Bovine blastocyst development in vitro: timing, sex, and viability following vitrification.
Selection of blastocysts based on their morphological characteristics and rate of development in vitro can skew the sex ratios. The aim of this study was to determine whether an embryo's developmental rate affects its survival after vitrification, and whether male and female embryos survive vitrification differently. In vitro fertilized bovine oocytes were cultured in potassium simplex optimized medium (KSOM) + 0.1% BSA for 96 h, and then into KSOM + 1% BSA (KSOM) or in sequential KSOM + 0.1% BSA for 96 h, and then into synthetic oviduct fluid (SOF) + 5% FBS (KSOM-SOF). In part 1 of this study, embryos cultured in each medium that had developed into blastocysts at approximately 144, 156, or 180 h were recovered from culture, graded, and then vitrified. After warming, blastocyst survival rates were immediately evaluated by reexpansion of the blastocoels. In the second part of the study, all blastocysts (n = 191) were sexed by polymerase chain reaction 48 h after warming. When cultured in KSOM medium, more 144-h blastocysts survived vitrification (68%) than blastocysts vitrified at 180 h (49%). Blastocysts derived at 156 h in KSOM-SOF survived vitrification better (87%) than blastocysts vitrified at either 144 h or 180 h, and subsequently hatched at a greater rate than those vitrified at 180 h. The overall blastocyst survival rates did not differ significantly whether embryos were cultured in KSOM or sequential KSOM-SOF. Blastocysts derived at 144 and 156 h in KSOM or KSOM-SOF were predominately male, and significantly more of them survived vitrification 48 h after warming. However, blastocysts cultured in KSOM-SOF, and then vitrified at 180 h were predominately female. Overall, blastocysts that survived vitrification, and subsequently hatched 48 h after warming, were male. In summary, embryos that reached the blastocyst stage earlier were predominantly males; these males had better morphology, endured vitrification, and subsequently hatched at a greater rate than did female blastocysts.


A Study on the N2O Reduction Rate According to Temperature and Residence Time in the Exhaust Gas Atmosphere Emitted on Combustion of Air and Oxygen.
N2O is a hazardous greenhouse gas. It should be reduced to solve global warming problems. In this study, experiments of N2O thermal decomposition were conducted by simulating the exhaust gas atmosphere emitted during the combustion of air and pure oxygen in an actual circulating fluidized bed system and incinerator system. As a result of comparing the N2O reduction rate in N2 and CO2 atmospheres, the N2O reduction rate in the CO2 atmosphere was 20% higher than that in the N2 atmosphere. It is judged that the N2O reduction rate is high in a CO2 atmosphere (exhaust gas from pure oxygen combustion) due to complex factors such as the reverse reaction, the diffusion coefficient, and static pressure-specific heat. Therefore, pure oxygen combustion increases the reduction rate of nitrous oxide. In addition, when operated with an appropriate residence time and temperature, a reduction effect of more than 95% can be expected, and the fuel consumption rate is also expected to improve.


Biomechanics of the knee during closed kinetic chain and open kinetic chain exercises.
Although closed (CKCE) and open (OKCE) kinetic chain exercises are used in athletic training and clinical environments, few studies have compared knee joint biomechanics while these exercises are performed dynamically. The purpose of this study was to quantify knee forces and muscle activity in CKCE (squat and leg press) and OKCE (knee extension). Ten male subjects performed three repetitions of each exercise at their 12-repetition maximum. Kinematic, kinetic, and electromyographic data were calculated using video cameras (60 Hz), force transducers (960 Hz), and EMG (960 Hz). Mathematical muscle modeling and optimization techniques were employed to estimate internal muscle forces. Overall, the squat generated approximately twice as much hamstring activity as the leg press and knee extensions. Quadriceps muscle activity was greatest in CKCE when the knee was near full flexion and in OKCE when the knee was near full extension. OKCE produced more rectus femoris activity while CKCE produced more vasti muscle activity. Tibiofemoral compressive force was greatest in CKCE near full flexion and in OKCE near full extension. Peak tension in the posterior cruciate ligament was approximately twice as great in CKCE, and increased with knee flexion. Tension in the anterior cruciate ligament was present only in OKCE, and occurred near full extension. Patellofemoral compressive force was greatest in CKCE near full flexion and in the mid-range of the knee extending phase in OKCE. An understanding of these results can help in choosing appropriate exercises for rehabilitation and training.


Correlation between in vitro and in vivo response to androgens in patients with aplastic anemia.
In an attempt to predict the clinical response to androgens, the effects of added fluoxymesterone (FMT) on the in vitro growth of marrow erythroid progenitors (CFU-E) were examined in 15 patients with aplastic anemia and 6 control subjects with normal marrow morphology. In the control group, the addition of FMT enhanced the growth of CFU-E in vitro, the maximum absolute increase being dependent on the basal number of CFU-E. In 10 out of the 11 aplastic anemia patients who responded to androgens in vivo. FMT enhanced the growth of CFU-E in vitro. In this group, there was a positive correlation between the basal number of CFU-E and the maximum absolute increase of CFU-E induced by FMT. In patients who did not respond to androgens in vivo, all except one showed no increase in the growth of CFU-E in vitro by the addition of FMT. Furthermore, in four out of seven patients examined prior to androgen therapy, the addition of FMT enhanced the in vitro growth of CFU-E. Androgen therapy in vivo resulted in marked hematological improvements in all of these four patients. Two out of the remaining three patients in whom FMT did not enhance the in vitro growh of CFU-E failed to respond in androgen therapy. These results indicate that an in vitro CFU-E culture system to the presence of FMT may be helpful in predicting the response to androgens in vivo in patients with aplastic anemia.


Opening up about Tissue Transglutaminase: When Conformation Matters More than Enzymatic Activity.
Tissue transglutaminase (tTG), also referred to as type 2 transglutaminase or Gαh, can bind and hydrolyze GTP, as well as function as a protein crosslinking enzyme. tTG is widely expressed and can be detected both inside cells and in the extracellular space. In contrast to many enzymes, the active and inactive conformations of tTG are markedly different. The catalytically inactive form of tTG adopts a compact "closed-state" conformation, while the catalytically active form of the protein adopts an elongated "open-state" conformation. tTG has long been appreciated as an important player in numerous diseases, including celiac disease, neuronal degenerative diseases, and cancer, and its roles in these diseases often depend as much upon its conformation as its catalytic activity. While its ability to promote these diseases has been traditionally thought to be dependent on its protein crosslinking activity, more recent findings suggest that the conformational state tTG adopts is also important for mediating its effects. In particular, we and others have shown that the closed-state of tTG is important for promoting cell growth and survival, while maintaining tTG in the open-state is cytotoxic. In this review, we examine the two unique conformations of tTG and how they contribute to distinct biological processes. We will also describe how this information can be used to generate novel therapies to treat diseases, with a special focus on cancer.


Radioisotopic measurements of coronary blood flow.
This review has presented a critical evaluation of current methods used in measurement of total and regional blood flow. Measurement of total coronary blood flow furnishes little evidence of pathological processes within the coronary vascular system. However, in conjunction with coronary vasodilators, measurement of total coronary blood flow may indicate the presence of coronary artery disease. Difficulties inherent in the determination of regional coronary blood flow lie primarily in defining clearance slopes and in the presence of accidental counts. More promising are methods using scintigraphs obtained with radioactive particles injected into the coronary artery. However, these procedures do not measure flow, only flow distribution. A procedure for measuring regional uptake of positron emitters by coincidence counting has been described; the method possesses definite advantages over other procedures, since it is noninvasive and appears to have good resolution. However, the method is dependent on higher decay density which necessitates the use of nuclides with a short half-life (e.g., 81rubidium).


The Cognitive Profile of Math Difficulties: A Meta-Analysis Based on Clinical Criteria.
Math difficulties (MD) manifest across various domain-specific and domain-general abilities. However, the existing cognitive profile of MD is incomplete and thus not applicable in typical settings such as schools or clinics. So far, no review has applied inclusion criteria according to DSM or ICD, summarized domain-specific abilities or examined the validity of response time scores for MD identification. Based upon stringent clinical criteria, the current meta-analysis included 34 studies which compared cognitive performances of a group with MD (n = 680) and a group without MD (n = 1565). Criteria according to DSM and ICD were applied to identify MD (percentile rank ≤ 16, age range 8-12 years, no comorbidities/low IQ). Effect sizes for 22 abilities were estimated and separated by their level and type of scoring (AC = accuracy, RT = response time). A cognitive profile of MD was identified, characterized by distinct weaknesses in: (a) computation (calculation [AC], fact retrieval [AC]), (b) number sense (quantity processing [AC], quantity-number linking [RT], numerical relations [AC]), and (c) visual-spatial short-term storage [AC]. No particular strength was found. Severity of MD, group differences in reading performance and IQ did not significantly moderate the results. Further analyses revealed that (a) effects are larger when dealing with numbers or number words than with quantities, (b) MD is not accompanied by any weakness in abilities typically assigned to reading, and (c) weaknesses in visual-spatial short-term storage emphasize the notion that number and space are interlinked. The need for high-quality studies investigating domain-general abilities is discussed.


A Novel Z-Ring Associated Protein ZapA-Like Protein (PA5407) From Pseudomonas aeruginosa Promotes FtsZ to Form Double Filaments.
Bacterial cell division is initiated by the assembly of the contraction ring (Z-ring), which consists of the self-assembled FtsZ protofilaments and dozens of other associate proteins. ZapA, a regulatory protein found in almost all bacteria, stabilizes FtsZ protofilaments to form bundles and enhances the Z-ring condensation. Here, we reported that another small protein from Pseudomonas aeruginosa, ZapA-Like protein (ZapAL; PA5407), is a new FtsZ associated protein. ZapAL exists in many Pseudomonas species and shares only 20% sequence identity to ZapA. ZapAL interacts with FtsZ and induces FtsZ to form long straight double filaments; in comparison, ZapA promotes long bundles with multiple FtsZ filaments. ZapAL has only a mild effect on GTPase activity of FtsZ, which is reduced by around 26% when 10 μM ZapAL is added in the solution. However, to study their assembly dynamics using light-scattering assay, we found that FtsZ-ZapAL double filament is stable and no depolymerization process is observed, which is different from ZapA. Further research found that ZapA and ZapL are likely to form heterodimers. The bundles formed by the mixture of FtsZ-ZapA-ZapAL will depolymerize after GTP is hydrolyzed. Consistent with ZapAL interaction with FtsZ in vitro, the expression of ZapAL-GFP was observed as a narrow band or spots in the middle of the cells, suggesting that it is a component of bacterial division machinery. Similar to ZapA, ZapAL is also not essential for bacterial cell division. Little changes were observed when zapAL gene was deleted, or overexpressed under normal conditions; however, overexpression of ZapAL caused zapA-deficient cells to grow approximately two times longer, showing a mild bacterial division defect. Although we still do not know the exact physiological roles of ZapAL, our results suggest that ZapAL is a novel Z-ring associate protein, which may work together with ZapA to stabilize the FtsZ protofilament and Z-ring structure.


