3D evaluation of the effect of disinfectants on dimensional accuracy and stability of two elastomeric impression materials.
The aim of this study was to determine and compare the dimensional changes of polyether and vinyl polyether siloxane impression materials under immersion disinfection with two different disinfectants in three time periods. Impressions were obtained from an edentulous master model. Sodium hypochlorite (5.25%) and glutaraldehyde (2%) were used for disinfection and measurements were done 30 min later after making impression before disinfection, after required disinfection period (10 min), and after 24 h storage at room temperature. Impressions were scanned using 3D scanner with 10 microns accuracy and 3D software was used to evaluate the dimensional changes with superimpositioning. Positive and negative deviations were calculated and compared with master model. There was no significant difference between two elastomeric impression materials (p>0.05). It was concluded that dimensional accuracy and stability of two impression materials were excellent and similar.


Phosphoglucomutase mutants of Escherichia coli K-12.
Bacteria with strongly depressed phosphoglucomutase (EC 126.96.36.199) activity are found among the mutants of Escherichia coli which, when grown on maltose, accumulate sufficient amylose to be detectable by iodine staining. These pgm mutants grow poorly on galactose but also accumulate amylose on this carbon source. Growth on lactose does not produce high amylose but, instead, results in the induction of the enzymes of maltose metabolism, presumably by accumulation of maltose. These facts suggest that the catabolism of glucose-1-phosphate is strongly depressed in pgm mutants, although not completely abolished. Anabolism of glucose-1-phosphate is also strongly depressed, since amino acid- or glucose-grown pgm mutants are sensitive to phage C21, indicating a deficiency in the biosynthesis of uridine diphosphoglucose or uridine diphosphogalactose, or both. All pgm mutations isolated map at about 16 min on the genetic map, between purE and the gal operon.


Hepatitis C Virus Replication Depends on Endosomal Cholesterol Homeostasis.
Similar to other positive-strand RNA viruses, hepatitis C virus (HCV) causes massive rearrangements of intracellular membranes, resulting in a membranous web (MW) composed of predominantly double-membrane vesicles (DMVs), the presumed sites of RNA replication. DMVs are enriched for cholesterol, but mechanistic details on the source and recruitment of cholesterol to the viral replication organelle are only partially known. Here we focused on selected lipid transfer proteins implicated in direct lipid transfer at various endoplasmic reticulum (ER)-membrane contact sites. RNA interference (RNAi)-mediated knockdown identified several hitherto unknown HCV dependency factors, such as steroidogenic acute regulatory protein-related lipid transfer domain protein 3 (STARD3), oxysterol-binding protein-related protein 1A and -B (OSBPL1A and -B), and Niemann-Pick-type C1 (NPC1), all residing at late endosome and lysosome membranes and required for efficient HCV RNA replication but not for replication of the closely related dengue virus. Focusing on NPC1, we found that knockdown or pharmacological inhibition caused cholesterol entrapment in lysosomal vesicles concomitant with decreased cholesterol abundance at sites containing the viral replicase factor NS5A. In untreated HCV-infected cells, unesterified cholesterol accumulated at the perinuclear region, partially colocalizing with NS5A at DMVs, arguing for NPC1-mediated endosomal cholesterol transport to the viral replication organelle. Consistent with cholesterol being an important structural component of DMVs, reducing NPC1-dependent endosomal cholesterol transport impaired MW integrity. This suggests that HCV usurps lipid transfer proteins, such as NPC1, at ER-late endosome/lysosome membrane contact sites to recruit cholesterol to the viral replication organelle, where it contributes to MW functionality.IMPORTANCE A key feature of the replication of positive-strand RNA viruses is the rearrangement of the host cell endomembrane system to produce a membranous replication organelle (RO). The underlying mechanisms are far from being elucidated fully. In this report, we provide evidence that HCV RNA replication depends on functional lipid transport along the endosomal-lysosomal pathway that is mediated by several lipid transfer proteins, such as the Niemann-Pick type C1 (NPC1) protein. Pharmacological inhibition of NPC1 function reduced viral replication, impaired the transport of cholesterol to the viral replication organelle, and altered organelle morphology. Besides NPC1, our study reports the importance of additional endosomal and lysosomal lipid transfer proteins required for viral replication, thus contributing to our understanding of how HCV manipulates their function in order to generate a membranous replication organelle. These results might have implications for the biogenesis of replication organelles of other positive-strand RNA viruses.


Is attendant at delivery associated with the use of interventions to prevent postpartum hemorrhage at home births? The case of Bangladesh.
Hemorrhage is the leading cause of maternal mortality in Bangladesh, the majority of which is due to postpartum hemorrhage (PPH), blood loss of 500 mL or more. Many deaths due to PPH occur at home where approximately 77% of births take place. This paper aims to determine whether the attendant at home delivery (i.e. traditional birth attendant (TBA) trained on PPH interventions, TBA not trained on interventions, or lay attendant) is associated with the use of interventions to prevent PPH at home births. Data come from operations research to determine the safety, feasibility, and acceptability of scaling-up community-based provision of misoprostol and an absorbent delivery mat in rural Bangladesh. Analyses were done using data from antenatal care (ANC) cards of women who delivered at home without a skilled attendant (N =66,489). Multivariate logistic regression was used to assess the likelihood of using the interventions. Overall, 67% of women who delivered at home without a skilled provider used misoprostol and the delivery mat (the interventions). Women who delivered at home and had a trained TBA present had 2.72 (95% confidence interval, 2.15-3.43) times the odds of using the interventions compared to those who had a lay person present. With each additional ANC visit (maximum of 4) a woman attended, the odds of using the interventions increased 2.76 times (95% confidence interval, 2.71-2.81). Other sociodemographic variables positively associated with use of the interventions were age, secondary or higher education, and having had a previous birth. Findings indicate that trained TBAs can have a significant impact on utilization of interventions to prevent PPH in home births. ANC visits can be an important point of contact for knowledge transfer and message reinforcement about PPH prevention.


The effect of cell fixation on the discrimination of normal and leukemia cells with laser tweezers Raman spectroscopy.
Laser tweezers Raman spectroscopy (LTRS) was used to characterize the effect of different chemical fixation procedures on the Raman spectra of normal and leukemia cells. Individual unfixed, paraformaldehyde-fixed, and methanol-fixed normal and transformed lymphocytes from three different cell lines were analyzed with LTRS. When compared to the spectra of unfixed cells, the fixed cell spectra show clear, reproducible changes in the intensity of specific Raman markers commonly assigned to DNA, RNA, protein, and lipid vibrations (e.g. 785, 1230, 1305, 1660 cm(-1)) in mammalian cells, many of which are important markers that have been used to discriminate between normal and cancer lymphocytes. Statistical analyses of the Raman data and classification using principal component analysis and linear discriminant analysis indicate that methanol fixation induces a greater change in the Raman spectra than paraformaldehyde. In addition, we demonstrate that the spectral changes as a result of the fixation process have an adverse effect on the accurate Raman discrimination of the normal and cancer cells. The spectral artifacts created by the use of fixatives indicate that the method of cell preparation is an important parameter to consider when applying Raman spectroscopy to characterize, image, or differentiate between different fixed cell samples to avoid potential misinterpretation of the data.


[A literature review of the concentrations of arsenic, lead, cadmium, and mercury in body fluids and tissues to establish normal levels and to detect exposure. 3. Mercury].
The 3rd of 4 communications on metal concentrations in human body media deals with mercury. Publications obtained from a comprehensive data bank search from January 1980 to April 1984 are listed in tables; study groups have been assigned to the following categories: normal, exposed; and exposed, exhibiting adverse health effects. Quality and strength of evidence of analytical procedures and reported data are discussed, particularly in light of adverse health effects. General methods and individual analytical procedures were presented in the 1st communication, a summary of mean exposure levels, critically elevated levels, and references will be published in the 4th communication.


Assessing arsenic exposure from consumption of seafood from Vieques-Puerto Rico: a pilot biomonitoring study using different biomarkers.
The various toxic effects associated with inorganic arsenic (iAs) warrants that exposure sources be identified. This pilot study evaluated if greater seafood consumption from Vieques-Puerto Rico is associated with increased exposure to iAs. Nail, hair, and urine samples were used as biomarkers of iAs exposure in adult women and men from Vieques classified as high (n = 31) and low (n = 21) seafood consumers, who reported eating fish and/or shellfish ≥1 time per week and once per month or less, respectively. The sum of urinary iAs (As III + As V), monomethylarsonic acid (MA[V]), and dimethylarsinic acid (DMA[V]), denoted as SumAs, fluctuated from 3.3 µg/g Cr (1.2 μg/L) to 42.7 μg/g Cr (42 μg/L) (n = 52). Levels of As in nail samples (n = 49) varied from 0.04 to 0.82 μg/g dry weight (dw), whereas in hair (n = 49) As was only detected in 49 % of the samples with a maximum value of 0.95 μg/g dw. None of the biomarkers of exposure to As exceeded exposure reference values for urine (50 μg/g Cr or 50 μg/L), nails (1 μg/g), or hair (1 μg/g). However, median (10.0 μg/g Cr; 10.6 μg/L) and 95th percentile (31.9 μg/g Cr; 40.4 μg/L) of urinary SumAs were higher in Vieques samples than in the those from the general population of other countries. Among the three biomarkers of exposure, nail samples reflected better the exposure to iAs from seafood consumption with significantly higher average As concentrations in high (0.24 μg/g) than low (0.12 μg/g) seafood consumers. Multivariate results for As in nail samples (R(2) = 0.55, p < 0.0001) showed a positive association with fish consumption, particularly for men, with levels increasing with years of residency in Vieques.


Phase II study of vinorelbine in heavily previously treated small cell lung cancer. Japan Lung Cancer Vinorelbine Study Group.
Twenty-four previously treated patients with refractory or relapsed small cell lung cancer (SCLC) were entered into a prospective, multicenter phase II study. All 24 patients had been pretreated with some form of cisplatin-based chemotherapy. The median time of chemotherapy was 4.2 months (range 1.4-9.4 months). Patients were treated with a dose of 25 mg/m2 of vinorelbine weekly. Twenty-four patients were eligible for response and for toxicity. Partial response was observed in 3 out of 24 eligible patients (12.5%; 95% confidence interval, 2.7-32.4%). All 3 patients who responded had previous chemotherapy including vincristine. The most common toxicity was leukopenia (91.7%, 66.7% in WHO 3-4 grade) and anemia (70.8%, 20.8% in WHO 3 grade). Nonhematological toxicities were moderate and mild. These results support a two-state sequential study design of previously untreated patients for further phase II study in SCLC.


