The neutralization effect of montelukast on SARS-CoV-2 is shown by multiscale in silico simulations and combined in vitro studies.

Small molecule inhibitors have previously been investigated in different studies as possible therapeutics in the treatment of severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2). In the current drug repurposing study, we identified the leukotriene (D4) receptor antagonist montelukast as a novel agent that simultaneously targets two important drug targets of SARS-CoV-2. We initially demonstrated the dual inhibition profile of montelukast through multiscale molecular modeling studies. Next, we characterized its effect on both targets by different in vitro experiments including the enzyme (main protease) inhibition-based assay, surface plasmon resonance (SPR) spectroscopy, pseudovirus neutralization on HEK293T/hACE2+TMPRSS2, and virus neutralization assay using xCELLigence MP real-time cell analyzer. Our integrated in silico and in vitro results confirmed the dual potential effect of montelukast both on the main protease enzyme inhibition and virus entry into the host cell (spike/ACE2). The virus neutralization assay results showed that SARS-CoV-2 virus activity was delayed with montelukast for 20 h on the infected cells. The rapid use of new small molecules in the pandemic is very important today. Montelukast, whose pharmacokinetic and pharmacodynamic properties are very well characterized and has been widely used in the treatment of asthma since 1998, should urgently be completed in clinical phase studies and, if its effect is proved in clinical phase studies, it should be used against coronavirus disease 2019 (COVID-19).

Structure-based engineering of an antiangiogenic scFv antibody for soluble production in E. coli without loss of activity.

Development of monoclonal antibody therapeutics against vascular endothelial growth factor receptor 2 (VEGFR-2) protein, which is the main regulator in angiogenesis, has been a major challenge for years. In the current study, we engineer an inclusion body forming single-chain variable fragment (scFv) against VEGFR-2 by using complementarity determining regions (CDR) grafting technique to improve its solubility and investigate the activity of the engineered molecule. CDR sequences of the target scFv were grafted into the framework of another intrinsically soluble scFv molecule. Based on the computational results, CDR grafting has increased the solubility of the grafted scFv molecule. Results confirmed that the grafting approach increased in vivo folding properties of the target scFv molecule compared with the original scFv molecule. Similar binding affinities to the VEGFR-2 were observed for the original and the grafted scFv by surface plasmon resonance assays. Biological activity assays, including human umbilical vein endothelial cells proliferation and wound healing assays, showed that grafted scFv molecule has an antiangiogenic property. This study suggests that an antiangiogenic scFv fully expressed as an inclusion body can be rescued by grafting its CDR regions to a scFv expressed in a soluble form without any loss in its binding property and its activity.

Serum Fetuin-A and Insulin Levels in Classic Congenital Adrenal Hyperplasia.

Androgens play a pivotal role in non-reproductive organs such as the kidney, heart, liver, and pancreas. As androgen receptors are expressed in pancreatic and liver cells, excess testosterone can result in hypersecretion of insulin and fetuin-A, a protein produced in the liver. The expression of fetuin-A, a natural inhibitor of tyrosine kinase activity in muscle and liver, leads to insulin resistance. In addition, insulin and fetuin-A levels are thought to be affected by drugs such as glucocorticoids (GCs) and fludrocortisone. However, whether fetuin-A and insulin levels are affected by androgens and GCs in patients with classic congenital adrenal hyperplasia (CAH) is unknown. This cross-sectional study included 56 CAH patients and 70 controls. Analyses were stratified by sex and prepubertal/pubertal status to control for potential changes in serum metabolic/inflammatory markers associated with the production of sex steroids. Fasting blood glucose, insulin, triglyceride, total cholesterol, high density lipoprotein-cholesterol, aspartate aminotransferase, alanine aminotransferase, fetuin-A, and high-sensitivity C-reactive protein (hs-CRP) levels were measured in blood samples. In addition, 17α-hydroxyprogesterone, androstenedione, total testosterone, free testosterone, and dehydroepiandrosterone sulfate levels were measured before medication was administered. Insulin and fetuin-A levels were significantly higher in CAH patients than in controls. The unfavourably high levels of these substances exhibited a positive correlation with total and free testosterone. Regression analysis revealed that fetuin-A and free testosterone were the only independent predictors of the insulin level, while insulin and free testosterone levels significantly predicted the fetuin-A level (R2=42.7% and 59.8%). Differences were also observed in triglyceride and hs-CRP levels between the pubertal and prepubertal groups. We conclude that serum fetuin-A and insulin levels may be associated with androgens in CAH patients.

