Inefficient Sox9 upregulation and absence of Rspo1 repression lead to sex reversal in the B6.XYTIR mouse gonad†.

Sry on the Y-chromosome upregulates Sox9, which in turn upregulates a set of genes such as Fgf9 to initiate testicular differentiation in the XY gonad. In the absence of Sry expression, genes such as Rspo1, Foxl2, and Runx1 support ovarian differentiation in the XX gonad. These two pathways antagonize each other to ensure the development of only one gonadal sex in normal development. In the B6.YTIR mouse, carrying the YTIR-chromosome on the B6 genetic background, Sry is expressed in a comparable manner with that in the B6.XY mouse, yet, only ovaries or ovotestes develop. We asked how testicular and ovarian differentiation pathways interact to determine the gonadal sex in the B6.YTIR mouse. Our results showed that (1) transcript levels of Sox9 were much lower than in B6.XY gonads while those of Rspo1 and Runx1 were as high as B6.XX gonads at 11.5 and 12.5 days postcoitum. (2) FOXL2-positive cells appeared in mosaic with SOX9-positive cells at 12.5 days postcoitum. (3) SOX9-positive cells formed testis cords in the central area while those disappeared to leave only FOXL2-positive cells in the poles or the entire area at 13.5 days postcoitum. (4) No difference was found at transcript levels of all genes between the left and right gonads up to 12.5 days postcoitum, although ovotestes developed much more frequently on the left than the right at 13.5 days postcoitum. These results suggest that inefficient Sox9 upregulation and the absence of Rspo1 repression prevent testicular differentiation in the B6.YTIR gonad.

Combined myoepithelial carcinoma and myoepithelioma in soft tissue: a case report and review of the literature.

INTRODUCTION: Soft tissue myoepithelial carcinoma and myoepithelioma are rare entities, part of myoepithelial tumors. They were incorporated into the World Health Organization classification of soft tissue tumors in 2002. Here we present an exceptional case of myoepithelial carcinoma and myoepithelioma association. To the best of our knowledge, such an association has never been reported in the literature. CASE PRESENTATION: We report a case of myoepithelial carcinoma combined with myoepithelioma occurring in the soft tissue of the right forearm of an 84-year-old Arabian man. We describe the clinical, radiological and pathological features dominated by histological polymorphism. We will also describe the proposed histological criteria of malignancy and the major role of immunohistochemistry in positive and differential diagnosis. We finally mention the therapeutic arsenal available. CONCLUSION: Through this work, we report that myoepithelioma of soft tissue can progress to malignant myoepithelioma.

Lysine-specific demethylase 1-mediated demethylation of histone H3 lysine 9 contributes to interleukin 1ß-induced microsomal prostaglandin E synthase 1 expression in human osteoarthritic chondrocytes.

INTRODUCTION: Microsomal prostaglandin E synthase 1 (mPGES-1) catalyzes the terminal step in the biosynthesis of PGE2, a critical mediator in the pathophysiology of osteoarthritis (OA). Histone methylation plays an important role in epigenetic gene regulation. In this study, we investigated the roles of histone H3 lysine 9 (H3K9) methylation in interleukin 1ß (IL-1ß)-induced mPGES-1 expression in human chondrocytes. METHODS: Chondrocytes were stimulated with IL-1ß, and the expression of mPGES-1 mRNA was evaluated using real-time RT-PCR. H3K9 methylation and the recruitment of the histone demethylase lysine-specific demethylase 1 (LSD1) to the mPGES-1 promoter were evaluated using chromatin immunoprecipitation assays. The role of LSD1 was further evaluated using the pharmacological inhibitors tranylcypromine and pargyline and small interfering RNA (siRNA)-mediated gene silencing. The LSD1 level in cartilage was determined by RT-PCR and immunohistochemistry. RESULTS: The induction of mPGES-1 expression by IL-1ß correlated with decreased levels of mono- and dimethylated H3K9 at the mPGES-1 promoter. These changes were concomitant with the recruitment of the histone demethylase LSD1. Treatment with tranylcypromine and pargyline, which are potent inhibitors of LSD1, prevented IL-1ß-induced H3K9 demethylation at the mPGES-1 promoter and expression of mPGES-1. Consistently, LSD1 gene silencing with siRNA prevented IL-1ß-induced H3K9 demethylation and mPGES-1 expression, suggesting that LSD1 mediates IL-1ß-induced mPGES-1 expression via H3K9 demethylation. We show that the level of LSD1 was elevated in OA compared to normal cartilage. CONCLUSION: These results indicate that H3K9 demethylation by LSD1 contributes to IL-1ß-induced mPGES-1 expression and suggest that this pathway could be a potential target for pharmacological intervention in the treatment of OA and possibly other arthritic conditions.

