A sensitive cell-based assay for testing potency of Botulinum neurotoxin type A.

Botulinum neurotoxin type A (BoNT/A) is a widely used biopharmaceutic for the treatment of neurological diseases and aesthetic medicine, allowing months-long paralysis of target muscles and glands. Large numbers of mice are used for multiple botulinum applications including batch release potency testing, antitoxin testing, countermeasure development and basic research. The mouse bioassay (MBA) has historically been the industry gold-standard in the botulinum field and is still heavily used for commercial product testing. BoNT/A intoxication causes severe suffering and application-specific, non-animal alternatives are urgently needed. It is widely accepted, that a cell-based assay (CBA) is the only way to faithfully replicate all the physiological steps of botulinum intoxication; comprising neuronal binding, internalization, endosomal escape, and cleavage of synaptosomal-associated protein of 25 kDa (SNAP25). However, it has not been straightforward to develop these assays and there are only a limited number of CBA currently in use. This is in part, due to the fact that very few cell lines have the appropriate levels of sensitivity to BoNT/A. In this study we have identified that LAN5 cells, a human neuroblastoma derived cell line, are sensitive to BoNT/A and can be engineered to express a recombinant NanoLuc luciferase tagged SNAP25 reporter molecule. On intoxication, the reporter molecule is cleaved and releases a NanoLuc-SNAP25 fragment which can be specifically captured on a 96-well plate for quantitative luminometry. Importantly, we demonstrate this new cell-based assay exhibits sensitivity comparable to the MBA.

Botulinum neurotoxin type A (BoNT/A) is extensively used in the treatment of neurological disorders and aesthetics. When the toxin enters cells, it targets a protein called SNAP25 and inhibits neurotransmitter release. Traditionally, the potency and safety of BoNT/A has been tested using the mouse bioassay, which causes significant distress to the animals being used. Our study introduces a new method for detecting BoNT/A activity based on LAN5 cells, which are a self-replicating, neuroblastoma-derived human cell line. We have engineered the cells to express a version of SNAP25 that allows the potency of BoNT/A to be measured using a luminescence assay. This new cell-based assay is as sensitive as the mouse bioassay and can be used for commercial product testing. This development could lead to fewer animals being used in research and commercial testing of BoNT/A, benefiting both scientific progress and animal welfare.

Could Birth Weight to Placental Weight Ratio Predict Postpartum Haemorrhage and Neonatal Intensive Care Unit Admission?

OBJECTIVE: To evaluate the usability of the ratio of birth weight to placental weight [fetoplacental ratio (FPR)] in predicting postpartum haemorrhage (PPH) and neonatal intensive care unit (NICU) admission. STUDY DESIGN: Prospective observational study. PLACE AND DURATION OF STUDY: Bursa Yuksek Ihtisas Training & Research Hospital, Bursa, Turkey, between July 2020 and July 2021. METHODOLOGY: Women who were supposed to have an uncomplicated delivery with a live, single, term pregnancy without any concomitant disease, were included in the study. Patients with PPH were accepted as the study group and patients without PPH were the control group. For NICU requirement, babies who were admitted to NICU were the study group, and babies who did not require NICU were the control group. The fetoplacental ratio was calculated by dividing the newborn weight to placental weight and evaluated in the prediction of NICU admission and PPH. RESULTS: The number of patients included in the study was 812. Approximately 7% of women had postpartum haemorrhage. The FPR was found as an independent predictor for PPH by nearly 3.5 fold. Women who experienced PPH had heavier placenta and lower fetoplacental ratio. Patients whose babies were admitted to NICU also had lower FPR with statistically significant differences. CONCLUSION: The fetoplacental ratio could be a promising predictor for PPH and NICU admission in the postpartum period. Since novel studies are needed using ultrasonographic measurements during antenatal surveillance to predict PPH or NICU admission. KEY WORDS: Birth weight, Neonatal intensive care unit, Placental weight, Postpartum haemorrhage, Cesarean birth, Vaginal birth, Fetoplacental ratio.

Short interdelivery interval in modern obstetrics: Maternal and neonatal outcomes.

