Potential confounders of bisphenol-a analysis in dental materials.
In the published literature, a variety of analytical methods have been used to quantify and report bisphenol A (BPA) release from dental resins. The objective of this study was to compare results obtained for quantification of BPA in dental resin extracts using an LC/UV analytical method and an LC/MS/MS method. A cured Bis-GMA-based resin representative of commercial dental products was extracted according to ISO 10993 guidelines for medical devices. d16BPA was included as an internal standard. Sample processing followed expert recommendations for minimizing BPA sample contamination. Extracts were separated using HPLC methods and analyzed for BPA using LC/UV and LC/MS/MS detection methods. The reported BPA concentrations were about 30-fold higher using LC/UV vs. LC/MS/MS. Full scan LC/MS/MS in both positive and negative modes showed that the apparent high BPA values seen with LC/UV were caused by co-elution of a previously unidentified chemical, thought to arise from one of the polymerization initiators. These results emphasize the potential difficulties in obtaining accurate analyses of BPA in complex mixtures such as dental resins and their extracts. Both good separation methodology and a detection method with high specificity and sensitivity are important to avoid incorrect identification of extractables, and consequent incorrect quantitative assignments for species of interest. Reliable methods are essential for accurate estimation of patient exposure to BPA and development of meaningful health risk assessments.


KM-01, a brassinolide inhibitor, its production, isolation and structure from two fungi Drechslera avenae and Pycnoporus coccineus.
A new brassinolide inhibitor, named KM-01, was isolated from the culture filtrates of two fungal species, Drechslera avenae and Pycnoporus coccineus, and the structure with absolute stereochemistry was elucidated as the fatty acid ester of an eremophilane sesquiterpene, bipolaroxin, based on spectroscopic analysis, chemical degradation, and synthesis of the fatty acid.


Measles encephalomyelitis: lack of evidence of viral invasion of the central nervous system and quantitative study of the nature of demyelination.
Measles encephalomyelitis appears to be an immune-mediated parainfectious disorder, but it is unclear whether viral invasion of brain is an obligate step in its development. Immunocytochemical methods were used to search for virus antigen in formalin-fixed, paraffin-embedded central nervous system (CNS) tissues from 10 patients with measles encephalomyelitis and 12 patients who had died of measles without CNS involvement. All the CNS tissues studied were viral antigen negative. Similarly fixed CNS tissues from all of 6 patients with subacute sclerosing panencephalitis were viral antigen positive and served as controls. The pattern of perivenular demyelination was also determined in 4 cases of measles encephalomyelitis using antibodies to myelin associated glycoprotein and myelin basic protein and a Luxol fast blue stain. Areas of demyelination in serial sections were quantitated, and no morphometrical differences were found among tissues stained with the three stains. The data suggest the lack of virus replication in the CNS during encephalomyelitis or fatal measles without CNS symptoms. The pattern of loss of myelin associated glycoprotein and myelin basic protein in regions of perivenular demyelination resembles that reported in experimental allergic encephalomyelitis. This pattern of demyelination has been proposed to result from a primary attack on the myelin sheath rather than from direct involvement of the oligodendroglial cell.


Functional capacity, muscle strength and falls in women with fibromyalgia.
Patients with fibromyalgia have difficulty with activities of daily living, they exhibit reduced muscle strength and high incidence of reported falls. The objective of this study was to evaluate the functional performance and lower limb muscle strength in women with fibromyalgia and determine the relationship between muscle strength and falls. Sixteen females with fibromyalgia and 16 healthy women participated in the study. Pain intensity, fibromyalgia impact on quality of life, physical activity level and fall prevalence were assessed. The peak torque and the rate of torque development were determined in maximal voluntary isometric contraction (hip, knee and ankle joints) using a load cell. The 30s chair stand, 8 ft up and go, sit and reach, and functional reach tests were used to characterize functional performance. Women with fibromyalgia showed deficits in lower limb muscle strength, balance and agility and exhibited decreased knee extension peak torque and rate of torque development. In addition, they showed lower hip adduction and extension peak torque in comparison to the control group (P>0.05). Hip extension rate of torque development, duration of fibromyalgia symptoms, overall pain, knee pain, and fibromyalgia impact were strong predictors of the number of falls in patients with fibromyalgia (R2=0.86; P<0.05), when considered collectively. Women with fibromyalgia showed reduced functional performance and lower limb muscle strength, mostly explained by pain. There was a high prevalence of falls in this population, as explained by hip extensors rate of torque development, duration of fibromyalgia symptoms and pain.