Evaluation of hemifield sector analysis protocol in multifocal visual evoked potential objective perimetry for the diagnosis and early detection of glaucomatous field defects.

PURPOSE: Multifocal visual evoked potential (mfVEP) is a newly introduced method used for objective visual field assessment. Several analysis protocols have been tested to identify early visual field losses in glaucoma patients using the mfVEP technique, some were successful in detection of field defects, which were comparable to the standard automated perimetry (SAP) visual field assessment, and others were not very informative and needed more adjustment and research work. In this study we implemented a novel analysis approach and evaluated its validity and whether it could be used effectively for early detection of visual field defects in glaucoma. METHODS: Three groups were tested in this study; normal controls (38 eyes), glaucoma patients (36 eyes) and glaucoma suspect patients (38 eyes). All subjects had a two standard Humphrey field analyzer (HFA) test 24-2 and a single mfVEP test undertaken in one session. Analysis of the mfVEP results was done using the new analysis protocol; the hemifield sector analysis (HSA) protocol. Analysis of the HFA was done using the standard grading system. RESULTS: Analysis of mfVEP results showed that there was a statistically significant difference between the three groups in the mean signal to noise ratio (ANOVA test, p < 0.001 with a 95% confidence interval). The difference between superior and inferior hemispheres in all subjects were statistically significant in the glaucoma patient group in all 11 sectors (t-test, p < 0.001), partially significant in 5 / 11 (t-test, p < 0.01), and no statistical difference in most sectors of the normal group (1 / 11 sectors was significant, t-test, p < 0.9). Sensitivity and specificity of the HSA protocol in detecting glaucoma was 97% and 86%, respectively, and for glaucoma suspect patients the values were 89% and 79%, respectively. CONCLUSIONS: The new HSA protocol used in the mfVEP testing can be applied to detect glaucomatous visual field defects in both glaucoma and glaucoma suspect patients. Using this protocol can provide information about focal visual field differences across the horizontal midline, which can be utilized to differentiate between glaucoma and normal subjects. Sensitivity and specificity of the mfVEP test showed very promising results and correlated with other anatomical changes in glaucoma field loss.

The role of hemifield sector analysis in multifocal visual evoked potential objective perimetry in the early detection of glaucomatous visual field defects.

OBJECTIVE: The purpose of this study was to examine the effectiveness of a new analysis method of mfVEP objective perimetry in the early detection of glaucomatous visual field defects compared to the gold standard technique. METHODS AND PATIENTS: Three groups were tested in this study; normal controls (38 eyes), glaucoma patients (36 eyes), and glaucoma suspect patients (38 eyes). All subjects underwent two standard 24-2 visual field tests: one with the Humphrey Field Analyzer and a single mfVEP test in one session. Analysis of the mfVEP results was carried out using the new analysis protocol: the hemifield sector analysis protocol. RESULTS: Analysis of the mfVEP showed that the signal to noise ratio (SNR) difference between superior and inferior hemifields was statistically significant between the three groups (analysis of variance, P < 0.001 with a 95% confidence interval, 2.82, 2.89 for normal group; 2.25, 2.29 for glaucoma suspect group; 1.67, 1.73 for glaucoma group). The difference between superior and inferior hemifield sectors and hemi-rings was statistically significant in 11/11 pair of sectors and hemi-rings in the glaucoma patients group (t-test P < 0.001), statistically significant in 5/11 pairs of sectors and hemi-rings in the glaucoma suspect group (t-test P < 0.01), and only 1/11 pair was statistically significant (t-test P < 0.9). The sensitivity and specificity of the hemifield sector analysis protocol in detecting glaucoma was 97% and 86% respectively and 89% and 79% in glaucoma suspects. These results showed that the new analysis protocol was able to confirm existing visual field defects detected by standard perimetry, was able to differentiate between the three study groups with a clear distinction between normal patients and those with suspected glaucoma, and was able to detect early visual field changes not detected by standard perimetry. In addition, the distinction between normal and glaucoma patients was especially clear and significant using this analysis. CONCLUSION: The new hemifield sector analysis protocol used in mfVEP testing can be used to detect glaucomatous visual field defects in both glaucoma and glaucoma suspect patients. Using this protocol, it can provide information about focal visual field differences across the horizontal midline, which can be utilized to differentiate between glaucoma and normal subjects. The sensitivity and specificity of the mfVEP test showed very promising results and correlated with other anatomical changes in glaucomatous visual field loss. The intersector analysis protocol can detect early field changes not detected by the standard Humphrey Field Analyzer test.

