Pathologic Lymph Node Regression After Neoadjuvant Chemotherapy Predicts Recurrence and Survival in Esophageal Adenocarcinoma: A Multicenter Study in the United Kingdom.

PURPOSE: There is limited evidence regarding the prognostic effects of pathologic lymph node (LN) regression after neoadjuvant chemotherapy for esophageal adenocarcinoma, and a definition of LN response is lacking. This study aimed to evaluate how LN regression influences survival after surgery for esophageal adenocarcinoma. METHODS: Multicenter cohort study of patients with esophageal adenocarcinoma treated with neoadjuvant chemotherapy followed by surgical resection at five high-volume centers in the United Kingdom. LNs retrieved at esophagectomy were examined for chemotherapy response and given a LN regression score (LNRS)-LNRS 1, complete response; 2, <10% residual tumor; 3, 10%-50% residual tumor; 4, >50% residual tumor; and 5, no response. Survival analysis was performed using Cox regression adjusting for confounders including primary tumor regression. The discriminatory ability of different LN response classifications to predict survival was evaluated using Akaike information criterion and Harrell C-index. RESULTS: In total, 17,930 LNs from 763 patients were examined. LN response classified as complete LN response (LNRS 1 ≥1 LN, no residual tumor in any LN; n = 62, 8.1%), partial LN response (LNRS 1-3 ≥1 LN, residual tumor ≥1 LN; n = 155, 20.3%), poor/no LN response (LNRS 4-5; n = 303, 39.7%), or LN negative (no tumor/regression; n = 243, 31.8%) demonstrated superior discriminatory ability. Mortality was reduced in patients with complete LN response (hazard ratio [HR], 0.35; 95% CI, 0.22 to 0.56), partial LN response (HR, 0.72; 95% CI, 0.57 to 0.93) or negative LNs (HR, 0.32; 95% CI, 0.25 to 0.42) compared with those with poor/no LN response. Primary tumor regression and LN regression were discordant in 165 patients (21.9%). CONCLUSION: Pathologic LN regression after neoadjuvant chemotherapy was a strong prognostic factor and provides important information beyond pathologic TNM staging and primary tumor regression grading. LN regression should be included as standard in the pathologic reporting of esophagectomy specimens.

Where Are We Now with Liver Transplantation in Neuroendocrine Neoplasms? The Place of Liver Transplantation for Grades 1 and 2 Well-Differentiated Unresectable Liver Metastatic Neuroendocrine Tumours.

PURPOSE OF REVIEW: This review outlines the role of liver transplantation in selected patients with unresectable neuroendocrine tumour liver metastases. It discusses the international consensus on eligibility criteria and outlines the efforts taking place in the UK and Ireland to develop effective national liver transplant programmes for neuroendocrine tumour patients. RECENT FINDINGS: In the early history of liver transplantation, indications included cancer metastases to the liver as well as primaries of liver origin. Often, liver transplantation was a salvage procedure. The early results were disappointing, including in patients with neuroendocrine tumours. These data discouraged the widespread adoption of liver transplantation for neuroendocrine tumour liver metastases (NET LM). A few centres persisted in performing liver transplantation for patients with NET LM and in determining parameters predictive of good outcomes. Their work has provided evidence for benefit of liver transplantation in a selected group of patients with NET LM. Liver transplantation for NET LM is now accepted as a valid indication by many professional bodies, including the European Neuroendocrine Tumour Society (ENETS) and the United Network for Organ Sharing (UNOS). It is nevertheless rarely utilised. The UK and the Republic of Ireland are commencing a pilot programme of liver transplantation in selected patients. This programme will help develop the expertise and infrastructure to make liver transplantation for NET LM a routine procedure.

The single cell transcriptional landscape of esophageal adenocarcinoma and its modulation by neoadjuvant chemotherapy.

