadenopathy, and 49% respiratory symptoms. Documented and presumed locations of H. capsulatum var capsulatum (Hcc) concerned almost all organs, with a particular affinity for the bone marrow and the digestive system. Co-infections were associated in 65%. Following treatment initiation, 10 patients (13%) died within 1 month and 17 patients (25%) died within a year. DH is a polymorphous systemic mycosis with haematological and digestive tropism. Co-infections are frequent and mortality rate is high.

Diel vertical migration of European hake Merluccius merluccius and associated temperature histories: insights from a pilot data-storage tagging (DST) experiment.

A pioneering experiment of archival tagging of European hake Merluccius merluccius provided evidence of a diel vertical migration pattern which was analysed using an automatic method, developed and validated through time-frequency analyses. Frequent vertical movements across the thermocline were observed and fish experienced rapid temperature changes >7° C. These tagging-recapture data also suggested a homing behaviour to feeding grounds.

External validation of Gout-calculator performance on a cohort of acute arthritis (SYNOLACTATE) sparing distal joints such as hallux and midfoot. A cross-sectional study of 170 patients.

OBJECTIVE: To evaluate the performance of the Gout-calculator in a cohort of consecutive acute arthritis affecting large and intermediate joints (without an attack on hallux or midfoot joints). METHODS: A retrospective study. Gout-calculator data were collected in medical records of patients included in the prospective consecutive cohort of acute arthritis called SYNOLACTATE. The diagnosis of gout was defined by the presence of sodium urate crystals in synovial fluid. The diagnostic performance of the Gout-calculator was studied by performing an ROC curve with the calculation of its AUC (95% CI) as well as the calculation of Sensitivity (Se), Specificity (Sp), and positive likelihood ratio (LR+). RESULTS: 170 patients with acute arthritis were included. Variables associated with the diagnosis of gout were as follows: serum uric acid > 350 µmol/L (OR 5.52 (2.52-12.1), p < 0.001), joint redness (OR 5.08 (1.85-14.0), p = 0.001), previous patient-reported arthritis attack (OR 4.04 (1.92-8.49), p < 0.001), male (OR 3.00 (1.17-7.69), p = 0.02), hypertension or cardiovascular disease (OR 2.33 (1.07-5.06), p = 0.03). The median (IQR) of Gout-calculator was significantly higher in gouty arthritis (7.0 [5.5-8.1]) than in associated-CPP acute arthritis (4.0 [2.0-5.8]), septic arthritis (3.0 [2.0-5.1]), or others arthritis (3.5 [2.0-5.5]). The AUC was 0.833 (0.768-0.897) with for the threshold ≥ 8, a Se at 27.5% (0.161-0.428), Sp 97.7% (0.934-0.992), and LR+ 11.9 (3.5-40). CONCLUSION: Despite diagnostic performances close to those published, the use of the Gout-calculator is not sufficient for the diagnosis of gout or to exclude the differential diagnosis of septic arthritis in the SYNOLACTATE cohort. KEY POINTS: • For a Gout-calculator threshold of ≤ 4, Sensitivity is 92.5%, Specificity 50.8% and LR- 0.15 to the gout diagnosis. • For a Gout-calculator threshold of > = 8, Sensitivity is 27.5%, Specificity 97.7% and LR+ 11.9 to the gout diagnosis. • In a population of acute arthritis affecting large joints, Gout-calculator is not sufficient to discriminate between gouty arthritis and septic arthritis.

Acute infectious pancreatitis due to Salmonella typhi: Case report and literature review.

Salmonella typhi remains an endemic disease in Madagascar. Acute pancreatitis remains a rare complication of S. typhi colitis. We presented the case of a 27-year-old male, admitted to febrile diarrhea, vomiting, and severe abdominal pain. Blood work-up revealed elevated plasma lipase level. Abdominal CT scan showed acute pancreatitis without necrosis. Blood and stool culture positivity for S. typhi. Patient was diagnosed as acute pancreatitis caused by S. typhi. The outcome was favorable under symptomatic medical treatment (rehydration and analgesic) combined with adapted antibiotic therapy. Acute pancreatitis is a possible complication of Salmonella infections. The presence of severe abdominal pain and febrile diarrhea should draw clinicians' attention to possible Salmonella acute pancreatitis.

