Vaginal Microbiota Profile in first-trimester miscarriages cases.

Due to its rising prevalence, first-trimester miscarriage is getting more attention. Abortion etiology and pathology, especially in non-pathological cases, are unknown. In this study, 435 vaginal swabs were collected from aborted women in Maternity Teaching Hospitals in Erbil and Shahid Dr. Khalid Hospital in Koya. We characterized the vaginal microbiota diversity and composition in first-trimester abortion and investigated the association between bacterial vaginosis and abortion before 12 weeks. Cultural, morphological, and biochemical characteristics for each bacterial genus were discovered, and VITEK-2 system was used to identify isolated bacteria. Samples from each type of bacteria were selected for sequencing utilizing 16 rRNA genes examining V4-V8 region for bacterial profiles. Bacterial vaginitis was found in 412 (94.7%) first-trimester miscarriages. Six Gram-positive and four Gram-negative bacteria were found in these 412 samples. Microorganism distribution varied Gram-positive bacteria Staphylococcus aureus (86) 20.87%, Enterococcus faecalis (31) 7.52%, Gardnerella vaginalis (24) 5.83%, Streptococcus agalactia (21) 5.1%, Lactobacillus equicursoris (14) 3.4% and Staphylococcus haemolyticus (12) 2.91%. Gram-negative bacteria including E. coli (107) 25.97%, Klebsiella pneumonia (76) 18.45%, Pseudomonas aeruginosa (29) 7.04% and Proteus mirabilis (12) 2.91%. Staphylococcus aureus had the highest rate of isolation at (86) 20.87%, while Lactobacillus equicursoris had the lowest rate at (14) 3.4%. Overall, the rate of isolation for Gram-negative bacteria (224) was 54.4%, while the rate for Gram-positive bacteria (188) was 45.6%.

Lupus-Induced Vasculitis and Multiple Organ Dysfunction Syndrome as the First Presentation of Systemic Lupus Erythematosus (SLE) in Pregnancy.

BACKGROUND Systemic lupus erythematosus (SLE) is an autoimmune disease characterized by autoantibody production leading to inflammation in multiple organs; it commonly affects young women in their child-bearing years. Clinical manifestations are diverse and range from mild arthritis to diffuse alveolar hemorrhage (DAH). DAH is a rare and devastating complication of SLE that carries a mortality rate of up to 50%, despite aggressive therapy. CASE REPORT A 21-year-old primigravida at 16 weeks gestation presents with a productive cough, rash, sore throat, and high-grade fever. Chest x-ray suggested multifocal pneumonia. Patient deteriorated despite antibiotics and intravenous (IV) fluids. She developed worsening anemia, leukopenia, and thrombocytopenia. Autoimmune workup was positive for Coombs, antinuclear antibody, anti-smith antibody, and hypocomplementemia. Skin biopsy was consistent with SLE. SLE vasculitis was suspected. She required mechanical intubation for rapid respiratory deterioration, with CT thorax suggesting ARDS. Bronchoscopy was done and confirmed DAH. Her course was further complicated with retinopathy and acute pancreatitis associated with SLE. She was treated with IV steroids, IV cyclophosphamide, and plasmapheresis, with significant clinical improvement and successful extubation. She delivered a healthy baby at 32 weeks gestation. CONCLUSIONS Early recognition and initiation of treatment is critical to survival in DAH and requires a high index of clinical suspicion. Treatment includes high-dose steroids, cyclophosphamide, and plasma exchange. Pregnancy increases the risk of adverse outcome in SLE. Seven cases of DAH in pregnant patients with SLE have been reported. Here, we report a catastrophic presentation of DAH, acute pancreatitis, and retinopathy in a pregnant patient with newly diagnosed SLE.

Transcriptional analysis of the cell division-related ssg genes in Streptomyces coelicolor reveals direct control of ssgR by AtrA.