Immune checkpoint blockade has recently proven effective in subsets of patients with esophageal adenocarcinoma (EAC) but little is known regarding the EAC immune microenvironment. We determined the single cell transcriptional profile of EAC in 8 patients who were treatment-naive (n = 4) or had received neoadjuvant chemotherapy (n = 4). Analysis of 52,387 cells revealed 10 major cell subsets of tumor, immune and stromal cells. Prior to chemotherapy tumors were heavy infiltrated by T regulatory cells and exhausted effector T cells whilst plasmacytoid dendritic cells were markedly expanded. Two dominant cancer-associated fibroblast populations were also observed whilst endothelial populations were suppressed. Pathological remission following chemotherapy associated with broad reversal of immune abnormalities together with fibroblast transition and an increase in endothelial cells whilst a chemoresistant epithelial stem cell population correlated with poor response. These findings reveal features that underlie and limit the response to current immunotherapy and identify a range of novel opportunities for targeted therapy.

Impact of neoadjuvant chemotherapy on nodal regression and survival in oesophageal adenocarcinoma.

BACKGROUND: The prognostic value of lymph node regression (LNR) following neoadjuvant chemotherapy (nCT) for oesophageal and gastro-oeosphageal adenocarcinoma remains unclear. This study aimed to characterise the long-term survival outcomes of LNR in patients having resectional surgery after nCT. METHODS: This study included patients undergoing oesophagectomy or extended total gastrectomy for oesophageal and junctional tumours (Siewert types 1,2,3) at the Queen Elizabeth Hospital Birmingham from 2012 to 2018. Lymph nodes retrieved at surgery were examined for evidence of a response to chemotherapy. Patients were classified as lymph node-negative (either negative nodes with no evidence of previous tumour involvement or negative with evidence of complete regression) or positive with either partial or no response. RESULTS: This study identified 183 patients who received nCT, of which 71% (130/183) had positive lymph nodes. Of these 130 patients, 44% (57/130) had a lymph node response and 56% (73/130) did not. The remaining 53 patients (29.0%) had negative lymph nodes with no evidence of tumour. Lymph node responders had a significant survival benefit compared to patients without lymph node response, but shorter than those with negative lymph nodes (median: 27 vs 18 vs NR months, p < 0·001). On multivariable analysis, lymph node responders had an improved overall (Hazard ratio (HR): 0.86, 95% CI: 0.80-0.92, p < 0.001) and recurrence-free (HR: 0.90, 95% CI: 0.82-0.98, p = 0.030) survival. CONCLUSION: Lymph node regression is an important prognostic factor, warranting closer evaluation over primary tumour response to help with planning further adjuvant therapy in these patients.

Survival and Nutritional Status of Male and Female Heart Transplant Patients Based on the Nutritional Risk Index.

Malnutrition among heart-transplant patients may affect survival. The aim was to investigate the survival and nutrition status among male and female heart transplant patients who underwent transplantation, before and 1 year after surgery based on the nutritional risk index (NRI). The medical records of ninety heart-transplant patients (2009-2014) from the King Faisal Specialist Hospital, Riyadh, were reviewed. The assessment included demographic data, anthropometric measurements, and NRI calculation. Moreover, postoperative data included the length of stay and survival. Paired t-test and survival analysis by Kaplan-Meier (KM) curves were used. A total of 90 patients (males 77.78%) were included. The prevalence of malnutrition in the preoperative phase by NRI was 60% (7.78% as severe; 40% as moderate, and 12.22% mild NRI scores). After 1 year, body mass index (BMI) and NRI increased significantly (p < 0.001). Furthermore, NRI was significantly different between men and women (p < 0.01), while KM survival curves were insignificantly different (p = 0.67). Recipients with postoperative moderate or severe nutritional risk (NRI < 97.5) had significantly shorter survival in the first-year post-transplantation (HR = 0.82; 95% CI, 0.75-0.89; p < 0.001). Our findings indicate that the NRI after 1 year of transplant correlated significantly with mortality. Besides, there was no significant gender difference regarding survival; however, malnutrition and low survival were more prominent among women.

High glucose load and endotoxemia among overweight and obese Arab women with and without diabetes: An observational study.