Cloning and sequencing of porcine LH-hCG receptor cDNA: variants lacking transmembrane domain.

Complementary DNA clones, encoding the LH-hCG (luteinizing hormone-human choriogonadotropic hormone) receptor were isolated by screening a lambda gt11 library with monoclonal antibodies. The primary structure of the protein was deduced from the DNA sequence analysis; the protein contains 696 amino acids with a putative signal peptide of 27 amino acids. Hydropathy analysis suggests the existence of seven transmembrane domains that show homology with the corresponding regions of other G protein-coupled receptors. Three other types of clones corresponding to shorter proteins were observed, in which the putative transmembrane domain was absent. These probably arose through alternative splicing. RNA blot analysis showed similar patterns in testis and ovary with a major RNA of 4700 nucleotides and several minor species. The messenger RNA was expressed in COS-7 cells, yielding a protein that bound hCG with the same affinity as the testicular receptor.

Paraquat poisoning in Western French Guyana: a public health problem persisting ten years after its withdrawal from the French market.

OBJECTIVE: Paraquat poisoning has almost disappeared from metropolitan France following its ban from the European market ten years ago. However, due to neighboring countries still authorizing paraquat use, French Guyana seems in a different situation. Here we aimed to report a series of paraquat-poisoned patients admitted to the emergency department of the Western French Guyana Hospital in Saint-Laurent du Maroni, to raise awareness of national health authorities on this persistent major issue. PATIENTS AND METHODS: We conducted a retrospective observational study describing the clinical features, the prognostic factors and the final outcome of paraquat-poisoned patients admitted to the emergency department between January 2008 and August 2014. RESULTS: Twenty-six paraquat-poisoned patients were included in the study. The median estimated paraquat dose intentionally ingested was 105 mg/kg (interquartile range, IQR: 359). Eighteen patients were treated with the cyclophosphamide/dexamethasone combination and seventeen with N-acetylcysteine in addition to the usual supportive care. Six patients survived and twenty died within a median 36h delay after admission (IQR: 130). Death was associated with cardiovascular (65%) and respiratory (35%) failure. Based on a bivariate analysis, predictive factors of death included (p≤0.05): advanced age, higher ingested paraquat dose, altered renal function, hypokalemia, acidosis, and dark blue dithionite test, observed on hospital admission. CONCLUSIONS: Paraquat poisoning still persists in French Guyana despite its withdrawal from the market. It is possible to determine the probability of death on patient admission based on routine clinical and biological parameters. There is an urgent need to request neighboring countries to ban paraquat with the aim of eradicating this dramatically life-threatening poisoning.

Monoclonal antibodies against native ant denatured forms of estrogen-induced breast cancer protein (BCEI/pS2) obtained by expression in Escherichia coli.

Several vectors were used to express the complementary DNA for breast cancer estrogen-induced protein BCEI (also called pS2) in Escherichia coli. The best results were obtained by using the pUR 290 expression vector after deletion of the sequence encoding the signal peptide of the protein. In these conditions, beta-galactosidase-BCEI/pS2 fusion protein accounted for approximately 20% of total proteins in bacterial extracts. It was purified by chromatography on DEAE-Trisacryl or by gel electrophoresis and electroelution. Polyclonal antibodies were obtained by immunization of rabbits and goats, and monoclonal antibodies were raised in mice. Two types of monoclonal antibodies were obtained: one class recognized the native protein and was very efficient for the immunoprecipitation and immunopurification of the protein from breast cancer cells; a second class recognized the denatured protein and was especially effective for immunoblot studies. BCEI/pS2 could be detected by immunocytochemistry in breast cancer biopsies using monoclonal antibodies on frozen or paraffin-embedded sections. One of the antibodies (mBCEI11) exhibited high affinity for the protein and could be used at 1.9 micrograms/ml concentration for immunolabeling of histological sections. The mBCEI11 antibody was used in immunoaffinity chromatography to purify the peptide in a single step from culture media of estrogen-treated MCF-7 cells.

