Endometrial evaluation with methylene blue staining in patients with normal hysteroscopy

Background: Normally, endometrium comprises of non-absorptive epithelium and does not take up stain. Conventional staining with methylene blue is explained on the basis of existence of apoptotic cells in endometrium.Methods: Of 50 patients of unexplained infertility, AUB, recurrent pregnancy loss were randomly selected and included in the study. Those with abnormal ultrasound and history of tuberculosis were excluded. Conventional hysteroscopy was performed using normal saline as distending medium and in those with grossly normal endometrium were subjected to staining with 5% methylene blue instilled trans cervically. After 5 min, irrespective of the size and pattern, focal dark blue stained areas were considered abnormal and randomly biopsied. Incidence of endometritis in both groups was compared after histopathological examination.Results: Of total 50 patients, histopathological report of only one patient with dark blue staining had evidence of endometritis. Rest had no evidence, of which 73.5% had light blue or unstained areas and 26.5% showed dark blue staining. No statistically significant difference was found between histopathological reports and light or dark blue staining (p=0.28). When percentage stained area was considered more than 50% only to be positive, sensitivity was 100%, specificity 94%, PPV 25% and NPV 100%. False positives were 75% and no false negatives were observed. Although p values improved but still statistically insignificant.Conclusions: Present study failed to establish any significant correlation between staining pattern and detection rate of endometritis. With no Indian studies published on chromohysteroscopy so far, role of methylene blue in detection of subtle endometrial changes in modern gynaecology in Indian subpopulation is yet to be established.

Granular cell tumour of clitoris: a case report

Granular cell tumour (GCT) is tumours showing neuroectodermal differentiation. It occurs mostly in the dermis and subcutaneous tissue and occurrence in the muscle layer is rare. The most common site is the head and neck region. It has been known to occur in the vulva, with clitoris being a very rare site. Granular cell tumours are slow growing tumours, which are mostly asymptomatic. We report a case of lump in the clitoris in a 42-year-old woman which was managed with surgical excision. Histopathology confirmed the diagnosis of granular cell tumour. Very few cases of granular cell tumour of clitoris have been reported in literature till date. Although mostly benign, the differential diagnosis of granular cell tumour should be borne in mind by the clinicians in cases of clitoral lump.

Exogenous application of putrescine at pre-anthesis enhances the thermotolerance of wheat (Triticum aestivum L.).

Antioxidant enzymes, besides being involved in various developmental processes, are known to be important for environmental stress tolerance in plants. In this study, the effect of treatment of 2.5 mM putrescine (Put), heat stress (HS -42°C for 2 h) and their combination on the expression and activity of antioxidant enzymes was studied at pre-anthesis in the leaves of two wheat (Triticum aestivum L.) cultivars — HDR77 (thermotolerant) and HD2329 (thermosusceptible). We observed that 2.5 mM Put before HS significantly enhanced the transcript levels of superoxide dismutase (SOD), catalase (CAT), cytoplasmic and peroxisomal ascorbate peroxidase (cAPX, pAPX) in both the cultivars. However, the activities of antioxidant enzymes (SOD, CAT, APX and GR), as well as accumulation of antioxidants (ascorbic acid and total thiol content) were higher in HDR77 than in HD2329 in response to the treatment 2.5 mM Put + HS. No significant change was observed in the proline accumulation in response to HS and combined treatment of 2.5 mM Put + HS. A decrease in the H2O2 accumulation, lipid peroxidation and increase in cell membrane stability (CMS) were observed in response to 2.5 mM Put + HS treatment, as compared to HS treatment alone in both the cultivars; HDR77 was, however, more responsive to 2.5 mM Put + HS treatment. Put (2.5 mM) treatment at pre-anthesis thus modulated the defense mechanism responsible for the thermotolerance capacity of wheat under the heat stress. Elicitors like Put, therefore, need to be further studied for temporarily manipulating the thermotolerance capacity of wheat grown under the field conditions in view of the impending global climate change.

Characterization of differentially expressed stress-associated proteins in starch granule development under heat stress in wheat (Triticum aestivum L.).

Abiotic stress causes abrupt increase in the expression of stress-associated proteins, which provide tolerance by modulating the defense mechanism of plants. Small heat shock proteins (sHSPs) and anti-oxidant enzymes are important for environmental stress tolerance of the plants. In this study, two full-length cDNAs encoding small heat shock protein (sHSP) and superoxide dismutase (SOD), designated as TasHSP and SODI were identified and characterized from C-306 (thermotolerant) and PBW343 (thermosusceptible) cultivars of wheat (Triticum aestivum L.). An alpha crystalline domain was observed in TasHSP and manganese/iron binding domain in case of SODI. Quantitative real-time PCR showed very high transcript level of TasHSP and SOD in C-306 compared to PBW343 at different stages of growth and against differential heat stress (HS). Under differential HS at milky-dough stage, the fold change in transcript of both TasHSP and SOD was observed maximum in C-306, compared to PBW343. Protein profiling and isoenzymes analysis showed the expression of several heat-stable proteins and prominent isoenzymes of SOD in C-306, compared to PBW343. Scanning electron microscopy (SEM) of starch granules showed globular, well-shaped and more numbers of endospermic cells in C-306, compared to defragmented, irregular shaped and shrunken granules in case of PBW343 under HS treatment (42°C for 2 h). Diurnal change in soluble starch synthase (SSS) activity showed an increase in the activity during afternoon (35°C), compared to morning (29°C) and evening (32°C) in both the cultivars. Under heat stress (42°C for 2 h), a drastic decrease in the SSS activity was observed, due to the thermal denaturation of the enzyme. Thermotolerance capacity analyzed using cell membrane stability (CMS) showed significantly higher CMS in case of C-306, compared to PBW343 at different stages of growth. Findings suggest that abundance of TasHSP and SODI during milky-dough stage plays a very important role in starch granule biosynthesis. The mechanism may be further exploited to develop tolerant wheat cultivar with high quality seeds.