Activation of Primary and Secondary Benzylic and Tertiary Alkyl (sp 3 )C-F Bonds Inside a Self-Assembled Molecular Container.
Alkyl fluorides are generally regarded as chemically inert. However, several literature examples describe the activation of alkyl (sp3)C-F bonds via strong Brønsted or Lewis acids under harsh conditions. We here report that catalytic amounts of the self-assembled resorcinarene capsule are able to activate alkyl (sp3)C-F bonds under mild conditions (40°C, no strong Brønsted or Lewis acid present). Kinetic measurements display a sigmoidal reaction progress after an initial induction period. Control experiments indicate that the presence of the supramolecular capsule is required for an efficient reaction acceleration.


[Statistic evaluation of prognostic risk factors in penetrating keratoplasty using Cox multiple regression model].
Preoperative risk factors that may influence the prognosis in penetrating keratoplasty were statistically analyzed on 698 consecutive eye that had been operated on at the University of Tokyo Hospital. A Cox multiple regression model (proportional hazards models) was constructed with simultaneous consideration of follow-up periods and prognostic factors. Six preoperative risk factors were identified: corneal endothelial damage, the presence of anterior synechiae, glaucoma, older donor age, the area of vascularization, and aphakia or pseudophakia. Simulations of the prognosis were attempted using the significant factors.


Depression in general practice.
The nature and course of depressive disorders in a group of general practice patients were assessed by screening 564 subjects who attended representative practices in Sydney by means of a self-report depression inventory. Of these, 25% of women and 17% of men reported significant depressive symptoms. An interview was sought with these "potential depressives" to determine, in particular, the extent to which they might be considered to have a psychiatric disorder, using the PSE case-finding procedure. Of those interviewed, 83% scored as psychiatric "cases", while the mean duration of their depression was nine months. A longitudinal component to the study suggested that the mental condition of the sample had improved minimally at six weeks' and at 20 weeks' follow-up assessments.


Cross-cultural adaptation, validity and reliability of the Korean version of the Kerlan-Jobe Orthopedic Clinic shoulder and elbow score.
The Kerlan-Jobe Orthopedic Clinic shoulder and elbow score (KJOC score) was developed for assessing performance and function in overhead athletes with shoulder and elbow injury and recently adapted as the standard score for overhead athletes in Major League Baseball. However, the Korean version of the KJOC score was not developed in the literature. The aim of the current study was to adapt the English version of the KJOC score to develop a Korean version (K-KJOC) and to evaluate its validity and reliability. A total of 52 professional baseball players in the Korean Baseball League completed the K-KJOC at two-week intervals during the off-season. The QuickDASH (11-point Disabilities of the Arm, Shoulder and Hand) score was also performed to evaluate the construct validity of the K-KJOC score. The internal consistency of reliability and test-retest reliability were assessed as well. The K-KJOC score was correlated with the Quick DASH - disability/symptom (-0.309, ~-0.268, p < 0.05), Quick DASH - work (r = -0.721 to -0.671, p = 0.000) and QuickDASH - sports (r = -0.721 to -0.714, p = 0.000). The internal consistency of the K-KJOC score was excellent (Cronbach's α: 0.917-0.966), and the intra-class correlation coefficients of test-retest reliability for the 10 items for the K-KJOC score were fair to excellent (ICC 0.505-0.937, p < 0.05). The K-KJOC score appeared to be a valid and reliable tool for assessing shoulder and elbow injuries in Korean overhead athletes.


Generation and characterization of function-blocking anti-ectodysplasin A (EDA) monoclonal antibodies that induce ectodermal dysplasia.
Development of ectodermal appendages, such as hair, teeth, sweat glands, sebaceous glands, and mammary glands, requires the action of the TNF family ligand ectodysplasin A (EDA). Mutations of the X-linked EDA gene cause reduction or absence of many ectodermal appendages and have been identified as a cause of ectodermal dysplasia in humans, mice, dogs, and cattle. We have generated blocking antibodies, raised in Eda-deficient mice, against the conserved, receptor-binding domain of EDA. These antibodies recognize epitopes overlapping the receptor-binding site and prevent EDA from binding and activating EDAR at close to stoichiometric ratios in in vitro binding and activity assays. The antibodies block EDA1 and EDA2 of both mammalian and avian origin and, in vivo, suppress the ability of recombinant Fc-EDA1 to rescue ectodermal dysplasia in Eda-deficient Tabby mice. Moreover, administration of EDA blocking antibodies to pregnant wild type mice induced in developing wild type fetuses a marked and permanent ectodermal dysplasia. These function-blocking anti-EDA antibodies with wide cross-species reactivity will enable study of the developmental and postdevelopmental roles of EDA in a variety of organisms and open the route to therapeutic intervention in conditions in which EDA may be implicated.


Polymerization transitions in two-dimensional systems of dipolar spheres.
We investigate the self-organization of dipolar spheres into polymer chains as a fundamental model of the self-assembly of particles having anisotropic interparticle interactions. Our study involves a combination of modeling with vertically vibrated magnetic beads simulating a quasi-two-dimensional fluid at equilibrium and corresponding Monte Carlo simulations of hard spheres with embedded extended dipoles. We find a transition from a gas-like phase to a polymerized phase upon cooling, in accord with the analytic theory of equilibrium polymerization.


STANDING COMPUTED TOMOGRAPHY IN NONANESTHETIZED LITTLE PENGUINS (EUDYPTULA MINOR) TO ASSESS RESPIRATORY SYSTEM ANATOMY AND MONITOR DISEASE.
Multidetector computed tomography (MDCT) scans were performed in clinically healthy, nonanesthetized, standing little penguins (Eudyptula minor) to determine reference ranges for air-sac and lung volumes, as well as lung density. Five of 15 clinically healthy birds were diagnosed with pulmonary granulomas on initial MDCT scans. Granulomas were not readily apparent on radiographs, even in cases where the entire normal pulmonary parenchymal architecture was effaced on the MDCT scan. Serial MDCT scans after antifungal and antimycobacterial therapies demonstrated a response to treatment. MDCT scanning in nonanesthetized little penguins proved to be a well-tolerated, non-invasive imaging modality for respiratory diseases that are otherwise difficult to diagnose, including aspergillosis and mycobacteriosis.


Retroviral reverse transcriptase: synthesis, structure, and function.
The virion particles of all retroviruses contain reverse transcriptase, a multifunctional enzyme required for the synthesis of a DNA copy of the viral RNA genome soon after entry into the infected cell. The enzyme is the target of the major antiviral drugs currently in use in the treatment of AIDS. This article reviews the strategies used by different retroviruses for the synthesis of the enzyme; the structure of the enzymes encoded by various viruses; the activities inherent in the enzyme; and the application of those activities to the process of reverse transcription of the viral RNA.


[Modifications of sleep "in utero" in guinea pig after maternal administration of alpha-methyl-L-DOPA (author's transl)].
Effects of maternal administration of alpha-methyl-L-DOPA on sleep in utero were studied by polygraphic method in Guinea Pig faetus from the 53th day of gestation. Maternal injection of alpha-methyl-L-DOPA (100 and 200 mg/kg) produces a typical sedative effect on the faetus from the first hour after drug administration. But, effects appear attenuated in comparison to those of the new-born Guinea Pig: a paradoxical sleep-deprivation of only 30% is observed during the 12 hours period after drug injection. Causes of the attenuation in utero of these drug effects are discussed.


Time Resolved Measurements of the Switching Trajectory of Pt/Co Elements Induced by Spin-Orbit Torques.
We report the experimental observation of spin-orbit torque induced switching of perpendicularly magnetized Pt/Co elements in a time resolved stroboscopic experiment based on high resolution Kerr microscopy. Magnetization dynamics is induced by injecting subnanosecond current pulses into the bilayer while simultaneously applying static in-plane magnetic bias fields. Highly reproducible homogeneous switching on time scales of several tens of nanoseconds is observed. Our findings can be corroborated using micromagnetic modeling only when including a fieldlike torque term as well as the Dzyaloshinskii-Moriya interaction mediated by finite temperature.


Diamides of cyclobutane-1,1-dicarbocylic acid. II.
A group of nine diamides of cyclobutane-1,1-dicarboxylic acid has been prepared and examined for general depressant and analgesic activity. Several showed activity in both respects but with little overlap. Several compounds were evaluated for anticonvulsant and myorelaxant activity. No activity was found.


[Tolerance and efficacy of proglumetacine in patients with rheumatoid arthritis].
The efficacy of proglumetacin , a new non-steroidal antiinflammatory drug, was assessed in 32 patients with rheumatoid arthritis. During treatment with 400-650 mg daily of proglumetacin over a period of 7-14 days, morning stiffness and side-effects were checked weekly or in severely ill patients daily. All patients but one completed the period of treatment. In spite of the short period of observation, a significant improvement was seen in the majority of cases (55%), while in 39% proglumetacin was not more effective than treatments before the admission to the study. In the group of patients treated for 14 days, morning stiffness parameters showed a significant improvement after 7 days and at the end of the period of study. Overall , only 3 patients referred side-effects: 1 case of transient headache and 2 cases of severe gastric pain. In our preliminary study, proglumetacin results to be effective as an antiinflammatory drug also in severe rheumatoid arthritis and safe for its low incidence of side-effects.


Antibiotic sensitivity pattern of Citrobacter isolated from various clinical specimens in a tertiary care hospital.
A total of 709 isolates of Citrobacter species were recovered from different clinical specimens at Kasturba Medical College, Mangalore. The antimicrobial susceptibility testing of the isolates was performed using Kirby-Bauer's method of disk diffusion. All the isolates were susceptible to Imipenem (100%) and 100% resistance to Ampicillin was recorded. Susceptibility to third generation cephalosporins varied between 29 - 43%. The beta-lactamase inhibitor combination used along with beta-lactam antibiotics increased their sensitivity. The minimum inhibitory concentration (MIC) of cephotaxime to Citrobacter species was determined by agar dilution method (MIC value = or < 8 microg/ml are considered susceptible). Two hundred and fifty three strains were found to be completely susceptible, 71 strains showed intermediate susceptibility and 385 strains were completely resistant to Cephotaxime.