SsgA-like proteins are a family of actinomycete-specific regulatory proteins that control cell division and spore maturation in streptomycetes. SsgA and SsgB together activate sporulation-specific cell division by controlling the localization of FtsZ. Here we report the identification of novel regulators that control the transcription of the ssgA-like genes. Transcriptional regulators controlling ssg gene expression were identified using a DNA-affinity capture assay. Supporting transcriptional and DNA binding studies showed that the ssgA activator gene ssgR is controlled by the TetR-family regulator AtrA, while the γ-butyrolactone-responsive AdpA (SCO2792) and SlbR (SCO0608) and the metabolic regulator Rok7B7 (SCO6008) were identified as candidate regulators for the cell division genes ssgA, ssgB and ssgG. Transcription of the cell division gene ssgB depended on the sporulation genes whiA and whiH, while ssgR, ssgA and ssgD were transcribed independently of the whi genes. Our work sheds new light on the mechanisms by which sporulation-specific cell division is controlled in Streptomyces.

A comparison of key aspects of gene regulation in Streptomyces coelicolor and Escherichia coli using nucleotide-resolution transcription maps produced in parallel by global and differential RNA sequencing.

Streptomyces coelicolor is a model for studying bacteria renowned as the foremost source of natural products used clinically. Post-genomic studies have revealed complex patterns of gene expression and links to growth, morphological development and individual genes. However, the underlying regulation remains largely obscure, but undoubtedly involves steps after transcription initiation. Here we identify sites involved in RNA processing and degradation as well as transcription within a nucleotide-resolution map of the transcriptional landscape. This was achieved by combining RNA-sequencing approaches suited to the analysis of GC-rich organisms. Escherichia coli was analysed in parallel to validate the methodology and allow comparison. Previously, sites of RNA processing and degradation had not been mapped on a transcriptome-wide scale for E. coli. Through examples, we show the value of our approach and data sets. This includes the identification of new layers of transcriptional complexity associated with several key regulators of secondary metabolism and morphological development in S. coelicolor and the identification of host-encoded leaderless mRNA and rRNA processing associated with the generation of specialized ribosomes in E. coli. New regulatory small RNAs were identified for both organisms. Overall the results illustrate the diversity in mechanisms used by different bacterial groups to facilitate and regulate gene expression.

Foster care as a viable alternative to institutional care in the Middle East: community acceptance and stigma across type of placement in Jordan.

OBJECTIVE: Utilizing an experimental vignette design, this study assessed attitudes in the Hashemite Kingdom of Jordan toward the implementation of foster care as an alternative to institutions for children in need of care and protection. METHODS: A sample of 111 adults were surveyed in Amman and presented with a vignette describing a 14-year-old boy who came into the care of the Ministry of Social Development when he was a baby after being placed by the grandfather because of shame surrounding the mother being unwed. The vignettes systematically varied as to whether the child was described as raised in an orphanage, with a relative in a kinship foster placement, or with a nonkin foster family. Participants were then asked a series of questions about their acceptance of the child, stigma that the community might attach to the child, and potential outcomes for the child. RESULTS: We found no differences across the acceptance and stigma questions between the kinship and non-kin foster conditions. The 2 foster care options were at least as acceptable as current institutional models across all domains, and participants were more likely to accept the child going to school with or being friends with their child if they were in foster care rather than an institution. CONCLUSIONS: These results represent the first evidence of public acceptance of foster care as a model of care in Jordan and may inform the process of local stakeholders implementing alternatives to institutional care on a meaningful and sustainable scale in the Kingdom and regionally.

Subcranial transnasal repair of cerebrospinal fluid rhinorrhea with free autologous grafts by the combined overlay and underlay techniques.