Dolichol biosynthesis in human malignant cells.
Cholesterol, ubiquinone and dolichol biosynthesis from mevalonic acid was measured in non-malignant and malignant cultured human lymphocytes, freshly isolated human mononuclear leucocytes and in cultured human hepatoma cells. The relative flux of mevalonate into ubiquinone, dilichol and cholesterol was not significantly different between malignant and non-malignant cells, although the extent of labelling of each product was an order of magnitude greater in the malignant cultured cells. The most prominent dolichol isolated from total cellular lipid and synthesized in short-term labelling of cultured leukaemic cells had a chain length one isoprene unit shorter than that observed in normal human cells. Cultured human hepatoma cells and mononuclear leucocytes isolated from the peripheral blood of individuals with lymphoblastic and myelogenic leukaemia similarly synthesized shorter-chain dolichols. The dolichols made in cultured non-tumorigenic cells, freshly isolated mononuclear leucocytes from a normal individual or a patient with non-haematological malignancy had normal chain length.


Regulation of adipocyte differentiation by the zinc finger protein ZNF638.
Zinc finger proteins constitute the largest family of transcription regulators in eukaryotes. These factors are involved in diverse processes in many tissues, including development and differentiation. We report here the characterization of the zinc finger protein ZNF638 as a novel regulator of adipogenesis. ZNF638 is induced early during adipocyte differentiation. Ectopic expression of ZNF638 increases adipogenesis in vitro, whereas its knockdown inhibits differentiation and decreases the expression of adipocyte-specific genes. ZNF638 physically interacts and transcriptionally cooperates with CCAAT/enhancer-binding protein (C/EBP) β and C/EBPδ. This interaction leads to the expression of peroxisome proliferator-activated receptor γ, which is the key regulator of adipocyte differentiation. In summary, ZNF638 is a novel and early regulator of adipogenesis that works as a transcription cofactor of C/EBPs.


[Echocardiographic changes in the elderly population].
Elderly population rate rises all around the world. Increased life time due to improvements in the treatment of diseases which have high mortality rates, getting easier to reach healthcare services and getting widespread of the antibiotic use is the main cause underlying the increase of geriatric population. Cardiovascular diseases are the most common cause of morbidity and mortality in the overall population. It is well known that the incidence of cardiovascular diseases increases with ageing. Anatomic and functional changes occur in the cardiovascular system as well as all other organ systems with senescence and only some of these changes can be detected clinically while others stay in the subclinical course. Echocardiography is an important option owing to be able to detect most of these changes.


Influence of β-CN Genotype on Physicochemical Properties and Functionality of Bovine Milk.
Several studies have been focused on the effect of milk protein genetic variants on milk physicochemical properties and functionality in recent years. β-casein, an important protein related to milk processibility, has been reported to have 2 main genetic variants A1 and A2, for which cows may be homozygous or heterozygous. In this study, several physicochemical properties of milk with β-casein variants A1A1, A1A2 and A2A2 from 3 collection occasions were analyzed. Higher manganese content and lower pH were found to be associated with the A1A1 variant compared with the other 2 genotypes. Better rennet and acid coagulation were found in A1A1 milk compared with A1A2 and A2A2 milk (although P > 0.05), while A2A2 milk was more stable to creaming compared with the other 2 genotypes, which may be linked to its smaller fat globule size. Thus, milk from cows with A1A1 genotype could be preferable for cheese making, while that with A2A2 variant can be used in formulations requiring good stability against creaming, and for, e.g., yogurt making, where the softer yogurt texture may be easier to digest.


A comparison of EUS features between CD-117 positive GI stromal tumors and CD-117 negative GI spindle cell tumors.
GI stromal tumors are spindle cell tumors that stain positive for immunohistochemical CD-117 (c-kit). Prognostic factors for malignancy include size (> or =4 cm), mitotic index (5 mitotic figures/50 high-powered fields), and ulcerated, cystic, or necrotic areas within the tumor. The purpose of this study was to compare these features in c-kit positive vs. c-kit negative tumors. All patients referred for EUS of submucosal lesions were identified, and histopathology, including immunohistochemical staining, was reviewed to determine all diagnoses of GI stromal tumors. Size, echo pattern, and presence of cystic spaces and ulceration were recorded as diagnosed by EUS. Histopathologic diagnoses were made by FNA or endoscopic submucosal-mucosal resection. If surgical resection followed, the surgical diagnosis, staining pattern, mitotic index, and presence of ulceration, necrosis, and nuclear atypia were recorded. Forty patients (21 men, 19 women; 38 white, 2 African American; mean age 58 +/- 2.6 years) had 46 EUS procedures performed for evaluation of spindle cell tumors. Seventeen stained positive for c-kit (mean age, 59 +/- 3.6 years; range 19 to 80 years) and 12 negative (mean age, 57 +/- 3.8 years; range 31 to 76 years); 11 were not stained for c-kit (excluded from analysis). On EUS, 7 were ulcerated, 3 cystic, and 6 were larger than 4 cm. This group of findings was observed in 12 patients, 11 of whom had c-kit positive tumors (11/17 vs. 1/12; p = 0.006). Tumors positive for c-kit were larger (42.4 +/- 5.5 mm vs. 19.0 +/- 5.9 mm; p = 0.005). There were 13 c-kit positive tumors in the stomach, 2 in the duodenum, and 1 each in the esophagus and at the gastroesophageal junction. Of the 12 c-kit negative tumors, 8 were located in the esophagus and 1 at the gastroesophageal junction (9/12 vs. 2/17; p < 0.01). Surgical resection was performed on 13 patients, 12 of whom had c-kit positive tumors, and 3 of these 12 tumors had greater than 5 mitoses per 50 high-powered field. If a GI stromal tumor is suspected, EUS-FNA with immunohistochemical staining should be performed for CD-117 (c-kit). C-kit tumors are more likely to have malignant features and should be resected or subjected to close clinical follow-up.


The effects of pumpkin seed oil supplementation on arterial hemodynamics, stiffness and cardiac autonomic function in postmenopausal women.
Postmenopausal women have a higher prevalence of hypertension than age-match men. Evidence from animal studies have demonstrated the antihypertensive effects of pumpkin seed oil (PSO). We examined the effects of PSO supplementation on vascular function and heart rate variability (HRV) in postmenopausal women with elevated blood pressure (BP). Participants were randomly assigned to either a PSO (n = 12) or a placebo group (n = 11). Participants in the PSO group consumed 3 g/day of PSO. Brachial and central BP, wave reflection (augmentation index, AIx), arterial stiffness (SI) and various HRV parameters were measured before and after 6 weeks. AIx, brachial and central systolic BP significantly (P < 0.05) decreased following PSO but not after placebo. SI and HRV parameters remained unchanged after PSO or placebo. PSO improved arterial hemodynamics in postmenopausal women and therefore might be effective in the prevention and treatment of hypertension in this population. CLINICAL TRIAL ID: (NCT03716960).


Regulation of respiration in myocardium in the transient and steady state.
1H/(31)P NMR has followed the metabolic response to increased work in the glucose- and pyruvate-perfused rat myocardium during a heart cycle and at the steady state. With electrical pacing and dobutamine, the heart O(2) consumption increases by 56%. The phosphocreatine (PCr) level initially declines, but recovers within 15 min to its control level; the oxymyoglobin (MbO(2)) saturation decreases by 15%. Because the MbO(2) signal reflects the intracellular PO(2), the capillary-to-cell O(2) gradient has increased to match the increased O(2) need. However, no transient metabolic fluctuation is observed in either PCr or MbO(2) throughout the entire cardiac cycle in both glucose and pyruvate-/glucose-perfused hearts. No systolic-diastolic variation is detectable under either high workload or hypoxic conditions. The results reveal that neither O(2) nor ADP is regulating respiration under increased energy demand in the steady or transient state.


Interaction among autoregulation, CO2 reactivity, and intracranial pressure: a mathematical model.
The relationships among cerebral blood flow, cerebral blood volume, intracranial pressure (ICP), and the action of cerebrovascular regulatory mechanisms (autoregulation and CO2 reactivity) were investigated by means of a mathematical model. The model incorporates the cerebrospinal fluid (CSF) circulation, the intracranial pressure-volume relationship, and cerebral hemodynamics. The latter is based on the following main assumptions: the middle cerebral arteries behave passively following transmural pressure changes; the pial arterial circulation includes two segments (large and small pial arteries) subject to different autoregulation mechanisms; and the venous cerebrovascular bed behaves as a Starling resistor. A new aspect of the model exists in the description of CO2 reactivity in the pial arterial circulation and in the analysis of its nonlinear interaction with autoregulation. Simulation results, obtained at constant ICP using various combinations of mean arterial pressure and CO2 pressure, substantially support data on cerebral blood flow and velocity reported in the physiological literature concerning both the separate effects of CO2 and autoregulation and their nonlinear interaction. Simulations performed in dynamic conditions with varying ICP underline the existence of a significant correlation between ICP dynamics and cerebral hemodynamics in response to CO2 changes. This correlation may significantly increase in pathological subjects with poor intracranial compliance and reduced CSF outflow. In perspective, the model can be used to study ICP and blood velocity time patterns in neurosurgical patients in order to gain a deeper insight into the pathophysiological mechanisms leading to intracranial hypertension and secondary brain damage.


Kinetic and molecular properties of the dihydrofolate reductase from pyrimethamine-sensitive and pyrimethamine-resistant clones of the human malaria parasite Plasmodium falciparum.
Dihydrofolate reductase (DHFR) (5,6,7,8-tetrahydrofolate: NADPH+-oxidoreductase; EC 184.108.40.206) was partially purified by affinity chromatography from three clones of the human malaria parasite Plasmodium falciparum. The three clones were representative of pyrimethamine-sensitive (clone 3D7) and pyrimethamine-resistant (clone HB3 and clone 7G8) parasites with ID50 values of 0.53 nM (3D7), 210 nM (HB3), and 540 nM (7G8), when tested in vitro against the drug. The specific activities of the partially purified DHFR differed by less than a factor of 2 between the sensitive clone 3D7 (442 +/- 39 nmol min-1 mg-1 protein) and the resistant clones HB3 (634 +/- 25 nmol min-1 mg-1 protein) and 7G8 (565 +/- 85 nmol min-1 mg-1 protein). The number of catalytic sites in partially purified DHFR from the three clones was similar and ranged from 151 to 194 pmol mg-1 protein. The Km value for NADPH was similar in all three clones (4.5-11.6 microM). The Km value for dihydrofolate was altered 13-fold comparing the sensitive clone 3D7 (3.2 +/- 0.6 microM) with the resistant clone HB3 (42.6 +/- 1.6 microM), with the Km for the resistant clone 7G8 falling in between (11.9 +/- 1.2 microM). The inhibition constants for pyrimethamine increased from 0.19 +/- 0.08 nM (3D7) to 2.0 +/- 0.3 nM (HB3) to 8.9 +/- 0.8 nM (7G8). The inhibition by pyrimethamine of the sensitive clone 3D7 was noncompetitive and competitive for the two other clones. The titration of partially purified DHFR with pyrimethamine revealed a 500-fold increase in the concentration of the drug needed to inhibit the DHFR activity by 50%, when the sensitive clone 3D7 (0.18 +/- 0.02 nM) was compared to the resistant clone 7G8 (95 +/- 16 nM). From the comparison of the specific activities and the catalytic center activities with the Km values for the substrate and the inhibition constants for pyrimethamine, both of which are altered in the resistant clones, we conclude that the molecular mechanism for pyrimethamine resistance in the three clones studied is not based on an overproduction of the DHFR but is due to a decreased affinity to antifolates by a structurally altered enzyme.