Pulmonary vein stenosis in patients with Smith-Lemli-Opitz syndrome.
To describe a group of children with co-incident pulmonary vein stenosis and Smith-Lemli-Opitz syndrome and to generate hypotheses as to the shared pathogenesis of these disorders. Retrospective case series. Five subjects in a pulmonary vein stenosis cohort of 170 subjects were diagnosed with Smith-Lemli-Opitz syndrome soon after birth. All five cases were diagnosed with Smith-Lemli-Opitz syndrome within 6 weeks of life, with no family history of either disorder. All cases had pathologically elevated 7-dehydrocholesterol levels and two of the five cases had previously reported pathogenic 7-dehydrocholesterol reductase mutations. Smith-Lemli-Opitz syndrome severity scores ranged from mild to classical (2-7). Gestational age at birth ranged from 35 to 39 weeks. Four of the cases were male by karyotype. Pulmonary vein stenosis was diagnosed in all cases within 2 months of life, earlier than most published cohorts. All cases progressed to bilateral disease and three cases developed atresia of at least one vein. Despite catheter and surgical interventions, all subjects' pulmonary vein stenosis rapidly recurred and progressed. Three of the subjects died, at 2 months, 3 months, and 11 months. Survival at 16 months after diagnosis was 43%. Patients with pulmonary vein stenosis who have a suggestive syndromic presentation should be screened for Smith-Lemli-Opitz syndrome with easily obtainable serum sterol tests. Echocardiograms should be obtained in all newly diagnosed patients with Smith-Lemli-Opitz syndrome, with a low threshold for repeating the study if new respiratory symptoms of uncertain etiology arise. Further studies into the pathophysiology of pulmonary vein stenosis should consider the role of cholesterol-based signaling pathways in the promotion of intimal proliferation.


Implant rehabilitation of edentulous jaws with predominantly monolithic zirconia compared to metal-acrylic prostheses: a 2-year retrospective clinical study.
Limited data are available on the clinical outcomes of patients with edentulism treated with predominantly monolithic zirconia fixed complete dentures (FCDs) compared to traditional restoration materials. The purpose of this study was to analyze the differences in terms of complications and failures of definitive full-arch implant rehabilitations made in metal-acrylic versus those made in monolithic zirconia with porcelain veneering limited to non-functional areas. This retrospective clinical study included 50 patients treated between January 2015 and December 2018, with 222 implants inserted in fifty edentulous jaws. All patients were treated with immediately loaded full-arch fixed prostheses (22 maxillary; 28 mandibular) each supported by four to six implants (two/four axial, two distally tilted). All 25 zirconia prostheses were predominantly monolithic with ceramic veneering limited to non-functional areas. The primary outcome measures were prosthetic success of the definitive restoration and implant survival. The secondary outcome measures were full mouth plaque score, full mouth bleeding score, peri-implant probing depths and periimplant keratinized tissue. All implants and prostheses analyzed had a minimum of 2 years of followup. No chipping of the veneered facial porcelain or other technical complication was observed over the study period achieving a prosthesis survival and success rate of 100%. No implants were lost, achieving a 100% survival rate. Bleeding on probing was positive in 33% and 13% of probing sites for metal-acrylic prosthesis and zirconia prosthesis, respectively (p = 0.0445). Plaque index was positive in 76% and 53% of probing sites for metal-acrylic prosthesis and zirconia prosthesis, respectively (p = 0.0491). Mean probing depth was 1.74mm (SD 0.89mm) for the 106 implants supporting metal-acrylic prosthesis and 1.52mm (SD 0.63mm) for the 116 implants supporting zirconia prosthesis (p=0.0412). No other statistically significant differences were found between the two groups. The results of this retrospective evaluation showed that predominantly monolithic zirconia is a feasible alternative to the conventional metal framework acrylic for full arch implant-supported prosthesis. The restauration material did not influence the failure rate and complication risk of both prosthesis and implants.


Characterization of a cellulose binding domain from Clostridium cellulovorans endoglucanase-xylanase D and its use as a fusion partner for soluble protein expression in Escherichia coli.
Different chimeric proteins combining the non-catalytic C-terminal putative cellulose binding domain of Clostridium cellulovorans endoglucanase-xylanase D (EngD) with its proline-threonine rich region PT-linker, PTCBD(EngD), cellulose binding domain of C. cellulovorans cellulose binding protein A, CBD(CbpA), cohesin domains Cip7, Coh6 and CipC1 from different clostridial species and recombinant antibody binding protein LG were constructed, expressed, purified and analyzed. The solubilities of chimeric proteins containing highly soluble domains Cip7, CipC1 and LG were not affected by fusion with PTCBD(EngD). Insoluble domain Coh6 was solubilized when fused with PTCBD(EngD). In contrast, fusion with CBD(CbpA) resulted in only a slight increase in solubility of Coh6 and even decreased solubility of CipC1 greatly. PTCBD(EngD) and Cip7-PTCBD(EngD) were shown to bind regenerated commercial amorphous cellulose Cuprophan. The purity of Cip7-PTCBD(EngD) eluted from Cuprophan was comparable to that purified by conventional ion exchange chromatography. The results demonstrated that PTCBD(EngD) can serve as a bi-functional fusion tag for solubilization of fusion partners and as a domain for the immobilization, enrichment and purification of molecules or cells on regenerated amorphous cellulose.


Persistent trigeminal artery associated with intracranial arterial aneurysm.
Two patients with persistent trigeminal artery associated with a ruptured arterial intracranial aneurysm arising from the ipsilateral internal carotid artery at the level of the posterior communicating artery are presented. One patient underwent successful clipping of the aneurysm. The other patient died on the x-ray table shortly after admission during the performance of emergency carotid angiography. In each patient, the anomalous vessel was considered to be incidental.


Malpositioned mechanical mitral valves causing left ventricular outflow tract obstruction.
Left ventricular outflow tract obstruction caused by a malpositioned prosthetic mechanical mitral valve has not been reported to date. In two cases of mitral valve replacement, the sewing ring was sutured to the mitral valve leaflet instead of the annulus, resulting in the ventricularization of a portion of the valve and partial obstruction of the outflow tract. Re-replacing the valve with the same type of mechanical valve, but within the annular plane, eliminated the obstruction.


Characterization of an induced pluripotent stem cell line (UMi040-A) bearing an auditory neuropathy spectrum disorder-associated variant in TMEM43.
Hearing loss is one of the most common sensory disorders. TMEM43 is expressed in cochlear glia-like supporting cells (GLSs) and is known to be associated with late-onset auditory neuropathy spectrum disorder (ANSD) and progressive hearing loss. Here, we describe the derivation of an induced pluripotent stem cell (iPSC) line from a patient lymphoblastoid cell line (LCL) carrying a single heterozygous nonsense variant (p.Arg372Ter (c.1114C > T)) in TMEM43 that leads to a truncated protein lacking the 4th transmembrane domain. This cell line can serve as a tool for disease modelling and development of therapeutic approaches to restore inner ear function.


The Use of DAO as a Marker for Histamine Intolerance: Measurements and Determinants in a Large Random Population-Based Survey.
Histamine intolerance (HIT) is a common adverse reaction to food where elimination and reintroduction of histamine-rich food is part of the investigation. Analysis of the enzyme diamine oxidase (DAO) is sometimes used as an additional tool for diagnosis. This study aimed to describe the distribution of DAO in a large representative cohort of adults and to determine the association between DAO activity and possible associated factors. The study is based on the population-based West Sweden Asthma Study and includes 1051 subjects. Subjects underwent structured interviews including questions on demography, asthma, allergy symptoms, and lifestyle factors. Subjects were assessed for specific-IgE-antibodies and measurement of DAO activity in serum. Previously suggested cut-off levels for low values (<3 U/mL), normal values (>10 U/mL), and median levels of DAO were used. In the group of 1051 subjects, only a few presented reactions upon histamine intake, whereas 44% presented DAO levels below the suggested normal cut-off levels. BMI and age were shown to have an impact on DAO activity among women with increasing activity of DAO with increasing BMI and age. Among men, only increasing age was seen to have an impact on DAO levels. There was no difference in DAO levels with different sensitization status to common foods or airborne allergens. No association between DAO levels and reported symptoms to histamine-rich foods could be found. In conclusion, the determination of the DAO enzyme needs to be re-evaluated and may not be used as a valuable tool for histamine intolerance using current cut-off values. Further studies are needed to improve the use of DAO as a biomarker for histamine intolerance.


Bronchoesophageal manifestations of acquired immunodeficiency syndrome.
Among the more common manifestations of acquired immunodeficiency syndrome (AIDS) are tumors and infections that occur in regions treated by the bronchoesophagologist. In reviewing our institutional experience in the diagnosis and treatment of 396 patients with AIDS in 1987, we have noted that 226 (57%) had some form of pneumonia and 133 (34%) had candidiasis. In this communication we discuss the various types of bronchopulmonary, oropharyngeal, and esophageal infections that have been reported among AIDS patients. We also review the universal precautions and specific guidelines recommended for safeguarding the bronchoesophagologist and other health care workers who treat these patients.


Studies of factors influencing alveolar lining layer formation in lung homogenate: further isolation and identification of interfering materials.
Materials which inhibit the formation of stable alveolar lining layer by lung surfactant were investigated with the pig lung homogenate. The inhibitory activity was assessed by the bubble stability test on the bubbles formed by mechanical agitation of a mixture of surface active fraction from lung homogenate and inhibitory materials. The most active inhibitory material was identified as free cholesterol. Free fatty acids had a remarkable inhibitory activity. By modified Wilhelmy balance, it was demonstrated that the addition of free cholesterol to the surface active fraction delayed the formation of the stable surface membrane and at a larger dosage it inhibited the lowering the surface tension at the minimum surface area. The significance of this phenomenon in pathological lungs was briefly discussed.


Sulfur vacancies on MoS2 enhanced the activation of peroxymonosulfate through the co-existence of radical and non-radical pathways to degrade organic pollutants in wastewater.
The enhancement of vacancies in catalysts involving Fenton-like reactions is a promising way to remove organic pollutants in wastewater, but sulfur vacancies are rarely involved. In this work, MoS2 containing defect sites were synthesized by a simple high-temperature treatment and then applied for activating peroxymonosulfate to eliminate organic pollutants in wastewater. The structure was characterized by several techniques such as XRD, BET, and XPS. Important influencing factors were systemically investigated. The results indicated that MoS2 with sulfur vacancies possessed a higher catalytic activity than that of the parent MoS2. The annealing temperature of the catalyst had a great effect on the removal of organic pollutants. Besides, the catalytic system had a wide pH range. Quenching and electron paramagnetic resonance (EPR) experiments indicated that the reaction system contained radical and non-radical species. The characterization results revealed that the defect sites in catalysts mainly strengthened the activity of catalysts. This study offers a new heterogeneous catalyst for the removal of organic pollutants via the peroxymonosulfate-based Fenton-like reactions.