Objective: To investigate the maternal, neonatal outcomes of the patients with short interdelivery interval (IDI) considering initial pregnancy outcomes. Materials and Methods: Women with two consecutive deliveries between 2016 and 2020 were included in the study. The maternal and neonatal outcomes of both pregnancies were reviewed. The time interval between consecutive deliveries was calculated. The patients were divided into two groups in terms of IDI either less or more than 24 months. Results: The number of patients with short IDI (≤24 months), and normal IDI was 1.915 and 1.370, respectively. About 15% of the women in both groups had at least one obstetric morbidity. The rates of uterine rupture, placenta previa, and peripartum hysterectomy were higher in women with short IDI. The number of patients with low birth weight, very low birth weight, and stillbirth was higher in the short IDI group. Conclusion: Patients with short interpregnancy intervals should be considered high-risk pregnancy. Adequate contraceptive methods should be used to prevent unintended pregnancies.

Are women diagnosed with early pregnancy loss at risk for anxiety, depression, and perinatal grief?

OBJECTIVES: To examine the effects of early pregnancy loss on emotions such as depression, grief, or a sense of hopelessness, while investigating different types of diagnoses, hospital stays, and treatments. METHODS: A prospective cohort epidemiological study was carried out in Bursa Yuksek Ihtisas Training and Research Hospital, Bursa, Turkey, between January and September 2019. The study included women diagnosed with early pregnancy loss classified into 3 groups: missed abortus, anembryonic pregnancy, and spontaneous abortion. The patients were screened via the Spielberger state-anxiety inventory (STAI-1) before initiating treatment. The Edinburgh postpartum depression scale (EPDS) and Perinatal Grief Scale (PGS) were also carried out in the first week of their hospital discharge. RESULTS: The study was carried out with a total of 116 patients. The median gestational week of the patients was calculated at 9, their median hospital stay was 2 days, and their median dose of misoprostol was 800 mcg. The STAI-1 revealed that median values computed for women in all groups indicated moderate anxiety. The EDPS also demonstrated depression-positive median values for women in all 3 groups (EPDS>13). However, no statistically significant difference was noted in comparisons of the 3 groups apropos STAI-1, EPDS, and PGS. CONCLUSION: Moderate anxiety, depressed mood, and perinatal grief were found in women diagnosed with early pregnancy loss, regardless of the type of abortion.

Comparison of ankle force, mobility, flexibility, and plantar pressure values in athletes according to foot posture index.

Objectives: This study aims to compare ankle force, mobility, flexibility, and plantar pressure distribution of athletes according to foot posture index (FPI). Patients and methods: Between September 2016 and May 2018, a total of 70 volunteer male athletes (mean age: 21.1±2.3 years; range, 18 to 25 years) were included. The athletes were divided into three groups according to their FPI as follows: having supinated feet (Group 1, n=16), neutral/normal feet (Group 2, n=36), or pronated feet (Group 3, n=18). Ankle range of motion (ROM), muscle flexibility, ankle joint strength, and plantar pressure distribution were measured. Results: There were significant differences among the three groups in both right and left ankle dorsiflexion ROM (p=0.009 and p=0.003, respectively). Group 1 had significantly smaller dorsiflexion ROM than the other groups. Group 1 also showed significantly less flexibility in the gastrocnemius and soleus muscles than the other foot posture groups. Groups 2 and 3 exhibited significant differences in the maximum torque (p=0.018), maximum work (p=0.008), and total work (p=0.008) of the right plantar flexor muscles at 60°/sec angular velocity. Peak pressure measurements of the right foot were higher in Group 1, compared to Groups 2 and 3 (p<0.001). Conclusion: The results of this study may help to enhance athletic performance by providing a guide for designing training programs appropriate for athletes with different foot types to address their specific muscle flexibility and strength deficiencies.

Relationship between body composition, multiple repeated sprint ability and vertical jump performance in elite badminton players / Relación entre la composición corporal, la capacidad de realizar múltiples sprints repetidos y el rendimiento del salto vertical en jugadores de bádminton de élite

