Human pluripotent stem cell-derived cells endogenously expressing follicle-stimulating hormone receptors: modeling the function of an inactivating receptor mutation.

Follicle-stimulating hormone (FSH) is crucial in the development and regulation of reproductive functions. The actions of human FSH and its receptor (FSHR) and mutations therein have mainly been studied using in vivo models, primary cells, cancer cells and cell lines ectopically expressing the FSHR. To allow studies of endogenous FSHR function in vitro, we differentiated FSHR-expressing cells from human pluripotent stem cells. FSH stimulation of the wild-type (WT), but not the inactivating Finnish founder mutant (A189V) receptor, activated the canonical cyclic adenosine monophosphate (cAMP)-dependent signaling pathway and downstream mediators. To investigate protein-protein interaction partners of FSHR at resting state and upon FSH stimulation, we expressed FSHR in HEK293 cells followed by affinity purification mass spectrometry analyses. We found 19 specific high-confidence interacting proteins for WT FSHR and 14 for A189V FSHR, several of which have been linked to infertility. Interestingly, while only WT FSHR interacted with FSH, insulin-like growth factor 1 receptor (IGF1R), for example, interacted with both WT and A189V FSHR upon FSH stimulation. In conclusion, our protocol allows detailed studies of FSH action and disease modeling in human cells endogenously expressing FSHR.

Myocardial Infarction in Non-obstructive Coronary Arteries (MINOCA) in the Perioperative Period can Epinephrine be Responsible?

Ephedrine, metaraminol, epinephrine and maneuvers like carotid sinus stimulation used during intraoperative period have been postulated to cause temporary spasm of the coronary vessels leading to decrease supply to the myocardium and precipitating myocardial infraction in non-obstructive coronary arteries (MINOCA). As an anaesthesiologists, we should be aware that even a dose as small as 25 mcg epinephrine infiltrated along with local anaesthetic in the subcutaneous plane may be responsible for coronary vessel spasm and thus myocardial infraction in nonobstructive coronary arteries. We report a case of 45 years old female with papillary carcinoma of thyroid who developed features of non-ST elevation myocardial infarction 5 minutes after the subcutaneous infiltration of 5 ml of 2% Xylocaine with 1:200000 Epinephrine. Patient was managed for acute Myocardial Infarction. Coronary angiogram done the next day revealed normal coronary arteries, hence the diagnosis Myocardial infraction in non-obstructive coronary arteries was made.

Displaced Intra-Articular Calcaneal Fractures: Evaluation of Clinical and Radiological Outcome Following Open Reduction and Internal Fixation with Locking Branched Calcaneal Plate.

Background An introduction of the World Health Organization Surgical Safety Checklist (WHO SSC) is essential to promote surgical safety. Objective To obtain country-specific information regarding the checklist in a leading medical institution in Nepal. Method The present research was a cross-sectional study with a survey conducted among healthcare professionals working in the operation theatre at the Tribhuvan University Teaching Hospital (TUTH) in Kathmandu, Nepal. A questionnaire was distributed to 150 healthcare professionals working in the operating theatre. Responses to the questionnaire were analysed descriptively and regression analyses used to identify factors associated with awareness of the checklist. Result In total, 127 healthcare professionals participated in the study, of whom 118 (92.9%) had been aware of the WHO SSC. A substantial proportion of participants (108, 91.5%) were not satisfied with the prevailing practice whereby the checklist was not routinely used during surgery. Lack of appropriate training was the most prevalent barrier to the checklist use (72, 67.9%), followed by unwillingness of staff to use the checklist (54, 50.9%), and lack of experience (42, 39.7%). The mean score on the survey was 6.0 out of 10. Regarding the results of the regression model on survey scores, surgeons had higher scores compared to nurses (unadjusted coefficient 0.80, 95% CI 0.20-1.40). Conclusion Most of the healthcare professionals were aware of the WHO SSC, however multiple barriers to the checklist use were identified. It is important to establish an effective use of WHO SSC in the operation theatre.

Ischemic Stroke as a Manifestation of Cholesterol Embolization Syndrome Following Percutaneous Coronary Intervention.

Cholesterol embolization syndrome (CES) is one of the major, yet underdiagnosed cause of morbidity and mortality following invasive coronary interventions. The major risk factors are elderly, male, atherosclerotic disease, anticoagulation and femoral access route. This multisystem disease affects skin, kidney, brain, eye and gastrointestinal tract. Only few cases of cholesterol embolization syndrome manifesting as an ischemic stroke are reported. We present a case of an elderly man, admitted to our ICU after percutaneous coronary intervention (PCI) who developed neurological deficits along with skin changes and renal failure. cholesterol embolization syndrome was suspected based upon the presence of cardiovascular risk factors, invasive cardiovascular intervention and clinical signs. The diagnosis of ischemic stroke made through plain MRI brain, revealed multiple areas of lacunar infarcts. He was treated with intermittent hemodialysis, statins and anti-platelet agents. On follow up, skin lesions and renal functions were improved; but slurring of speech and paresis persisted.