Green synthesis of N,S-doped carbon dots for tartrazine detection and their antibacterial activities.
A simple, green and low-cost method was developed for the synthesis of highly fluorescent N,S-doped carbon dots (N,S-CDs) via the hydrothermal treatment of Gandha Prasarini (GP) leaves as a natural source of carbon, nitrogen and sulfur. The as-prepared N,S-CDs exhibited excitation-dependent green fluorescence emission (λex = 450 nm, λem = 525 nm) with excellent stability, and were used as a fluorescent probe for the selective detection of tartrazine with a limit of detection of 0.18 μM. The fluorescence quenching of N,S-CDs was due to the inner filter effect. The developed method has been employed for the determination of tartrazine in honey and soft drinks with satisfactory recovery ranging from 92 to 110.2%. In addition, the antibacterial activity of the N,S-CDs was explored against both Gram-negative bacteria, Escherichia coli (E. coli) and Pseudomonas aeruginosa (P. aeruginosa), and Gram-positive bacteria, Staphylococcus aureus (S. aureus). The antibacterial mechanism of the N,S-CDs was investigated. The results indicated that the antibacterial activity was due to the membrane damage of the bacteria by the N,S-CDs. Besides, the N,S-CDs showed negligible lytic effects on human erythrocytes. These findings will inspire further exploitation of CD-based nano-bactericides in biomedical applications.


Noise-induced coherence in bistable systems with multiple time delays.
We study the correlation properties of noise-driven bistable systems with multiple time-delay feedbacks. For small noisy perturbation and feedback magnitude, we derive the autocorrelation function and the power spectrum based on the two-state model with transition rates depending on the earlier states of the system. A comparison between the single and double time delays reveals that the auto correlation functions exhibit exponential decay with small undulation for the double time delays, in contrast with the remarkable oscillatory behavior at small time lags for the single time delay.


Endogenous immunoreactive digitalis-like substance in neonatal serum and placental extracts.
Therapeutic levels of digoxin in the serum of untreated neonates delivered to mothers who had not received the drug prenatally were detected by radio-immunoassay. Digoxin levels in neonates should be interpreted with care because of the unknown contribution by the endogenous digitalis-like substance (DLS) to the level of the drug. Three commercially available radio-immunoassay kits were compared with regard to their sensitivity and reproducibility in detecting the endogenous DLS. The kit from Clinical Assays (Cambridge, Mass., USA) was selected for further investigations. In a series of 35 paired samples of maternal and cord blood the average DLS values in terms of digoxin were 0,52 +/- 0,07 and 0,81 +/- 0,27 ng/ml respectively. This difference is statistically highly significant. In the case of infants with DLS values of 1-1,5 ng/ml in terms of digoxin, approximately 1 week was required to reach nontherapeutic digoxin levels, i.e. below 0,5 ng/ml. Gel chromatography showed that the DLS in neonatal serum was more closely associated with protein than is authentic digoxin. In placental extracts it followed the elution profile of the protein completely, but it shifted to fractions with a lower molecular weight than haemoglobin after trypsinization. The level of DLS in neonatal serum was also increased by more than half its original value by trypsinization. Proteolysis therefore seems to have a releasing effect on DLS. The molecular size of this substance is probably in the same range as that of polypeptides, since it was not dialysable from trypsinized and untreated samples through a membrane with a 22 000 dalton molecular weight cut-off point.


Simulation of the electron ionization mass spectra of the Novichok nerve agent.
Novichok is one of the most feared and controversial nerve agents, which existence was confirmed only after the Salisbury attack in 2018. A new attack on August 2020, in Russia, was confirmed. After the 2018 attack, the agent was included in the list of the most dangerous chemicals of the Chemical Weapons Convention (CWC). However, information related to its electron ionization mass spectrometry (EI/MS), essential for unambiguous identification, is scarce. Therefore, investigations about Novichok EI/MS are urgent. In this work, we employed Born-Oppenheimer molecular dynamics through the Quantum Chemistry Electron Ionization Mass Spectrometry (QCEIMS) method to simulate and rationalize the EI/MS spectra and fragmentation pathways of 32 Novichok molecules recently incorporated into the CWC. The comparison of additional simulations with the measured EI spectrum of another Novichok analog is very favorable. A general scheme of the fragmentation pathways derived from simulation results was presented. The present results will be useful for elucidation and prediction of the EI spectra and fragmentation pathways of the dangerous Novichok nerve agent.


Atypical patterns of respiratory sinus arrhythmia index an endophenotype for depression.
Can atypical patterns of parasympathetic nervous system activity serve as endophenotypes for depression? Using respiratory sinus arrhythmia (RSA) as an index of parasympathetic nervous system function, we examined this question in two studies: one involving mothers with and without depression histories and their offspring (at high and low risk for depression, respectively), and a further study of adolescent sibling pairs concordant and discordant for major depression. In both studies, subjects were exposed to sad mood induction; subjects' RSA was monitored during rest periods and in response to the mood induction. We used Gottesman and Gould's (2003) criteria for an endophenotype and a priori defined "atypical" and "normative" RSA patterns (combinations of resting RSA and RSA reactivity). We found that atypical RSA patterns (a) predicted current depressive episodes and remission status among women with histories of juvenile onset depression and healthy controls, (b) predicted longitudinal trajectories of depressive symptoms among high- and low-risk young offspring, (c) were concordant across mothers and their juvenile offspring, (d) were more prevalent among never-depressed youth at high risk for depression than their low-risk peers, and (e) were more concordant across adolescent sibling pairs in which both versus only one had a history of major depression. Thus, the results support atypical RSA patterns as an endophenotype for depression. Possible mechanisms by which RSA patterns increase depression risk and their genetic contributors are discussed.


Automated red cell exchange in sickle cell disease.
Red cell exchange is important in the care of acutely ill sickle-cell patients, and may be life-saving. An automated red cell exchange technique has been developed using a Baxter blood cell separator, enabling an isovolaemic exchange to be performed within 2.5 h. A total of 20 procedures have been performed in 15 patients, including one woman in the third trimester of pregnancy, with a mean decrease of 72% in the circulating sickle haemoglobin (HbS) level. This method enables almost all adult patients with sickle cell anaemia to have their HbS reduced to safe levels by only one procedure. The procedure was well tolerated by all patients, including those who were acutely ill. This technique provides an effective procedure for reducing the percentage of circulating HbS rapidly in acutely ill patients with complications of sickle cell anaemia.


[The effect of curiozin gel and adhesive plaster application on the healing and blood flow dynamics in skin autografts from various areas of body in rats].
The stimulating effect of curiozin gel and plaster application on the engraftment of skin autografts from various areas of body was studied in rats. Curiozin gel in a single dose of 75 mg was used for the daily treatment of autografts over the entire postoperation period of time. The adhesive plaster was applied onto skin autografts for 5 days after skin transplantation. The skin autografts in the test group exhibited normalized volume blood flow dynamics, reduced duration of the inflammatory reaction, and improved the quality of engraftment as compared to the untreated control group. The best healing and blood-flow-normalizing effect was achieved by combined application of curiozin gel and adhesive plaster.


Microwave-assisted synthesis and tyrosinase inhibitory activity of chalcone derivatives.
A series of chalcones and their derivatives were synthesized, and their inhibitory effects on the diphenolase activity of mushroom tyrosinase were evaluated. The results showed that some of the synthesized compounds exhibited significant inhibitory activity, and four compounds exhibited more potent tyrosinase inhibitory activity than the reference standard inhibitor kojic acid (5-hydroxy-2-(hydroxymethyl)-4H-pyran-4-one). Specifically, 1-(-1-(4-methoxyphen- yl)-3-phenylallylidene)thiosemicarbazide (18) exhibited the most potent tyrosinase inhibitory activity with IC₅₀ value of 0.274 μM. The inhibition mechanism analysis of 1-(-1-(2,4-dihydroxyphenyl)-3-phenylallylidene) thiosemicarbazide (16) and 1-(-1-(4-methoxyphenyl)-3-phenylallylidene) thiosemicarbazide (18) demonstrated that the inhibitory effects of the two compounds on the tyrosinase were irreversible. Preliminary structure activity relationships' analysis suggested that further development of such compounds might be of interest.


Role of opioid ligands in the irritable bowel syndrome.
Endogenous opioid peptides - enkephalins, beta-endorphin and dynorphins - are located in specific sites of the brain, the spinal cord, the autonomic ganglia and the enteric nervous system. Endogenous opioids participate in the regulation of nervous visceral afference and sensitivity as well as of several visceral motor function induced by the central nervous system and through the enteroenteric and the myoenteric reflexes. Their final effect on gut physiology is the net and harmonically balanced result of their binding to mu, delta and kappa opioid receptor subtypes. Exogenous opioid receptor ligands with different affinities for the opioid receptor subtypes have been effectively used to modify and normalize altered gut functions. The mu receptor agonists - morphine and, to a greater extent, the meperidine congeners diphenoxylate and loperamide - have been shown to slow gastrointestinal transit by their effects on the circular and longitudinal muscle of the intestine. Diphenoxylate and, more efficiently, loperamide, for the lack of any effect on the central nervous system, have been usefully employed in the treatment of diarrhea in irritable bowel syndrome (IBS) patients. Unlike the mu receptor agonists morphine and loperamide, which invariably stimulate colonic motility, trimebutine, which has almost equal affinity for mu, delta and kappa receptors, has no effect on normal colonic activity but reduces the abnormal increase in postprandial motor activity in IBS patients and accelerates slow large bowel transit in constipated patients. Opioid ligands can be usefully employed to normalize altered visceral sensitivity in IBS patients. The kappa receptor agonist fedotozine exerts its antinociceptive effect by acting on peripheral nerve endings of sensory vagal and nonvagal afferent pathways. Fedotozine has been shown to increase the threshold of perception to colonic distension in experimental conditions and to affect favourably symptoms of IBS in clinical trials.


Mating and nutritional state affect the reproduction of Aedes albopictus mosquitoes.
Mated female Aedes albopictus mosquitoes that were maintained after emergence without carbohydrate were more likely to develop eggs after a small blood meal than were unmated females maintained on the same diet. The effect was due to male accessory gland substances transferred to the female during mating. Neither the endogenous reserves of protein and lipid nor the number of eggs developed per volume of ingested blood differed between mated and unmated females, suggesting that the utilization of existing reserves was altered by mating. Methoprene administered to both mated and unmated females that ingested small blood meals reduced the likelihood that egg development would occur. Small volumes of blood were more likely to trigger oogenesis in both mated and unmated females if their abdomens were additionally distended.