SUMMARY: The purpose of this study was to assess the correlation between the body composition, multiple repeated sprint ability (MRSAB) test, and vertical jump performance. Fifteen voluntary elite Turkish badminton players participated in the study. The MRSAB test consisted of 2 repetitions of 4 movements (4x3m) separated by 30 seconds of passive recovery. The best time (BT), meantime (MT), total time (TT), and fatigue index (FI) were measured. Body composition was evaluated through dual-energy X-ray absorptiometry (DEXA). Squat jump (SJ) test for explosive power and countermovement jump (CMJ) test for elastic power were used. The main findings were that there was a significant correlation between MRSAB MT and TT with lean body mass, lean arm mass, and trunk lean mass (kg) of male badminton players. However, no significant correlation was observed among MRSAB MT, BT, and TT with another total/regional body composition of female badminton players (p>0.05). Moreover, the results of male badminton players showed that there was a significant negative correlation between FI and percentage ofbody fat (%BF), percentage of leg fat (%LF), percentage of trunk fat (%TF), and trunk mass. In addition, there was a significant negative correlation between FI with %BF, body fat mass kg, percentage of arm fat (%AF), arm fat mass, and %LF of female badminton players. It was also found in the study that there was a significant correlation between SJ and %LF; CMJ and %BF, body fat mass kg, %AF, %LF and leg fat mass kg. However, no significant correlation was observed among CMJ and SJ with other total/regional body compositions of male badminton players (p>0,05). Finally, changes in body composition are important issues for the physical performance level of badminton players as regional excess body fat may cause deterioration, especially in repeated-sprint ability and jumping performance.

RESUMEN: El propósito de este estudio fue evaluar la correlación entre la composición corporal, la prueba de capacidad de sprint repetido (MRSAB) y el rendimiento del salto vertical. Quince jugadores voluntarios de bádminton turcos de élite participaron en el estudio. El test MRSAB consistió en 2 repeticiones de 4 movimientos (4x3m) separados por 30 segundos de recuperación pasiva. Se midieron el mejor tiempo (BT), el tiempo medio (MT), el tiempo total (TT) y el índice de fatiga (FI). La composición corporal se evaluó mediante absorciometría de rayos X de energía dual (DEXA). Se utilizó la prueba de salto desde la sentadilla (SJ) para la potencia explosiva y la prueba de salto con contramovimiento (CMJ) para la potencia elástica. Los principales hallazgos fueron una correlación significativa entre MRSAB MT y TT con la masa corporal magra, la masa magra del brazo y la masa magra del tronco (kg) de los jugadores de bádminton hombres. Sin embargo, no se observó una correlación significativa entre MRSAB MT, BT y TT con otra composición corporal total/regional de jugadoras de bádminton (p>0,05). Además, los resultados de los jugadores hombres de bádminton mostraron que había una correlación negativa significativa entre FI y el porcentaje de grasa corporal (% BF), porcentaje de grasa en las piernas (% LF), porcentaje de grasa en el tronco (% TF) y masa del tronco. Además, hubo una correlación negativa significativa entre FI con %GC, masa de grasa corporal en kg, porcentaje de grasa en el brazo (%AF), masa de grasa en el brazo y %LF de las jugadoras de bádminton. También se descubrió en el estudio que había una correlación significativa entre SJ y %LF; CMJ y %BF, masa grasa corporal kg, %AF, %LF y masa grasa de piernas kg. Sin embargo, no se observó una correlación significativa entre CMJ y SJ con otras composiciones corporales totales/regionales de los hombres jugadores de bádminton (p>0,05). Finalmente, los cambios en la composición corporal son cuestiones importantes para el nivel de rendimiento físico de los jugadores de bádminton, debido a que el exceso de grasa corporal regional, puede causar un deterioro, especialmente en la capacidad de repetir sprints y en el rendimiento de los saltos.

Transvaginal natural orifice transluminal endoscopic surgery for ectopic pregnancy.

AIMS: This study aims to describe our transvaginal natural orifice transluminal endoscopic surgery (vNOTES) technique for ectopic pregnancy that can be performed using conventional laparoscopic equipment with the addition of a self-constructed pessary port and to evaluate the safety, feasibility, and outcomes of the technique. METHODS: This is a retrospective study evaluating outcomes of patients who underwent vNOTES for ectopic pregnancy (n = 21) between August 2019 and April 2021. RESULTS: Twenty-one patients underwent vNOTES as intended without any intraoperative complications. Three patients (14.3%) were nulliparous, 16 patients (76.2%) had no history of vaginal delivery, and 13 patients (61.9%) had a history of at least one cesarean delivery. Eight patients (38.1%) had a history of previous abdominal surgery other than cesarean section. The mean duration of surgery was 43.4 ± 12.6 min. The mean visual analog scale scores for pain were 2.45 ± 1.13 at 2 h after surgery, 0.45 ± 0.83 at 12 h after surgery, and 0.18 ± 0.36 at 24 h after surgery. The median duration of postoperative hospital stay was 1 day (range, 1-2). There were no postoperative complications within 30 days after surgery. CONCLUSIONS: vNOTES can be performed in an existing laparoscopy setup with the addition of a self-constructed pessary port. vNOTES is a safe and effective surgical treatment option for ectopic pregnancy, even in patients who have not had a vaginal delivery and have had multiple abdominal surgeries.