Adherence with a low-FODMAP diet in irritable bowel syndrome: are eating disorders the missing link?
The low-FODMAP diet has emerged as an option for the treatment of irritable bowel syndrome (IBS). This diet is very restrictive, and compliance is usually low. Preliminary findings suggest an association between eating disorders (EDs) and the risk of developing IBS. The aim of this study was to assess the correlation between compliance with a low-FODMAP diet and the risk of ED behaviours among patients with IBS. A single-centre prospective study was carried out among 233 IBS patients (79.8% females) at University College London Hospital, who commenced a low FODMAPs group programme for IBS (Rome III or IV). Self-reported diet adherence at the end of the 6-week programme was measured. At baseline, and at the 6-week follow-up visit, participants completed the validated IBS-Symptom Severity Score, the SCOFF ED screening questionnaire and the Hospital Anxiety and Depression Scale. Adherence with a low-FODMAP diet was found in 95 (41%) patients. Overall, 54 (23%) patients were classified to be at risk for ED behaviour. Adherence was 57% in the ED group (31/54) versus 35% in the non-ED group (64/179); P<0.05. Adherence with a low-FODMAP diet was highest (51%) in the IBS with diarrhoea subtype and lowest (10%) in IBS with constipation. There was no significant correlation between IBS-Symptom Severity Score and either adherence (P=0.39) or ED behaviour (P=0.28). In this IBS cohort, greater adherence to a low-FODMAP diet is associated with ED behaviour. The implications of our study are important in clinical practice for a clinician to have a high index of suspicion of EDs in IBS patients when a high level of low-FODMAP diet achieved.


Arterial stiffening influence of sympathetic nerve activity: evidence from hand transplantation in humans.
Studies in animals and humans suggest that sympathetic activity exerts a stiffening influence on large and middle-sized artery walls. We sought to obtain further evidence on this issue by measuring radial artery distensibility in an allotransplanted and thus denervated hand using the contralateral artery as control. In 2 men, blood pressure was measured by a semiautomatic device (Dinamap). Diastolic diameter, systo-diastolic diameter excursion (ultrasound Wall Track system), and distensibility (Reneman formula) of both radial arteries were measured at a level corresponding to 4 cm below the suture of the transplanted hand 40 days after surgery and every 4 weeks for the next 6 months. After surgery, systo-diastolic diameter excursion and distensibility were much greater in the transplanted radial artery than in the contralateral vessel, reaching values similar to the contralateral ones after 4 months, when signs of reinnervation of the transplanted hands had appeared. Radial deinnervation was accompanied by an increased arterial distensibility, which provides further evidence of the sympathetic stiffening effect on arterial wall in humans.


Adverse childhood experiences, adult anxiety and social capital among women in rural Kenya.
Hundreds of millions of people suffer anxiety disorders globally, demonstrating need for scalable and effective interventions. Adverse childhood experiences contribute to this mental health burden. The stress-buffering hypothesis, which posits social factors moderate prior adversity and subsequent mental health outcomes, provides one theoretical avenue to consider observations that group-based microfinance programs improve social capital. We investigate associations between adverse childhood experiences, generalized anxiety among adults and social capital associated with participation in a group-based microfinance program in rural Kenya. Adult participants (n = 400 women) responded to standardized measures of childhood adversity in June 2018, group-affiliated social capital and generalized anxiety in June 2019. Cumulative adverse childhood experiences predicted higher anxiety, which was statistically moderated by the presence of group-affiliated interpersonal trust. This study is the first to find social capital associated with participation in a group-based microfinance program statistically moderates expected associations between adverse childhood experiences and adult generalized anxiety. Future study should be conducted using a cluster-randomized control design to further assess the potential of this intervention method to ameliorate associations between past adversity and current mental health.


Deep Brain Stimulation for Essential Vocal Tremor: A Technical Report.
Essential vocal tremor (EVT) is the presence of a tremulous voice that is commonly associated with essential tremor. Patients with EVT often report a necessary increase in vocal effort that significantly worsens with stress and anxiety and can significantly impact quality of life despite optimal medical and behavioral treatment options. Deep brain stimulation (DBS) has been proposed as an effective therapy for vocal tremor, but very few studies exist in the literature that comprehensively evaluate the efficacy of DBS for specifically addressing EVT. We present a technical report on our multidisciplinary, comprehensive operative methodology for treatment of EVT with frameless, awake deep brain stimulation (DBS).


The Lys 280 → Gln mutation mimicking disease-linked acetylation of Lys 280 in tau extends the structural core of fibrils and modulates their catalytic properties.
Amyloid fibrillar aggregates isolated from the brains of patients with neurodegenerative diseases invariably have post-translational modifications (PTMs). The roles that PTMs play in modulating the structures and polymorphism of amyloid aggregates, and hence their ability to catalyze the conversion of monomeric protein to their fibrillar structure is, however, poorly understood. This is particularly true in the case of tau aggregates, where specific folds of fibrillar tau have been implicated in specific tauopathies. Several PTMs, including acetylation at Lys 280, increase aggregation of tau in the brain, and increase neurodegeneration. In this study, tau-K18 K280Q, in which the Lys 280 → Gln mutation is used to mimic acetylation at Lys 280, is shown, using HX-MS measurements, to form fibrils with a structural core that is longer than that of tau-K18 fibrils. Measurements of critical concentrations show that the binding affinity of monomeric tau-K18 for its fibrillar counterpart is only marginally more than that of monomeric tau-K18 K280Q for its fibrillar counterpart. Quantitative analysis of the kinetics of seeded aggregation, using a simple Michaelis-Menten-like model, in which the monomer first binds and then undergoes conformational conversion to β-strand, shows that the fibrils of tau-K18 K280Q convert monomeric protein more slowly than do fibrils of tau-K18. In contrast, monomeric tau-K18 K280Q is converted faster to fibrils than is monomeric tau-K18. Thus, the effect of Lys 280 acetylation on tau aggregate propagation in brain cells is expected to depend on the amount of acetylated tau present, and on whether the propagating seed is acetylated at Lys 280 or not.


Correlation of animal brain tumor models with human neuro-oncology.
The paper reviews the histopathology and cytopathology of a number of experimental animal brain tumor models to determine their relevance to human neuro-oncology. The models discussed include brain tumors produced by local implants of carcinogenic hydrocarbons and by the administration of resorptive N-nitroso compounds, some of the cell lines derived from the latter, tumors resulting from the intracerebral inoculation of oncogenic viruses, and the differentiating neuroepithelial component of a transplantable mouse teratoma. The structural and biochemical criteria that can be applied for the identification of the various stages of neuronal and glial cell maturation are highly precise. They reveal a wide diversity of tumor cell types that are now available in experimental neuro-oncogenesis.


Effect of bone marrow-derived stem cells on chondrocytes from patients with osteoarthritis.
Increasing numbers of individuals are suffering from osteoarthritis every year, and the directed intra-articular injection of bone marrow stem cells has provided a promising treatment strategy for osteoarthritis. Although a number of studies have demonstrated that intra-articular injection of bone marrow stem cells produced desirable results, the mechanism underlying this effect has not been elucidated. In the current study, the effect of bone marrow stem cells on chondrocytes from patients with osteoarthritis was observed in a co-culture system. Human chondrocytes were obtained from patients with osteoarthritis who underwent surgical procedures and bone marrow stem cells were obtained from bone marrow aspirates, and then the chondrocytes were then cultured alone or cocultured with bone marrow stem cells in 0.4-µm Transwell inserts. The differentiation and biological activity of chondrocytes in the culture system were measured, and the inflammatory factors and OA-associated markers were also measured. The results indicated that coculture with human bone marrow stem cells increases cell proliferation of chondrocytes and inhibits inflammatory activity in osteoarthritis.


The antibiotic resistome: what's new?
The antibiotic resistome is dynamic and ever expanding, yet its foundations were laid long before the introduction of antibiotics into clinical practice. Here, we revisit our theoretical framework for the resistome concept and consider the many factors that influence the evolution of novel resistance genes, the spread of mobile resistance elements, and the ramifications of these processes for clinical practice. Observing the trends and prevalence of genes within the antibiotic resistome is key to maintaining the efficacy of antibiotics in the clinic.


Posttraumatic Stress Disorder and Risk for Stroke in Young and Middle-Aged Adults: A 13-Year Cohort Study.
Background and Purpose- In older populations, transient ischemic attack (TIA) and ischemic stroke have been linked to psychological factors, including posttraumatic stress disorder (PTSD). Whether PTSD also increases risk for early incident stroke in young adults is unknown. Methods- We prospectively assessed the incidence of TIA and ischemic stroke in a cohort of 987 855 young and middle-aged Veterans (mean age of 30.29±9.19 years; 87.8% men, 64.4% white) who first accessed care through the Veterans Health Administration from October 2001 to November 2014 and were free of TIA and ischemic stroke at baseline. For each outcome, time-varying multivariate Cox models were constructed to examine the effect of PTSD on incident stroke. We also assessed for effect modification by sex. Additional sensitivity analyses controlled for healthcare utilization. Results- Over a 13-year period, TIA and ischemic stroke were diagnosed in 766 and 1877 patients, respectively. PTSD was diagnosed in 28.6% of the sample during follow-up. In unadjusted analyses, PTSD was significantly associated with new-onset TIA (hazard ratio [HR], 2.02; 95% CI, 1.62-2.52) and ischemic stroke (HR, 1.62; 95% CI, 1.47-1.79). In fully adjusted models, the association between PTSD and incident TIA (HR, 1.61; 95% CI, 1.27-2.04) and ischemic stroke (HR, 1.36; 95% CI, 1.22-1.52) remained significant. The effect of PTSD on ischemic stroke risk was stronger in men than in women (HR, 0.63; 95% CI, 0.47-0.86; P=0.003), but no effect of sex was found for TIA. Conclusions- PTSD is associated with a significant increase in risk of early incident TIA and ischemic stroke independent of established stroke risk factors, coexisting psychiatric disorders, and healthcare utilization. Sex moderated the relationship for adults with ischemic stroke but not TIA. These findings suggest that psychological factors, including PTSD, may be important targets for future age-specific prevention strategies for young adults.