Expression of peroxisome proliferator-activated receptors α, ß, γ, and H- and L-prostaglandin D synthase during osteoarthritis in the spontaneous hartley guinea pig and experimental dog models.

OBJECTIVE: To investigate the expression of peroxisome proliferator-activated receptors (PPAR) α, ß, and γ, and hematopoietic and lipocalin-type prostaglandin D synthase (H- and L-PGDS) over the course of osteoarthritis (OA) in the spontaneous Hartley guinea pig and the anterior cruciate ligament transection dog models. METHODS: Guinea pigs were sacrificed at 2 (control group), 4, 8, and 12 months of age (n = 5 per group). Non-operated (control) and operated dogs were sacrificed at 4, 8, and 12 weeks postsurgery. Cartilage was evaluated histologically using the Osteoarthritis Research Society International (OARSI) guidelines. The expression of PPAR-α, ß, γ, and H- and L-PGDS was evaluated by real-time PCR and immunohistochemistry. The nonparametric Spearman test was used for correlation analysis. RESULTS: PPAR-α, ß, and γ were detected in medial tibial plateau from control animals in both the spontaneous and surgical models. Levels of PPAR-α and ß did not change over the course of OA, whereas PPAR-γ levels decreased during progression of disease. We also observed that the expression of H-PGDS remained unchanged, whereas L-PGDS increased over the course of OA. PPAR-γ levels correlated negatively, whereas L-PGDS levels correlated positively, with the histological score of OA. CONCLUSION: The level of PPAR-γ decreased, whereas level of L-PGDS increased during the progression of OA. These data suggest that reduced expression of PPAR-γ may contribute to the pathogenesis of OA, whereas enhanced expression of L-PGDS may be part of a reparative process.

Egr-1 contributes to IL-1-mediated down-regulation of peroxisome proliferator-activated receptor γ expression in human osteoarthritic chondrocytes.

INTRODUCTION: Peroxisome proliferator-activated receptor (PPAR)γ has been shown to exhibit anti-inflammatory and anti-catabolic properties and to be protective in animal models of osteoarthritis (OA). We have previously shown that interleukin-1ß (IL-1) down-regulates PPARγ expression in human OA chondrocytes. However, the mechanisms underlying this effect have not been well characterized. The PPARγ promoter harbors an overlapping Egr-1/specificity protein 1 (Sp1) binding site. In this study, our objective was to define the roles of Egr-1 and Sp1 in IL-1-mediated down-regulation of PPARγ expression. METHODS: Chondrocytes were stimulated with IL-1 and the expression levels of Egr-1 and Sp1 mRNAs and proteins were evaluated using real-time reverse transcriptase-polymerase chain reaction (RT-PCR) and Western blotting, respectively. The role of de novo protein synthesis was evaluated using the protein synthesis inhibitor cycloheximide (CHX). The recruitment of Sp1 and Egr-1 to the PPARγ promoter was evaluated using chromatin immunoprecipitation (ChIP) assays. The PPARγ promoter activity was analyzed in transient transfection experiments. The roles of Egr-1 and Sp1 were further evaluated using small interfering RNA (siRNA) approaches. The level of Egr-1 in cartilage was determined using immunohistochemistry. RESULTS: Down-regulation of PPARγ expression by IL-1 requires de novo protein synthesis and was concomitant with the induction of the transcription factor Egr-1. Treatment with IL-1 induced Egr-1 recruitment and reduced Sp1 occupancy at the PPARγ promoter. Overexpression of Egr-1 potentiated, whereas overexpression of Sp1 alleviated, the suppressive effect of IL-1 on the PPARγ promoter, suggesting that Egr-1 may mediate the suppressive effect of IL-1. Consistently, Egr-1 silencing prevented IL-1-mediated down-regulation of PPARγ expression. We also showed that the level of Egr-1 expression was elevated in OA cartilage compared to normal cartilage. CONCLUSIONS: Our results indicate that induction and recruitment of Egr-1 contributed to the suppressive effect of IL-1 on PPARγ expression. They also suggest that modulation of Egr-1 levels in the joint may have therapeutic potential in OA.