On the lipid granularity of renomedullary interstitial cells in benign and malignant courses of renal hypertension.
The number of lipid droplets of the renal medullary interstitial cells was registered in 40 rats with "two-kidney Goldblatt hypertension" and in 27 sham operated normotensive controls. A strong degranulation in the unclaimed kidneys was always associated with the malignant course of hypertension, characterized by a disturbance in the sodium and water balance. Giving 2 per cent saline in addition to demineralized water as a drinking fluid the decrease in the number of granules was prevented in most of the malignant cases. Significant changes in the granule count were never registered at a benign course of hypertension. Degranulation of the medullary interstitial cells in the unclamped kidneys does not unequivocally represent the loss of medullary antihypertensive function. It seems to be directly determined by the disturbance of sodium and water balance and indirectly by the hypertension.


[Effect of the hodja (magic healer) on Turkish psychiatric patients in West Germany--an evaluation of clinical examples].
Turkish magic healers called hodjas are working in West-Germany today. Our knowledge about their influence on Turkish patients is poor. They are consulted at least from patients with psychogenous diseases. The degree and the multiple dimensions of their influence and the possibility for the doctor to master the conflicting cultural identity of these patients is demonstrated on the base of four clinical case-histories.


Chemical crosslinking of histone H1o to histone neighbours in nuclei and chromatin.
Crosslinking of histones in mouse liver nuclei and extended chromatin with a bifunctional reagent leads to the formation of H1-H1o heterodimers as well as H1o-H1o homodimers. H1o can be also crosslinked to the core histones. Thus, the location of histone H1o within the basic repeating chromatin structure seems to be analogous to that of H1 histone.


Initial experience of single-incision plus one port total laparoscopic pancreaticoduodenectomy.
The use of single-incision plus one-port laparoscopic pancreaticoduodenectomy (SILPD + 1) has been never reported, and its safety and efficacy remain unknown. This study aimed to evaluate the short-term outcomes of SILPD + 1 compared with those of conventional laparoscopic pancreaticoduodenectomy (CLPD). Fifty-seven cases of laparoscopic pancreaticoduodenectomy (LPD) were performed between November 2021, and March 2022. Among them, 10 cases of LPD were performed using a single-incision plus one-port device. Based on the same inclusion and exclusion criteria, 47 cases of LPD performed using traditional 5-trocar were included as a control group. The patient's demographic characteristics, intraoperative, and postoperative variables were prospectively collected and retrospectively analyzed. Three men and seven women were included in the SILPD + 1 group. All baseline parameters of both groups were comparable, except for age. Patients were younger in the SILPD + 1 group (47.2 ± 18.3 years vs. 60.6 ± 11.7 years, P = 0.05) than that in the CLPD group. Compared with the CLPD group, median operation time (222.5 (208.8-245.0) vs. 305.0 (256.0-37.0) min, P < 0.001) was shorter, median postoperative VAS scores on days 1-3 were lower, and median cosmetic score (21.0 (19.0-23.5) vs. 17.0 (16.0-20.0), P = 0.026) was higher one month after the surgery in the SILPD + 1 group. The estimated blood loss, conversion rate, blood-transfusion rate, exhaust time, time of drainage tube removal, postoperative hospital stays, and perioperative complications were comparable between the two groups. In a high-volume LPD center, SILPD + 1 is safe and feasible for well-selected patients without increasing the operation time and complications. It even has the advantages of reduced postoperative pain and improved cosmetic results.


Localisation of low molecular weight sperm antigens during capacitation and acrosome reaction.
FITC-labelled sperm-specific antibodies against hamster spermatozoa were utilized as probes in acrosome reaction assays. An indirect immunofluorescence test demonstrated the localisation of two sperm proteins of 19 kDa and 23 kDa on the anterior acrosomal cap region of washed cauda epididymal sperm. These proteins were not detected in reacted acrosome or on immature or immotile sperm. Antisperm agglutinating antibodies specific to these two low molecular weight sperm antigens could be useful probes for evaluating the acrosomal status of mammalian spermatozoa.