Luteinizing hormone/human chorionic gonadotropin receptors in breast cancer.

Recent studies have suggested that human choriogonadotropin (hCG), in addition to its function in regulating steroidogenesis, may also play a role as a growth factor. Immunocytochemistry using two different monoclonal antibodies (LHR29 and LHR1055) raised against the human luteinizing hormone/human chorionic gonadotropin (LH/hCG) receptor allowed us to detect this receptor in breast cancer cell lines (T47D, MCF7, and ZR75) in individual cancer biopsies and in benign breast lesions. The receptor was also present in epithelial cells of normal human and sow breast. In the latter, its concentration increased after ovulation. The presence of LH/hCG receptor mRNA was confirmed by reverse transcription-PCR using primers extending over exons 2-4, 5-11, and 9-11. The proportion of LH/hCG-receptor positive cells and the intensity of the immunolabeling varied in individual biopsies, but there was no obvious correlation with the histological type of the cancer. These results are compatible with previous studies suggesting that during pregnancy, hCG is involved in the differentiation of breast glandular epithelium and that this hormone may play an inhibitory role in mammary carcinogenesis and in the growth of breast tumors.

Immunocytochemical study with monoclonal antibodies to progesterone receptor in human breast tumors.

Mouse hybridomas secreting monoclonal antibodies against rabbit uterine progesterone receptor (PR) have been prepared. Several of these immunoglobulins exhibited high affinity towards human progesterone receptor and two (LET 126 and LET 64) were selected as giving the best immunoperoxidase staining of human progesterone target organs. Using the indirect peroxidase-antiperoxidase method of Sternberger, optimal conditions for demonstrating PR involved brief fixation of frozen sections with formaldehyde-containing fixatives, among them picric acid-paraformaldehyde. This method allowed us to detect the receptor in breast carcinoma epithelial cells, T47D cell line, and uterine endometrium and myometrium. No staining was observed in intestine and muscle. Specific staining for PR was confined to the nucleus, irrespective of the concentration of progesterone in the blood of the patient. In a preliminary study of 27 human breast cancers by the immunocytochemical method, the presence or absence of nuclear staining for PR correlated well with the concentration of cytosolic progesterone receptor determined by a steroid-binding assay on tumor extracts. Differences in the intensity and distribution of staining within a section were observed, suggesting heterogeneity of the PR content of breast cancer cells. In 19 tumors, the immunocytochemical method for PR localization was also used in combination with a slightly modified Abbott ER-ICA staining for estrogen receptor to compare the distribution of both receptors within the same biopsy on adjacent frozen sections. Various combinations of estrogen receptor and PR contents that have been determined by steroid-binding assay have also been detected by the double immunocytochemical assay.

[Placental abruption: Background and revisited pronostic factors about a series of 171 cases]. / Hématome rétroplacentaire : terrain et facteurs pronostiques revisités à propos d'une série de 171 cas en Guyane francaise.

OBJECTIVE: To describe the epidemiological, clinical and prognostic factors of placental abruption and fetal death in utero and to investigate possible risk factors for their occurrence. PATIENTS AND METHODS: Observational retrospective study including the women having presented a placental abruption between January 2001 and January 2012, in a IIB maternity. Women's sociodemographic characteristics, clinical symptoms and the method used to detect placental abruption were collected. Patient data of those whose pregnancy resulted in fetal death were compared to those with more favorable outcomes. RESULTS: There were 171 cases of placental abruption among 21,913 patients having delivered, which represents a 0.78% incidence. Diagnosis was rarely based on clinical data (30%). The rate of fetal death in utero represented 25% of the pregnancy's outcomes. A history of fetal death in utero increased the risk of placental abruption (P<0.001). This complication was more frequent for patients who did not have pregnancy monitoring (P=0.054) and before 37 weeks of amenorrhoea (P=0.005). CONCLUSION: Placental abruption is an important cause of perinatal mortality and maternal morbidity. Among the observed risk factors, only regular pregnancy monitoring can be an easy way to prevent these complications.