Association of World Trade Center (WTC) Occupational Exposure Intensity with Chronic Obstructive Pulmonary Disease (COPD) and Asthma COPD Overlap (ACO).
Reported associations between World Trade Center (WTC) occupational exposure and chronic obstructive pulmonary disease (COPD) or asthma COPD overlap (ACO) have been inconsistent. Using spirometric case definitions, we examined that association in the largest WTC occupational surveillance cohort. We examined the relation between early arrival at the 2001 WTC disaster site (when dust and fumes exposures were most intense) and COPD and ACO in workers with at least one good quality spirometry with bronchodilator response testing between 2002 and 2019, and no physician-diagnosed COPD before 9/11/2001. COPD was defined spirometrically as fixed airflow obstruction and ACO as airflow obstruction plus an increase of ≥ 400 ml in FEV1 after bronchodilator administration. We used a nested 1:4 case-control design matching on age, sex and height using incidence density sampling. Of the 17,928 study participants, most were male (85.3%) and overweight or obese (84.9%). Further, 504 (2.8%) and 244 (1.4%) study participants met the COPD and ACO spirometric case definitions, respectively. In multivariable analyses adjusted for smoking, occupation, cohort entry period, high peripheral blood eosinophil count and other covariates, early arrival at the WTC site was associated with both COPD (adjusted odds ratio [ORadj] = 1.34, 95% confidence interval [CI] 1.01-1.78) and ACO (ORadj = 1.55, 95%CI 1.04-2.32). In this cohort of WTC workers, WTC exposure intensity was associated with spirometrically defined COPD and ACO. Our findings suggest that early arrival to the WTC site is a risk factor for the development of COPD or of fixed airway obstruction in workers with pre-existing asthma.


Patients' understanding, management practices, and challenges regarding hypertension: A qualitative study among hypertensive women in a rural Bangladesh.
Hypertension, as one of the main predisposing factors of many non-communicable diseases, is generally underdiagnosed among women with a significant uncontrolled rate. This study explores the understanding, management practice and challenges related to hypertension among hypertensive women in rural Bangladesh. A qualitative study was conducted among hypertensive rural women at Kumarkhali Upazilla, Kushtia, Bangladesh, using purposive and snowball sampling technique. Data was collected through in-depth interviews among twenty-three hypertensive women until they reached saturation. Data were analyzed thematically. Findings of the study found that a small number of participants perceived the symptoms, risk factors, management and treatment of hypertension based on biomedical understanding. Also, their awareness level and adherence to preventive practices reflected a significant gap between biomedical preventive practices and local practices. A substantial number of participants preferred home management and alternative treatment for hypertension over the medication adherence and hospital treatment. This investigation revealed that poor socio-economic conditions, such as financial insufficiency, and, gender-based negligence impacted women's perception of and practice for hypertension and resulted in risky hypertension management behaviors. Based on the study, formulation of a comprehensive health education program for creating awareness, provisioning of significant interventions services related to hypertensive care are needed. Further intensive research is needed at the community-level to manage this chronic disease.


Direct In Vivo MRI Discrimination of Brain Stem Nuclei and Pathways.
The brain stem is a complex configuration of small nuclei and pathways for motor, sensory, and autonomic control that are essential for life, yet internal brain stem anatomy is difficult to characterize in living subjects. We hypothesized that the 3D fast gray matter acquisition T1 inversion recovery sequence, which uses a short inversion time to suppress signal from white matter, could improve contrast resolution of brain stem pathways and nuclei with 3T MR imaging. After preliminary optimization for contrast resolution, the fast gray matter acquisition T1 inversion recovery sequence was performed in 10 healthy subjects (5 women; mean age, 28.8 ± 4.8 years) with the following parameters: TR/TE/TI = 3000/2.55/410 ms, flip angle = 4°, isotropic resolution = 0.8 mm, with 4 averages (acquired separately and averaged outside k-space to reduce motion; total scan time = 58 minutes). One subject returned for an additional 5-average study that was combined with a previous session to create a highest quality atlas for anatomic assignments. A 1-mm isotropic resolution, 12-minute version, proved successful in a patient with a prior infarct. The fast gray matter acquisition T1 inversion recovery sequence generated excellent contrast resolution of small brain stem pathways in all 3 planes for all 10 subjects. Several nuclei could be resolved directly by image contrast alone or indirectly located due to bordering visualized structures (eg, locus coeruleus and pedunculopontine nucleus). The fast gray matter acquisition T1 inversion recovery sequence has the potential to provide imaging correlates to clinical conditions that affect the brain stem, improve neurosurgical navigation, validate diffusion tractography of the brain stem, and generate a 3D atlas for automatic parcellation of specific brain stem structures.


The stress and fear levels of microwave toe-treated broiler chickens grown with two photoperiod programs.
The current study was conducted to investigate the influence of microwave toe treatment and an increasing photoperiod on stress and fear levels in broiler chickens. Upon delivery from the hatchery, the toe tips of 364 male and 364 female broilers were exposed to microwave energy to restrict claw growth, whereas the same numbers of birds retained intact toes. Birds from each sex and toe treatment were grown under 23 h of light or increasing photoperiods, with 4 replicates of each treatment combination. Elevated activity of plasma creatine kinase (CK) and increased heterophil to lymphocyte (H/L) ratios were employed as indicators of stress. Tonic immobility (TI) was conducted on d 10, 22, and 36 and used as an index of fear response. Males had lower plasma CK activity and H/L ratios (P < 0.05). Birds on 23 h of light had a higher activity of plasma CK (P < 0.05) but similar H/L ratios compared with birds on the increasing photoperiod. Microwave toe treatment did not affect the activity of plasma CK or H/L ratios. Photoperiod, microwave toe treatment, or sex did not affect the number of attempts required to induce TI. The increasing lighting program decreased the duration of TI on d 10 (P < 0.05), had no effect on d 22, and increased duration of TI on d 36 (P < 0.05). There was no microwave toe treatment or sex effect on the duration of TI at any ages tested.


Synthesis of New Abscisic Acid (ABA) Analogs Possessing a Geometrically Rigid Cyclized Side Chain.
In a solution, cis-abscisic acid (ABA) isomerizes into the trans-isomer which is physiologically inactive, and this structural instability is regarded as a reason for insuitability of ABA in agricultural applications. The side chain of ABA, the 2-cis-4-trans-3-methyl-2,4-pentadienoic acid moiety, was replaced with various substituted phenyl groups to examine biological effects of the inflexibility of the part. Construction of the phenyl moiety was achieved by cyclizing the ionone derivatives and subsequent aromatization. Some of those new compounds showed ABA-like activity in both seed germination and transpiration assays.


A review of hepatitis C virus (HCV) vertical transmission: risks of transmission to infants born to mothers with and without HCV viraemia or human immunodeficiency virus infection.
Hepatitis C virus (HCV) vertical transmission studies have reported conflicting findings, possibly due to differences in HCV transmission risk factors among maternal populations, or to methodological differences. Systematic review of worldwide published and unpublished HCV vertical transmission studies. Standardized diagnostic criteria were applied to minimize methodological differences, and transmission rates recalculated according to maternal HCV viraemic and human immunodeficiency virus (HIV) infection status. In all, 976 eligible infants from 28 studies were followed up sufficiently for recalculation of transmission rates. Overall transmission rates were less than 10% in 8/12 studies of HIV negative mothers, compared with 2/7 studies comprising at least 50% HIV-coinfected mothers. Rates from 409 viraemic mothers in 15 studies ranged from 0% to 41%, being less than 10% from HIV negative mothers in 6/13 studies and from HIV positive mothers in 1/6 studies. Nine studies measured maternal viraemia levels, with only 2/30 transmitting mothers having < 10(6) copies/ml of HCV RNA. Eight transmissions were identified overall from non-viraemic mothers. Significant transmission rate variation remained after accounting for maternal viraemia and HIV coinfection, possibly due to differences in other vertical transmission risk factors, in frequencies of postnatal transmission, or residual differences in study methodologies. Overall, HCV transmission is largely restricted to infants born to HCV viraemic mothers, and low risks among most HIV negative mothers may be due to lower HCV viraemia levels. International agreement on standardized diagnostic criteria for HCV vertical transmission would facilitate pooling of individual findings, to allow more precise transmission estimates and further investigation of risk factors.


Inhibition of DNA primase by 9-beta-D-arabinofuranosyladenosine triphosphate.
9-beta-D-Arabinofuranosyladenosine triphosphate (araATP) is a potent inhibitor of DNA primase. Primase readily incorporates araATP into primers, and primers containing araAMP are then elongated by DNA polymerase alpha (pol alpha) upon addition of dNTPs. AraATP did not inhibit utilization of primers under conditions where the ability of pol alpha to elongate primers was independent of the dATP concentration. The fraction of primers elongated by pol alpha was reduced by araATP only when elongation was dependent upon the dATP concentration. When the Ki for primase was measured in terms of the inhibition of the synthesis of primers that can be utilized by pol alpha, we obtained Ki = 2.7 microM (37 degrees C) and 2.0 microM (25 degrees C). Inhibition was competitive with ATP. Inhibition of pol alpha activity by araATP was measured under conditions where primase-catalyzed primer synthesis was required for the pol alpha activity. The decreased pol alpha activity was due to primase inhibition, and at constant dATP, araATP inhibition was competitive with ATP and gave Ki = 1.2 microM, similar to the Ki for primase alone. Increasing the dATP concentration had no effect on inhibition. In combination with previously reported in vivo data, we conclude that DNA primase is the primary in vivo target of the arabinofuranosyl nucleotides, not pol alpha.


The effects of cosmic particle radiation on pocket mice aboard Apollo XVII: appendix I. Condition of flight animals on recovery; food intake; observations on hypothalamus, pituitary, and adrenal glands.
The rationale for studying certain hypothalamic nuclei and the pituitary and adrenal glands of the pocket mice that flew on Apollo XVII was the need to evaluate the effects of the potentially severe stress on these animals in the foreign environment of flight canister, weightlessness, increased G forces, and other unnatural conditions. Decrease in body weight and variability of food intake were significant among the four flight animals that were recovered alive. The mean nuclear diameter of neurons in the arcuate and ventromedial hypothalamic nuclei did not differ significantly from the values obtained in the control animals. On the other hand, the mean nuclear diameter of neurons in the supraoptic nucleus of the flight mice was significantly greater than in the control groups. Comparisons of the adeno- and neuropypophysis revealed no significant differences among the three groups. Insofar as they were studied, the adrenals were similar in all groups.