Strength through adversity: bereaved cancer carers' accounts of rewards and personal growth from caring.
Many studies have identified negative and distressing consequences experienced by informal cancer carers, but less attention has been given to positive and beneficial aspects of caring. This qualitative study examined the positive aspects of caring as subjectively constructed by bereaved informal cancer carers, a group of individuals who are in a position to make sense of their caring experiences as a coherent whole. Twenty-three bereaved informal cancer carers were interviewed, and their accounts were analyzed using a thematic analytical approach from a phenomenological perspective. The participants were able to identify positive and beneficial aspects of caring. These included the discovery of personal strength, through adversity, acceptance, and necessity; the deepening of their relationship with the person for whom they cared; and personal growth through altered relationships with others and altered perspectives on living. Many participants gave accounts of focusing on these positive benefits when they reflected on their caring experiences. We concluded that benefit finding in the face of adverse events serves an important function in allowing individuals to incorporate difficult experiences into their worldview in a meaningful way, thus maintaining positive beliefs about the world. This has implications for the development of interventions for informal cancer carers and for those who are bereaved following caring.


Bedside Shift-to-Shift Handoffs: A Systematic Review of the Literature.
Multiple institutions identify communication and shift-to-shift handoffs as a source of clinical errors. Conducting handoffs at the bedside has been proposed as a potential solution to improve the quality of communication at change of shift. This article reports a systematic review of English-language research articles, published between January 1, 2008, and October 31, 2014, focused on bedside shift-to-shift handoffs. Forty-one articles were identified and analyzed in this systematic review.


Agonist-Specific Conformations of the M2 Muscarinic Acetylcholine Receptor Assessed by Molecular Dynamics.
Binding of muscarinic ligands, both antagonists and agonists, and their effects on the conformation of the M2 acetylcholine receptor were modeled in silico and compared to experimental data. After docking of antagonists to the M2 receptor in an inactive conformation (3UON, 5ZK3, 5ZKB, or 5ZKB) and agonists in an active conformation (4MQS), 100 ns of conventional molecular dynamics (MD) followed by 500 ns of accelerated MD was run. Conventional MD revealed ligand-specific interactions with the receptor. Antagonists stabilized the receptor in an inactive conformation during accelerated MD. The receptor in complex with various agonists attained different conformations specific to individual agonists. The magnitude of the TM6 movement correlated with agonist efficacy at the non-preferential Gs pathway. The shape of the intracellular opening where the receptor interacts with a G-protein was different for the classical agonist carbachol, super-agonist iperoxo, and Gi/o-biased partial agonists JR-6 and JR-7, being compatible with experimentally observed agonist bias at the G-protein level. Moreover, a wash-resistant binding of the unique agonist xanomeline associated with interactions with membrane lipids was formed during accelerated MD. Thus, accelerated MD is suitable for modeling of ligand-specific receptor binding and receptor conformations that is essential for the design of experiments aimed at identification of the secondary binding sites and understanding molecular mechanisms underlying receptor activation.


[Legal liability of the radiologist in missed breast cancer diagnosis].
There have been few lawsuits alleging missed breast cancer diagnosis in Belgium so far. However, since the introduction of the Flemish organised breast cancer screening program in June 2001, one might expect an increase in the years to come. In the present paper, we focus on risk management pointers that minimize the risk of incurring a malpractice lawsuit.


In vitro and in vivo conjugation of dietary hydroxycinnamic acids by UDP-glucuronosyltransferases and sulfotransferases in humans.
Hydroxycinnamic acids are a class of phenolic antioxidants found widely in dietary plants. Their biotransformation in the human organism primarily involves Phase II conjugation reactions. In this study, activities of UDP-glucuronosyltransferases (UGTs) and sulfotransferases (SULTs) towards major dietary hydroxycinnamic acids (caffeic, dihydrocaffeic, dihydroferulic, ferulic and isoferulic acids) were investigated. Conjugate formation was evaluated using human liver and intestinal S9 homogenates, and in vitro characterization was carried out using recombinant human UGTs and SULTs. Analysis of the kinetics of hydroxycinnamic acid conjugation in human S9 homogenates revealed that intrinsic clearance (V(max)/K(m)) is much greater for sulfation than for glucuronidation. Assessment of activity using a panel of recombinant human SULTs showed that SULT1A1 is most active in the sulfation of caffeic, dihydrocaffeic and isoferulic acids, while SULT1E1 is most active in the sulfation of ferulic and dihydroferulic acids. Only isoferulic acid was significantly glucuronidated by human liver S9 homogenates, explained by the high activity of liver-specific UGT1A9. Studies on the kinetics of active SULTs and UGTs demonstrated a markedly lower K(m) for SULTs. To further corroborate our findings, we carried out an intervention study in healthy humans to determine the hydroxycinnamic acid conjugates in urine after consumption of hydroxycinnamate-rich coffee (200 ml). Analysis showed that sulfates are the main conjugates in urine, with the exception of isoferulic acid, which is mainly glucuronidated. These data suggest that sulfates are the predominant hydroxycinnamic acid conjugates in humans, and that SULT mediated sulfation is a major factor determining the bioavailability of hydroxycinnamic acids in vivo.


Short rest interval lengths between sets optimally enhance body composition and performance with 8 weeks of strength resistance training in older men.
To determine if 8 weeks of periodized strength resistance training (RT) utilizing relatively short rest interval lengths (RI) in between sets (SS) would induce greater improvements in body composition and muscular performance, compared to the same RT program utilizing extended RI (SL). 22 male volunteers (SS: n = 11, 65.6 ± 3.4 years; SL: n = 11, 70.3 ± 4.9 years) were assigned to one of two strength RT groups, following 4 weeks of periodized hypertrophic RT (PHRT): strength RT with 60-s RI (SS) or strength RT with 4-min RI (SL). Prior to randomization, all 22 study participants trained 3 days/week, for 4 weeks, targeting hypertrophy; from week 4 to week 12, SS and SL followed the same periodized strength RT program for 8 weeks, with RI the only difference in their RT prescription. Following PHRT, all study participants experienced increases in lean body mass (LBM) (p < 0.01), upper and lower body strength (p < 0.001), and dynamic power (p < 0.001), as well as decreases in percentage body fat (p < 0.05). Across the 8-week strength RT phase, SS experienced significantly greater increases in LBM (p = 0.001), flat machine bench press 1-RM (p < 0.001), bilateral leg press 1-RM (p < 0.001), narrow/neutral grip lat pulldown (p < 0.01), and Margaria stair-climbing power (p < 0.001), compared to SL. This study suggests 8 weeks of periodized high-intensity strength RT with shortened RI induces significantly greater enhancements in body composition, muscular performance, and functional performance, compared to the same RT prescription with extended RI, in older men. Applied professionals may optimize certain RT-induced adaptations, by incorporating shortened RI.


Intraspinal tumours in the Kenya African.
Thirty-one cases of intraspinal tumours in the African have been described, with age, sex incidence, frequency, site and histopathology shown. Intraspinal tumours in this series are compared with the larger series. Extradural and intramedullary tumours together with cervical spine tumours appear to be more frequent in this series. There is a high incidence of dumbell tumours in the neurinomas. Sarcomas are the most common type of tumours and mainly affect the thoracic spine.


Ent-kaurene synthase from the fungus Phaeosphaeria sp. L487. cDNA isolation, characterization, and bacterial expression of a bifunctional diterpene cyclase in fungal gibberellin biosynthesis.
ent-Kaurene is the first cyclic diterpene intermediate of gibberellin biosynthesis in both plants and fungi. In plants, ent-kaurene is synthesized from geranylgeranyl diphosphate via copalyl diphosphate in a two-step cyclization catalyzed by copalyl diphosphate synthase and ent-kaurene synthase. A cell-free system of the fungus Phaeosphaeria sp. L487 converted labeled geranylgeranyl diphosphate to ent-kaurene. A cDNA fragment, which possibly encodes copalyl diphosphate synthase, was isolated by reverse transcription-polymerase chain reaction using degenerate primers based on the consensus motifs of plant enzymes. Translation of a full-length cDNA sequence isolated from the fungal cDNA library revealed an open reading frame for a 106-kDa polypeptide. The deduced amino acid sequence shared 24 and 21% identity with maize copalyl diphosphate synthase and pumpkin ent-kaurene synthase, respectively. A fusion protein produced by expression of the cDNA in Escherichia coli catalyzed the two-step cyclization of geranylgeranyl diphosphate to ent-kaurene. Amo-1618 completely inhibited the copalyl diphosphate synthase activity of the enzyme at 10(-6) M, whereas it did not inhibit the ent-kaurene synthase activity even at 10(-4) M. These results indicate that the fungus has a bifunctional diterpene cyclase that can convert geranylgeranyl diphosphate into ent-kaurene. They may be separate catalytic sites for the two cyclization reactions.


Ultrasensitive surface-enhanced Raman scattering detection in common fluids.
Detecting target analytes with high specificity and sensitivity in any fluid is of fundamental importance to analytical science and technology. Surface-enhanced Raman scattering (SERS) has proven to be capable of detecting single molecules with high specificity, but achieving single-molecule sensitivity in any highly diluted solutions remains a challenge. Here we demonstrate a universal platform that allows for the enrichment and delivery of analytes into the SERS-sensitive sites in both aqueous and nonaqueous fluids, and its subsequent quantitative detection of Rhodamine 6G (R6G) down to ∼75 fM level (10(-15) mol⋅L(-1)). Our platform, termed slippery liquid-infused porous surface-enhanced Raman scattering (SLIPSERS), is based on a slippery, omniphobic substrate that enables the complete concentration of analytes and SERS substrates (e.g., Au nanoparticles) within an evaporating liquid droplet. Combining our SLIPSERS platform with a SERS mapping technique, we have systematically quantified the probability, p(c), of detecting R6G molecules at concentrations c ranging from 750 fM (p > 90%) down to 75 aM (10(-18) mol⋅L(-1)) levels (p ≤ 1.4%). The ability to detect analytes down to attomolar level is the lowest limit of detection for any SERS-based detection reported thus far. We have shown that analytes present in liquid, solid, or air phases can be extracted using a suitable liquid solvent and subsequently detected through SLIPSERS. Based on this platform, we have further demonstrated ultrasensitive detection of chemical and biological molecules as well as environmental contaminants within a broad range of common fluids for potential applications related to analytical chemistry, molecular diagnostics, environmental monitoring, and national security.


