Role of Neurokinin B and Dynorphin A in pituitary gonadotroph and somatolactotroph cell lines

The role of Neurokinin B (NKB) and Dynorphin A (Dyn) in the regulation of the hypothalamic pituitary axis is an important area of recent investigation. These peptides are critical for the rhythmic release of GnRH, which subsequently stimulates the secretion of the gonadotropins, luteinizing hormone (LH) and follicle-stimulating hormone (FSH). The present study utilized the gonadotroph cell line LβT2 and the somatolactotroph GH3 cell line to examine the possible role of these peptides in pituitary hormone secretion. The NKB receptor (NK3R) and the Dyn receptor (the κ-opiate receptor (KOR)) were both detected in LβT2 cells and GH3 cells. NKB, by itself, failed to increase gonadotropin LHβ and FSHβ promoter activities and did not modulate the effects of GnRH on gonadotropin promoter activity. In GH3 cells, NKB significantly increased TRH-induced PRL promoter activity although NKB alone did not have an effect on basal PRL promoter activity. Dyn had no effect on gonadotropin promoters alone or in combination with GnRH stimulation. PRL promoters stimulated by TRH were not significantly changed by Dyn. TRH-induced PRL promoter activity was further increased in the presence of higher concentrations of NKB, whereas Dyn did not have a significant effect on the PRL promoter even at a high concentration. In addition, TRH-induced ERK (Extracelluar signal-regulated kinase) activation was enhanced in the presence of NKB. Our current study demonstrated that NKB had a stimulatory effect on PRL expression in a PRL-producing cell, but had no effect on gonadotropin secretion from a gonadotroph cell line.     

Effects of metformin administration on plasma gonadotropin levels in women with infertility,with an in vitro study of the direct effects on the pituitary gonadotrophs

Changes in LH and FSH levels were evaluated before and after metformin administration. In all 25 patients, plasma LH levels were significantly reduced after 3 months of metformin administration (500–1,500 mg/day). When patients were classified into a PCOS group (n = 12) or a non-PCOS group (n = 13), the reduction in LH levels only remained significant in the PCOS group. Plasma FSH levels were unchanged following metformin treatment when all patients were considered collectively and when patients were classified based on PCOS. LH/FSH ratio was significantly reduced only in the PCOS group. To examine the direct effect of metformin on gonadotropin-secreting cells, gonadotroph cell line, LβT2 was used for in vitro studies. Treatment of LβT2 cells with metformin modified neither the LHβ nor the FSHβ subunit promoter activity. The GnRH-induced LHβ promoter activity was not modulated in the presence of metformin. In contrast, GnRH-induced FSHβ promoter activity was significantly potentiated in the presence of metformin. Our results suggest that metformin does indeed modulate the basal level of LH and the LH/FSH ratio, albeit indirectly, particularly in the patients with PCOS. Additionally our results suggest that metformin does directly regulate FSH gene expression.     

Expression of the bric-a-brac tramtrack broad complex protein NAC-1 in cervical carcinomas seems to correlate with poorer prognosis.

PURPOSE: Recent studies have suggested a novel oncogenic role of a bric-a-brac tramtrack broad complex (also known as POZ) domain gene, NAC-1, in ovarian carcinomas. The aim of this study was to clarify the functional role of NAC-1 in human cervical carcinomas. EXPERIMENTAL DESIGN: NAC-1 expression in cervical cancer was assessed by immunohistochemistry, and data on clinical variables were collected by retrospective chart review. NAC-1 gene knockdown using small interfering RNA and a NAC-1 gene transfection system were used to asses NAC-1 function in cervical cancer in vivo. RESULTS: Immunohistochemical and gene expression analysis revealed that NAC-1 is significantly overexpressed in cervical adenocarcinomas and adenosquamous carcinomas compared with squamous cell carcinomas. Patients with squamous cell carcinomas positive for NAC-1 expression who received radiotherapy had significantly shorter overall survival than peers whose tumors did not express NAC-1, and multivariate analysis showed that NAC-1 expression was an independent prognostic factor for overall survival after radiotherapy. Overexpressions of the NAC-1 gene stimulated cell proliferation in cervical carcinoma cells of the TCS, CaSki, and HeLa P3 lines, which do not have endogenous NAC-1 expression. NAC-1 gene knockdown inhibited cell growth and induced apoptosis in HeLa, HeLa TG, and ME180 cells, all of which overexpressed NAC-1. CONCLUSIONS: Our findings suggest that NAC-1 may play an important role in cervical carcinomas; moreover, these findings provide a rationale for future development of NAC-1-based therapy for cervical carcinomas that overexpress this candidate oncogene.     