Forced migration-related traumatic experiences and collective memory in Ezidi asylum-seekers coming to Diyarbakir province from Shingal region.

PURPOSE: The purpose of this study was to investigate the collective memory of Ezidi asylum-seekers who migrated from Iraq to Turkey in 2014. METHODS: Kurdish-speaking Ezidi participants of 25-65 years of age (ten male and ten female) who have arrived to Fidanlik Camp in Diyarbakir, Turkey from the Iraqi Shingal region have been included in the study. Data were collected through semi-structured in depth interviews and qualitative content analysis was conducted on the collected data. RESULTS: The word "edict", which was used to define violent attacks, had a significant importance in all participants' memories. The collective memory observed in the Ezidi society leads a collective fear. Homelessness and religious discrimination are other concerns observed in the population. CONCLUSION: In conclusion, due to the depth and strength of the oral narrative tradition in the Kurdish culture, a significant collective memory is formed within the Ezidi society. Accumulation of previous negative events may prevent Ezidis to form a healthy identity. Traumas of the Ezidis should be investigated in more detail, and studies with larger scopes should be conducted in the future.

Dipeptidyl peptidase-4 and adenosine deaminase enzyme levels in polycystic ovary syndrome.

Polycystic ovary syndrome (PCOS) is one of the most common endocrinopathies in reproductive age women and insulin resistance (IR) and hyperinsulinism play a critical role in the pathogenesis. Glucagon-like peptide-1 (GLP-1), promotes insulin secretion, inhibits glucagon secretion. GLP-1 is degraded by dipeptidyl peptidase-4 (DPP-4). DPP-4, also interacts with adenosine deaminase (ADA). Therefore, IR may have a significant connection with ADA activity. The aim of this study is to compare levels of DPP-4 and ADA enzymes in PCOS and infertile patients. Forty-four patients with PCOS and 44 infertile patients with normal ovarian reserve were enrolled in the study. Serum ADA, DPP-4, AMH, glucose and insulin levels were measured. HOMA-IR method was used to assess insulin sensitivity. ADA, DPP-4, AMH, HOMA-IR (p < .05) and insulin levels (p < .01) were found to be increased in PCOS patients. Considering all study participants AMH levels were found to be positively correlated with ADA (r: 0.734) and DPP-4 (r: 0.449) levels. Also ADA levels were found to be positively correlated with DPP-4 (r: 0.472), insulin (r: 0.216) and HOMA-IR (r: 0.223). Our findings about the elevation of DPP-4 levels in patients with PCOS suggest that the use of DPP-4 inhibitors may be beneficial in treatment of these patients.

The protective effects of caffeic acid phenethyl ester against toluene-induced nephrotoxicity in rats.

Caffeic acid phenethyl ester (CAPE) has antioxidant and anti-inflammatory properties. The aim of this study is to examine the negative effects of toluene on kidney tissues and functions and to investigate the protective effects of CAPE against toluene-induced nephrotoxicity in rats. A total of 21 male Wistar rats were divided into three groups of equal number in each. The rats in group I were the controls. Toluene was intraperitoneally injected into the rats in group II with a dose of 500 mg/kg. Rats in group III received CAPE daily while exposed to toluene. After 14 days of experimental period, all rats were killed by decapitation. Enzymatic activities of superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), and catalase (CAT) and malondialdehyde (MDA) levels were studied in the rat kidneys. Blood urea nitrogen (BUN) and serum creatinine levels were measured for renal function. The CAT and SOD enzyme activities and serum creatinine levels were significantly increased in rats treated with toluene when compared with the controls. But GSH-Px activity, MDA, and BUN levels showed statistically nonsignificant changes. However, increased CAT and SOD enzyme activities and decreased serum creatinine levels were detected in the rats that received CAPE while exposed to toluene. The GSH-Px activity and MDA and BUN levels in the same group did not show statistically significant changes. The results of our study demonstrated that toluene damages kidney tissue and is a nephrotoxic substance. CAPE was able to prevent the renal damage as antioxidant, antitoxic, and nephroprotective agent.