Dietary intake influences gut microbiota activity. Nevertheless, there is a lack of evidence available that illustrates the acute effects of high glucose meal on metabolic endotoxemia. The present study assessed the acute impact of high glucose meal on endotoxemia and other clinical parameters in Saudi females with varying degrees of glycemia.The subjects were 64 consenting pre-menopausal women, grouped into 3: control [n = 14 lean, non-T2DM, BMI = 22.2 ±â€Š2.2 kg/m]; overweight [n = 16, non-T2DM, BMI = 28.5 ±â€Š1.5 kg/m] and T2DM [n = 34, BMI = 35.2 ±â€Š7.7 kg/m]. After an overnight fast, all subjects were given a standardized high-glucose (75 g) meal. Anthropometrics were taken and blood samples were withdrawn at baseline and postprandial (0, 2 and 4-hours), serum glucose, endotoxin and lipid profile were quantified.At baseline, total cholesterol, LDL-cholesterol, triglycerides and serum glucose levels were significantly higher (P values <.01) whereas significantly lower HDL-cholesterol levels (P < .01) were observed in T2DM subjects compared to other groups. Baseline endotoxin levels were highest in the overweight group (3.2 ±â€Š1.1 mmol/L) as compared to control (2.0 ±â€Š0.5 mmol/L) and T2DM (2.7 ±â€Š1.2 mmol/L) (P = .046). HDL-cholesterol, LDL-cholesterol and triglycerides, significantly decreased in the T2DM group after 2 hours (P values <.05), whereas unremarkable changes observed in other groups. Lastly, endotoxin levels significantly increased only in the overweight group (3.2 ±â€Š1.1 vs 4.2 ±â€Š1.4 mmol/L; P < .05), 4 hours postprandial.High glucose meal elevates endotoxemia only among overweight subjects and impairs dysbiosis.

Unusual Presentation of Mammary Calciphylaxis in a Patient on Long-Standing Renal Dialysis.

INTRODUCTION: Calciphylaxis is a condition which involves ectopic deposition of calcium in arterioles of various organ systems. Here, we present a rare case of mammographic calcifications in a patient on long-standing renal dialysis. CASE: This is a 57-year-old female who presented to breast clinic with unilateral severe pain and lumpiness of her breast. On mammography, calcifications were identified which were associated with focal lesions on ultrasound. An image-guided core biopsy showed numerous foci of histological calcification within a large area of fat necrosis. The location within arterioles was confirmed by CD31 immunohistochemistry. The diagnosis of fat necrosis with benign calcifications due to mammary calciphylaxis was made. CONCLUSION: The hallmark of calciphylaxis is the deposition of calcium within small- to medium-sized vessel walls. This leads to ischaemia and necrosis of tissue. In the breast, only a few cases were reported in which patients presented with a necrotic lesion resembling carcinoma. These are reviewed in this report.

Effect of Housing System and Rosemary and Cinnamon Essential Oils on Layers Performance, Egg Quality, Haematological Traits, Blood Chemistry, Immunity, and Antioxidant.

Housing system and nutrition are non-genetic factors that can improve the well-being of animals to obtain higher quality products. A better understanding of how different housing systems and essential oils can influence the performance of layers is very important at the research and commercial levels. The current study aimed to investigate the effects of a housing system and dietary supplementation of rosemary and cinnamon essential oils on layers' performance and egg quality. A factorial arrangement (2 × 3) was performed include two housing systems (floor and cage) and three different types of essential oils (0, 300 mg/kg diet of rosemary and 300 mg/kg diet of cinnamon essential oils) to study their effects on the productive performance, egg quality, immunity, oxidative stress and haematology of ISA brown laying hens during the production stages (from 28 to 76 weeks of age). Birds were randomly divided into two groups each comprising of 1500 birds; the first group was moved from the litter to reared laying cages while the second group was floor reared. Each group was randomly divided into three groups, the first was considered as a control group, the second treated with rosemary essential oil, and the third with cinnamon essential oil. The differences in egg production and weight, egg quality, feed intake and conversion, blood picture and chemistry, immunity, and antioxidant parameters between the different housing systems (floor and cage) were not significant at (p < 0.05 or 0.01). On the other hand, the egg production and weight, Haugh unit, feed intake and conversion, blood cholesterol, Alanine Aminotransferase (ALT), Aspartate Aminotransferase (AST), urea, Ca, P, immunity, and antioxidant parameters were significantly (p < 0.05 or 0.01) better in rosemary and cinnamon groups than in the control group. Furthermore, the results of dietary supplementation with rosemary and cinnamon were very close. Regarding egg production and weight, there were no significant differences due to the interactions. The differences in egg mass among the interactions were also not significant except at 68-76 weeks, where the cage × cinnamon group was the highest. Under the floor rearing system, birds that were fed a diet supplemented with or without essential oils (EOs) consumed more feed than those raised under the cage system. Regarding feed conversion rate (FCR), the differences among the interactions were not significant except at 44-52, 52-60 and 68-76 weeks, where the cage × cinnamon group was the lowest. Excluding glutathione peroxidase (GPx) activity (p < 0.001), all immunity and antioxidant indices were not statistically different as a consequence of the interaction among EOs and housing systems. Additionally, the highest levels of phosphorus were observed for layers fed diets enriched with cinnamon oil with the cage or floor system. In conclusion, the data suggested that supplementation of rosemary and cinnamon essential oils in laying hen diet showed significantly positive effects on hen performance and egg production. Cholesterol, liver and kidney functions, immunity, and antioxidant parameters improved with rosemary and cinnamon supplementation when compared to the control. Additionally, the different housing systems did not result in any positive or negative impact on these traits.