Comparative evaluation between two nutritional supplements in the improvement of telogen effluvium.
Telogen effluvium (TE) is defined as a diffuse hair loss characterized by shortening of the anagen phase and precipitation of the telogen phase, with a consequent reduction of total hair volume. Nutritional supplementation is indicated under conditions in which TE is related to dietary disorders. The aim of this clinical study was to compare the efficacy of two different supplements in monotherapy for TE. Female adult patients were randomized to receive two oral nutritional supplements (group 1: a supplement composed of zinc, biotin, iron, vitamins A, C, E, and B complex, folic acid, magnesium, and amino acids of keratin and collagen and group 2: calcium pantothenate cystine, thiamine nitrate, medicinal yeast, keratin, and aminobenzoic acid) to treat hair loss for 180 days. They were evaluated clinically and by digital trichoscopy. Clinical evaluation showed significant clinical improvement (P<0.05) for the evaluated parameters: hair loss, hair volume, density of hair (scalp cover), hair shine, hair strength, in 180 days. At 90 days evaluation, group 1 showed significant improvement for all parameters, while group 2 did not show any significant improvement for hair shine and hair strength. In the digital trichoscopy, there was a significant improvement only in group 1 (11.09%×7.76%) after 180 days. In idiopathic TE, the nutritional component should be suspected; the supplementation of an association of nutrients in recommended daily intake can lead to significant improvement of the condition from the first trimester of use. The use of an association with proven efficacy and a safety profile and posologic convenience facilitate its indication and patient adherence.


Incidence and age-standardized rates of pancreatic cancer in Sri Lanka from 2001 to 2010: An analysis of national cancer registry data.
Varying trends in the incidence of pancreatic cancer (PC) are observed in many Asian countries. This study aimed at describing the incidence and age-standardized rates of PC in Sri Lanka from 2001-2010. A retrospective cohort evaluation of patients with PC from 2001 to 2010 was performed using the population-based data published by the Sri Lanka National Cancer Registry. The trends in the incidence of PC was analyzed by age and sex using joinpoint regression analysis. A total of 808 PC patients studied from 2001-2010, of which males were 438 (54.2%). The mean (±standard deviation) age of the total population was 55.7 (±13.8) years [males = 56.5 (±13.3) vs. females = 54.8 (±14.3), P=0.07]. The World Health Organization (WHO) age-standardized incidence of PC in Sri Lanka increased marginally from 0.44 per 100,000 in 2001 (95% confidence interval (CI) = 0.34-0.54) to 0.58 per 100,000 in 2010 (95% CI = 0.46-0.69) which is a 1.3-fold increase (P < 0.05 for trend) with an estimated annual percentage change (EAPC) of 3.5 (95% CI = 0.5-6.6). The proportional increase in incidence was more significant in females compared to males. The analysis of the overall cohort rates of PC in Srilanka between 2001-2010 showed the highest rates in the 60-70-year category with an EAPC of 5.06 (95% CI = 1.3-9.0). In our study, we found that there was a marginal rise in the incidence of PC in Sri Lanka with a higher proportional increase in females compared to males.


The relationship between brown adipose tissue activity and neoplastic status: an (18)F-FDG PET/CT study in the tropics.
Brown adipose tissue (BAT) has thermogenic potential. For its activation, cold exposure is considered a critical factor though other determinants have also been reported. The purpose of this study was to assess the relationship between neoplastic status and BAT activity by 2-deoxy-2-[(18)F]fluoro-D-glucose (18F-FDG) positron emission tomography/computed tomography (PET/CT) in people living in the tropics, where the influence of outdoor temperature was low. (18)F-FDG PET/CT scans were reviewed and the total metabolic activity (TMA) of identified activated BAT quantified. The distribution and TMA of activated BAT were compared between patients with and without a cancer history. The neoplastic status of patients was scored according to their cancer history and (18)F-FDG PET/CT findings. We evaluated the relationships between the TMA of BAT and neoplastic status along with other factors: age, body mass index, fasting blood sugar, gender, and outdoor temperature. Thirty of 1740 patients had activated BAT. Those with a cancer history had wider BAT distribution (p = 0.043) and a higher TMA (p = 0.028) than those without. A higher neoplastic status score was associated with a higher average TMA. Multivariate analyses showed that neoplastic status was the only factor significantly associated with the TMA of activated BAT (p = 0.016). Neoplastic status is a critical determinant of BAT activity in patients living in the tropics. More active neoplastic status was associated with more vigorous TMA of BAT.


[Identification of tetracenomycin X from a marine-derived Saccharothrix sp. guided by genes sequence analysis].
The crude extracts of the fermentation broth from a marine sediment-derived actinomycete strain, Saccharothrix sp. 10-10, showed significant antibacterial activities against drug-resistant pathogens. A genome-mining PCR-based experiment targeting the genes encoding key enzymes involved in the biosynthesis of secondary metabolites indicated that the strain 10-10 showed the potential to produce tetracenomycin-like compounds. Further chemical investigation of the cultures of this strain led to the identification of two antibiotics, including a tetracenomycin (Tcm) analogs, Tcm X (1), and a tomaymycin derivative, oxotomaymycin (2). Their structures were identified by spectroscopic data analysis, including UV, 1D-NMR, 2D-NMR and MS spectra. Tcm X (1) showed moderate antibacterial activities against a number of drug-resistant pathogens, including methicillin-resistant Staphylococcus aureus (MRSA) and vancomycin-resistant Enterococci (VRE) pathogens, with the MIC values in the range of 32-64 microg x mL(-1). In addition, 1 also displayed significant cytotoxic activities against human cancer cell lines, including HL60 (leukemia), HepG2 (liver), and MCF-7 (breast) with the IC 50 values of 5.1, 9.7 and 18.0 micromol x L(-1), respectively. Guided by the PCR-based gene sequence analysis, Tcm X (1) and oxotomaymycin (2) were identified from the genus of Saccharothrix and their 13C NMR data were correctly assigned on the basis of 2D NMR spectroscopic data analysis for the first time.


Experimental Evidence of Dioxole Unimolecular Decay Pathway for Isoprene-Derived Criegee Intermediates.
Ozonolysis of isoprene, one of the most abundant volatile organic compounds emitted into the Earth's atmosphere, generates two four-carbon unsaturated Criegee intermediates, methyl vinyl ketone oxide (MVK-oxide) and methacrolein oxide (MACR-oxide). The extended conjugation between the vinyl substituent and carbonyl oxide groups of these Criegee intermediates facilitates rapid electrocyclic ring closures that form five-membered cyclic peroxides, known as dioxoles. This study reports the first experimental evidence of this novel decay pathway, which is predicted to be the dominant atmospheric sink for specific conformational forms of MVK-oxide (anti) and MACR-oxide (syn) with the vinyl substituent adjacent to the terminal O atom. The resulting dioxoles are predicted to undergo rapid unimolecular decay to oxygenated hydrocarbon radical products, including acetyl, vinoxy, formyl, and 2-methylvinoxy radicals. In the presence of O2, these radicals rapidly react to form peroxy radicals (ROO), which quickly decay via carbon-centered radical intermediates (QOOH) to stable carbonyl products that were identified in this work. The carbonyl products were detected under thermal conditions (298 K, 10 Torr He) using multiplexed photoionization mass spectrometry (MPIMS). The main products (and associated relative abundances) originating from unimolecular decay of anti-MVK-oxide and subsequent reaction with O2 are formaldehyde (88 ± 5%), ketene (9 ± 1%), and glyoxal (3 ± 1%). Those identified from the unimolecular decay of syn-MACR-oxide and subsequent reaction with O2 are acetaldehyde (37 ± 7%), vinyl alcohol (9 ± 1%), methylketene (2 ± 1%), and acrolein (52 ± 5%). In addition to the stable carbonyl products, the secondary peroxy chemistry also generates OH or HO2 radical coproducts.


Production of hybrid cell lines secreting antibodies to herpes simplex virus type 2.
Following different immunization schedules we have produced 102 myeloma--spleen cell hybrid lines (30.8% of the total) secreting antibodies to HSV-2 infected cells. The highest yield of hybrids was derived from fusion of myeloma cells with spleen cells from mice sensitized by footpad injection of formalin-inactivated virus and boosted intravenously with the same antigen. To date, twelve of the antibody-producing hybrids have been cloned by limiting dilutions to generate over 100 monoclonal lines reacting specifically with HSV-infected but not with mock-infected cells. Several hybrid lines have also been injected into mice to generate tumors and high titers of anti-HSV-specific antibodies have been detected in ascitic fluids. All of the hybrids so far tested have retained the capacity to secrete anti-HSV antibodies after long-term in culture or repeated passages through mice. Antibodies reacting with cytoplasmic or nuclear antigens in infected cells as well as type-specific and type common antibodies have been detected.


Preparation of monoclonal antibodies to hirudin and hirudin peptides. A method for studying the hirudin--thrombin interaction.
A panel of four monoclonal antibodies was obtained against hirudin, a potent and specific inhibitor of thrombin, by immunizing three groups of mice with protein conjugates made of recombinant desulfatohirudin (group I) or two synthetic peptides representing the C-terminal sequences 40-65 (group II) and 52-65 (group III) of hirudin. Only the monoclonal antibody 4049-83-12, obtained from the group I of mice, showed high affinity for hirudin (Kd of 0.6 nM) and in vitro neutralizing properties. The anti-peptide monoclonal antibodies bound hirudin with lower affinity (Kd of 1.5-7 nM) and showed lower neutralizing capacities. An epitope analysis performed by competitive ELISA using various hirudin analogues and by limited proteolysis of the hirudin-antibody complex revealed that the binding domains of all the anti-peptide antibodies were located close to the C-terminus of hirudin, since the bond between Glu-61 and Glu-62 was not cleaved by the V8 staphylococcal protease in the presence of these antibodies. The epitope of the antibody 4049-83-12 was strictly conformation-dependent, it recognized neither S-carboxymethylated hirudin nor any peptides of hirudin. The cleavage of the bond between Glu-43 and Gly-44 by V8 protease, as well as the cleavage of the bond between Lys-47 and Pro-48 by lysyl endopeptidase, was prevented by the binding of the antibody 4049-83-12 to hirudin. The possibility that this epitope overlapped with a region of hirudin involved in the binding to thrombin is discussed.


Racial Misclassification and Disparities in Neonatal Abstinence Syndrome Among American Indians and Alaska Natives.
Maternal substance misuse can result in neonatal abstinence syndrome (NAS), a drug withdrawal process in newborns exposed in utero to drugs. This study aimed to examine the effect of racial misclassification of American Indians and Alaska Natives (AI/AN) on rates of NAS in two hospital discharge datasets in the Pacific Northwest. We conducted probabilistic record linkages between the Northwest Tribal Registry and Oregon and Washington hospital discharge datasets to correct racial misclassification of AI/AN people. We assessed outcomes using International Classification of Disease, Ninth Revision/Tenth Revision, Clinical Modification (ICD-9-CM or ICD-10-CM) diagnosis codes. Linkage increased ascertainment of NAS cases among AI/AN by 8.8% in Oregon and by 18.1% in Washington. AI/AN newborns were 1.5 and 3.9 times more likely to be diagnosed with NAS than NHW newborns in Oregon and Washington, respectively. The results showed that newborns residing in rural Washington were 1.4 times more likely to be diagnosed with NAS than those living in urban areas. Correct racial classification is an important factor in improving data quality for AI/AN populations and establishing accurate surveillance to help address the disproportionate burden of neonatal abstinence syndrome among AI/AN. The results highlight the need for programing efforts tailored by insurance status and rurality for pregnant women using substances.