Quantitative evaluation of elderly skin based on digital image analysis

Background: The evaluation of the skin state when it is healthy at the time of examination, but predisposed to disease, is based solely on the subjective assessment of clinicians. This assessment may vary from moment to moment and from rater to rater. Purpose: We focused on skin texture and aimed to develop a new method to evaluate skin integrity of the elderly using a digital skin image, by verifying the reliability of the clinical evaluation of elderly skin image by specialists to create a ‘gold standard,’ and by analyzing a digital skin image to identify the indices that explain the skin condition as evaluated by specialists. Methods: A total of 208 skin images were collected from 34 subjects. And then we processed and analyzed images. Modified Kobayashi's method and texture analysis method were used in image analysis. Kobayashi's method included the process of density transformation, cross binarization, short straight line matching and extraction of main sulcus, and calculation of each index. Texture analysis was performed on cross‐binarized images for quantification of uniformity. On the other hand, skin textures in images were scored by six plastic surgeons, using a 10‐point Likert scale, where 1 represented ‘very bad regularity’ and 10 represented ‘very good regularity.’ The inter‐rater reliability was verified by means of the intraclass correlation coefficients (ICC). Finally, stepwise multiple regression analysis was used to extract useful indices; where the clinical evaluation of the physicians (gold standard) was considered to be a dependent variable, and indices obtained from digital skin image analysis to be independent variables. Results: The ICC of raters was 0.92 (95% confidence interval; 0.91–0.94) when including all raters, therefore the score of all raters was used. As a result of stepwise multiple regression, the index of interval (L), thickness (T_ave), energy 0°, and entropy 45° independently explained the clinician evaluation. R² in multiple regression equation was 0.59. Conclusions: It is shown that quantitative evaluation using skin images and their analysis is one method of determining skin integrity in the elderly. The indices to explain the clinical evaluation of specialists were defined and these indices were obtained simply by using a skin image.     

Pharmacokinetics of L-arginine in adults with moderately severe malaria

Severe malaria is associated with decreased nitric oxide (NO) production and low plasma concentrations of L-arginine, the substrate for NO synthase. Supplementation with L-arginine has the potential to improve NO bioavailability and outcomes. We developed a pharmacokinetic model for L-arginine in moderately severe malaria to explore the concentration-time profile and identify important covariates. In doses of 3, 6, or 12 g,L-arginine was infused over 30 min to 30 adults with moderately severe malaria, and plasma concentrations were measured at 8 to 11 time points. Patients who had not received L-arginine were also assessed and included in the model. The data were analyzed using a population approach with NONMEM software. A two-compartment linear model with first-order elimination best described the data, with a clearance of 44 liters/h (coefficient of variation [CV] = 52%) and a volume of distribution of 24 liters (CV = 19%). The natural time course of L-arginine recovery was described empirically by a second-order polynomial with a time to half recovery of 26 h. The half-life of exogenous L-arginine was reduced in patients with malaria compared with that for healthy adults. Weight and ethnicity were significant covariates for clearance. MATLAB simulations of dosing schedules for use in future studies predicted that 12 g given over 6, 8, or 12 h will provide concentrations above the K(m) of endothelial cell CAT-1 transporters in 90%, 75%, and 60% of patients, respectively.     

Human artificial chromosome (HAC) vector with a conditional centromere for correction of genetic deficiencies in human cells

Human artificial chromosome (HAC)-based vectors offer a promising system for delivery and expression of full-length human genes of any size. HACs avoid the limited cloning capacity, lack of copy number control, and insertional mutagenesis caused by integration into host chromosomes that plague viral vectors. We previously described a synthetic HAC that can be easily eliminated from cell populations by inactivation of its conditional kinetochore. Here, we demonstrate the utility of this HAC, which has a unique gene acceptor site, for delivery of full-length genes and correction of genetic deficiencies in human cells. A battery of functional tests was performed to demonstrate expression of NBS1 and VHL genes from the HAC at physiological levels. We also show that phenotypes arising from stable gene expression can be reversed when cells are "cured" of the HAC by inactivating its kinetochore in proliferating cell populations, a feature that provides a control for phenotypic changes attributed to expression of HAC-encoded genes. This generation of human artificial chromosomes should be suitable for studies of gene function and therapeutic applications.     