Deletion of PAX9 and oligodontia: a third family and review of the literature.
This study was conducted to report a family affected by benign hereditary chorea in which a large deletion including TTF1, PAX9, and other genes was identified and results in oligodontia. Clinical and radiological studies of the two affected members (mother and daughter) were used to describe the oligodontia present in both of them. The missing teeth in both patients are described in detail, and these data are compared with the dental anomalies observed in the only two other families with deletions of PAX9 and with the data available for 12 previously reported families carrying different types of PAX9 mutations. There is a clinical relevance for recognizing such families, and offering available therapies since childhood is stressed. Some genotype-phenotype correlations between PAX9 mutations and dental anomalies can be drawn.


Strong optical modulation of surface plasmon polaritons in metal/semiconductor nanostructures.
We demonstrate strong modulation of the transmission around the surface plasmon polariton (SPP) resonance in metal/semiconductor hybrid nanostructures based on Ag film on top of InGaAs. The change in the real and imaginary parts of the refractive index due to photoexcited carriers in InGaAs generates a shift in the SPP resonance and enhanced transmission near the SPP resonance. Temporal evolution of the complex refractive index was traced by comparing the transient transmission with finite-difference time-domain (FDTD) simulations.


Changes in frailty among ICU survivors and associated factors: Results of a one-year prospective cohort study using the Dutch Clinical Frailty Scale.
Frailty is an important predictor for the prognosis of intensive care unit (ICU) patients. This study examined changes in frailty in the year after ICU admission, and its associated factors. Prospective cohort study including adult ICU patients admitted between July 2016-December 2017. Frailty was measured using the Clinical Frailty Scale (CFS), before ICU admission, at hospital discharge, and three and 12 months after ICU admission. Multivariable linear regression was used to explore factors associated with frailty changes. Frailty levels changed among 1300 ICU survivors, with higher levels at hospital discharge and lower levels in the following months. After one year were 42% of the unplanned, and 27% of the planned patients more frail. For both groups were older age, longer hospital length of stay, and discharge location associated with being more frail. Male sex, higher education level and mechanical ventilation were associated with being less frail in the planned patients. One year after ICU admission, 42% and 27% of the unplanned and planned ICU patients, respectively, were more frail. Insight in the associated factors will help to identify patients at risk, and may help in informing patients and their family members. ClinicalTrials.gov database (NCT03246334).


The PPARδ ligand L-165041 inhibits VEGF-induced angiogenesis, but the antiangiogenic effect is not related to PPARδ.
Peroxisome proliferator-activated receptor (PPAR)δ is known to be expressed ubiquitously and involved in lipid and glucose metabolism. Recent studies have demonstrated that PPARδ is expressed in endothelial cells (ECs) and plays a potential role in endothelial survival and proliferation. Although PPARα and PPARγ are well recognized to play anti-inflammatory, antiproliferative, and antiangiogenic roles in ECs, the general effect of PPARδ on angiogenesis in ECs remains unclear. Thus, we investigated the effect of the PPARδ ligand L-165041 on vascular EC proliferation and angiogenesis in vitro as well as in vivo. Our data show that L-165041 inhibited VEGF-induced cell proliferation and migration in human umbilical vein ECs (HUVECs). L-165041 also inhibited angiogenesis in the Matrigel plug assay and aortic ring assay. Flow cytometric analysis indicated that L-165041 reduced the number of ECs in the S phase and the expression levels of cell cycle regulatory proteins such as cyclin A, cyclin E, CDK2, and CDK4; phosphorylation of the retinoblastoma protein was suppressed by pretreatment with L-165041. We confirmed whether these antiangiogenic effects of L-165041 were PPARδ-dependent using GW501516 and PPARδ siRNA. GW501516 treatment did not inhibit VEGF-induced angiogenesis, and transfection of PPARδ siRNA did not reverse this antiangiogenic effect of L-165041, suggesting that the antiangiogenic effect of L-165041 on ECs is PPARδ-independent. Together, these data indicate that the PPARδ ligand L-165041 inhibits VEGF-stimulated angiogenesis by suppressing the cell cycle progression independently of PPARδ. This study highlights the therapeutic potential of L-165041 in the treatment of many disorders related to pathological angiogenesis.