Studies in burns: XV. Use of a topical lipid in treating human burns.
Sixty patients were selected from the 24-month burn population admitted to the Medical College of Georgia. Selection was on the basis of age, size of burn, and survival beyond 120 hours. Thirty-one of these patients were treated with ethyl linoleate agent (hELate) in addition to the modalities we routinely use for burn care. hELate was applied in a dose of 25 mg/kg only once in the period after burn. Significant pain relief and enhancement of wound healing (with return of hair and pigment) occurred in the hELate-treated patient. Less narcotic was needed, fewer grafts and reconstructive procedures were required, and hospital stay was reduced significantly. It is suggested that hELate is a safe and useful adjunctive to burn care.


Rhizobacterial exopolysaccharides elicit induced resistance on cucumber.
The role of exopolysaccharides (EPSs) from a plant growth-promoting rhizobacterium, Burkholderia gladioli IN26, on elicitation of induced systemic resistance was investigated. A purified EPS induced expression of PR- 1a::GUS on tobacco and elicited induced resistance against Colletotrichum orbiculare on cucumber. The maximum level of disease protection was noted when seeds were soaked in 200 ppm of the EPS. Our results indicate that EPS from specific rhizobacteria can elicit induced resistance and suggest that bacterial EPS might be a useful elicitor of resistance under field conditions.


Incidence of bacterial transmission and transfusion reactions by blood components.
Successful reduction of the risk of viral transmission via blood components has focused attention on the risk of transfusion-associated bacterial sepsis. The incidence of transfusion reactions due to bacteria is currently estimated at between 1 per 100,000 and 1 per 1,000,000 units in the case of packed red blood cells and between 1 per 900 and 1 per 100,000 units in the case of platelet concentrates. For autologous transfusion, only isolated case reports are available which do not permit a quantitative risk assessment. No reliable data on the risk of morbidity and mortality due to transfusion-associated bacterial infection are available for Germany at present. As platelet concentrates provide more favourable conditions for bacterial growth by virtue of being stored at room temperature, these blood components are thought to bear a considerably greater risk of bacterial contamination than red blood cells. The rate of contamination of platelet concentrates is cited at between 0.02% and 1.2%, depending on the production and bacterial culture techniques used, whereas the rate for packed red blood cells is between 0.1% and 0.2%. Risk reduction strategies include careful selection of blood donors, optimisation of donation and production techniques, and detection of bacteria in samples of blood components prior to transfusion. Also of importance for patient safety are quality assurance measures in the preparation for transfusion, in measures of patient monitoring, and investigation of transfusion reactions.


CUB-domain-containing protein 1 overexpression in solid cancers promotes cancer cell growth by activating Src family kinases.
The transmembrane glycoprotein, CUB (complement C1r/C1s, Uegf, Bmp1) domain-containing protein 1 (CDCP1) is overexpressed in several cancer types and is a predictor of poor prognosis for patients on standard of care therapies. Phosphorylation of CDCP1 tyrosine sites is induced upon loss of cell adhesion and is thought to be linked to metastatic potential of tumor cells. Using a tyrosine-phosphoproteomics screening approach, we characterized the phosphorylation state of CDCP1 across a panel of breast cancer cell lines. We focused on two phospho-tyrosine pTyr peptides of CDCP1, containing Tyr707 and Tyr806, which were identified in all six lines, with the human epidermal growth factor 2-positive HCC1954 cells showing a particularly high phosphorylation level. Pharmacological modulation of tyrosine phosphorylation indicated that, the Src family kinases (SFKs) were found to phosphorylate CDCP1 at Tyr707 and Tyr806 and play a critical role in CDCP1 activity. We demonstrated that CDCP1 overexpression in HEK293 cells increases global phosphotyrosine content, promotes anchorage-independent cell growth and activates several SFK members. Conversely, CDCP1 downregulation in multiple solid cancer cell lines decreased both cell growth and SFK activation. Analysis of primary human tumor samples demonstrated a correlation between CDCP1 expression, SFK and protein kinase C (PKC) activity. Taken together, our results suggest that CDCP1 overexpression could be an interesting therapeutic target in multiple solid cancers and a good biomarker to stratify patients who could benefit from an anti-SFK-targeted therapy. Our data also show that multiple tyrosine phosphorylation sites of CDCP1 are important for the functional regulation of SFKs in several tumor types.


[Atrophic rhinitis A.D. 1991].
In the Netherlands the Central Veterinary Institute (CVI) and the Animal Health Services carried out investigations on aetiology, pathogenesis, therapy and prevention of Atrophic Rhinitis (AR) since 1970. Since the discovery of AR toxin in AR pathogenic P. multocida in SPF piglets, quicker and cheaper tests were developed e.g. the mouse test and guinea pig skin test. Today ELISA-tests with poly- and monoclonals are also available. The CVI carried out DNA probes and is developing a PCR-test. Tests to detect a natural infection were difficult to develop. The SN test by Bechmann, the skin test by Schimmelpfenning and the ELISA test by Foged give some perspectives. From investigations with B. bronchiseptica in piglets, it appears that the role of this germ is overestimated in the development of AR. On 16-week-old pigs an infection with AR toxigenic P. multocida caused conchae atrophy and septum deviation within 4 weeks. This fact has to be taken into account concerning the preventive medications in piglets. Only high maternal antibody titres against the AR toxin are able to protect the piglets for a sufficiently long period. Conchae atrophy and brachygnathia superior are indicators for suspicion of AR but do not prove the diagnosis of AR. By means of selective agars the isolation of P. multocida from nose, tonsils and lungs could be improved. The incubation period of clinical AR may last longer than two years. The AR pathogenic P. multocida is predominantly transmitted by pigs, incidentally also by rabbits, cattle, sheep, goat, poultry, dogs and cats. Humans can get infected and diseased. The AR pathogenic P. multocida has to be classified as a zoonosis. By way of stamping out and air filtering the transmission can be prevented. Vaccination cannot avoid an infection with the AR pathogenic P. multocida. The Animal Health Services developed a program to certify breeding herds 'free of AR pathogenic P. multocida'.


Hematopoietic effect of echinochrome on phenylhydrazine-induced hemolytic anemia in rats.
Hemolytic anemia (HA) is a serious health condition resulting from reduced erythrocytes' average life span. Echinochrome (Ech) is a dark-red pigment found in shells and spines of sea urchins. Studying the potential therapeutic effect of Ech on phenylhydrazine (PHZ)-induced HA in rats. Eighteen rats were divided into three groups (n = 6): the control group, the phenylhydrazine-induced HA group and the Ech group, injected intraperitoneally with PHZ and supplemented with oral Ech daily for 6 days. Ech resulted in a considerable increase in RBCs, WBCs, and platelets counts, hemoglobin, reduced glutathione, catalase, and glutathione-S-transferase levels, and a significant decrease in aspartate & alanine aminotransferases, alkaline phosphatase, gamma-glutamyl transferase, bilirubin, creatinine, urea, urate, malondialdehyde & nitric oxide levels in anemic rats. Histopathological examination of liver and kidney tissue samples showed marked improvement. Ech ameliorated phenylhydrazine-induced HA with a hepatorenal protective effect owing to its anti-inflammatory and antioxidant properties.


Effect of bifocal and prismatic bifocal spectacles on myopia progression in children: three-year results of a randomized clinical trial.
Myopia is a significant public health problem, making it important to determine whether a bifocal spectacle treatment involving near prism slows myopia progression in children. To determine whether bifocal and prismatic bifocal spectacles control myopia in children with high rates of myopia progression and to assess whether the treatment effect is dependent on the lag of accommodation and/or near phoria status. This 3-year randomized clinical trial was conducted in a private practice. A total of 135 (73 female and 62 male) Chinese-Canadian children (aged 8-13 years; mean [SE] age, 10.29 [0.15] years; mean [SE] myopia, -3.08 [0.10] D) with myopia progression of at least 0.50 D in the preceding year were randomly assigned to 1 of 3 treatments. A total of 128 (94.8%) completed the trial. Single-vision lenses (control, n = 41), +1.50-D executive bifocals (n = 48), and +1.50-D executive bifocals with 3-Δ base-in prism in the near segment of each lens (n = 46). MAIN OUTCOMES AND MEASURES Myopia progression (primary) measured using an automated refractor following cycloplegia and increase in axial length (secondary) measured using ultrasonography at intervals of 6 months for 36 months. RESULTS Myopia progression over 3 years was an average (SE) of -2.06 (0.13) D for the single-vision lens group, -1.25 (0.10) D for the bifocal group, and -1.01 (0.13) D for the prismatic bifocal group. Axial length increased an average (SE) of 0.82 (0.05) mm, 0.57 (0.07) mm, and 0.54 (0.06) mm, respectively. The treatment effect of bifocals (0.81 D) and prismatic bifocals (1.05 D) was significant (P < .001). Both bifocal groups had less axial elongation (0.25 mm and 0.28 mm, respectively) than the single-vision lens group (P < .001). For children with high lags of accommodation (≥ 1.01 D), the treatment effect of both bifocals and prismatic bifocals was similar (1.1 D) (P < .001). For children with low lags (<1.01 D), the treatment effect of prismatic bifocals (0.99 D) was greater than of bifocals (0.50 D) (P = .03). The treatment effect of both bifocals and prismatic bifocals was independent of the near phoria status. Bifocal spectacles can slow myopia progression in children with an annual progression rate of at least 0.50 D after 3 years. These results suggest that prismatic bifocals are more effective for myopic children with low lags of accommodation. TRIAL REGISTRATION clinicaltrials.gov Identifier: NCT00787579.


Serum holotranscobalamine, vitamin B12, folic acid and homocysteine levels in patients with vitiligo.
Few studies have investigated the role of vitamin B12 metabolism in vitiligo. We tested the hypothesis that vitamin B12 and folate metabolism might have an influence on the pathogenesis of vitiligo. Full blood count and levels of folic acid, vitamin B12, homocysteine and holotranscobalamine were examined for 69 patients with vitiligo and 52 controls. The vitiligo group had higher levels of homocysteine (P < 0.01) and haemoglobin (P < 0.01) levels, and lower levels of vitamin B12 (P < 0.01) and holotranscobalamine (P < 0.0001) than the control group. Folic acid levels were similar for both groups. In a risk analysis, hyperhomocysteinaemia (≥ 15 μmol/L, P < 0.01) and vitamin B12 deficiency (< 200 pg/mL, P < 0.01) were significant risk factors for vitiligo. Patients with holotranscobalamine levels in the lowest quartile had an increased risk for co-occurrence of vitiligo (P < 0.005). Vitamin B12 deficiency and hyperhomocysteinaemia may share a common genetic background with vitiligo.


