Transfection and cloning of genes for membrane antigens using the FACS

In order to facilitate cloning of genes for cell surface molecules, we cotransfected LTK^? mouse fibroblasts with thymidine kinase (TK) genes and total human or mouse DNA. TK⁺ cells, selected by growth in HAT medium, were stained with fluorochrome conjugated monoclonal antibodies or other fluorescent ligands which bind to one or another membrane differentiation antigen or receptor. We isolated fluorescent transfectants expressing these molecules using a fluorescence activated cell sorter (FACS). For some antigens, spontaneous gene amplification occurred. By repeated cycles of FACS sorting and regrowth we obtained high expressing clones. We then isolatedcDNA and genomic clones using selectedcDNA probes to screen phage withcDNA inserts. DNA from virtually any tissue source transfected equally well for the various molecules except for DNA from a trophoblast derived choriocarcinoma cell line which did not transfect for Leu-2.     

Cross-Sectional Serosurvey of Companion Animals Housed with SARS-CoV-2–Infected Owners,Italy

We conducted a serologic survey among dogs and cats in Italy to detect antibodies against severe acute respiratory syndrome virus 2 (SARS-CoV-2). We found that SARS-CoV-2 seroprevalence was higher among cats (16.2%) than dogs (2.3%). In addition, seroprevalence was higher among animals living in close contact with SARS-CoV-2–positive owners.     

Metformin Decreases Circulating Androgen and Estrogen Levels in Nondiabetic Women With Breast Cancer

IntroductionDiabetic patients treated with metformin have a lower risk of developing BC or a better BC prognosis. Metformin might reduce cancer growth through direct antiproliferative effects or through indirect mechanisms, particularly the reduction of insulin. In a randomized study on nondiabetic BC patients in natural menopause with high testosterone levels, we observed a significant decrease in insulin and in testosterone levels with metformin 1500 mg/d compared with 1000 mg/d. We present the results of a new analysis of our study on the effect of metformin on the bioavailability of sex hormones.Patients and MethodsOne hundred twenty-four eligible women were initially invited to take metformin 500 mg/d for 3 months. The 108 women who completed the first 3 months continued the study using 1000 mg/d for 1 month. The women were then randomized into 2 groups, and, for the subsequent 5 months, 1 group increased the dose to 1500 mg/d, and the other group continued with 1000 mg/d.ResultsNinety-six women completed the study, 43 receiving metformin 1500 mg/day, and 53 receiving 1000 mg/day. The women receiving 1500 mg/d showed a greater and significant reduction of free testosterone (?29%) and estradiol (?38%), a borderline significant reduction of estrone and insulin-like growth factor-1, and a nonsignificant reduction of androstenedione. They also showed a nonsignificant increase of dehydroepiandrosterone sulfate.ConclusionMetformin does not interfere with the production of dehydroepiandrosterone sulfate. Besides, it decreases estradiol levels, basically through the reduction of testosterone. These hormonal changes might have clinical relevance.     

Foetal and neonatal ovarian cysts: a 5-year experience

Background Abdominal cystic formations in newborns are relatively common and often diagnostic suspicion arises in this regard even before birth as a result of ultrasound scans carried out during pregnancy. The aim of this study is to highlight the problems posed by the prenatal diagnosis of abdominal cysts in order to outline the most appropriate therapeutic approach in case of suspected ovarian cysts. Materials and methods Between January 2003 and January 2007, 57 women were enrolled in this study for a prenatal ultrasound (US) that revealed the presence of an echo-rare or echo-free area in the foetal abdomen. After birth all babies underwent blood tests and abdominal US scans in order to confirm or identify the nature of the cyst. If abdominal US could not show the nature of the cystic formation, magnetic resonance imaging with sedation was performed. When the radiological tests were not useful to identify the nature of the cysts and surgery was then necessary, surgical procedures were performed with laparoscopy. Results Ultrasounds were useful to identify the diameter of the cysts but not all their origins; also MRI confirmed the morphology and volume of the cysts, but could not give further details about their origin. Discussion Abdominal ultrasound and finally laparoscopy used to treat and remove the cysts were useful to monitor all simple abdominal cysts. MRI seemed not to be useful for the treatment of this condition, especially in the pediatric age when mild sedation is required.     