Impact of neuroendocrine morphology on cancer outcomes and stage at diagnosis: a UK nationwide cohort study 2013-2015.

BACKGROUND: The diagnosis of neuroendocrine neoplasms (NENs) is often delayed. This first UK population-based epidemiological study of NENs compares outcomes with non-NENs to identify any inequalities. METHODS: Age-standardised incidence rate (ASR), 1-year overall survival, hazard ratios and standardised mortality rates (SMRs) were calculated for all malignant NENs diagnosed 2013-2015 from UK national Public Health records. Comparison with non-NENs assessed 1-year overall survival (1YS) and association between diagnosis at stage IV and morphology. RESULTS: A total of 15,222 NENs were identified, with an ASR (2013-2015 combined) of 8.6 per 100,000 (95% CI 8.5-8.7); 4.6 per 100 000 (95% CI, 4.5-4.7) for gastro-entero-pancreatic (GEP) NENs. The 1YS was 75% (95% CI, 73.9-75.4) varying significantly by sex. Site and morphology were prognostic. NENs (predominantly small cell carcinomas) in the oesophagus, bladder, prostate, and female reproductive organs had a poorer outcome and were three times more likely to be diagnosed at stage IV than non-NENs. CONCLUSION: Advanced stage at diagnosis with significantly poorer outcomes of some NENs compared with non-NENs at the same anatomical site, highlight the need for improved access to specialist services and targeted service improvement.

Retrospective study on mixed neuroendocrine non-neuroendocrine neoplasms from five European centres.

BACKGROUND: Mixed neuroendocrine non-neuroendocrine neoplasm (MiNEN) is a rare diagnosis, mainly encountered in the gastro-entero-pancreatic tract. There is limited knowledge of its epidemiology, prognosis and biology, and the best management for affected patients is still to be defined. AIM: To investigate clinical-pathological characteristics, treatment modalities and survival outcomes of a retrospective cohort of patients with a diagnosis of MiNEN. METHODS: Consecutive patients with a histologically proven diagnosis of MiNEN were identified at 5 European centres. Patient data were retrospectively collected from medical records. Pathological samples were reviewed to ascertain compliance with the 2017 World Health Organisation definition of MiNEN. Tumour responses to systemic treatment were assessed according to the Response Evaluation Criteria in Solid Tumours 1.1. Kaplan-Meier analysis was applied to estimate survival outcomes. Associations between clinical-pathological characteristics and survival outcomes were explored using Log-rank test for equality of survivors functions (univariate) and Cox-regression analysis (multivariable). RESULTS: Sixty-nine consecutive patients identified; Median age at diagnosis: 64 years. Males: 63.8%. Localised disease (curable): 53.6%. Commonest sites of origin: colon-rectum (43.5%) and oesophagus/oesophagogastric junction (15.9%). The neuroendocrine component was; predominant in 58.6%, poorly differentiated in 86.3%, and large cell in 81.25%, of cases analysed. Most distant metastases analysed (73.4%) were occupied only by a poorly differentiated neuroendocrine component. Ninety-four percent of patients with localised disease underwent curative surgery; 53% also received perioperative treatment, most often in line with protocols for adenocarcinomas from the same sites of origin. Chemotherapy was offered to most patients (68.1%) with advanced disease, and followed protocols for pure neuroendocrine carcinomas or adenocarcinomas in equal proportion. In localised cases, median recurrence free survival (RFS); 14.0 mo (95%CI: 9.2-24.4), and median overall survival (OS): 28.6 mo (95%CI: 18.3-41.1). On univariate analysis, receipt of perioperative treatment (vs surgery alone) did not improve RFS (P = 0.375), or OS (P = 0.240). In advanced cases, median progression free survival (PFS); 5.6 mo (95%CI: 4.4-7.4), and median OS; 9.0 mo (95%CI: 5.2-13.4). On univariate analysis, receipt of palliative active treatment (vs best supportive care) prolonged PFS and OS (both, P < 0.001). CONCLUSION: MiNEN is most commonly driven by a poorly differentiated neuroendocrine component, and has poor prognosis. Advances in its biological understanding are needed to identify effective treatments and improve patient outcomes.