Protection of nicotinic acid against oxidative stress-induced cell death in hepatocytes contributes to its beneficial effect on alcohol-induced liver injury in mice.
Oxidative stress plays a pathological role in the development of alcoholic liver disease. In this study, we investigated the effects of nicotinic acid (NA) supplementation on H2O2-induced cell death in hepatocytes and alcohol-induced liver injury in mice. Hepatocytes were exposed to H2O2 (0-0.4 mM) for 16 h after a 2-h pretreatment with NA (0-100 μM). Cell viability, intracellular glutathione and total NAD contents were determined. In animal experiments, male C57BL/6 mice were exposed to Lieber-De Carli liquid diet [+/- ethanol with/without NA supplementation (0.5%, w/v) for 4 weeks]. Nicotinic acid phosphoribosyltransferase (NaPRT) is the first enzyme participated in the NA metabolism, converting NA to nicotinic acid mononucleotide (NaMN). In NaPRT-expressing Hep3B cells, H2O2-induced cell death was attenuated by NA, whereas in NaPRT-lost HepG2 cells, only NaMN conferred protective effect, suggesting that NA metabolism is required for its protective action against H2O2. In Hep3B cells, NA supplementation prevented H2O2-inudced declines in intracellular total NAD and GSH/GSSG ratios. Further mechanistic investigations revealed that conservation of Akt activity contributed to NA's protective effect against H2O2-inudced cell death. In alcohol-fed mice, NA supplementation attenuated liver injury induced by chronic alcohol exposure, which was associated with alleviated hepatic lipid peroxidation and increased liver GSH concentrations. In conclusion, our findings indicate that exogenous NA supplementation may be an ideal choice for the treatment of liver diseases that involve oxidative stress.


Paleomicrobiology of Bartonella infections.
Studying ancient infectious diseases is a challenge, as written contemporary descriptions, when available, are often imprecise and do not allow for accurate discrimination among the pathogens endemic at that time. Paleomicrobiology offers a unique access to the history of these infections by identifying precisely the causative agents. Body louse-transmitted infections are amongst the most epidemic diseases in history, especially in war and famine periods. Of these, Bartonella quintana was detected by suicide PCR in 4000-year-old human remains, thus representing the oldest evidence to date of an arthropod-transmitted infection to human beings. This species has also been detected in human specimens from the 11th to 15th, 18th and 19th centuries. In addition, Bartonella henselae, a cat- and flea-associated pathogen, was detected in cat specimens from the 13th to 18th centuries, therefore demonstrating an association of the bacterium and its reservoir for over 800 years. Therefore, pathogenic Bartonella species have been involved in several outbreaks in the past millennia and should systematically be investigated in human remains from suspected epidemics.


First evidence of the clinical impact of a general practitioner with a special interest in epilepsy.
The purpose of the study was to review the clinical outcomes of people with epilepsy (PWE) attending a primary care-based specialist epilepsy service. The case notes of 355 people attending the service and subsequent follow-up from 2005 to 2013 were reviewed. There had been 37 deaths (all nonattributable to epilepsy), and 38 people had left the area, leaving 280 people who completed the audit. Positive outcomes could be attributed in 94% still attending the service at the end of follow-up. Seventy-five percent of people on treatment, referred with poor seizure control, achieved seizure remission with antiepilepsy drug (AED) changes initiated by the service. This study suggests that the majority of people who attended the service had a positive outcome and provides the first evidence for the clinical effectiveness of a general practitioner (GP) with special interest in epilepsy (GPwSIe) and provides support for the recommendations in earlier government reports to promote the use of such a service. Clinical Commissioning Groups (CCGs) and Government should consider investment in this intermediate tier of care as a means to both improving the quality of care and potentially reducing costs.