OBJECTIVE: To determine the efficacy of subcranial transnasal repair of cerebrospinal fluid rhinorrhea with free autologous grafts by the combined overlay and underlay techniques using the surgical microscope and/or endoscope. PATIENTS AND METHODS: Twenty patients with CSF rhinorrhea were included in this retrospective study. They were 13 males and 7 females. Their age ranged from 7 to 62 years (mean: 39.35). The etiologies of the leak were iatrogenic in 10 cases, spontaneous in 5 cases, traumatic in 4 cases and one case was associated with meningeo-encephalocele. Preoperative nasal endoscopic examination, computed tomography (CT) with intrathecal non-ionic contrast and magnetic resonance imaging (MRI) were done when indicated. Endoscopic and/or microscopic repair of the CSF fistula was done by a combination of both underlay and overlay repair with free autologous grafts as follows: Gelfoam with fibrin glue, strips of fat, facia lata, Gelfoam with fibrin glue (underlay), septal cartilage, Gelfoam with fibrin glue and strips of fat (overlay). RESULTS: Complete closure of the leak was achieved in all patients. In one case of spontaneous CSF leak which was operated endoscopically, the leak recurred 6 months postoperatively and ceased spontaneously after a month with conservative medical treatment. No major complications were seen and no patients developed meningitis or postoperative anosmia. CONCLUSION: Subcranial transnasal repair with free autologous grafts by the combined overlay and underlay techniques using the endoscope or surgical microscope is a safe and successful method of treating CSF leaks, provided that the CSF leak is precisely located and the site can be reached with the endoscope or surgical microscope.

Sellar floor reconstruction with nasal turbinate tissue after endoscopic endonasal transsphenoidal surgery for pituitary adenomas.

BACKGROUND: The emerging endoscopic endonasal transsphenoidal approach for pituitary adenomas is safe and effective. An intraoperative sellar floor reconstructive method after this approach is challenging.[nl] OBJECTIVE: To describe a simple method of sellar reconstruction after endoscopic endonasal transsphenoidal surgery for pituitary adenomas by nasal turbinate tissue.[nl] MATERIALS AND METHODS: Thirty patients with defects in the floor of the sella turcica, after endoscopic endonasal transsphenoidal surgery for pituitary adenomas, underwent reconstruction with nasal turbinate tissue. The surgical technique is described.[nl] RESULTS: Patients who underwent this sellar reconstruction did not show postoperative cerebrospinal leak or other complications.[nl] CONCLUSION: Nasal turbinate tissue is an excellent source of donor material for successful reconstruction of the sellar floor. It is costless, safe, soft, malleable and easy to obtain in the same field of surgery with suitable size without inducing side effects or complications.

Transgenic tilapia and the tilapia genome.

The tilapia fish (Oreochromis niloticus) has an important place in the aquaculture of the developing world. It is also a very useful laboratory animal, and readily lends itself to the transgenic technology. Through the use of reporter genes, a range of potential gene promoters have been tested in tilapia, both through transient and stable expression of the reporter construct. Using the transgenic technology, growth enhanced lines of tilapia have been produced. These fish have no abnormalities and offer a considerable growth advantage for future exploitation. It is however crucial that transgenic fish, to be exploited in aquaculture, be sterile, and various methods of achieving sterility are considered. These include triploidy, gene knock out of crucial hormone encoding genes via homologous recombination, and knock down of the function of the same genes via ribozyme or antisense technologies. Transgenic tilapia also offer the potential for exploitation as biofactories in the production of valuable pharmaceutical products, and this is also discussed.

Expression of a novel piscine growth hormone gene results in growth enhancement in transgenic tilapia (Oreochromis niloticus).