Amyand's hernias in childhood (a report on 21 patients): a single-centre experience.

PURPOSE: Amyand's hernia is a very rare type of hernia and is characterised by the presence of a vermiform appendix in the inguinal hernia sac. It may present as a tender inguinal or inguinoscrotal swelling, and is usually misdiagnosed as an irreducible or strangulated inguinal hernia. This is a presentation of our experiences with these unusual hernias. METHODS: This study conducted a retrospective analysis of 21 patients with Amyand's hernias operated on at our institution between April 2007 and February 2011. The age and sex distributions of the patients, the types of hernias, and preoperative diagnostic tests were evaluated. RESULTS: Twenty patients were male (95.3 %) and one was female (4.7 %). The median age was 20.3 months (ranging from 2 months to 10 years). In nine patients, the conditions were diagnosed using ultrasonographic (USG) imaging preoperatively. Nine of the 21 patients underwent emergency operations. The 12 remaining patients were operated on after preparations were completed. The operative findings included 12 normal appendixes, five inflamed appendixes, one perforated appendix, and three hernias whose inner hernia sac surface was adhered to the appendix. Nine patients underwent inguinal hernia repair with appendectomy, and 12 patients had hernia repair without an appendectomy. Recurring hernias and appendicitis were not detected in any patients within the follow-up time. CONCLUSIONS: Although a routine appendectomy is not required for a normal-looking appendix, in a case where the inflammatory status of the appendix adheres to the surface of the hernia sac, an appendectomy is required. We believe that USG examination in the diagnosis of Amyand's hernias can provide additional contribution.

[Percutaneous closure of secundum atrial septal defects in pediatric and adult patients: short- and mid-term follow-up results]. / Eriskin ve çocuklardaki sekundum tip atriyal septal defektlerin perkütan yaklasim ile kapatilmasi: Kisa-orta dönem izlem sonuçlarimiz.

OBJECTIVES: We aimed to evaluate the short- and mid-term results of patients with atrial septal defect (ASD) who were treated with percutaneous closure. STUDY DESIGN: Seventy-nine patients with secundum ASD (54 female and 25 male; mean age 26.2±17.2; range 3 to 71] years) were included in this study. Patients were evaluated by transthoracic (TTE) and/or transesophageal echocardiography (TEE). Amplatzer septal occluder (ASO) was used in all patients. In 76 patients, the procedure was performed under local anesthesia with TTE, while in the other 3 patients, it was performed with general anesthesia with TEE. Patients were followed up at the 1st, 3rd, 6th and 12th months and annually thereafter. Mean follow-up time was 13.6±6.6 months. RESULTS: Mean diameter of ASDs was 18.2±7.5 mm and 20.7±8.04 mm during balloon dilatation, and mean diameter of implanted devices was 22.7±8.5 mm. Procedural time was 40.2±12.6 minutes and fluoroscopy time was 10.9±4.1 minutes. The procedure was successfully performed in all patients (100%). One patient with cardiac tamponade died seven days after cardiac surgery. In two patients, the implanted devices embolized to the pulmonary circulation. Residual flow was found in three patients immediately after the procedure, without residual shunts one month after closure. Mild pericardial effusion in one patient and significant residual shunt due to device malposition in another were discovered during the follow-up at 1 and 6 months, respectively, after the procedure. CONCLUSION: Our findings showed that percutaneous closure of ASDs is successful in most patients with a low complication rate, and demonstrated that residual shunts do not develop in the majority of patients in the short- and mid-term.

A Novel Peptide-Based Detection of SARS-CoV-2 Antibodies.