New functional zonation in the ovary as shown by immunohistochemistry of luteinizing hormone receptor.

Monoclonal anti-LH receptor antibodies were used to study receptor distribution in the various structures of porcine ovary. Primordial and primary follicles were not labeled. Immunoreactivity appeared on the thecal cells of secondary follicles of approximately 100-200 microns in diameter. In large antral follicles and preovulatory follicles granulosa cells were also labeled, whereas two zones were observed in the theca interna. The region close to the lamina basalis (approximately 1/3 of the thecal cells) appeared devoid of LH receptors, whereas a strong labeling was observed on the most external region. Thecal cells were also labeled in atretic follicles. There was no labeling of the oocyte. In cyclic corpora lutea only the most external cells, probably of thecal origin, were stained. The most abundant cells, of granulosa origin, were not labeled. In the interstitium few cells were immunolabeled; they proceeded either from remnants of atretic follicles or existed as groups of isolated cells. Thus both the theca interna of preovulatory follicles and the corpus luteum are heterogenous structures composed of two zones of which only one is directly responsive to LH. No LH immunoactivity could be detected in nontarget organs (liver, lung, thyroid gland, small intestine, fallopian tube, and uterus).

Monoclonal antibodies for immunocytochemistry of progesterone receptors (PR) in various laboratory rodents, livestock, humans, and chickens: identification of two epitopes conserved in PR of all these species.

Over 90 mouse monoclonal antibodies have been raised against rabbit and human uterine progesterone receptor (PR). These antibodies, because of their specificity, are powerful tools with which to examine the localization, structure, and function of PR. A selection of 22 well characterized mABs was made to test their ability to give the best immunocytochemical staining of PR in various species. Comparative analysis of the antibodies led to the following conclusions. Li 417 (and, to a lesser extent, Let 126) was the best monoclonal in humans; Let 126 and Mi 60 were the most sensitive monoclonals in guinea pigs, rabbits, and monkeys. In sheep, sows, cows and mares as well as in rats and chickens Let 81 or Let 548 gave the best results (Let 126 was also effective in sows and mares, while Li 169 was also effective in sheep and cows). Two antibodies (Li 169 and Let 548) cross-reacted with PR in all of the species tested, including mammals and birds, and appeared to recognize two highly conserved antigenic sites. Remarkably, these conserved sequences are located in the highly variable N-terminal part of the receptor; they may, thus, be related to the still poorly understood function of this domain of the receptor.

Monoclonal antibodies against luteinizing hormone receptor. Immunochemical characterization of the receptor.

Human CG (hCG)-receptor complexes were solubilized from porcine testicular membranes. They were chromatographed on an immunomatrix of Affi-Gel 10-D1E8 anti beta-hCG monoclonal antibody (this antibody has been shown not to interfere with hCG binding to receptor). Elution was performed at alkaline pH, a condition in which hCG-receptor complexes are relatively stable. Immunization of a mouse with these partially (approximately 15%) purified hormone-receptor complexes allowed the preparation of 20 different hybridomas, each secreting antireceptor antibodies. The latter were used for receptor characterization. Immunoblot of testicular membrane extracts or of purified receptor preparations showed the presence of a major band at approximately 85,000 mol wt and minor bands at approximately 68,000 mol wt and approximately 48-45,000 mol wt. The width of all these bands suggested some microheterogeneity possibly due to glycosylation. The same approximately 85,000 mol wt receptor was seen in ovarian membranes, but no detectable antigen was observed in liver, muscle, and kidney membranes. An immunoaffinity method (using antibody LHR 38) was devised to purify the receptor in a single step. This demonstrated that the purified receptor preparation did not contain any protein component other than those detected by immunoblot. Comparison of receptors purified by immunoaffinity chromatography using either antireceptor or antihormone monoclonal antibodies showed in both cases the presence of the 85,000 mol wt and 48-45,000 mol wt species, but the absence, in the latter case, of the 68,000 mol wt species. This suggests that the 68,000 mol wt receptor cannot bind hormone and does not form oligomers with other receptor species.