Act of fibulin-1 in preeclamptic patients: can it be a predictive marker?

Preeclampsia is one of the leading causes of maternal-neonatal morbidity and mortality, especially in developed and developing countries. Incidence of preeclampsia differs in accordance with parity, race, age, geography, and concomitant diseases. The role of placental implantation and risk factors was elucidated precisely. Antenatal care, use of medications, change in lifestyle, and nutritional supplementation were investigated for the prevention or decrease the complications; however, to date, there has not exposed a proper approach for prevention and prediction. The trigger mechanism or circumstance is still debate. Placental development especially spiral artery remodeling might be supposed to be the accused primary site of preeclampsia. Extracellular matrix proteins play a crucial role in implantation. Fibulin is one of these proteins which represents an association with matrix proteins, basement membranes, and elastic fibers. Fibulins are mainly functioning in the remodeling of tissues especially blood vessels, endocardial cushion, the mesenchymal, and connective tissue of several organs including heart, lung, intestine, kidneys, and liver. Several diseases were associated with altered fibulin levels. We aimed to examine fibulin-1 levels in preeclamptic patients and to focus on the possible role of fibulin-1 in preeclampsia. MATERIAL AND METHOD: A prospective observational, case-control study was achieved. Patients diagnosed with preeclampsia and healthy controls were recruited in the study. Patients' demographic features, perinatal outcomes, complications, obstetrics doppler ultrasonographic evaluations, laboratory results, and serum fibulin-1 levels were reviewed. The comparison of the groups was determined statistically. Correlation analysis and multivariate logistic analysis were calculated. The receiver operating characteristic (ROC) curve was used to indicate fibulin-1 levels for the prediction of preeclampsia. RESULTS: A total of 36 healthy pregnant and 38 preeclamptic patients were included in the study. Comparison of the groups with age, gravidity, BMI, APGAR scores, birth weight did not differ significantly. Kidney and liver function tests and complete hemogram parameters did not have a clinically important difference. Fibulin-1 levels were significantly lower in patients with preeclampsia. The ROC curve for fibulin-1 for predicting the preeclampsia risk was analyzed. The area under the ROC curves was 0.682 (95% CI [0.560-0.804, p < .007) for fibulin-1. The optimal cutoff value of fibulin-1 for detecting preeclampsia was ≤ 27.81 ng/ml, at which the sensitivity was 61.1% and specificity was 63.2 %. CONCLUSION: Fibulin-1 levels could be a beneficial marker for preeclampsia diagnosis and prediction. It might have a role in the etiopathology of preeclampsia, due to its function in the extracellular matrix.

Do patients with a single abnormal OGTT value need a globally admitted definition such as "borderline GDM"? Pregnancy outcomes of these women and the evaluation of new inflammatory markers.