The need for rapidly developed diagnostic tests has gained significant attention after the recent pandemic. Production of neutralizing antibodies for vaccine development or antibodies to be used in diagnostic tests usually require the usage of recombinant proteins representing the infectious agent. However, peptides that can mimic these recombinant proteins may be rapidly utilized, especially in emergencies such as the recent outbreak. Here, we report two peptides that mimic the receptor binding domain of the spike protein of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and investigate their binding behavior against the corresponding human immunoglobulin G and immunoglobulin M (IgG and IgM) antibodies in a clinical sample using a quartz crystal microbalance (QCM) sensor. These peptides were immobilized on a QCM sensor surface, and their binding behavior was studied against a clinical serum sample that was previously determined to be IgG and IgM-positive. It was determined that designed peptides bind to SARS-CoV-2 antibodies in a clinical sample. These peptides might be useful for the detection of SARS-CoV-2 antibodies using different methods such as enzyme-linked immunosorbent assay (ELISA) or lateral flow assays. A similar platform might prove to be useful for the detection and development of antibodies in other infections.

Decreased echogenicity of the embryo is correlated with absence of cardiac activity.

PURPOSE: The purpose of this study was to investigate whether the sonographic echogenicity of embryos is associated with cardiac activity in utero. METHODS: The present study reviewed a total of 164 embryos having a gestational age between 6 and 8 weeks. These embryos were examined by transvaginal ultrasonography and a comparison of their echogenicity was made with respect to those of the placenta and the myometrium. Grade II embryos were less echogenic than the placenta or had similar echogenicity with the myometrium, whereas grade I embryos shared the same echogenicity as the placenta. In contrast, grade III embryos were less echogenic than the myometrium. RESULTS: Most of the embryos with cardiac activity were detected to have grade II echogenicity (78/130, 60.0%), whereas the remaining embryos had grade I echogenicity (52/130, 40.0%). In contrast, most of the embryos without cardiac activity had grade III echogenicity (20/34, 58.8%), whereas the remaining embryos had either grade II (8/34, 23.5%) or grade I (6/34, 17.7%) echogenicity. CONCLUSIONS: Decreased echogenicity of embryos on grayscale ultrasound in the early first trimester is correlated with an absence of cardiac activity.

Evaluation of fetal nasofrontal angle in the second trimester in normal pregnancies.

PURPOSE: The nasal bone is one of the important fetal structures to be radiologically evaluated in the first and second trimesters. In this study, we aimed to evaluate the nasal root and to determine the normal range of the naso-frontal angle, which may potentially be used as a predictive or diagnostic parameter in the diagnosis of the above-mentioned syndromes or other congenital abnormalities with associated nasal features using obstetric ultrasonography in normal fetuses. METHODS: We evaluated the naso-frontal angle using transabdominal ultrasonography in 195 healthy fetuses between the 18(th) and 21(st) weeks of gestation. The measurements were performed with the fetal neck in mild flexion and three echogenic lines (nasal end, skin on the nasal bone, nasal bone) visualized. RESULTS: The minimum, maximum and mean naso-frontal angle values were 110°, 143° and 128°, respectively, between the 18(th) and 21(st) weeks of gestation. Linear regression analysis showed no association between NFA and gestational week (Beta = 0.020 p = 0.786). CONCLUSIONS: The normal range of NFA between the 18(th) and 21(st) weeks of gestation was determined in this study. The association between NFA measurements and various congenital anomalies or fetal outcomes remains to be investigated.

Evaluation of the chemopreventive effects of Hypericum perforatum L on DMBA-applied rat oral mucosa.