Negative pressure wound therapy versus conventional wound care in cancer surgical wounds: A meta‐analysis of observational studies and randomised controlled trials

The application of negative pressure wound therapy (NPWT) in cancer surgical wounds is still controversial, despite its promising usage, because of the risks of increased tumorigenesis and metastasis. This study aimed to review the risks and benefits of NPWT in surgical wounds with the underlying malignant disease compared with conventional wound care (CWC). The first outcome was wound complications, divided into surgical site infection (SSI), seroma, hematoma, and wound dehiscence. The secondary outcome was hospital readmission. We performed a separate meta‐analysis of observational studies and randomised controlled trials (RCTs) with CI 95%. Thirteen observational studies with 1923 patients and seven RCTs with 1091 patients were included. NPWT group showed significant decrease in the risk of SSI (RR = 0.45) and seroma (RR = 0.61) in observational studies with P value <0.05, as well as RCTs but were not significant (RR = 0.88 and RR = 0.68). Wound dehiscence (RR = 0.74 and RR = 1.15) and hospital readmission (RR = 0.90 and RR = 0.62) showed lower risks in NPWT group but were not significant. Hematoma (RR = 1.08 and RR = 0.87) showed no significant difference. NPWT is not contraindicated in cancer surgical wounds and can be considered a beneficial palliative treatment to promote wound healing.     

Mental Health Disorders of the Indonesian People in the Early Stages of the COVID-19 Pandemic: Who is Vulnerable to Experiencing it?

The extraordinary situation related to COVID-19 makes people worry about their health, family health, work, finances, and other daily activities. This condition can lead to social unrest, which has consequences for mental health problems. This study aims to determine the mental health consequences at the beginning of the COVID-19 pandemic in Indonesia. This is a cross-sectional study involving a target population aged 18 years and over who had access to electronic communication devices. An online questionnaire was randomly distributed and snowballed throughout 34 provinces in Indonesia. The study was conducted from 2 to 4 May 2020. Non-parametric and multivariate linear regression analyses were performed to identify factors associated with anxiety and depression. Two thousand seven hundred forty-three participants were involved in this study, with 69.16% female. In sum, 6.92% of participants had General Anxiety Disorder-7 (GAD-7) scores ≥ 10 for moderate-severe anxiety symptoms, and 8.57% had Patient Health Questionnaire-9 (PHQ-9) scores ≥ 10 for moderate-severe depressive symptoms. The multivariate linear regression analyses showed that the strongest factors influencing anxiety and depression were a history of mental illnesses, chronic illnesses, the group affected by layoffs or job seekers, unemployed, students, younger age group, living in a rented house, single, and female. In contrast, the lower and secondary education level seems to reduce the risk of depression compared to those with higher education levels. Anxiety and depression occur during the periods of activity restriction during the COVID-19 pandemic and are influenced by several modifiable and non-modifiable factors. There is an urgent need to emphasize vulnerable groups such as those with a history of illness, those affected by layoffs/looking for work, and the younger age group.     

Adverse drug reaction reporting in Australian hospitals

Objective To determine the status of adverse drug reaction (ADR) reporting in Australian hospitals. Method Postal questionnaire to all 299 directors of pharmacy in Australian hospitals. Key findings The response rate was 49.5%. Hospitals of different size were well represented: <100 beds (23%), 100–199 beds (35%) and 200–499 beds (30%). ADR policies were reported by 67% of pharmacy departments. In most hospitals, doctors or pharmacists were responsible for ADR submissions to the Australian ADR Advisory Committee (ADRAC). Follow‐up action included reports to Drug and Therapeutics Committees (57%) and drug bulletins (37%). Advice on ADRs was forwarded to the patient, the general practitioner (GP) and the community pharmacist by 96%, 89% and 11% of hospitals. Methods of informing patients were verbal (91%), card (17%) or letter (13%). Methods used to notify GPs included discharge summary (70%), letter (26%) or via the patient (15%). Data from ADRAC indicated a median 2.5 (range 0–362) reports from participating hospitals in 2000. The median ADR reporting rate was 0.02% (range 0–1.09) of patient admissions. There was no association between ADR reporting rates and the existence of an ADR policy or a centralised ADR reporting system. There was an association between hospital ADR reporting rates and provision of feedback or a reward for ADR reporting (P<0.001). Conclusions Procedures or policies for ADR reporting are available in most Australian hospitals but this has no direct effect on ADR reporting rates. These findings suggest a gap between policy and implementation of ADR reporting. Strategies to improve ADR reporting could include improved feedback and electronic submissions to ADRAC from a centralised service.