INTRODUCTION: One of the approaches to diagnose Gestational Diabetes Mellitus (GDM) is to detect two or more elevated values in 3-h Glucose Tolerance Test (OGTT) after an abnormal 50 gr Glucose Challenge Test (GCT). Patients with single elevated OGTT generally postulated as healthy; however, these patients could experience adverse perinatal and maternal issues more frequently. We aimed to investigate the maternal and neonatal outcomes of women with single abnormal OGTT primarily by comparing these women with healthy controls and GDM patients. Secondarily; Mean Platelet Volume (MPV), Platelet Distribution Width (PDW), Neutrophil to Lymphocyte Ratio (NLR) and Platelet to Lymphocyte Ratio (PLR) which were defined as novel inflammatory markers recently, were evaluated among these women within the first trimester and before delivery values whether these markers could use as a predictive marker of GDM. MATERIALS AND METHODS: A retrospective cohort study was achieved in Bursa Yuksek Ihtisas Education and Training Hospital between January 2016 and April 2020. Patients who had GCT and OGTT at 24th-28th weeks of gestation were reviewed. Patients with GDM, women with single elevated OGTT value, and women with normal OCT values were recruited at the study as groups 1, 2, and 3 respectively. Maternal-neonatal outcomes and postpartum complications were reviewed from hospital registry system. Each complication were accumulated in a group entitled peripartum complication (a patient who had more than 1 complication for example preeclampsia and acute fetal distress was added in the peripartum complication group as one patient).The novel inflammatory markers were evaluated as NLR and PLR, and thrombocyte parameters as MPV and PDW were compared within the groups, and between the groups individually in the time period of first trimester and before delivery. RESULTS: A total of 10,579 patients were screened with OCT, of these a total of 1718 patients' results were between 140 mg/dl and 199 mg/dl. The numbers of the women who diagnosed GDM and who had single elevated OGTT were 508 and 469 respectively. Numbers of the patients who gave birth in our hospital and whose data were reviewed adequately were 464 in groups 1, 406 in group 2, and 768 in group 3.Patients with single elevated OGTT had increased rates of peripartum complication, acute fetal distress (AFD), IUGR, preterm delivery, cesarean delivery rate, macrosomia, labor arrest, blood component transfusion, post-partum complication and stillbirth than healthy controls. Statistical analysis of comparison between group 2 and 3 has revealed that; patients with single elevated OGTT had more peripartum complication (p = .032; odds ratio [OR] = 1.2, 95% CI: 1.02-1.54), had more babies with macrosomia (p < .001; [OR] = 1.7, 95% CI: 1.2-2.4), had more postpartum complication (p = .040; [OR] = 3, 95% CI: 0.997-9.1), and had higher cesarean rates (p < .001; [OR] = 1.29, 95% CI: 1.1-1.4).Evaluating the first trimester CBC parameters between groups; only PLR differed statistically significant in GDM patients. These parameters before delivery were also analyzed PLR and NLR values did not differ between all groups, on the other hand; MPV values were higher and PDW values were lower in healthy controls comparing GDM and single elevated OGTT group. CONCLUSION: Patients with single elevated OGTT had a higher risk of maternal and neonatal consequences than women with normal OCT, which was comparable levels to patients with GDM. These patients should not be underestimated and could be classified as an individual diagnose such as "Borderline GDM." To intervene in these patients with dietary advice and lifestyle changes like exercise could decrease neonatal and maternal adverse outcomes.

Recurrent sigmoid volvulus: Cause of colon perforation, sepsis, and fetal death.

Sigmoid volvulus is one of the most common reasons for intestinal obstruction. Recurrent sigmoid volvulus occurred in only a few cases. Torsion of the sigmoid colon around the mesenteric portion initiates ischemia, gangrene, and perforation. Here, we presented a case with recurrent sigmoid volvulus. A 19-year-old Syrian refugee has been admitted to emergency room during 30th gestation week with acute abdomen, constipation, and lack of gas passage. She had a medical history of rectosigmoidoscopy and detorsion of sigmoid volvulus 6 days ago. Her vital signs showed tachycardia, hypotension, and tachypne, and her body temperature was 35.6°C. Sonography revealed nonviable fetus. Emergency laparotomy has been performed. Sigmoid volvulus and contaminated intraperitoneal area with feces was evaluated. A cesarean section, total colectomy, and end-ileostomy have been performed. Patient stayed 3 days in intensive care unit and was discharged on the 17th day. Sigmoid volvulus is a real emergency. Delayed treatment of sigmoid volvulus is related to maternal and fetal mortality.

Determinants of cesarean-related complications: high number of repeat cesarean, operation type or placental pathologies?