Posterolateral dislocation of the elbow joint. Relationship to medial instability.
Dislocation of the elbow joint is the second most common dislocation in the upper extremity, dislocation of the shoulder being the most common. It has been reported that uncomplicated dislocation of the elbow joint may be associated with a decreased range of motion, degenerative changes in the elbow joint, ectopic calcification, or neurological deficits. As the medial collateral ligament complex can be completely disrupted during dislocation, we evaluated the association between the long-term results of treatment of simple posterolateral dislocation of the elbow and the presence of persistent medial or valgus elbow instability. Fifty patients who had a mean age of thirty-three years (range, eighteen to fifty-eight years) had closed reduction of a posterolateral dislocation of the elbow without associated fractures. The extremity was immobilized in an above-the-elbow plaster cast for three weeks. After a mean duration of follow-up of nine years (range, six to thirteen years), forty-one patients were evaluated with an interview, a physical examination, and radiographs made while a valgus load was applied to the elbow. The average score according to the system of The Hospital for Special Surgery was 91 points (range, 49 to 100 points), and thirty-one patients described their elbow function as good or excellent. Twenty-four patients had evidence of medial instability on radiographs made while a valgus load was applied to the elbow. Twenty-one patients had signs of degeneration of the joint, and twenty-five patients had ectopic ossification. Magnetic resonance imaging combined with arthrography was performed for the first twenty patients; eight had evidence of rupture of the medial collateral ligament, seven had generalized degenerative changes of the cartilage, and four had a chondral defect of the capitellum. (The study could not be completed for the remaining patient.) Medial instability on radiographs was correlated with signs of degeneration (p = 0.001), ectopic ossification (p = 0.01), a worse score according to the system of The Hospital for Special Surgery (p = 0.002), and persistent pain (p = 0.04). Posterolateral dislocation of the elbow joint can lead to persistent valgus instability that is associated with a worse overall clinical and radiographic result.


Implant survival and patient-reported outcomes after total hip arthroplasty in young patients with developmental dysplasia of the hip.
Developmental dysplasia of the hip (DDH) is a common cause of hip pain in young patients, and may require treatment with total hip arthroplasty (THA). This study evaluates implant survival and describes patient-reported outcomes after primary THA in DDH patients aged 35 or younger. A retrospective study with prospective follow-up was conducted at a major academic medical centre. Patient charts were reviewed to identify young THA patients with DDH, and follow-up surveys were conducted to determine implant survival and patient-reported outcomes. Kaplan-Meier survival analysis was performed to evaluate implant survival, and the hip disability and osteoarthritis outcome score (HOOS) was used to describe patient-reported outcomes. This study included 61 patients (75 THAs, 75% follow-up), and the mean time to follow-up was 13 years (range 3-25 years). The 10-year and 20-year implant survival was 87% (95% CI, 78%-94%) and 55% (95% CI, 37%-72%), respectively, and implant survival was significantly better in patients over the age of 25 at the time of surgery (p value <0.01). The mean HOOS patient scores were 83 for pain (SD 20.29), 78 for symptoms (SD 19.72), 83 for ADLs (SD 20.89), and 74 for sports (SD 25.92). Patients that were younger at the time of surgery or required custom implants reported worse HOOS-Symptom scores at follow-up (p value = 0.02). Young patients with DDH have good outcomes after surgery. Patient factors and implant characteristics should be considered when predicting implant survival and outcomes after THA in young patients with DDH.


The safety of treatments for tobacco use disorder.
Tobacco continues to be a leading cause of preventable morbidity and mortality in the world. First-line pharmacotherapies for the treatment of tobacco use disorder include nicotine replacement therapy, bupropion sustained-release (SR), and varenicline. We provide an overview of current evidence on the safety of first-line pharmacotherapies for the treatment of tobacco use disorder. Randomized clinical trials published in English up to July 2015 were identified and reviewed through searches of PUBMED using the terms nicotine replacement therapy, bupropion SR, varenicline, smoking, and tobacco cessation. Nicotine replacement has few contraindications and side effects and can be recommended to almost all tobacco users. Bupropion SR should be used with caution in patients with bipolar disorder or liver or kidney disease, and alternative treatments should be considered for patients with a history of seizures or who are at risk for seizures. The only contraindication for varenicline is an allergy to the medication, and nausea is the most common side effect. Varenicline can be used safely in patients with cardiovascular disease. Varenicline can be used in patients with stable psychiatric disease and safety can be ensured through close clinical monitoring.


Irrigation Device Successfully Tests Valve Competence During Minimally Invasive Mitral Valve Repair.
Minimally invasive mitral valve repair has become standard. We hypothesized that reproducible water testing with less air contaminations may help achieve successful repair and prevent air embolism. We used a long shaft irrigation device connected to an infuser pump to water test mitral valve repair in 104 consecutive patients with severe mitral regurgitation due to type II mitral valve dysfunction. All patients underwent mitral valve repair via small right thoracotomy. Water testing was performed by simultaneously monitoring root pressure to confirm valve competence and adequate leaflet coaptation with suitable pressure. Irrigation was stopped when root pressure reached 60 mm Hg. Simultaneous transesophageal echocardiography showed a negligible amount of air introduction. Predischarge echocardiography revealed residual mitral regurgitation of less than or equal to 1 in all patients, and no patient experienced any major adverse event such as stroke or perioperative myocardial infarction. This novel water test allowed us to correctly check mitral valve competence and reduce air contamination.


Expression of beta-galactoside alpha 2,6-sialyltransferase does not alter the susceptibility of human colon cancer cells to NK-mediated cell lysis.
The extent of processing of N-linked oligosaccharides and the sialylation of the target cell membranes has been positively correlated with resistance to lysis mediated by NK cells, but a conclusive evidence has never been reached. Colon cancer tissues express an increased activity of beta-galactoside alpha 2,6-sialyltransferase (EC 126.96.36.199, alpha 2,6ST), which catalyzed the addition of sialic acid in alpha 2,6-linkage to Gal beta 1,4GlcNAc (N-acetyllactosamine) sequences of glycoprotein N-linked chains. The resulting increased level of membrane alpha 2,6-sialylation appears to be related with a more invasive behavior of cancer cells. This phenomenon may depend on a decreased sensitivity of colon cancer cells to NK cells. To obtain conclusive evidence on the role played by sialylation of N-linked chains in determining the target cell susceptibility to NK-mediated lysis, human colon cancer cell lines not expressing sialyltransferases acting on N-linked chains were transfected with a rat alpha 2,6ST cDNA. Stable transfectants expressed different levels of alpha 2,6ST activity, were reactive with the Sambucus nigra lectin, specific for alpha 2,6-linked sialic acid, and compared with control transfectants, showed a remarkable decrease in the number of unsubstituted Gal beta 1,4GlcNAc terminal sequences. The NK susceptibility of these clones was found to be identical to that of control transfectants, either when unstimulated- or IL-2-stimulated lymphocytes were used as effectors. Neuraminidase treatment of target cells does not result in significant changes to NK susceptibility. Our data demonstrate that sialic acid alpha 2,6-linked to N-linked chains of target cell glycoproteins does not play a major role in recognition of the target by human NK cells.


Is there a role for magnetic resonance imaging in the evaluation of non-traumatic intraparenchymal haemorrhage in children?
In contrast to adults, intraparenchymal haemorrhage (IPH) is at least as common as ischaemic stroke in children. There is often uncertainty about the most appropriate modality for imaging in the acute stage. To examine the diagnostic value of MRI and MR angiography (MRA) in the detection of underlying pathology in children with non-traumatic IPH. A retrospective review was conducted of children with IPH from January 1997 to March 2003. After exclusion of patients with traumatic IPH or previously diagnosed vascular malformation, aneurysm, or brain tumour, 50 children were identified. Case notes and imaging studies were reviewed. An underlying lesion was demonstrated with MR in two-thirds of children (25/38) with IPH. A vascular lesion was the commonest cause, followed by tumour. Three children had false-negative MR scans. MR is a valuable non-invasive imaging modality for detection of both vascular and non-vascular causes of paediatric IPH. The high rate of the latter in childhood makes incorporation of MR into paediatric IPH imaging protocols especially important. Clinical guidelines regarding the optimum sequence of investigations in non-traumatic IPH would be helpful to standardize practice and enable critical appraisal.


AMPKα, C/EBPβ, CPT1β, GPR43, PPARγ, and SCD Gene Expression in Single- and Co-cultured Bovine Satellite Cells and Intramuscular Preadipocytes Treated with Palmitic, Stearic, Oleic, and Linoleic Acid.
We previously demonstrated that bovine subcutaneous preadipocytes promote adipogenic gene expression in muscle satellite cells in a co-culture system. Herein we hypothesize that saturated fatty acids would promote adipogenic/lipogenic gene expression, whereas mono- and polyunsaturated fatty acids would have the opposite effect. Bovine semimembranosus satellite cells (BSC) and intramuscular preadipocytes (IPA) were isolated from crossbred steers and cultured with 10% fetal bovine serum (FBS)/Dulbecco's Modified Eagle Medium (DMEM) and 1% antibiotics during the 3-d proliferation period. After proliferation, cells were treated for 3 d with 3% horse serum/DMEM (BSC) or 5% FBS/DMEM (IPA) with antibiotics. Media also contained 10 μg/mL insulin and 10 μg/mL pioglitazone. Subsequently, differentiating BSC and IPA were cultured in their respective media with 40 μM palmitic, stearic, oleic, or linoleic acid for 4 d. Finally, BSC and IPA were single- or co-cultured for an additional 2 h. All fatty acid treatments increased (p = 0.001) carnitine palmitoyltransferase-1 beta (CPT1β) gene expression, but the increase in CPT1β gene expression was especially pronounced in IPA incubated with palmitic and stearic acid (6- to 17- fold increases). Oleic and linoleic acid decreased (p = 0.001) stearoyl-CoA desaturase (SCD) gene expression over 80% in both BSC and IPA. Conversely, palmitic and stearic acid increased SCD gene expression three fold in co-cultured in IPA, and stearic acid increased AMPKα gene expression in single- and co-cultured BSC and IPA. Consistent with our hypothesis, saturated fatty acids, especially stearic acid, promoted adipogenic and lipogenic gene expression, whereas unsaturated fatty acids decreased expression of those genes associated with fatty acid metabolism.