Valproic acid suppresses interleukin-1ß-induced microsomal prostaglandin E2 synthase-1 expression in chondrocytes through upregulation of NAB1.

OBJECTIVE: Microsomal prostaglandin E(2) synthase-1 (mPGES-1) catalyzes the terminal step in the biosynthesis of PGE(2). Early growth response factor-1 (Egr-1) is a key transcription factor in the regulation of mPGES-1, and its activity is negatively regulated by the corepressor NGF1-A-binding protein-1 (NAB1). We examined the effects of valproic acid (VA), a histone deacetylase inhibitor, on interleukin 1ß (IL-1ß)-induced mPGES-1 expression in human chondrocytes, and evaluated the roles of Egr-1 and NAB1 in these effects. METHODS: Chondrocytes were stimulated with IL-1 in the absence or presence of VA, and the level of mPGES-1 protein and mRNA expression were evaluated using Western blotting and real-time reverse-transcription polymerase chain reaction (PCR), respectively. mPGES-1 promoter activity was analyzed in transient transfection experiments. Egr-1 and NAB1 recruitment to the mPGES-1 promoter was evaluated using chromatin immunoprecipitation assays. Small interfering RNA (siRNA) approaches were used to silence NAB1 expression. RESULTS: VA dose-dependently suppressed IL-1-induced mPGES-1 protein and mRNA expression as well as its promoter activation. Treatment with VA did not alter IL-1-induced Egr-1 expression, or its recruitment to the mPGES-1 promoter, but prevented its transcriptional activity. The suppressive effect of VA requires de novo protein synthesis. VA induced the expression of NAB1, and its recruitment to the mPGES-1 promoter, suggesting that NAB1 may mediate the suppressive effect of VA. Indeed, NAB1 silencing with siRNA blocked VA-mediated suppression of IL-1-induced mPGES-1 expression. CONCLUSION: VA inhibited IL-1-induced mPGES-1 expression in chondrocytes. The suppressive effect of VA was not due to reduced expression or recruitment of Egr-1 to the mPGES-1 promoter and involved upregulation of NAB1.

Contribution of H3K4 methylation by SET-1A to interleukin-1-induced cyclooxygenase 2 and inducible nitric oxide synthase expression in human osteoarthritis chondrocytes.