BACKGROUND: Cesarean delivery (CD) is one of the most common operations worldwide. Vaginal birth after cesarean (VBAC) could be a solution to decrease increased CD rates. On the other hand, risks of VBAC on maternal and neonatal outcomes drifts physicians and patients to a scheduled CD. Successive CDs, especially after the 3rd operation, increase complications for the fetus and the mother. Operation type (emergency or elective CD) could be a risk factor of increased morbidities, like placental implantation anomalies. Evaluation of these conditions related to complications and morbidities were investigated. Material and methods: Women who underwent the fourth and more repeat CD in Bursa Yuksek Ihtisas Training Research Hospital between March 2016 and December 2019 were retrospectively reviewed. Pre-operative characteristics, per-operative and post-operative complications were reviewed. Patients were separated into groups as operational type, repeat cesarean number, and major morbidities. A comparison between groups was evaluated. Results: A total of 46.048 women gave birth, of which 17,721 underwent CDs with a rate of 38%. The rate of primary CD was 18%. The number of the fourth or more CD performed was 854. The number of patients who underwent fourth and fifth or more CD and of these operational data could be accessed was 599 and 145, respectively. The overall complications were detected as severe adhesions (n: 220), preterm delivery (n: 91), stillbirth (n: 9), admission to NICU (n: 98), bladder injury (n: 10), uterine scar dehiscence (n: 6), uterine rupture (n: 6), uterine atony (n: 26), blood transfusion requirement (n: 68), preterm delivery (n: 91), placenta previa totalis (n: 24), morbidly adherent placenta (n: 14), hysterectomy (n: 12), partial uterine resection (n: 2), uterus-conserving interventions (n: 26). The number of patients with major morbidity was 105. Emergency cesarean performed in 339 of 744 patients. A comparison of the emergency cesarean group with elective repeat cesarean group revealed no significant difference in operative adverse outcomes. Comparing patients between 4th repeat CD with 5th and more CD revealed a significant difference in severe adhesion, morbidly adherent placenta and hysterectomy. Previa totalis were detected in 24 patients. All of them experienced major morbidity with 12 of them underwent hysterectomy. The rest of them performed Uterus-conserving treatments (B-Lynch Suture, Bacri Balloon, Hypogastric artery ligation ) and a total of 51 units of packed red blood cells and 32 units of Fresh Frozen Plasma were transfused to 9 (37%) of 24 patients. Conclusion: The major risk factor of the morbidity is placenta previa whose incidence has dramatically increased after 3rd cesarean. Emergency cesarean did not increase the complication rate in the present study. Fourth and more repeat CDs ought to be performed by experienced obstetricians in high-equipped tertiary hospitals.

Double-Binding Botulinum Molecule with Reduced Muscle Paralysis: Evaluation in In Vitro and In Vivo Models of Migraine.

With a prevalence of 15%, migraine is the most common neurological disorder and among the most disabling diseases, taking into account years lived with disability. Current oral medications for migraine show variable effects and are frequently associated with intolerable side effects, leading to the dissatisfaction of both patients and doctors. Injectable therapeutics, which include calcitonin gene-related peptide-targeting monoclonal antibodies and botulinum neurotoxin A (BoNT/A), provide a new paradigm for treatment of chronic migraine but are effective only in approximately 50% of subjects. Here, we investigated a novel engineered botulinum molecule with markedly reduced muscle paralyzing properties which could be beneficial for the treatment of migraine. This stapled botulinum molecule with duplicated binding domain-binary toxin-AA (BiTox/AA)-cleaves synaptosomal-associated protein 25 with a similar efficacy to BoNT/A in neurons; however, the paralyzing effect of BiTox/AA was 100 times less when compared to native BoNT/A following muscle injection. The performance of BiTox/AA was evaluated in cellular and animal models of migraine. BiTox/AA inhibited electrical nerve fiber activity in rat meningeal preparations while, in the trigeminovascular model, BiTox/AA raised electrical and mechanical stimulation thresholds in Aδ- and C-fiber nociceptors. In the rat glyceryl trinitrate (GTN) model, BiTox/AA proved effective in inhibiting GTN-induced hyperalgesia in the orofacial formalin test. We conclude that the engineered botulinum molecule provides a useful prototype for designing advanced future therapeutics for an improved efficacy in the treatment of migraine.

Developmental differences of kinematic and muscular activation patterns in instep soccer kick.

Kinematic and neuromuscular activity differences amongst soccer players in different age groups were examined in this study. Thirty male soccer players evenly divided into three age groups (Group 1: age 12-13; Group 2: age 14-15; Group 3: age 16-17) were asked to perform instep kicks towards a target 11 m away. Their anthropometrics, instep kick kinematics, resultant ball velocities, both legs isokinetic strength, and electromyography (EMG) during kicking were compared amongst the three age groups. There were significant differences in height, body mass, body mass index, ball velocities, and isokinetic strength values amongst three age groups. Also, kicking kinematics including angular and linear velocities of hip, knee, ankle, and toe were significantly different (p < 0.05) amongst groups in several kicking phases. Furthermore, the activities of m. rectus femoris, m. vastus medialis, m. biceps femoris were significantly different amongst groups (p < 0.05). The ball velocities and leg strength parameters increased with age, neuromuscular activations, and kinematic parameters differed especially in leg-coking and forward swing phase of instep soccer kick. It should be concluded that an increase of resultant ball velocity of the instep kick is closely associated with chronical age, the development of leg muscle strength, and the neuromuscular activity of the kicking leg.