Several lines of transgenic G1 and G2 tilapia fish (Oreochromis niloticus) have been produced following egg injection with gene constructs carrying growth hormone coding sequences of fish origin. Using a construct in which an ocean pout antifreeze promoter drives a chinook salmon growth hormone gene, dramatic growth enhancement has been demonstrated, in which the mean weight of the 7 month old G2 transgenic fish is more than three fold that of their non transgenic siblings. Somewhat surprisingly G1 fish transgenic for a construct consisting of a sockeye salmon metallothionein promoter spliced to a sockeye salmon growth hormone gene exhibited no growth enhancement, although salmon transgenic for this construct do show greatly enhanced growth. The growth enhanced transgenic lines were also strongly positive in a radio-immuno assay for the specific hormone in their serum, whereas the non growth enhanced lines were negative. Attempts to induce expression from the metallothionein promoter by exposing fish to increased levels of zinc were also unsuccessful. Homozygous transgenic fish have been produced from the ocean pout antifreeze/chinook salmon GH construct and preliminary trials suggest that their growth performance is similar to that of the hemizygous transgenics. No abnormalities were apparent in the growth enhanced fish, although minor changes to skull shape and reduced fertility were noted in some fish. There is also preliminary evidence for improved food conversion ratios when growth enhanced transgenic tilapia are compared to their non-transgenic siblings. The long term objective of this study is to produce lines of tilapia which are both growth enhanced and sterile, so offering improved strains of this important food fish for aquaculture.

Tobacco budworm P-glycoprotein: biochemical characterization and its involvement in pesticide resistance.

Since pesticides have been shown to interact with P-glycoprotein (P-gp), the purpose of this study was to examine the possible role of P-gp in pesticide resistance in the tobacco budworm (Heliothis virescens). Using three P-gp antibodies, P-gp expression in various resistant populations of tobacco budworms was found to be 2-6-times that of the susceptible larvae. Tobacco budworm P-gp was glycosylated and localized primarily in the cuticle and fat body with little expression in the mid gut. To determine the role of P-gp in pesticide resistance, resistant tobacco budworm larvae were treated with a P-gp inhibitor, quinidine, and challenged with various doses of thiodicarb. Inhibition of P-gp decreased the LD50 for thiodicarb by a factor of 12.5. Quinidine treatment did not result in a significant inhibition of the P-450 system nor did it alter the feeding of the larvae, suggesting the potential involvement of P-gp in pesticide resistance. An age-dependent increase in P-gp expression was detected in resistant larvae as compared to control, susceptible larvae. This correlates with the reported age-dependent increase in resistance and is further evidence supporting the role of P-gp in the development of pesticide resistance.

Comparison of the activity of carp and rat beta-actin gene regulatory sequences in tilapia and rainbow trout embryos.

A comparative study on the level of expression of lacZ reporter constructs driven by equivalent carp and rat beta-actin regulatory sequences was carried out in embryos of tilapia and rainbow trout. DNA was microinjected into fertilised tilapia and rainbow trout eggs and the embryos/fry were assayed at various developmental stages for beta-galactosidase expression. We provide evidence to demonstrate that the carp beta-actin promoter/ lacZ reporter gene is expressed at higher levels than the equivalent rat beta-actin construct in both species.

P-glycoprotein involvement in cuticular penetration of [14C]thiodicarb in resistant tobacco budworms.

Pesticides have been shown to interact with the multidrug resistance protein associated with cancer chemotherapy, P-glycoprotein (P-gp). P-gp, therefore, has also been implicated in the development of pesticide resistance. The purpose of this study was to characterize the effect P-gp has on the accumulation of the carbamate pesticide, thiodicarb. For these studies, resistant tobacco budworm larvae, expressing four times the P-gp as susceptible larvae, were pretreated with the P-gp inhibitor, quinidine, and challenged topically with thiodicarb. Quinidine enhanced thiodicarb toxicity in a dose-dependent manner, with mortality in the presence of P-gp inhibition increased up to 33%. Quinidine treatment increased [14C]thiodicarb accumulation 2- to 3-fold as compared to thiodicarb treatment alone. This study suggests that P-gp contributes to quinidine synergism of thiodicarb toxicity and suggests that P-gp may be involved in cuticular resistance to pesticides.

Distally based medial island septocutaneous flap for repair of soft-tissue defects of the lower leg.

A medial island septocutaneous flap, based on the lower septocutaneous vessels from the posterior tibial artery and vein, has been designed. This flap was rotated 90 degrees in 5 cases and 180 degrees in 2 cases to cover soft-tissue defects on the lower third of the leg.