Microfluidic aptasensor POC device for determination of whole blood potassium.
An integrated microfluidic Au nanoparticle (AuNP) aptasensor device is proposed for monitoring the concentration of potassium (K+) ions in the bloodstream of patients with chronic kidney disease (CKD). In the proposed detection device, the AuNPs in the AuNP/aptamer complex are displaced by the serum K+ ions and react with NaCl to produce a color change in the detection region from which the K+ ion concentration is then inversely derived. The microfluidic device comprises two main components, namely an AuNP aptasensor PMMA (Poly(methyl methacrylate))/paper-microchip and a colorimetric analysis system for the quantitative detection of K+ ion concentration in whole blood. The functions of PMMA/paper microchips include reagent storage, K+ ion/aptamer reaction, and separation of serum from whole blood samples (blood filter). Experimental results show that the microfluidic device provides a linear response over the K+ ion concentration in range of 0.05-9 mM in artificial serum and has a detection limit (LOD) of 0.01 mM. Moreover, the detection results obtained for the 137 whole blood and 287 serum samples of CKD patients are very consistent (R2 = 0.968 and R2 = 0.980) with the measurement results obtained using an ion-selective electrodes (ISE) method. Results confirm that the current microfluidic aptasensor device provides a highly-sensitive and convenient method for performing the point-of-care (POC) monitoring of the whole blood K+ ion concentration.


Reaching for the stars: SLPs shine on literacy teams.
Speech-language pathologists (SLPs) who work with school-age children have the opportunity to contribute to the literacy development of specific students as well as the literacy instruction and intervention programs in their district. These efforts are made in collaboration with other professionals and parents and may be referred to as participation on literacy teams. This article describes successful literacy teams in elementary schools including the variety of possible members as well as common characteristics of successful literacy teams. We describe how SLPs participate by developing collegial relationships with their team, using best practices for literacy, and supporting children as they progress in the general curriculum. Finally, we discuss some strategies for SLPs to take stock of their own literacy team participation, identify new goals, and map out a plan to reach new heights in literacy as integral members of literacy teams.


Vitamin D production depends on ultraviolet-B dose but not on dose rate: a randomized controlled trial.
Ultraviolet-B (UV-B) radiation increases serum vitamin D level expressed as 25-hydroxyvitamin D(3) (25(OH)D), but the dose-response relationship and the importance of dose rate is unclear. Of 172 fair-skinned persons screened for 25(OH)D, 55 with insufficient baseline 25(OH)D≤50 nm (mean 31.2 nm) were selected and randomized to one of 11 groups of five participants. Each group was exposed to one of four different UV-B doses: 0.375, 0.75, 1.5 or 3.0 standard erythema dose (SED) for 1, 5, 10 or 20 min. All participants had four UV-B sessions with 2- to 3-day interval with 24% of their skin exposed. Skin pigmentation and 25(OH)D were measured before and after the irradiations. The increase in 25(OH)D after UV-B exposure (adjusted for baseline 25(OH)D) was positively correlated with the UV-B dose (P=0.001; R(2) =0.176) but not to dose rate (1-20 min). 25(OH)D increased in response to four UV-B treatments of 3 SED with 24.8 nm on average and 14.2 nm after four UV-B treatments of just 0.375 SED. In conclusion, the increase in 25(OH)D after UV-B exposure depends on the dose but not on the dose rate (1-20 min). Further, a significant increase in 25(OH)D was achieved with a very low UV-B dose.


Role of ICAM-1 in neutrophil-mediated lung vascular injury after occlusion and reperfusion.
We examined the role of intracellular adhesion molecule 1 (ICAM-1 or CD54) in the development of pulmonary edema in rabbits after pulmonary artery occlusion and reperfusion using a monoclonal antibody (MAb) RR1/1 directed against ICAM-1, a ligand for the CD18 leukocyte adhesion glycoprotein complex. A vascular clamp was placed around the left pulmonary artery for 24 h and then released to allow reperfusion for 2 h. Lungs subjected to 24 h of unilateral pulmonary artery occlusion showed increased binding of 125I-labeled RR1/1 and immunocytochemical evidence of ICAM-1 expression in pulmonary vascular endothelial cells compared with the contralateral lung. MAbs RR1/1 (0.5 mg/kg) or IB4 (1.0 mg/kg) (MAb directed against an epitope on the CD18 adhesion glycoprotein) was infused 45-60 min before the start of reperfusion to assess the roles of ICAM-1 and CD18 in the response. After reperfusion, the lungs were removed, suspended from one end of a weighing balance, and perfused with Ringer-albumin (0.5 g/100 ml), and the changes in lung weight were monitored for 60 min. Lung tissue myeloperoxidase (MPO) content (a marker of neutrophil sequestration) was determined after reperfusion. The increases in lung weight gain in the RR1/1- and IB4-treated groups of 960 +/- 100 and 865 +/- 110 mg, respectively, were less (P less than 0.05) than in untreated controls (3,550 +/- 725 mg).(ABSTRACT TRUNCATED AT 250 WORDS)


Analysis of the effectiveness of emergency countermeasures in the 30-km zone during the early phase of the Chernobyl accident.
Some radiation-emergency countermeasures, including evacuation, were implemented in the settlements of the 30-km zone during the early phase of the accident at the Chernobyl Nuclear Power Plant. These countermeasures are described and compared with the international recommendations. An analysis of the effectiveness of the emergency countermeasures was conducted based upon the results of a wide-scale public survey. Quantitative assessments of the effectiveness (dose reduction) of the countermeasures were derived.


[Staining of carious dentin with Kariesdetektor].
In a histological study of the staining effect of the staining solution Kariesdetektor 60 human teeth with carious lesions of varying sizes were separated into one control half and one test half each immediately following their extraction. Non-decalcified thin ground sections were prepared with the aid of the saw-cut-and-grind technique. The test halves were stained with Kariesdetektor, the control preparations with toluidine blue. The staining effect of the disclosing dye was evaluated under the light microscope and compared with the control preparations. In enamel the disclosing dye stained all carious areas except the dark and the more peripheral transparent zones. In dentinal caries the zones of necrosis and penetration were safely stained. Zones of demineralization, dead tracts, and transparency zones were not stained. Kariesdetektor is a reliable optical aid in removing infected carious hard tooth structures.


Selection of aptamers specific for adipose tissue.
Obesity has reached epidemic proportions, affecting more than one tenth of the world's population. As such, adipose tissue is being increasingly recognized as an important therapeutic target for obesity and related metabolic disorders. While many potential targets of adipose tissue have been established and drugs developed, very few of those drugs specifically target adipose tissue without affecting other tissue. This results from a limited knowledge of both cell-surface markers and physicochemical traits specific to adipocytes that might otherwise be exploited by circulating drugs. Here we report the use of cell-SELEX technology to select two aptamers that can specifically recognize mature adipocytes: adipo-1 and adipo-8. Adipo-8 shows high affinity for differentiated, mature 3T3-L1 adipocytes with a K(d) value of 17.8±5.1 nM. The binding was sustained upon incubation at 37°C and insulin stimulation, but was lost upon trypsin treatment. The binding ability was also verified on frozen tissue slides with low background fluorescence and isolated adipocytes. Aptamer adipo-8 selected from a random library appears to bind to mature differentiated adipocytes specifically. This aptamer holds great promise as a molecular recognition tool for adipocyte biomarker discovery or for targeted delivery of molecules to adipocytes.


Antiischemic and antiarrhythmic activities of some novel alinidine analogs in the rat heart.
The antiischemic and antiarrhythmic effects of alinidine and a number of novel alinidine analogs were examined by using perfused rat-heart models. In the isolated working rat heart, the alinidine analog TH91:21 (10 microM; a butyl derivative) significantly increased the postischemic recovery of the heart in terms of both power and efficiency when compared with the control group. In the in situ perfused heart model, this same compound, along with TH91:22 (10 microM; a pentyl derivative) also significantly reduced the severity of both ischemia- and reperfusion-induced arrhythmias in both paced and unpaced hearts. Thus this study is the first to demonstrate the potent antiarrhythmic efficacy of two novel alinidine analogs TH91:21 and TH91:22, with TH91:21 also demonstrated to be a potent antiischemic agent in the isolated working rat heart. Although the mode of action of these compounds remains unclear, results from this study suggest that it is not simply a result of bradycardia or blockade of KATP channels, two actions these compounds possess. These compounds thus possess a novel and beneficial pharmacologic profile worthy of further study.


Anti-beta2-glycoprotein I (beta2GPI) monoclonal antibodies with lupus anticoagulant-like activity enhance the beta2GPI binding to phospholipids.
beta2-Glycoprotein I (beta2GPI), a plasma glycoprotein with phospholipid-binding property, is known to be the actual target antigen for autoimmune type anticardiolipin antibodies (aCLs). Certain groups of aCLs (anti-beta2GPI antibodies) exert lupus anticoagulant (LA) activity and perturb the function of vascular endothelial cells. This investigation aimed at highlighting some insights into the molecular basis by which aCLs exert their biological effects by using anti-beta2GPI mAbs with well-characterized epitopes from mice and from patients with antiphospholipid syndrome. Anti-beta2GPI mAbs directed against the third domain (Cof-20 and Cof-22) and fourth domain (Cof-21, EY1C8, and EY2C9) of beta2GPI inhibited the thrombin generation induced by Russell's viper venom in diluted plasma and that induced by the prothrombinase complex reconstituted with purified clotting factors. This anticoagulant activity was abrogated in the presence of an excess amount of phospholipids, thus resembling the LA activity. In stark contrast, anti-beta2GPI mAbs directed against the fifth domain and the carboxy-terminal region of the fourth domain showed no LA-like activity. These findings suggest that the LA activity of anti-beta2GPI antibodies depends on their epitope specificity. Experiments carried out to clarify the mechanism of the LA activity showed that anti-beta2GPI mAbs with LA-like activity, but not those without this effect, enhance the beta2GPI binding to phospholipids. In addition, the F(ab')2 fragment, but not the Fab' fragment, of the anti-beta2GPI mAbs was found to enhance the LA activity and the beta2GPI binding to phospholipids, suggesting that anti-beta2GPI antibodies induce formation of multiple complexes of beta2GPI on the surface of phospholipids because of their bivalent property. This clustering of beta2GPI molecules induced by anti-beta2GPI antibodies, probably because of their multivalent property and epitope specificity, might hinder the lateral mobility and activation of clotting factors on the surface of phospholipids and thus exert LA activity. Clustering of beta2GPI molecules may also explain the molecular mechanism by which anti-beta2GPI antibodies alter the function of leukocytes and endothelial cells. The well-documented heterogeneous LA activity of aCLs (anti-beta2GPI antibodies) may also be explained by their epitope specificity.