Duplication of clostridial binding domains for enhanced macromolecular delivery into neurons.

Neurological diseases constitute a quarter of global disease burden and are expected to rise worldwide with the ageing of human populations. There is an increasing need to develop new molecular systems which can deliver drugs specifically into neurons, non-dividing cells meant to last a human lifetime. Neuronal drug delivery must rely on agents which can recognise neurons with high specificity and affinity. Here we used a recently introduced 'stapling' system to prepare macromolecules carrying duplicated binding domains from the clostridial family of neurotoxins. We engineered individual parts of clostridial neurotoxins separately and combined them using a strong alpha-helical bundle. We show that combining two identical binding domains of tetanus and botulinum type D neurotoxins, in a sterically defined way by protein stapling, allows enhanced intracellular delivery of molecules into neurons. We also engineered a botulinum neurotoxin type C variant with a duplicated binding domain which increased enzymatic delivery compared to the native type C toxin. We conclude that duplication of the binding parts of tetanus or botulinum neurotoxins will allow production of high avidity agents which could deliver imaging reagents and large therapeutic enzymes into neurons with superior efficiency.

A genetic approach to study polyubiquitination in Saccharomyces cerevisiae.

Ubiquitin has seven lysines, all of which are used to generate polyubiquitin chains in the yeast Saccharomyces cerevisiae. While the biology associated with chains formed through lysines 48 and 63 is well studied, other chain types are more poorly characterized. We outline a methodology for using synthetic genetic analysis to examine ubiquitin mutants. Ubiquitin is encoded by four loci, necessitating several alterations to standard protocols, including the use of the SK1 strain background, which sporulates with very high efficiency. The methods described here could be used to examine other ubiquitin mutants, including those that do not support viability.

Prediction of neonatal respiratory distress syndrome via pulmonary artery Doppler examination.

OBJECTIVE: We aimed with this study to evaluate the role of pulmonary artery acceleration time to ejection time ratio (PATET) in the prediction of respiratory distress syndrome (RDS) in preterm neonates. MATERIALS AND METHODS: In this prospective cohort study, 105 singleton pregnant women with no congenital abnormalities and pregnancy complications who delivered before 37 weeks of gestational age were included. All the patients underwent ultrasound examination to obtain fetal pulmonary artery Doppler. 15 patients were excluded from the study as they did not give birth within 3 days subsequent to ultrasound examination, or inadequate Doppler measurements. After delivery the neonates were grouped according to diagnosis of RDS as RDS + and RDS-. RESULTS: One hundred five women met the inclusion criteria. Regarding the Doppler findings; only the PATET ratio was significantly different between the groups (0.2965 ± 0.042 versus 0.386 ± 0.068 p < .001, Z = -5.206). There was an inverse correlation between the diagnosis of RDS in the neonates and the PATET values, even after adjusted for gestational age estimated fetal weight and fetal gender (r = -0.52 and p = .0017). A cut-off value of 0.327 provided optimal specificity of 77.1%, a sensitivity of 90.9%, a negative predictive value of 95.4%, and a positive predictive value 52.7%. CONCLUSIONS: In consideration of these results fetal PATET ratio is a promising noninvasive tool to predict RDS in cases of preterm deliveries.

Genetic analysis reveals functions of atypical polyubiquitin chains.

Although polyubiquitin chains linked through all lysines of ubiquitin exist, specific functions are well-established only for lysine-48 and lysine-63 linkages in Saccharomyces cerevisiae. To uncover pathways regulated by distinct linkages, genetic interactions between a gene deletion library and a panel of lysine-to-arginine ubiquitin mutants were systematically identified. The K11R mutant had strong genetic interactions with threonine biosynthetic genes. Consistently, we found that K11R mutants import threonine poorly. The K11R mutant also exhibited a strong genetic interaction with a subunit of the anaphase-promoting complex (APC), suggesting a role in cell cycle regulation. K11-linkages are important for vertebrate APC function, but this was not previously described in yeast. We show that the yeast APC also modifies substrates with K11-linkages in vitro, and that those chains contribute to normal APC-substrate turnover in vivo. This study reveals comprehensive genetic interactomes of polyubiquitin chains and characterizes the role of K11-chains in two biological pathways.

The protective effect of Gingko biloba in a rat model of ovarian ischemia/reperfusion injury: Improvement in histological and biochemical parameters.