OBJECTIVE: To investigate the role of histone H3 lysine 4 (H3K4) methylation in interleukin-1ß (IL-1ß)-induced cyclooxygenase 2 (COX-2) and inducible nitric oxide synthase (iNOS) expression in human osteoarthritic (OA) chondrocytes. METHODS: Chondrocytes were stimulated with IL-1, and the expression of iNOS and COX-2 messenger RNA and proteins was evaluated by real-time reverse transcriptase-polymerase chain reaction analysis and Western blotting, respectively. H3K4 methylation and the recruitment of the histone methyltransferases SET-1A and MLL-1 to the iNOS and COX-2 promoters were evaluated using chromatin immunoprecipitation assays. The role of SET-1A was further evaluated using the methyltransferase inhibitor 5'-deoxy-5'-(methylthio)adenosine (MTA) and gene silencing experiments. SET-1A level in cartilage was determined using immunohistochemistry. RESULTS: The induction of iNOS and COX-2 expression by IL-1 was associated with H3K4 di- and trimethylation at the iNOS and COX-2 promoters. These changes were temporally correlated with the recruitment of the histone methyltransferase SET-1A, suggesting an implication of SET-1A in these modifications. Treatment with MTA inhibited IL-1-induced H3K4 methylation as well as IL-1-induced iNOS and COX-2 expression. Similarly, SET-1A gene silencing with small interfering RNA prevented IL-1-induced H3K4 methylation at the iNOS and COX-2 promoters as well as iNOS and COX-2 expression. Finally, we showed that the level of SET-1A expression was elevated in OA cartilage as compared with normal cartilage. CONCLUSION: These results indicate that H3K4 methylation by SET-1A contributes to IL-1-induced iNOS and COX-2 expression and suggest that this pathway could be a potential target for pharmacologic intervention in the treatment of OA and possibly other arthritic diseases.

Primary solitary fibrous tumor of the retroperitoneum.

Solitary fibrous tumor is an uncommon neoplasm affecting adults and typically located in the pleura and can also occur in a large number of other extra thoracic sites. We present the case of a solitary fibrous tumor (SFT) of the retroperitoneum and describe their histopathological and immunohistochemical features. The identification of SFT in the retroperitoneum is of importance because its clinico-pathological behaviour is still unclear. The pathologist plays a fundamental role in establishing both the positive and differential diagnosis.

Prevalence and causes of blindness, low vision and status of cataract in 50 years and older citizen of Qatar-a community based survey.

BACKGROUND: Rapid Assessment for the Avoidable Blindness (RAAB) was conducted in Qatar during 2009. We present the prevalence and determinants of visual disabilities and status of cataract among citizens aged 50 years and older. METHODS: Residents of randomly selected houses and clusters participated in the survey. Opticians noted the presenting and the best corrected vision of participants from 49 clusters. Ophthalmologists examined participants with additional instruments like bio-microscope, digital camera, auto-perimeter and auto-refractor in a mobile van. World Health Organization recommended principal cause of blindness (Visual acuity [VA] < 3/60 in better eye), Severe visual impairment (SVI) (<6/60), low vision (VA < 6/18) and unilateral blindness (VA < 3/60) were designated. Persons with VA < 6/18 and cataract were interviewed to calculate coverage and barriers for cataract surgeries. Age sex adjusted prevalence of visual disabilities and their 95% Confidence Intervals (CI) were estimated. RESULTS: We examined 2,433 (97.3%) participants. The age sex adjusted prevalence of bilateral blindness was 1.28% [95% CI 1.22-1.35], SVI (1.67%), low vision (3.66%) and unilateral blindness (3.61%) in 50 years and older population. Female and older age groups were significant risk factors of visual disabilities. Cataract and glaucoma were the main causes of visual disabilities. The coverage of cataract services was 68.2%. Believing that cataract as an aging process (25) and adequate vision in the fellow eye (15) were the reasons for delay in surgery. CONCLUSIONS: To reduce avoidable blindness, un-operated cataract should be addressed. Primary and secondary eye care systems should be strengthened to improve the care of blinding eye diseases in Qatar.

Acrokeratoelastoidosis.

Acrokeratoelastoidosis (AKE) is a rare skin disorder characterized by a papular eruption and fragmentation of elastic tissue that primarily involves the margins of the hands and feet. The most common histopathologic findings of AKE are hyperkeratosis and degeneration of elastic fibers. Various treatment options have been tried with little help or without success. We report the third case of acrokeratoelastoidosis in an Arabic woman localized exclusively to the feet.

[Predisposition to infection in patients with pemphigus. Retrospective study of 141 cases]. / Prédisposition aux infections des malades ayant un pemphigus Etude rétrospective de 141 cas.