Parenteral nutrition and fasting reduces mucosal addressin cellular adhesion molecule-1 (MAdCAM-1) mRNA in Peyer's patches of mice.
Mucosal addressin cellular adhesion molecule-1 (MAdCAM-1) in Peyer's patches (PP) is the gateway molecule for cellular migration into the mucosal immune system. Lack of enteral feeding during parenteral nutrition (PN) rapidly decreases PP MAdCAM-1, leading to drops in mucosal T and B cells and intestinal and respiratory IgA. We determined the molecular events associated with MAdCAM-1 mRNA and protein during PN (short and long term) and fasting (1 and 2 days). Experiment 1: Cannulated mice received PN for 8 hours (short-term PN, n = 6) or chow + saline (chow, n = 6). Experiment 2: Cannulated mice received PN (long-term PN, n = 4) or chow (n = 3) for 5 days. Experiment 3: Noncannulated chow mice were fasted for 1 and 2 days (n = 2/time). Total cellular RNA from the PP was quantified for MAdCAM-1 mRNA by real-time polymerase chain reaction (PCR). MAdCAM-1 protein was measured by Western blot. PN rapidly down-regulated MAdCAM-1 gene expression. After 8 hours of PN with lack of enteral feeding, MAdCAM-1 mRNA levels dropped 20% (0.8-fold vs chow, p > .05); 5 days of PN reduced MAd-CAM-1 levels 64% (0.34-fold vs chow, p < .05). PN reduced MAdCAM-1 protein levels by 30% (chow: 329 +/- 14 vs PN: 230 +/- 35, p < .05) after 5 days. Fasting of uncannulated mice decreased MAdCAM-1 mRNA levels by 16% (0.84-fold, p < .05) at day 1 and 30% (0.7-fold, p < .05) by day 2 compared with chow. Both PN with lack of enteral feeding and fasting down-regulate MAdCAM-1 mRNA and protein levels in PP. The MAdCAM-1 changes are due to lack of enteral stimulation rather than toxic effects of PN.


The non-Hodgkin's lymphomas: old and new thinking.
Knowledge of non-Hodgkin's lymphomas has increased enormously in the last 10-15 years. The various types are considered to reflect the normal immunological processes that take place in the lymphoid tissue after antigenic challenge. This concept, which states that non-Hodgkin's lymphomas are neoplastic counterparts of normal processes, has implications for morphology, immunology and clinical behaviour. Based on this concept, as well as on immunohistological and functional data, a hypothetical scheme of B-cell development is proposed. The relation to normal physiology also holds for lymphomas localized outside the lymph nodes. Normal structure and function in two important extranodal lymphoid tissues, the gastrointestinal mucosa and the skin-associated lymphoid tissues, are related to the lymphomas arising in these sites. This relation and some of its implications are discussed, with special reference to the important processes of homing and recirculation of lymphocytes.


Vector-meson-dominance model for radiative decays involving light scalar mesons.
We study a vector-dominance model which predicts quite a large number of currently interesting decay amplitudes of the types S-->gammagamma, V-->Sgamma, and S-->Vgamma, where S and V denote scalar and vector mesons, in terms of three parameters. As an application, the model makes it easy to study in detail a recent proposal to boost the ratio Gamma(phi-->f(0)gamma)/Gamma(phi-->a(0)(0)gamma) by including the isospin violating a(0)(0)-f(0) mixing. However, we find that this effect is actually small in our model.


[Echo-guided percutaneous drainage of renal cysts].
Treatment of renal cysts in the ultrasound guided percutaneous drainage, associated with intra-cavital sclerosis-inducing agents to avoid recurrence, is often proposed. This report describes the technique and the most suitable sclerosis inducing agents. The best results are obtained with alcohol. No increase in complications or discomfort was observed.


[Angiography in vasculitis].
The method of angiography in vasculitis has recently been changed due to the development of digital subtraction angiography (DSA). Intra-venous DSA is widely used in the diagnosis of vasculitis, although intra-arterial DSA is needed for depicting small vessel lesions. Low-osmolality contrast medium is another advancement in the angiography of vasculitis, which can make peripheral angiography painless. We discussed angiographic features in vasculitis diseases, including Takayasu arteritis, Buerger disease, moyamoya disease, and vasculo-Behçet disease. Angiography is the most important in order to diagnose vasculitis and evaluate the degree and extent of disease. Further, interventional radiology, which uses the technic of angiography for the purpose of therapy, is employed for the vascular treatment of vasculitis.


Mature adipocytes inhibit in vitro differentiation of human preadipocytes via angiotensin type 1 receptors.
Recent studies suggest that angiotensin II (Ang II) plays a role in the adipogenesis of murine preadipocytes. Here, we examined the role of Ang II for the differentiation of primary cultured human preadipocytes. Preadipocytes were isolated from human adipose tissue and stimulated to differentiate. Quantitation of gene expression during adipogenesis was performed for renin-angiotensin system (RAS) genes. The influence of the RAS on adipogenic differentiation was investigated by addition of either angiotensinogen (AGT), Ang II, or angiotensin receptor antagonists to the differentiation medium. We also examined the influence of adipocytes on adipogenesis by co-culture experiments. Expression of the RAS genes AGT, renin, angiotensin-converting enzyme, and Ang II type 1 receptor increased during adipogenesis. Stimulation of the Ang II type 1 receptor by Ang II reduced adipose conversion, whereas blockade of this receptor markedly enhanced adipogenesis. Adipocytes were able to inhibit preadipocyte differentiation in the co-culture, and this effect was abolished by blockade of the Ang II type 1 receptor. This finding points to a functional role of the RAS in the differentiation of human adipose tissue. Because AGT secretion and Ang II generation are characteristic features of adipogenesis, we postulate a paracrine negative-feedback loop that inhibits further recruitment of preadipocytes by maturing adipocytes.


The Association of Reproductive Hormone Levels and All-Cause, Cancer, and Cardiovascular Disease Mortality in Men.
Testosterone (T) levels have been associated with mortality, but controversy exists. Our objective was to investigate associations between serum levels of total T, SHBG, free T, estradiol, LH and FSH, and subsequent mortality with up to 30 years of follow-up. This was a prospective cohort study consisting of men participating in four independent population-based surveys (MONICA I-III and Inter99) from 1982 to 2001 and followed until December 2012 with complete registry follow-up. A total of 5350 randomly selected men from the general population aged 30, 40, 50, 60, or 70 years at baseline participated. All-cause mortality, cardiovascular disease (CVD) mortality, and cancer mortality were the main outcomes. A total of 1533 men died during the follow-up period; 428 from CVD and 480 from cancer. Cox proportional hazard models revealed that men in highest LH quartile had an increased all-cause mortality compared to lowest quartile (hazard ratio [HR], 1.32; 95% confidence interval [CI], 1.14-1.53). Likewise, increased quartiles of LH/T and estradiol increased the risk of all-cause mortality (HR, 1.23; 95% CI, 1.06-1.43; HR, 1.23; 95% CI 1.06-1.43). No association to T levels was found. Higher LH levels were associated with increased cancer mortality (HR, 1.42; 95% CI, 1.10-1.84) independently of smoking status. Lower CVD mortality was seen for men with T in the highest quartile compared to lowest (HR, 0.72; 95% CI, 0.53-0.98). Furthermore, negative trends were seen for SHBG and free T in relation to CVD mortality, however insignificant. The observed positive association of LH and LH/T, but not T, with all-cause mortality suggests that a compensated impaired Leydig cell function may be a risk factor for death by all causes in men. Our findings underpin the clinical importance of including LH measurement in the diagnostic work-up of male patients seeking help for possible androgen insufficiency.


Predictors of work-related injuries and illnesses: national survey findings.
This study used national survey data of working adults (aged 33-41) to identify factors associated with the occurrence of occupational injuries and illnesses. Data from the National Longitudinal Survey of Youth were used to compare selected employment and personal characteristics for respondents who reported experiencing a work-related injury or illness with those who did not. Multivariate analyses were performed to calculate nationally representative odds ratios reflecting the likelihood for specific individual attributes and job characteristics to be associated with the reporting of a work-related injury or illness, while controlling for relevant covariates. In this study the incidence of occupational injuries was related to several demographic factors, including low family income and rural residence, and several job characteristics, including working in a high-hazard occupation, job dissatisfaction, and exposure to six specific hazardous job activities: (1) performing lots of physical effort on the job, (2) lifting or carrying more than 10 lbs, (3) using stairs and inclines, (4) kneeling or crouching, (5) reaching, and (6) hearing special sounds. These results suggest targeted prevention strategies for decreasing the incidence of work-related injuries and illnesses, such as worker self-assessment of the total physical effort demanded by a job and periodic monitoring of workforce job satisfaction.


Mesogonadal shunts for extrahepatic portal vein thrombosis and variceal hemorrhage.
Extrahepatic portal vein thrombosis (EHPVT) may occur in children or adults and usually comes to clinical attention due to complications of portal hypertension such as variceal hemorrhage. A variety of standard surgical techniques exist to manage these patients, but when these fail surgical options are limited. We describe two novel portosystemic shunts that utilize the gonadal vein as an autologous conduit. Four patients were evaluated for EHPVT with variceal bleeding. None of the patients were candidates for a standard splenorenal shunt due to prior surgical procedures. The first patient underwent a left mesogonadal shunt and the remaining 3 patients underwent a right mesogonadal shunt. Postoperative ultrasound or computed tomography (CT) scan confirmed early patency of the shunt in each patient. There have been no further episodes of variceal hemorrhage with follow-up of 3.5 years in the child who underwent the left mesogonadal shunt, and 17, 19, and 20 months in the patients who underwent the right mesogonadal shunt. Three of the 4 shunts remain patent. One shunt thrombosis occurred in a patient homozygous for the Factor V Leiden mutation despite anticoagulation with coumadin. This is the first report of the successful use of the gonadal vein as an in situ conduit for constructing a portosystemic shunt. In conclusion, the right and left mesogonadal shunts may be useful as salvage operations for patients with EHPVT who have failed standard surgical shunt procedures.


[Successful gene therapy of mice with congenital erythropoietic porphyria].
Porphyrias are a group of disorders due to a genetic deficiency in one of the heme biosynthetic pathway enzymes. Congenital erythropoietic porphyria (CEP) is the most severe type characterized by a deficiency in uroporphyrinogen III synthase (UROS) activity. Bone marrow transplantation represents a curative treatment for patients, as long as human leucocyte antigen-compatible donor is available. We used a recently obtained murine model to check the feasibility of gene therapy in this disease. Lentivirus-mediated transfer of the human UROS cDNA into hematopoietic stem cells (HSCs) from Uros(mut 248) mice resulted in a complete and long-term enzymatic, metabolic and phenotypic correction of the disease, favored by a survival advantage of corrected red blood cells. These results demonstrate for the first time that the cure of this mouse model of CEP at moderate transduction level supports the proof of concept of a gene therapy in this disease by transplantation of genetically modified HSCs.