BACKGROUND: Ovarian torsion is one of the most common gynecological emergencies, which especially affects women of reproductive age. OBJECTIVES: We aimed to evaluate the effect of Ginkgo biloba (GB) supplementation in ovarian ischemia/ reperfusion injury in an experimental torsion/de-torsion rat model. MATERIAL AND METHODS: This study was carried out in the Ege University Faculty of Medicine in Izmir, Turkey. Thirty mature female Sprague-Dawley albino rats were randomly divided into 5 groups: in Group 1 (control), the abdominal wall was only opened and closed; in the torsion group (Group 2), ischemia was induced for 3 h, using atraumatic vascular clips to create a torsion model; in the torsion/GB group (Group 3), the rats were given 80 mg/kg (oral gavage) of GB 30 min before torsion was induced and the torsion model was formed; in the torsion/de-torsion group (Group 4), the rats underwent 3 h of ischemia and then the vascular clips were removed and reperfusion took place for 3 h; in the torsion/de-torsion/GB group (Group 5), the rats underwent 3 h of ischemia followed by GB (oral gavage) 30 min prior to a 3-h reperfusion period. Ovarian tissue damage was evaluated by a histopathological scoring system. Ovarian tissue malondialdehyde (MDA) and plasma pentraxin-3 were measured. RESULTS: In comparison with the sham group, both the torsion and torsion/de-torsion groups had significantly higher scores for follicular degeneration, vascular congestion, edema, hemorrhage, and leukocyte infiltration. Ginkgo biloba significantly decreased these scores in both groups. Ovarian malondialdehyde and plasma pentraxin 3 were significantly higher both in the torsion and torsion/de-torsion groups compared with the sham group. Ginkgo biloba decreased these levels significantly both in the torsion/GB and torsion/de-torsion/GB groups. CONCLUSIONS: Supplementing GB during a surgical procedure decreases ischemia/reperfusion injury to an ovary in an experimental rat model based on histopathological parameters, tissue malondialdehyde, and plasma pentraxin-3 levels.

Relationship between first trimester aneuploidy screening test serum analytes and placenta accreta.

OBJECTIVE: The aim of this study is to determine whether there is a relationship between first trimester serum pregnancy-associated plasma protein A (PAPP-A) and free beta human chorionic gonadotropin (fßhCG) MoM values and placenta accreta in women who had placenta previa. STUDY DESIGN: A total of 88 patients with placenta previa who had first trimester aneuploidy screening test results were enrolled in the study. Nineteen of these patients were also diagnosed with placenta accreta. As probable markers of excessive placental invasion, serum PAPP-A and fßhCG MoM values were compared in two groups with and without placenta accreta. RESULTS: Patients with placenta accreta had higher statistically significant serum PAPP-A (1.20 versus 0.865, respectively, p = 0.045) and fßhCG MoM (1.42 versus 0.93, respectively, p = 0.042) values than patients without accreta. CONCLUSIONS: Higher first trimester serum PAPP-A and fßhCG MoM values seem to be associated with placenta accreta in women with placenta previa. Further studies are needed to use these promising additional tools for early detection of placenta accreta.

Different fatigue-resistant leg muscles and EMG response during whole-body vibration.

The purpose of this study was to determine the effects of static whole-body vibration (WBV) on the Electromyograhic (EMG) responses of leg muscles, which are fatigue-resistant in different manner. The study population was divided into two groups according to the values obtained by the Fatigue Index [Group I: Less Fatigue Resistant (LFR), n=11; Group II: More Fatigue Resistant (MFR), n=11]. The repeated electromyographic (EMG) activities of four leg muscles were analyzed the following determinants: (1) frequency (30 Hz, 35 Hz and 40 Hz); (2) stance position (static squat position); (3) amplitude (2 mm and 4 mm) and (4) knee flexion angle (120°), (5) vertical vibration platform. Vibration data were analyzed using Minitab 16 (Minitab Ltd, State College, PA, USA). The significance level was set at p<.05. The study results showed that static WBV stimuli given at different frequencies and amplitudes resulted in a significant increase (p<.05) in compared, the LFR group showed significantly (1) higher rates of quadriceps femoris and hamstring muscle fatigue (p<.05), (2) higher levels of knee extensor and flexor torque (p<.05) and (3) higher percentage increases in EMG activation at higher frequencies (max at 40 Hz) and amplitudes (4 mm) (p<.05). The present study can be used for the optimal prescription of vibration exercise and can serve to guide the development of training programs.