Binding of steroids by proteins in follicular fluid of the human ovary.

Steroid-protein interactions were studied in the antral fluid of preovulatory follicles of the human ovary. Two high affinity binding systems are described. These steroid-binding proteins do not differ from those identified in the plasma of women (transcortin and sex binding protein), as indicated by similar specificity, binding affinity, and electrophoretic pattern. Their concentrations in the fluid are either similar to or lower than those in plasma. Since the steroid concentrations in antral fluid are approximately 20- to 100-fold higher than the binding capacities of the specific steroid binders, the buffer role that many authors attribute to the proteins which bind steroids is unlikely in antral fluid.

Comparison of immunocytochemical and molecular features with the phenotype in a case of incomplete male pseudohermaphroditism associated with a mutation of the luteinizing hormone receptor.

We report the case of an infant who presented at birth with a hypoplastic phallus associated with hypospadias. Low testosterone production, normal serum levels of steroid precursors, and increased LH in response to LH-releasing hormone supported a defect in Leydig cell differentiation or function. Conventional microscopic study of the testes showed fibroblastic cells in the interstitium. However immunocytochemical analysis using anti-LH receptor and anti-P450c17 antibodies demonstrated that about one third of these cells were Leydig cells or precursors of Leydig cells. No histological feature could distinguish the latter cells from fibroblasts. A homozygous substitution of cysteine 133 for arginine was found in the extracellular domain of the receptor. This is the first naturally occurring missense mutation found in the extracellular domain of the LH receptor. COS-7 cells transfected with the mutant receptor exhibited a marked impairment of hCG binding, whereas some cAMP production could be observed at high hCG concentrations. We propose that the partial impairment of LH receptor function, as reflected by the presence of Leydig cells, was responsible for the incomplete male pseudohermaphroditism observed in our patient.

Nitric oxide synthase in the guinea pig preoptic area and hypothalamus: distribution, effect of estrogen, and colocalization with progesterone receptor.

Nitric oxide (NO) may function as an intercellular messenger in the hypothalamus and may play a role in the control of gonadotropin-releasing hormone (GnRH) secretion and sexual behavior. Progesterone also plays an important role in the regulation of reproductive functions. Recent experiments have shown that progesterone-induced sexual behavior in ovariectomized, estrogen-primed rats was caused by the release of NO from nitric oxide synthase (NOS)-containing neurons and the subsequent stimulation of the release of GnRH. To provide further neuroanatomical support for the role of NO in these gonadal steroid-dependent behavioral and physiological processes, we determined (1) the distribution of the nicotinamide-adenosine-dinucleotide phosphate-diaphorase (NADPHd) and NOS enzymes in the guinea pig preoptic area and hypothalamus, regions that contain steroid receptors; (2) the effect of estrogen on NADPHd activity in these regions; and (3) the neuroanatomical relationship between NOS and the progesterone receptor (PR). For this purpose, single-(NADPHd) and double- (NADPHd with NOS or NADPHd with PR or NOS with PR) staining techniques were applied to sections of brains of guinea pigs. The studies showed scattered NADPHd-positive neurons in most parts of the preoptic area and heavily stained cells in the hypothalamus. In these regions, the pattern and density of NOS immunoreactivity closely corresponded to the pattern of NADPHd staining. Quantitative analysis showed an increase in the number of NADPHd-positive neurons in the ventrolateral nucleus of ovariectomized animals primed with estradiol. Approximately 16% of the NOS-immunoreactive (IR) cells in the rostral preoptic area and 55% of NOS-IR cells in the ventrolateral nucleus displayed PR immunoreactivity. These results suggest that NOS may be regulated by gonadal steroids and provide neuroanatomical evidence that progesterone may exert its effect directly on more than half of NOS-synthesizing cells in the ventrolateral nucleus, a key region in the control of sexual behavior.