Plasma cell-free DNA methylation marks for episodic memory impairment: a pilot twin study.

Decline in episodic memory performance usually causes the first clinical symptoms of Alzheimer's disease. At present, Alzheimer's disease can only be diagnosed at a very late stage when neurodegeneration and cognitive impairment is already irreversible. New early disease markers are needed for earlier and more efficient Alzheimer's disease intervention. To identify early disease markers, we implemented a genome-wide bisulphite sequencing method for the analysis of plasma cell-free DNA methylation profiles and compared differences associated with episodic memory performance in Finnish twin pairs. A noticeable amount of cell-free DNA was present in plasma, however, the amounts as well as the genomic coverage of these fragments varied substantially between individuals. We found no significant markers associated with episodic memory performance in the twins' plasma cell-free DNA methylation profiles. Furthermore, our results indicate that due to the low genomic coverage of cell-free DNA fragments and the variety in these fragments between individuals, the implemented genome-wide bisulphite sequencing method is not optimal for comparing cell-free DNA methylation differences between large groups of individuals.

Post operative diagnosis of early gastric cancer in a low risk population and the possibility of risk stratified screening.

BACKGROUND: Gastric cancer is the second commonest cause of cancer related mortality worldwide. Though its incidence is more in Eastern Asia, it is increasing in the South Asian subcontinent. The diagnosis of early gastric cancer (EGC) confined to the mucosa or submucosa, is an important concern due to a better outcome at this stage where five year survival rates could increase by 90 percent. Though mass screening is done in few countries, it has not been applied in developing countries like Nepal. Preoperative diagnosis of EGC is rare in Nepal. The aim of this study is to analyze the clinicopathological features of postoperative cases of gastric cancer managed in a tertiary care university hospital of Nepal. METHODS: All patients with histological diagnosis of gastric cancer admitted in the Department of Surgery, Tribhuvan University Teaching Hospital, Kathmandu, Nepal during the three year period (September 2010 to August 2013) were analyzed retrospectively. RESULTS: Ninety two patients with endoscopic diagnosis of gastric cancers were admitted during the past three years. The mean age was 60 years ranging from 28 years to 85 years with the male to female ratio of 2.8:1. Five patients were younger than 40 years and all were in advanced stage. Thirty five percent of the patients belonged to Janajatis (Hill) community though they comprise only 23% of the population and about 65% of them belonged to an area involving 25% of the country. Seventy six cases were operated. Out of 92 patients, 4 patients were diagnosed as early gastric cancer postoperatively. All patients with early gastric cancer were above 50 years with CT Scan abdomen revealing focal thickening without lymphadenopathy. CONCLUSION: Over the years, the incidence of gastric cancer is increasing in Nepal. Though 92% are advance gastric cancers, few have been diagnosed and treated early. A screening program in a country like Nepal with diverse ethnicity and difficult terrain might be helpful if it targets high risk people in high risk areas.

Accuracy of triple test score in the diagnosis of palpable breast lump.

Breast lump is a very sensitive issue for the patient so a reliable, non invasive and prompt diagnosis helps to lessen the associated anxiety and leads to early definitive treatment. The aim of this study was to evaluate the accuracy of Triple Test Score (TTS) as a clinical tool for the diagnosis of a palpable breast lump. This diagnostic test study was carried out in the Department of Surgery of Tribhuvan University Teaching Hospital. Of the 117 patients admitted with breast diseases from the breast clinic over thirteen months, 87 had breast lump. Fifty patients underwent Triple Test Score ( physical examination, mammography and fine needle aspiration cytology) and were categorized into benign, suspicious and malignant. This was later correlated with the histopathological findings. Nineteen patients with breast lumps interpreted by TTS as benign correlated with the histopathological findings whereas of 31 malignant lumps, 30 turned out to be malignant and one benign. This gives TTS an over all accuracy of 98% with 100% sensitivity, 95.2% specificity and positive predictive value of 96.7%. Carcinoma was seen in 29 (58%) cases in the age group of 35 to 70 years. The mean age at diagnosis of benign and malignant disease was 41.8 and 45.1 years respectively. In conclusion, TTS is an accurate and least invasive diagnostic test based on which definitive treatment can be initiated.