Ragaglitazar: the pharmacokinetics, pharmacodynamics, and tolerability of a novel dual PPAR alpha and gamma agonist in healthy subjects and patients with type 2 diabetes.
Ragaglitazar is a novel dual peroxisome proliferator-activated receptor (PPAR) alpha and gamma agonist intended to restore insulin sensitivity and correct diabetic dyslipidemia. These studies assessed single-dose pharmacokinetics and tolerability of ragaglitazar in healthy subjects, as well as multiple-dose pharmacokinetics, pharmacodynamics, and tolerability of ragaglitazar in healthy subjects and in patients with type 2 diabetes. Healthy subjects received a single oral dose (1-120 mg), and healthy subjects and type 2 diabetic patients received a loading dose and thereafter once-daily doses (0.5-16 mg) of ragaglitazar for 6 and 20 days, respectively. Ragaglitazar was rapidly absorbed (tmax: 1.5-1.7 h), with mean AUC0-24 h and Cmax proportional to dose after single and multiple dosing; t1/2 was 80 hours following a single dose and 104 hours in healthy subjects and 122 hours in patients after multiple dosing. Administration of 4 mg ragaglitazar to patients (n = 4) for 21 days resulted in mean decreases from baseline in fasting levels of plasma glucose (18%), C-peptide (18%), fructosamine (6%), triglycerides (36%), free fatty acids (49%), total cholesterol (11%), low-density lipoprotein (LDL) cholesterol (21%), and very low-density lipoprotein (VLDL) cholesterol (15%), as well as an increase in high-density lipoprotein (HDL) cholesterol (33%). Overall, ragaglitazar was well tolerated; with multiple dosing, there was a higher incidence of adverse events for patients that, at the highest dose level (16 mg), included peripheral edema and anemia.


A Novel Focal Seizure Pattern Generated in Superficial Layers of the Olfactory Cortex.
Seizure patterns identified in focal epilepsies caused by diverse etiologies are likely due to different pathogenic mechanisms. We describe here a novel, region-specific focal seizure pattern that mimics seizure activity observed in a subpopulation of patients submitted to presurgical monitoring with intracerebral electrodes. Distinctive seizure-like events (SLEs) are induced in the olfactory regions by acute treatment of both tangential brain slices and the isolated guinea pig brain with the potassium channel blocker 4-aminopyridine. Analysis of field potentials, intracellular activities, and extracellular potassium changes demonstrates that SLEs in the piriform cortex initiate in the superficial layer 1 lacking principal neurons with an activity-dependent increase of extracellular potassium. SLE progression (but not onset) does not require the participation of synaptic transmission and is mediated by diffusion of potassium to deep cortical layers. The novel seizure pattern here described is not observed in other cortical regions; it is proposed to rely on the peculiar organization of the superficial piriform cortex layers, which are characterized by unmyelinated axons and perisynaptic astroglial envelopes. This study reveals a sequence of ictogenic events in the olfactory cortex that were never described before in other cortical structures and supports the notion that altered potassium homeostasis and unmyelinated fibers may represent a potential vehicle for focal ictogenesis.SIGNIFICANCE STATEMENT We describe a novel seizure pattern peculiar of the olfactory cortex that resembles focal seizures with low-voltage fast activity at onset observed in humans. The findings suggest that network mechanisms responsible for seizure onset can be region specific.


Magnitude of Turnover Intention and Associated Factors among Health Professionals Working in Public Health Institutions of North Shoa Zone, Amhara Region, Ethiopia.
Health workers are one of the most important building blocks of the health system. High turnover rate contribute to the shortfalls and unbalanced distribution of health personnel in the health workforce. Turnover intention is the strongest predictor of actual turnover. To assess the magnitude of turnover intention and associated factors among health professionals working in public health institutions of North Shoa Zone, Amhara region, Ethiopia. A health facility based cross-sectional study was conducted from March to April 2016, on 612 health professionals working in public health institutions of North Shoa Zone, Amhara region, Ethiopia, using a multistage stratified sampling technique. Data were collected using a pretested self-administered structured questionnaire. The data were entered using Epidata version 3.1 and analyzed using SPSS version 22 software. Descriptive statistics were conducted to summarize the sample characteristics. Backward stepwise logistic regression model was fitted and AOR with 95% CI was calculated to identify the associated factors. P-value <0.05 was taken as a cut-off point for statistical significance. Ethical issues were addressed. Among the 568 health professionals who responded to the questionnaire, 348 (61.3%) of them reported to have the intention to leave their current workplaces. The results showed that being a male [AOR = 1.496 (95% CI: 1.016, 2.204)], medical doctor [AOR = 0.318 (95% CI: 0.122, 0.824)], unsatisfied with the work nature [AOR = 1.822 (95% CI: 1.206, 2.753)], unsatisfied with the incentives [AOR = 1.668 (95% CI: 1.105, 2.517)], and unsatisfied with the supervision [AOR = 1.916 (95% CI: 1.274, 2.881)] and having low normative commitment [AOR = 2.176 (95% CI: 1.482, 3.196)] were significantly associated with turnover intention of the health professionals. The magnitude of turnover intention was high among health professionals working in public health institutions of North Shoa Zone. Health service managers and policymakers should develop evidence based retention strategies considering the determinants of health professionals' intention to leave.


The simian T-lymphotropic virus STLV-PP1664 from Pan paniscus is distinctly related to HTLV-2 but differs in genomic organization.
We have isolated a highly divergent simian T-lymphotropic virus, STLV-PP1664, from a wild-caught bonobo (Pan paniscus). Previous phylogenetic analysis suggested that this virus represents an additional type of STLV but this has now become a matter of discussion. We have now obtained and analyzed the entire genome of STLV-PP1664. All major genes and their corresponding viral messengers were identified. Sequence comparison and phylogenetic analysis indicated that this virus, together with the closely related panp isolate, belongs to an early lineage within the PTLV-2 clade, differing from HTLV-2 by about 25%. In contrast to the HTLV-1 and HTLV-2 LTR, only two 21-bp repeats instead of three were found in the STLV-PP1664 LTR. Additional messengers, resulting from alternative splicing, potentially encode five different accessory proteins from open reading frames in the pX region: prorfI, porfII, ptorfV', and two isoforms of Rex. The amino acid sequences of these proteins are only distinctly related to the accessory proteins from HTLV-2. These data suggest a different genomic organization of the STLV-PP1664 pX region than that of HTLV-2. We conclude that STLV-PP1664, although related to HTLV-2, has some distinct features in the LTR and the pX regions, the impact of which needs further investigation. Although arguments pro and contra a distinct classification are nearly equally balanced, we propose to classify this virus as an STLV-2, designated STLV-2PP1664.


Conformational Heterogeneity in the Activation Mechanism of Bax.
Bax is known for its pro-apoptotic role within the mitochondrial pathway of apoptosis. However, the mechanism for transitioning Bax from cytosolic to membrane-bound oligomer remains elusive. Previous nuclear magnetic resonance (NMR) and electron paramagnetic resonance (EPR) studies defined monomeric Bax as conformationally homogeneous. Yet it has recently been proposed that monomeric Bax exists in equilibrium with a minor state that is distinctly different from its NMR structure. Here, we revisited the structural analysis of Bax using methods uniquely suited for unveiling "invisible" states of proteins, namely, NMR paramagnetic relaxation enhancements and EPR double electron-electron resonance (DEER). Additionally we examined the effect of glycerol, the co-solvent of choice in DEER studies, on the structure of Bax using NMR chemical-shift perturbations and residual dipolar couplings. Based on our combined NMR and EPR results, Bax is a conformationally homogeneous protein prior to its activation.


Lactococcus garvieae: where is it from? A first approach to explore the evolutionary history of this emerging pathogen.
The population structure and diversity of Lactococcus garvieae, an emerging pathogen of increasing clinical significance, was determined at both gene and genome level. Selected lactococcal isolates of various origins were analyzed by a multi locus sequence typing (MLST). This gene-based analysis was compared to genomic characteristics, estimated through the complete genome sequences available in database. The MLST identified two branches containing the majority of the strains and two branches bearing one strain each. One strain was particularly differentiated from the other L. garvieae strains, showing a significant genetic distance. The genomic characteristics, correlated to the MLST-based phylogeny, indicated that this "separated strain" appeared first and could be considered the evolutionary intermediate between Lactococcus lactis and L. garvieae main clusters. A preliminary genome analysis of L. garvieae indicated a pan-genome constituted of about 4100 genes, which included 1341 core genes and 2760 genes belonging to the dispensable genome. A total of 1491 Clusters of Orthologous Genes (COGs) were found to be specific to the 11 L. garvieae genomes, with the genome of the "separated strain" showing the highest presence of unique genes.


Diacylglycerol acyltransferase-1 inhibition enhances intestinal fatty acid oxidation and reduces energy intake in rats.
Acyl CoA:diacylglycerol acyltransferase-1 (DGAT-1) catalyzes the final step in triacylglycerol (TAG) synthesis and is highly expressed in the small intestine. Because DGAT-1 knockout mice are resistant to diet-induced obesity, we investigated the acute effects of intragastric (IG) infusion of a small molecule diacylglycerol acyltransferase-1 inhibitor (DGAT-1i) on eating, circulating fat metabolites, indirect calorimetry, and hepatic and intestinal expression of key fat catabolism enzymes in male rats adapted to an 8 h feeding-16 h deprivation schedule. Also, the DGAT-1i effect on fatty acid oxidation (FAO) was investigated in enterocyte cell culture models. IG DGAT-1i infusions reduced energy intake compared with vehicle in high-fat diet (HFD)-fed rats, but scarcely in chow-fed rats. IG DGAT-1i also blunted the postprandial increase in serum TAG and increased β-hydroxybutyrate levels only in HFD-fed rats, in which it lowered the respiratory quotient and increased intestinal, but not hepatic, protein levels of Complex III of the mitochondrial respiratory chain and of mitochondrial hydroxymethylglutaryl-CoA synthase. Finally, the DGAT-1i enhanced FAO in CaCo2 (EC50 = 0.3494) and HuTu80 (EC50 = 0.00762) cells. Thus, pharmacological DGAT-1 inhibition leads to an increase in intestinal FAO and ketogenesis when dietary fat is available. This may contribute to the observed eating-inhibitory effect.


Optimal wireless receiver structure for omnidirectional inductive power transmission to biomedical implants.
In order to achieve omnidirectional inductive power transmission to biomedical implants, the use of several orthogonal coils in the receiver side (Rx) has been proposed in the past. In this paper, the optimal Rx structure for connecting three orthogonal Rx coils and the power management is found to achieve the maximum power delivered to the load (PDL) in the presence of any Rx coil tilting. Unlike previous works, in which a separate power management has been used for each coil to deliver power to the load, different resonant Rx structures for connecting three Rx coils to a single power management are studied. In simulations, connecting three Rx coils with the diameters of 3 mm, 3.3 mm, and 3.6 mm in series and resonating them with a single capacitor at the operation frequency of 100 MHz led to the maximum PDL for large loads when the implant was tilted for 45o. This optimal Rx structure achieves higher PDL in worst-case scenarios as well as reduces the number of power managements to only one.