Prognostic significance of circumferential resection margin involvement in patients receiving potentially curative treatment for oesophageal cancer.

INTRODUCTION: The utility of Circumferential Resection Margin (CRM) status in predicting prognosis in oesophageal cancer is controversial, with different definitions used by the College of American Pathologists and the Royal College of Pathologists. We aimed to determine prognostic significance of CRM involvement and evaluate which system is the best predictor of prognosis. METHODS: A cohort of 390 patients who had potentially curative oesophagectomy (- + neoadjuvant chemotherapy) were analysed. Associations between CRM involvement and patient outcome were assessed for the whole cohort, and for pre-specified subgroups of T3 tumours and those who received neo-adjuvant chemotherapy. RESULTS: CRM-involvement was associated with higher T and N stage, tumour differentiation, increased tumour length and both lymphovascular and perineural invasion. Overall Survival (OS) and Recurrence Free Survival (RFS) significantly worsened with CRM-involvement (p = 0.001, p < 0.001). R1a (<1 mm but no macroscopic involvement) resulted in significantly improved OS (p = 0.037) and RFS (P = 0.026) compared to R1b (macroscopic involvement), but did not differ significantly from R0 (≥1 mm). The association between CRM-involvement and both OS and RFS remained significant regardless of whether neoadjuvant chemotherapy was given. However, CRM-involvement was not a significant prognostic marker in T3 patients (p = 0.148). Multivariable analysis found N stage, lymphovascular invasion, patient age and neoadjuvant chemotherapy to be significantly predictive of patient outcome. CRM-involvement was not a significant independent prognostic marker. CONCLUSIONS: CRM-involvement was not found to be independently predictive of prognosis, after accounting for other prognostic markers. As such, CRM should not be considered a major prognostic factor in patients with oesophageal cancer.

Impact of the socioeconomic status on the severity and outcome of community-acquired pneumonia among Egyptian children: a cohort study.

BACKGROUND: Community-acquired pneumonia (CAP) is one of the five leading causes of death among children in developing countries, accounting for approximately three million deaths per year. Identification of the modifiable risk factors of CAP may help to reduce the burden of this disease. In this study, the impact of the socioeconomic status (SES) on the severity and outcome of CAP among Egyptian children was studied. METHODS: This was a prospective longitudinal cohort study which included 1,470 children diagnosed with CAP, aged two to 15 years (median age 5.4 years). The diagnosis of CAP was based on clinical and radiological findings. A structured questionnaire and the patients' medical records were used for the data collection. The subjects were divided into two groups: mild and severe CAP. Social and demographic variables were compared, and a multivariate logistic regression analysis was performed. RESULTS: THE MULTIVARIATE ANALYSIS SHOWED THAT A LOW MATERNAL EDUCATION LEVEL (OR: 3.8; 95% CI: 2.12 -6.70; P = .0001), unavailability of adequate medical care (OR: 3.1; 95% CI: 1.99 -4.88; P = .0001), a low family income (OR: 2.2; 95% CI: 0.99 -4.78; P = .047), and parents' smoking habits (OR: 2.0; 95% CI: 1.15 -3.55; P = .014) were significant independent predictive risk factors for severe CAP among Egyptian children. CONCLUSION: Public health measures against these socio-demographic risk factors should be identified as priorities in order to help reduce the disease burden of deaths from severe CAP among Egyptian children.