The influence of protease hydrolysis of lactic acid bacteria on the fermentation induced soybean protein gel: Protein molecule, peptides and amino acids.
The purpose explored the influence of protease hydrolysis of lactic acid bacteria and its hydrolysates on the fermentation induced soybean protein gel. The changes of protein molecule, peptides and amino acids after hydrolysis were analyzed. With the extension of fermentation time, the molecules (≤30 kDa) of L. paracasei-gel and L. plantarum-gel increased to 68.8% and 63%, the surface hydrophobicity of protein was increased, internal sulfhydryl groups and soluble proteins were reduced. These results indicated physicochemical properties of protein molecules were changed, small molecular proteins produced by hydrolysis were aggregated through intermolecular forces to promote gel formation. Peptide profiles and amino acid profiles showed that the peptides (≤10AA) of L. casei-gel increased by 3.32%, the peptides derived from 7S protein increased by more than 5%, the hydrophobic peptides and hydrophobic amino acids increased, these enzymatic hydrolysates may form insoluble aggregate through intermolecular forces to promote gel formation.


BLIMP1 transcriptionally induced by EGFR activation and post-translationally regulated by proteasome and lysosome is involved in keratinocyte differentiation, migration and inflammation.
B lymphocyte-induced maturation protein-1 (BLIMP1) is a transcriptional repressor, and plays a crucial role in the regulation of development and functions of various immune cells. Currently, there is limited understanding about the regulation of BLIMP1 expression in keratinocytes and crosstalk between EGFR and BLIMP1 in skin homeostasis. The aim of the study was to investigate the regulation and functional link between EGFR and BLIMP1 in human epidermal keratinocytes. Immunoblotting and Q-PCR were used to determine the molecular mechanism of BLIMP1 expression induced by EGFR in primary human epidermal keratinocytes (NHEK) and HaCaT cells. In functional assay, effects of BLIMP1 knockdown on EGFR-mediated cytokine production, differentiation, and migration in NHEK were evaluated by Q-PCR, ELISA, immunoblotting, and/or wound-healing assay. EGFR activation by EGFR ligands could upregulate the protein and mRNA levels of BLIMP1 in NHEK and HaCaT cells. This effect was dependent on PKC, p38, and ERK activation. Additionally, the stability of BLIMP1 protein was under the control of the proteasome and lysosome degradation systems. EGF could also upregulate BLIMP1 expression in skin squamous cell carcinomas. In addition, BLIMP1 knockdown enhanced the EGFR-mediated IL8, CXCL5 and IL6 gene expression and keratinocyte migration, but reduced the EGFR-mediated suppression of differentiation marker K10. Our findings shed new insights into the regulation of BLIMP1 expression by EGFR-mediated gene transcription and proteasome/lysosome-mediated degradation in keratinocytes. Functionally, BLIMP1 is a negative regulator of EGF-induced inflammation and migration in keratinocytes, and exerts a gene-specific regulation on keratinocyte differentiation.


Morphometry of lumbar muscles in the seated posture with weight-bearing MR scans.
Conventional imaging studies of human spine are done in a supine posture in which the axial loading of the spine is not considered. Upright images better reveal the interrelationships between the various internal structures of the spine. The objective of the current study is to determine the cross-sectional areas, radii, and angulations of the psoas, erector spinae, and multifidus muscles of the lumbar spine in the sitting posture. Ten young (mean age 31 ± 4.8 years) asymptomatic female subjects were enrolled. They were seated in an erect posture and weight-bearing T1 and T2 MRIs were obtained. Cross-sectional areas, radii, and angulations of the muscles were measured from L1-L5. Two observers repeated all the measurements for all parameters, and reliability was determined using the inter- and intra-class coefficients. The Pearson product moment correlation was used for association between levels, while level differences were used using a linear regression model. The cross-sectional areas of the psoas and multifidus muscles increased from L1 to L5 (1.9 ± 1.1 to 12.1 ± 2.5 cm2 and 1.8 ± 0.3 to 5.7 ± 1.4 cm2). The cross-sectional area of the erector spinae was greatest at the midlevel (13.9 ± 2.2 cm2) and it decreased in both directions. For the angle, the range for psoas muscles was 75-105°, erector spinae were 39-46° and multifidus was 11-19°. Correlations magnitudes were inconsistent between levels and muscle types. These quantitated data improve our understanding of the geometrical properties in the sitting posture. The weight-bearing MRI-quantified morphometrics of human lumbar spine muscles from this study can be used in biomechanical models for predicting loads on spinal joints under physiological and traumatic situations.