A Low-Cost Wireless Bite Force Measurement Device

Assessing maximum voluntary bite force is important to characterize the functional state of the masticatory system. Due to several factors affecting the estimation of the maximum bite force, a unique solution combining desirable features such as reliability, accuracy, precision, usability, and comfort is not available. The aim of the present study was to develop a low-cost bite force measurement device allowing for subject-specific customization, comfortable bite force expression, and reliable force estimation over time. The device was realized using an inexpensive load cell, two 3D printed ergonomic forks hosting reusable subject-specific silicone molds, a read-out system based on a low-cost microcontroller, and a wireless link to a personal computer. A simple model was used to estimate bite force taking into account individual morphology and device placement in the mouth. Measurement reliability, accuracy, and precision were assessed on a calibration dataset. A validation procedure on healthy participants was performed to assess the repeatability of the measurements over multiple repetitions and sessions. A 2% precision and 2% accuracy were achieved on measurements of forces in the physiological range of adult bite forces. Multiple recordings on healthy participants demonstrated good repeatability (coefficient of variation 11%) with no significant effect of repetition and session. The novel device provides an affordable and reliable solution for assessing maximum bite force that can be easily used to perform clinical evaluations in single sessions or in longitudinal studies.     

标准同轴晶状体乳化与微小切口白内障手术:一项角膜内皮的研究

目的:评估新的微小切口白内障手术(MICS)是否较标准同轴晶状体乳化引起较少的内皮损害方法:年龄相关性白内障患者 200 例随机分配接受标准同轴晶状体乳化(SCP)或 MICS 晶状体乳化。术前及术后1a 测量角膜中央及 12 点内皮细胞计数、细胞大小变异系数、六角形细胞百分比以及角膜中央厚度。结果:患者 173 例完成了试验。在 1a 的随访检查中,MICS 组内皮细胞损失为 6.5204%,而 SCP 组为 8.726%(P<0.00005)。两组在细胞大小变异系数、六角形细胞百分比、术后角膜厚度以及在切口部位和角膜中央测得的内皮细胞密度变异方面无显著差异。在硬性白内障患者中,与手术步骤无关,内皮细胞密度损失的增加量较高。结论:比较 SCP 与 MICS,本研究表明后者引起的内皮细胞密度损失明显地较低。     

Clinical outcomes of primary arterial embolization in severe hepatic trauma: A systematic review

Purpose

This purpose of this systematic review was to determine the safety and efficacy of arterial embolization as the primary treatment for grade III-V liver trauma, excluding the postoperative use of arterial embolization.

Effects of resynchronization therapy on sympathetic activity in patients with depressed ejection fraction and intraventricular conduction delay due to ischemic or idiopathic dilated cardiomyopathy

This study assesses the effect of biventricular pacing on sympathetic nerve activity (SNA) in patients with depressed ejection fraction and intraventricular conduction delay (IVCD). Biventricular pacing has been shown to result in hemodynamic improvement in patients with depressed ejection fraction and IVCD. The effect of biventricular pacing on SNA, however, remains unclear. A total of 15 men with a mean ejection fraction of 25 +/- 4% were enrolled. Arterial pressure, central venous pressure and SNA were recorded during 3 minutes of right atrial (RA) pacing and RA-biventricular pacing. Pacing was performed at a rate 5 to 10 beats faster than sinus rhythm, with an atrioventricular interval equal to 100 ms during RA-biventricular pacing. RA-biventricular pacing resulted in greater arterial pressures (p <0.05) than RA pacing (146 +/- 15/83 +/- 11 vs 141 +/- 15/80 +/- 10 mm Hg). There were no differences in central venous pressures between the 2 pacing modes (p = 0.76). SNA was significantly less during RA-biventricular pacing (727 +/- 242 U) than during RA pacing (833 +/- 332 U) (p <0.02). Furthermore, there was a positive correlation between baseline QRS duration and the decrease in SNA noted with RA-biventricular pacing (r = 0.58, p = 0.03). Biventricular pacing results in improved hemodynamics and a decrease in SNA compared with intrinsic conduction in patients with left ventricular dysfunction and IVCD. If the current findings are also present with chronic biventricular pacing, then this form of therapy may have a positive impact on mortality.