Serum trace elements in obese Egyptian children: a case-control study.

BACKGROUND: To date, only a few studies on child obesity concerned Trace Elements (TE). TE is involved in the pathogenesis of obesity and obesity related diseases. We tried to assess trace elements status [zinc (Zn), copper (Cu), selenium (Se), iron (Fe), and chromium (Cr)] in obese Egyptian children and their relationships with serum leptin and metabolic risk factors of obesity. METHODS: This was a case-control study performed with 80 obese children (BMI ≥ 95thcentile for age and gender) and 80 healthy non-obese children with comparable age and gender as the control group. For all subjects, serum Zn, Cu, Se, Fe, ferritin and Cr as well as biochemical parameters including lipid profile, serum glucose and homeostasis model assessment of insulin resistance (HOMA-IR) were assessed. Levels of serum leptin were measured by (enzyme-linked immunosorbent assay [ELISA] method), and serum insulin was measured by an electrochemiluminesce immunoassay. RESULTS: Compared to the control group, serum Zn, Se, and Fe levels were significantly lower (all P < 0.01) and serum Cu level was significantly higher (P < 0.01) in the obese children. Meanwhile, no significant differences were observed in serum ferritin or Cr levels (P > 0.05). A significant negative correlation was found between serum leptin and zinc levels in the obese children (r = -0.746; P < 0.01). Further, serum Zn showed significant negative correlations with total cholesterol TC levels (P < 0.05) and were positively correlated with high density lipoprotein- cholesterol HDL-C levels (P < 0.01) in the obese children. In addition, serum Se levels showed significant positive correlations with HOMA-IR values in the obese children (P < 0.01). CONCLUSION: The obese children may be at a greater risk of developing imbalance (mainly deficiency) of trace elements which may be playing an important role in the pathogenesis of obesity and related metabolic risk factors.

Müllerian adenosarcoma arising in a subserosal adenomyoma.

Müllerian adenosarcomas most commonly arise in the uterine corpus from the eutopic endometrium. Occasionally, they arise in the cervix, vagina, broad and round ligaments, and ovaries and rarely in extragenital sites, usually in association with endometriosis. The authors report a rare case of extraendometrial, intramural adenosarcoma of low grade arising from a subserosal adenomyoma in a 46-year-old woman who presented with vaginal bleeding. No evidence of eutopic endometrial origin was identified; the adenosarcoma showed only limited myometrial invasion and no serosal involvement. Because of its subserosal location, the tumor would be overstaged as IC (deeply myoinvasive) in the new FIGO staging system. Stage IC tumors would normally be considered for adjuvant treatment, which would be inappropriate in this case. To avoid overstaging and inappropriate treatment, it is important to recognize that adenosarcoma can, rarely, arise in adenomyomas (or adenomyosis).

Identification of lesions indicating rejection in kidney transplant biopsies: tubulitis is severely under-detected by conventional microscopy.

BACKGROUND: In the current international Banff classification of kidney transplant rejection, tubulitis and intimal arteritis are regarded as the key histological features of acute rejection. Grade 1 tubulitis can sometimes be seen in biopsies that do not represent acute rejection; but in the case of intimal arteritis, just one lymphocyte can justify anti-rejection treatment. Our aim was to audit reliability and accuracy of recognizing tubulitis and intimal arteritis using the approach recommended by the Banff classification and correlate any discrepancies with subsequent graft function. METHODS: This is a retrospective review of all kidney transplant biopsies reported as negative for rejection from 1 January 2009 to 31 December 2009 to assess the presence or absence of occult tubulitis and arteritis. Lymphocytes were immunostained with CD3, using Periodic Acid Schiff as a counterstain. Sections were reviewed to detect missed intimal arteritis and tubulitis. Discrepancies between the report and the immunostain results were analysed by biopsy type and broken down by the reporting pathologist. The graft function of any patient with missed lesions was checked to test for adverse impact on the patient. RESULTS: 'Missed' tubulitis was found in 68% of biopsies, but only two such cases subsequently developed biopsy-proven acute rejection. Only one case of missed intimal arteritis was found (1%) and the subsequent clinical course suggested that this was probably early rejection. There was no significant difference between the reporting pathologists. CONCLUSIONS: We conclude that tubulitis is missed very frequently, but the Banff classification seems to be 'calibrated' to allow for this and it does not seriously affect the identification of clinically significant acute rejection. Immunostaining is therefore not indicated in routine practice because (by Banff criteria) it would result in over-diagnosis of rejection. Intimal arteritis can indicate acute rejection even if extremely mild.