GATM and GAMT synthesize creatine locally throughout the mammalian body and within oligodendrocytes of the brain.
The enzymes glycine amidinotransferase, mitochondrial (GATM also known as AGAT) and guanidinoacetate N-methyltransferase (GAMT) function together to synthesize creatine from arginine, glycine, and S-Adenosyl methionine. Deficiency in either enzyme or the creatine transporter, CT1, results in a devastating neurological disorder, Cerebral Creatine Deficiency Syndrome (CCDS). To better understand the pathophysiology of CCDS, we mapped the distribution of GATM and GAMT at single cell resolution, leveraging RNA sequencing analysis combined with in vivo immunofluorescence (IF). Using the mouse as a model system, we find that GATM and GAMT are coexpressed in several tissues with distinct and overlapping cellular sources, implicating local synthesis as an important mechanism of creatine metabolism in numerous organs. Extending previous findings at the RNA level, our analysis demonstrates that oligodendrocytes express the highest level of Gatm and Gamt of any cell type in the body. We confirm this finding in the mouse brain by IF, where GATM localizes to the mitochondria of oligodendrocytes, whereas both oligodendrocytes and cerebral cortical neurons express GAMT. Interestingly, the latter is devoid of GATM. Single nucleus assay for transposase-accessible chromatin sequencing (snATAC-seq) analysis of 4 brain regions highlights a similar primacy of oligodendrocytes in the expression of GATM and GAMT in the human central nervous system. Importantly, an active putative regulatory element within intron 2 of human GATM is detected in oligodendrocytes but not neurons.


stf1: non-wee mutations epistatic to cdc25 in the fission yeast Schizosaccharomyces pombe.
In Schizosaccharomyces pombe, cdc25 is a cell cycle regulated inducer of mitosis. wee1 and phenotypically wee alleles of cdc2 are epistatic to cdc25. Mutant alleles of a new locus, stf1 (suppressor of twenty-five), identified in a reversion analysis of conditionally lethal cdr1-76 cdc25-22 and cdr2-96 cdc25-22 double mutant strains, also suppress both temperature-sensitive and gene disruption alleles of cdc25. These mutants, by themselves, are phenotypically indistinguishable from wild type strains; hence they represent the first known mutations that are epistatic to cdc25 and do not display a wee phenotype. stf1 genetically interacts with other elements of mitotic control in S. pombe. stf1-1 is additive with wee1-50, cdc2-1w and cdc2-3w for suppression of cdc25-22. Also, like wee1- and cdc2-w, stf1- suppression of cdc25 is reversed by overexpression of the putative type 1 protein phosphatase bws1+/dis2+. Interaction with various mutants and plasmid overexpression experiments suggest that stf1 does not operate either upstream or downstream of wee1. Similarly, it does not operate through cdc25 since it rescues the disruption. stf1 appears to encode an important new element of mitotic control.


Evidence for a persistent, environment-dependent and deteriorating subtype of subclinical psychotic experiences: a 6-year longitudinal general population study.
Research suggests that subclinical psychotic experiences during adolescence represent the behavioral expression of liability for psychosis. Little is known, however, about the longitudinal trajectory of liability in general population samples. Growth mixture modeling was used to examine longitudinal trajectories of self-reported positive psychotic experiences in the Youth Self Report (YSR), completed three times over a period of 6 years by a general population cohort of adolescents aged 10-11 years at baseline (n=2230). Four groups with distinct developmental trajectories of low, decreasing, increasing and persistent levels of mild positive psychotic experiences were revealed. The persistent trajectory was associated strongly with cannabis use, childhood trauma, developmental problems and ethnic minority status, and consistently displayed strong associations with factors known to predict transition from subclinical psychotic experience to clinical psychotic disorder (severity of and secondary distress due to psychotic experiences, social and attentional problems and affective dysregulation) and also with high levels of parental-reported psychotic experiences and use of mental health care at the end of the follow-up period. Progressively weaker associations were found for the increasing, decreasing and low trajectories respectively. The results suggest that the outcome of early developmental deviation associated with later expression of psychotic experiences is contingent on the degree of later interaction with environmental risks inducing, first, persistence of psychotic experiences and, second, progression to onset of need for care and service use. Insight into the longitudinal dynamics of risk states in representative samples may contribute to the development of targeted early intervention in psychosis.


Vaginal metastases in gestational trophoblastic neoplasia.
To evaluate the clinical experience and outcomes of patients with gestational trophoblastic neoplasia (GTN) complicated by vaginal metastases. A review of patients with vaginal metastases from GTN treated at a regional trophoblastic disease center from 1962 to 2006. Vaginal metastases were present in 36 (4.5%) of the 804 patients treated for GTN. FIGO stage was II in 13 patients (36%), III in 22 patients (61%) and IV in 1 patient (3%). Twenty-three patients (65%) were low-risk by modified WHO criteria. The vaginal metastases were most frequently single lesions (61%) on the anterior vaginal wall (49%) with a histologic classification of choriocarcinoma (67%). Significant bleeding necessitated blood transfusion (median, 7 units; range, 1-26 units) in 13 patients (36%). Seven patients (19%) required 1 or more procedures for control of bleeding, including excision, suturing and/or hypogastric artery ligation/embolization. Twenty-three patients (64%) received single-agent chemotherapy with methotrexate and/or actinomycin-D, while 13 patients (36%) received multiagent chemotherapy regimens. Overall, 29 (81%) of 36 patients with vaginal metastases were cured. Vaginal metastasis from GTN does not uniformly confer a worse prognosis or necessitate multiagent chemotherapy, although procedures for control of bleeding may be required.


Towards a more rational approach to the alcohol problem.
Alcohol is the favoured intoxicant of European man and his descendants overseas. As such, its dangers are generally glossed over. The "disease model" of the last forty years owes much of its popularity to the implication that there is something psychologically or biochemically abnormal about those who become "alcoholics" and that the rest of us can happily drink in comparative safety. Unfortunately, the facts do not fit these convenient assumptions. In the last twenty years worldwide recorded production of alcoholic beverages has doubled and the epidemiological evidence that has accumulated during this period makes it clear that, when the per capita consumption of a country or community rises, all the easily measurable ill effects, medical and social, also rise. Conversely, if per capita consumption falls, even modestly, these ill effects also fall. Per capita consumption is thus the crucial factor determining the scale of the whole gamut of adverse effects which are, by any standards, intolerable in many different countries at the present time. This has very inconvenient implications--for individual "social drinkers", for governments and for the alcohol industry--which have not yet been faced.


Effect of fibrin glue on the biomechanical properties of human Descemet's membrane.
Corneal transplantation has rapidly evolved from full-thickness penetrating keratoplasty (PK) to selective tissue corneal transplantation, where only the diseased portions of the patient's corneal tissue are replaced with healthy donor tissue. Descemet's membrane endothelial keratoplasty (DMEK) performed in patients with corneal endothelial dysfunction is one such example where only a single layer of endothelial cells with its basement membrane (10-15 µm in thickness), Descemet's membrane (DM) is replaced. It is challenging to replace this membrane due to its intrinsic property to roll in an aqueous environment. The main objective of this study was to determine the effects of fibrin glue (FG) on the biomechanical properties of DM using atomic force microscopy (AFM) and relates these properties to membrane folding propensity. Fibrin glue was sprayed using the EasySpray applicator system, and the biomechanical properties of human DM were determined by AFM. We studied the changes in the "rolling up" tendency of DM by examining the changes in the elasticity and flexural rigidity after the application of FG. Surface topography was assessed using scanning electron microscopy (SEM) and AFM imaging. Treatment with FG not only stabilized and stiffened DM but also led to a significant increase in hysteresis of the glue-treated membrane. In addition, flexural or bending rigidity values also increased in FG-treated membranes. Our results suggest that fibrin glue provides rigidity to the DM/endothelial cell complex that may aid in subsequent manipulation by maintaining tissue integrity.


Characterization of Porphyromonas gingivalis sialidase and disruption of its role in host-pathogen interactions.
Key to onset and progression of periodontitis is a complex relationship between oral bacteria and the host. The organisms most associated with severe periodontitis are the periodontal pathogens of the red complex: Tannerella forsythia, Treponema denticola and Porphyromonas gingivalis. These organisms express sialidases, which cleave sialic acid from host glycoproteins, and contribute to disease through various mechanisms. Here, we expressed and purified recombinant P. gingivalis sialidase SiaPG (PG_0352) and characterized its activity on a number of substrates, including host sialoglycoproteins and highlighting the inability to cleave diacetylated sialic acids - a phenomenon overcome by the NanS sialate-esterase from T. forsythia. Indeed SiaPG required NanS to maximize sialic acid harvesting from heavily O-acetylated substrates such as bovine salivary mucin, hinting at the possibility of interspecies cooperation in sialic acid release from host sources by these members of the oral microbiota. Activity of SiaPG and P. gingivalis was inhibited using the commercially available chemotherapeutic zanamivir, indicating its potential as a virulence inhibitor, which also inhibited sialic acid release from mucin, and was capable of inhibiting biofilm formation of P. gingivalis on oral glycoprotein sources. Zanamivir also inhibited attachment and invasion of oral epithelial cells by P. gingivalis and other periodontal pathogens, both in monospecies but also in multispecies infection experiments, indicating potential to suppress host-pathogen interactions of a mixed microbial community. This study broadens our understanding of the multifarious roles of bacterial sialidases in virulence, and indicates that their inhibition with chemotherapeutics could be a promising strategy for periodontitis therapy.


Active intensity noise suppression for a broadband mid-infrared laser source.
Excess relative intensity noise (RIN) constitutes one of the major limitations of most spectroscopic methods involving lasers. Here, we present an active RIN suppression scheme for a coherent mid-infrared (MIR) light source (8.4-11 µm), based on intra-pulse difference frequency generation (DFG). Three different stabilization concepts that rely on modulating the intensity of the driving near-infrared (NIR) pulse train with an acousto-optic modulator are investigated and compared. By using the wings of the NIR spectrum to generate the error signal, a RIN suppression of the MIR pulse train of up to a factor of 20 was achieved in the band between 1 Hz and 100 kHz, resulting in a total integrated RIN of 0.07%.


Impact of the Induction of Labor on Hemophilia Carriers and Their Newborn Infants.
Hemophilia is a risk for severe hemorrhage in newborns during the perinatal period and excessive postpartum hemorrhage (PPH) in hemophilia carriers. Vacuum extraction or use of forceps should be avoided to prevent neonatal intracranial hemorrhage (ICH). Optimal modes of delivery such as vaginal or cesarean section are open to debate. The safety of the induction of labor is also worthy of investigation. Here we ask if labor induction is a safe delivery mode for pregnant women who are hemophilia carriers and their infants. We looked at 13 deliveries by hemophilia carriers at our hospital from 2005 to 2018. Two of the five male neonates complicated by hemophilia suffered ICH complications (40%). Both were delivered by induced labor. No deliveries by carriers had PPH which required treatment. Our data indicate that the induction of labor may provoke ICH in infants with hemophilia. We suggest that induction of labor is not a preferable delivery method for hemophilia carriers to avoid neonatal ICH.