OBJECTIVE: Hypericum perforatum L also known as St. John's wort is known to have many beneficial properties for the organism including its antioxidant and anticancer activities. It is also known to have shown antiproliferative and cytotoxic effects against various cancer cell lines. The purpose of this study was to investigate the effects of Hypericum perforatum L on 7,12-dimethylbenz(a)anthracene-induced rat oral squamous cell carcinoma model. DESIGN: The in vitro antioxidant properties of Hypericum perforatum L was determined and an extract was prepared. Thirty Wistar male rats were divided randomly into 4 groups (Control group, DMBA group, HP + DMBA group, HP group). The antioxidant defense mechanisms in tissue and blood samples were evaluated biochemically and immunohistochemically, the carcinomatous changes in connective tissue were investigated immunohistochemically and epithelial changes in the tissue samples were evaluated histopathologically. RESULTS: The extract revealed inhibitory effects on some antioxidant enzymes (catalase, glutathione peroxidase). Immunohistochemical evaluations revealed no invasive changes in the connective tissue. Hypericum perforatum L demonstrated chemopreventive effects although it did not prevent carcinomatous changes altogether. CONCLUSIONS: Hypericum perforatum L is a promising chemopreventive agent and further studies are needed in order to evaluate the full potential of this plant.

Evaluation of the pathophysiological role of Fetuin A levels in adolescents with polycystic ovary syndrome.

OBJECTIVES: Polycystic ovary syndrome (PCOS) is an endocrinopathy, in which hyperandrogenism and hyperinsulinism have both occurred. Fetuin-A, a natural inhibitor of tyrosine kinase, leads to insulin resistance. The aim was to evaluate the relationship between fetuin-A and hyperandrogenism and hyperinsulinism and the role of fetuin-A in the pathophysiology of PCOS. METHODS: Thirty-eight cases with PCOS and 40 healthy adolescents were included in the study. PCOS and controls were divided into obese/non-obese subgroups. LH, FSH, total and free testosterone (TT, FT), SHBG, androstenedione, DHEAS were measured in patients with PCOS. Fasting glucose, insulin, lipid profile, AST, ALT, HsCRP, and fetuin levels of PCOS patients and healthy controls were also measured. RESULTS: Fetuin-A levels were higher in PCOS patients than in controls. In the obese-PCOS group, when compared to non-obese PCOS patients; the levels of SHBG and HDL were low while cholesterol, LDL, triglyceride, HOMA-IR, FT, FAI, and HSCRP levels were high, but Fetuin-A levels were similar. In the obese-PCOS group, fetuin-A levels were higher than in obese-controls. HOMA-IR and fetuin-A levels were higher in non-obese PCOS patients than in non-obese controls. In the PCOS group, fetuin-A was positively correlated with TT, FT, FAI and androstenedione and negatively correlated with SHBG. Regression analysis demonstrated that FT, SHBG, and androstenedione significantly predicted fetuin-A levels (R2=54%). In non-obese PCOS patients and controls, fetuin-A was positively correlated with insulin and HOMA-IR. CONCLUSIONS: These results suggest a relationship between androgen levels and fetuin-A in PCOS cases, independent of insulin resistance, and may shed light on further studies.

SARS-CoV-2 neutralizing antibody development strategies.

In December 2019 a novel coronavirus was detected in Wuhan City of Hubei Province-China. Owing to a high rate of transmission from human to human, the new virus called SARS-CoV-2 differed from others by its unexpectedly rapid spread. The World Health Organization (WHO) described the most recent coronavirus epidemic as a global pandemic in March 2020. The virus spread triggered a health crisis (the COVID-19 disease) within three months, with socioeconomic implications. No approved targeted-therapies are available for COVID-19, yet. However, it is foreseen that antibody-based treatments may provide an immediate cure for patients. Current neutralizing antibody development studies primarily target the S protein among the structural elements of SARS-CoV-2, which mediates the cell entry of the virus through the angiotensin converting enzyme 2 (ACE2) receptor of host cells. This review aims to provide some of the neutralizing antibody development strategies for SARS-CoV-2 and in vitro and in vivo neutralization assays.

Can major amputation rates be decreased in diabetic foot ulcers with hyperbaric oxygen therapy?