The effect of leptin on maturation of the ovarian compartments of albino rats: an ultrastructural and histological study.

BACKGROUND: Leptin serves as a signal to the central nervous system with information on the critical amount of adipose tissue stores that is necessary for activation of the hypothalamic-pituitary-ovarian axis. OBJECTIVES: To document the histological and ultrastructural changes that occur in the ovarian follicles of immature albino rats treated with leptin when compared with controls. Furthermore, the endometrial histological and immunohistochemical, and vaginal cytological changes suggestive of ovulation were assessed. ANIMALS AND METHODS: The study was carried out on 50 immature female albino rats aged 22 days; 24 of them were injected with 5 microg leptin daily and 26 rats were taken as controls. Vaginal smears were taken daily, three animals were sacrificed every 2-4 days from each group, ovaries and uteri were dissected and specimens were prepared for electron microscopic, histological and/or immunohistochemical assessment. The research project was approved by The Histology Department Committee of Alexandria Medical School, which is licensed for animal care and use. RESULTS: Electron microscopic and histological examination confirmed the occurrence of maturational changes in various ovarian components from 26 days of age in leptin-treated rats, with ovulation occurring from the age of 30 days. The granulosa, theca and stroma cells showed signs of steroidogenesis, with increased mitosis within granulosa cells. The ooplasm showed an increased number of organelles, and annulate lamellae were demonstrated. The zona pellucida revealed microvilli, adhering junctions and gap junctions. Similarly, the endometrial histological and vaginal cytological maturational changes were detected in leptin-treated rats from 26 days of age. Furthermore, there was high expression of estrogen receptor-alpha in almost all columnar and stroma cells of the endometrium. However, the control rats ovulated around the normal age of maturation, i.e. 42 days. CONCLUSION: We documented ultrastructural, histological, immunohistochemical and cytological evidence that leptin accelerates the onset of puberty in female albino rats. The potential role of exogenous leptin, in cases of impaired reproductive function in humans, needs to be elucidated.

Changes in transcription profile and cytoskeleton morphology in pelvic ligament fibroblasts in response to stretch: the effects of estradiol and levormeloxifene.

Failure of ligamentous support of the genital tract to resist intra-abdominal pressure is a plausible underlying mechanism for the development of pelvic organ prolapse, but the nature of the molecular response of pelvic tissue support remains unknown. We hypothesized that the expression of genes coding for proteins involved in maintaining the cellular and extracellular integrity would be altered as a result of mechanical stretch. Therefore, cDNA microarrays were used to examine the difference in transcriptional profile in RNA of primary culture fibroblasts subjected to mechanical stretch and those that remained static. Out of 34 mechano-responsive genes identified (P < 0.05), four were coding for regulation of actin cytoskeleton remodelling, and its interaction with the extracellular matrix proteins; these are phosphatidyl inositol-4-phosphate 5-kinase (PIP5K1C), the human signal-induced proliferation associated gene-1 (SIPA-1), TNFRSF1A-associated via death domain (TRADD) and deoxyribonuclease 1-like 1 (DNase 1-L1). The transcriptosomal changes led us to investigate the phenotypic consequences of stretch, levormeloxifene and estradiol (E(2)) on the cytoskeleton of cultured fibroblasts. The percentage of cells with abnormal F-actin configuration was significantly higher in fibroblasts subjected to stretch compared with the static model (P < 0.0001). Levormeloxifene caused similar significant alterations in actin morphology of the static fibroblasts. The use of E(2) did not reverse the process or protect the cells from the effect of stretch, but significantly increased the rate of fibroblast proliferation, suggestive of a role in healing process. Mechanical stretch and/or levormeloxifene disturb the fibroblasts ability to maintain the cytoskeleton architecture and we speculate that they may disrupt ligamentous integrity and result in clinical prolapse.