INTRODUCTION: The relation between pemphigus and infection is complex. The aim of this work was to determine the frequency and impact of infection as well as the factors associated with it among our patients with pemphigus. METHODS: This retrospective case series examined records of patients with pemphigus admitted to the dermatology unit of Ibn Sina University Hospital of Rabat between 1989 and 2004. We compared the patients with and without infections as well as the patients with and without severe bacterial infections according to patient profile and outcome. The principal outcome measure was death and the secondary measure, duration of hospitalization. RESULTS: Of the 141 patients with pemphigus included in our study, 68% developed an infection. Infections were bacterial in 52% of cases, fungal in 50%, herpetic in 19% and parasitic in 1.5%. They were associated with diabetes mellitus and immunosuppressive drugs. Severe bacterial infection was frequent in patients with diabetes and rare in those treated with corticosteroids or with pemphigus foliaceous. Death occurred significantly more often among infected subjects (p=0.01), especially those with severe bacterial infections (p<0.001). Hospitalization was slightly longer among patients with infections (p=0.01) but shorter in cases of severe bacterial infections (p=0.004). DISCUSSION: Our study confirms the predisposition to all types of infection among patients with pemphigus, as well as the undeniable role its treatment plays in promoting infection. Severe bacterial infections were most often contracted before hospitalization. The literature reports a wide variety of infections, with pemphigus-herpes documented most often. Prevention and management require hygienic measures as well as early diagnosis and treatment of pemphigus and infections. Preventive treatment may be considered, but studies are needed to define the patients who might benefit from it.

Dexmedetomidine during local anesthesia.

The objective of our study was to assess the efficacy and safety of dexmedetomidine given in a small dose for a 1-h infusion as an adjuvant to local analgesia in ophthalmic operations. The study was double-blind prospective, randomized, and placebo controlled. We studied the effects of a small dose of dexmedetomidine (0.5 micro.kg(-1).h(-1) for 10 min followed by 0.2 micro.kg(-1).h(-1) for 50 min. Patients were divided randomly into two groups with 20 patients in each: group A was the study group and group B was the placebo group. Heart rate, systolic blood pressure, and diastolic blood pressure were significantly lower in the dexmedetomidine group than the placebo group. Bispectral index values were significantly lower in the dexmedetomidine group than the placebo group. Also, intraocular pressure significantly decreased in the dexmedetomidine group compared to the placebo group. The study revealed that dexmedetomidine in the studied dose has a sedative effect, provides safe control of heart rate and blood pressure, and also decreases intraocular pressure during ophthalmic surgery under local anesthesia.

Determinants of women's choice of their obstetrician and gynecologist provider in the UAE.

BACKGROUND: The objective of this study is to evaluate women's priorities and preferences in selecting their obstetrician and gynecologist in a non-Western society. METHODS: Consecutive sample of 508 patients attending the obstetric and gynecologic services of AL-Ain Hospital; AL-Ain; United Arab Emirates during 4 months was recruited. Participants were interviewed by using a structured 26-item questionnaire administered by means of face-to-face interview within 24 h of admission in the ward or before consultation in the outpatient clinic. Respondents were asked about their preferences, priorities, determinants of choice of their obstetrician and gynecologist, their view of his/her role as a medical provider, and their perception of importance of each of provider's characteristics and roles on a five-point Likert scale. RESULTS: Four hundred thirty-nine (86.4%) participants preferred female physicians. Sixty-one (12%) had no preference and eight (1.6%) preferred male physicians. Reasons for female selection were privacy during intimate examination (89.1%) or counseling (68.8%), religious beliefs (74.3%), and cultural traditions (45.3%). Female preference was significantly associated with higher parity (P = 0.002), religion (P = 0.005), nationality (P = 0.01), occupation (P = 0.02), education (P = 0.04), and poor recognition of physician's role as professional/expert (P < 0.00001). Male preference was significantly associated with experience at previous encounter (P = 0.03), obstetric consultation (P = 0.04), and perceiving physician's role as skilled communicator (P = 0.01) or health educator (P = 0.04). Other physician characteristics affecting choice were professionalism, bedside manners, empathy, communication, competence, availability, and religion. CONCLUSIONS: Physician's attitude, professional profile, sex, and religious faith determine women's choice of obstetricians and gynecologists. Most women prefer female providers because of embarrassment during pelvic examination and reproductive counseling, religious beliefs, and sociocultural values.