Although hyperbaric oxygen therapy has been used for diabetic foot ulcer since the 1980s, there is little information on its efficacy. The aim of this study is to evaluate whether hyperbaric oxygen can decrease major amputation rates and to determine the predictive factors. A total of 184 consecutive patients were treated with hyperbaric oxygen therapy as an adjunct to standard treatment modalities for their diabetic foot ulcer. Of these patients, 115 were completely healed, 31 showed no improvement and 38 underwent amputation. Of the amputations, nine (4.9%) were major amputations (below knee) and 29 were minor. Major amputations were associated with the Wagner grade (p < 0.0001), with the age of the patients (p = 0.028) and with the age of the wounds (p = 0.018). Hyperbaric oxygen therapy can help to reduce the major amputation rates in diabetic foot ulcer. However, further large, multicentre, randomised controlled studies are needed to make more accurate conclusions.

Studies on alkaline serine protease produced by Bacillus clausii GMBE 22.

An alkali tolerant Bacillus strain having extracellular serine alkaline protease activity was newly isolated from compost and identified as Bacillus clausii GMBE 22. An alkaline protease (AP22) was 4.66-fold purified in 51.5% yield from Bacillus clausii GMBE 22 by ethanol precipitation and DEAE-cellulose anion exchange chromatography. The purified enzyme was identified as serine protease by LC-ESI-MS analysis. Its complete inhibition by phenylmethanesulfonylfluoride (PMSF) also justified that it is a serine alkaline protease. The molecular weight of the enzyme is 25.4 kDa. Optimal temperature and pH values are 60 degrees C and 12.0, respectively. The enzyme showed highest specificity to N-Suc-Ala-Ala-Pro-Phe-pNA. The K(m) and k(cat) values for hydrolysis of this substrate are 0.347 mM and 1141 min(-1) respectively. The enzyme was affected by surface active agents to varying extents. The enzyme is stable for 2 h at 30 degrees C and pH 10.5. AP22 is also stable for 5 days over the pH range 9.0-11.0 at room temperature. AP22 has good pH stability compared with the alkaline proteases belonging to other strains of Bacillus clausii reported in the literature.

Periosteal osteoblastoma of the distal femur.

Periosteal osteoblastoma is a rare benign osteoblastic bone tumour located on the cortical bone. We report a case of periosteal osteoblastoma located at the distal femur in a 31-year-old man. Clinical, radiological and histopathologic findings are described and differential diagnosis is discussed. Despite its rarity, periosteal osteoblastoma should be considered as a possible diagnosis together with periostitis ossificans, periosteal chondroma, periosteal osteosarcoma and parosteal osteosarcoma when confronted with a superficial bone lesion.

Leg length discrepancies in adult femoral shaft fractures treated with intramedullary nailing.

BACKGROUND: To evaluate the leg length discrepancy (LLD) retrospectively in adult femoral shaft fractures treated with intramedullary nailing (IMN). METHODS: Sixty-three patients (58 male, 5 female; mean age 29.9+/-12.4; range 15 to 77 years) were included in the study. Fractures were identified according to the Winquist-Hansen (W) system and AO classification. 16 W0, 18 WI, 16 WII, 7 WIII, and 6 WIV fractures and 35 type A, 22 type B, and 6 type C fractures were repaired. Thirty-one (49.2%) patients had multiple injuries. Fourteen patients sustained an open fracture. LLDs were measured on physical examination and using orthoroentgenography. RESULTS: The mean follow-up was 90.2+/-29.9 (39-193) months. The mean LLD was 12.3+/-15.2 [12-(-60)] mm using orthoroentgenography and 12.9+/-13.7 [10-(-60)] mm according to manual measurement. In seven cases, no LLD was observed. Twenty-seven shortenings and one lengthening were observed in the 28 femurs with a discrepancy greater than 10 mm (44.4%). There was no statistical correlation between LLD and open or closed fracture (r=0.02, p=0.86), polytrauma (r=-0.09, p=0.47), or delayed surgery (p=0.31), but there was a tendency to a greater discrepancy in comminuted fractures (WIII, IV) (r=0.33, p=0.007). CONCLUSION: LLD may be seen in high rates in adult femoral shaft fracture cases treated with IMN. Static IMN following absolute restoration of the length may prevent this problem in femoral diaphysis fractures, especially comminuted WIII and IV types.