Comparison of Beneficial Metabolic Effects of Liraglutide and Semaglutide in Male C57BL/6J Mice.
Semaglutide and liraglutide are glucagon-like peptide-1 (GLP-1)-based diabetes drugs. Semaglutide possesses a longer half-life. Utilizing relatively lower doses, we compared the beneficial metabolic effects of these 2 drugs in mice fed a high-fat diet (HFD), aiming to deepen our mechanistic understanding on their energy homeostatic functions. Male C57BL/6J mice were fed an HFD for 10 weeks, followed by daily phosphate-buffered saline (PBS, as control); liraglutide (150 μg/kg body weight); or semaglutide (12 μg/kg body weight, low dose [LD]; or 60 μg/kg body weight, high dose [HD]) injection for 4 weeks. Metabolic tolerance and other tests were conducted within the 4-week period. Expression of metabolism-related genes, including Fgf21 in the liver and adipose tissues, was assessed after mice were euthanized. HFD-induced body weight gain, increasing inguinal fat tissue mass, glucose defects and insulin intolerance were effectively and comparably attenuated in the 3 experimental groups. HD semaglutide showed an even better effect on attenuating hyperleptinemia. Liraglutide but not semaglutide treatment enhanced hepatic fibroblast growth factor 21 (FGF21) protein level. All 3 experimental groups showed elevated expression of genes that encode pyruvate dehydrogenase kinase 4 and enoyl-CoA hydratase and 3-hydroxyacyl-coenzyme A dehydrogenase, associated with reduced plasma triglyceride levels. Finally, the plasma "GLP-1" level in HD semaglutide-treated mice was 14-fold higher than in HFD-fed control mice. Liraglutide, but not semaglutide, increased hepatic FGF21 protein level, whereas semaglutide had a greater effect on attenuating hyperleptinemia. Thus, these 2 GLP-1-based diabetes drugs may target metabolic organs, including liver and adipose tissue, with differing levels of efficacy.


Pediatric reference data for bone mineral density in the calcaneus for healthy children 2, 4, and 7 years of age by dual-energy x-ray absorptiometry and laser.
Dual-energy X-ray absorptiometry and laser (DXL) Calscan measures bone mineral density (BMD) in the calcaneus. In the present study, the DXL Calscan device has been modified for use in pediatric practice. It includes a function for measuring calcaneal height, which makes it possible to calculate volumetric bone mineral apparent density (BMAD). The aims of the present study were to evaluate the method when used in children, to create pediatric reference values in healthy Swedish 2-, 4-, and 7-yr-old children for BMD, bone mineral content (BMC), and BMAD, and to study whether these parameters were related to auxological data. The method was well tolerated by all children. Intraindividual coefficients of variation for BMC and BMD decreased with increasing age. The mean BMD was 0.17+/-0.003 g/cm2 in 2-yr-old children, 0.22+/-0.003 g/cm2 in 4-yr-old children, and 0.30+/-0.005 g/cm2 in 7-yr-old children. This study provides normative data as percentile values for BMD, BMC, and BMAD in young children measured with DXL Calscan. BMD was significantly correlated with age (p<0.001), height (p=0.001), weight (p<0.001), and body mass index standard deviation score (p<0.001). Seven-year-old girls showed significantly higher BMD than boys.


Preliminary experience of maintenance treatment with intravenous ustekinumab as a rescue treatment for loss of response to subcutaneous doses.
It has been demonstrated that ustekinumab (UST) is effective as an induction and maintenance therapy in patients with Crohn's disease (CD). However, a significant number of patients experience an insufficient response or a secondary non-response. We report six cases from our center that underwent a rescue treatment by changing maintenance treatment to weight-adjusted intravenous UST, obtaining a subsequent clinical improvement.


Facile discovery of red blood cell deformation and compromised membrane/skeleton assembly in Prader-Willi syndrome.
Prader-Willi syndrome (PWS) is a rare congenital disease with genetic alterations in chromosome 15. Although genetic disorders and DNA methylation abnormalities involved in PWS have been investigated to a significant degree, other anomalies such as those in erythrocytes may occur and these have not been clearly elucidated. In the present study, we uncovered slight anemia in children with PWS that was associated with increased red blood cell (RBC) distribution width (RDW) and contrarily reduced hematocrit (HCT) values. Intriguingly, the increased ratio in RDW to HCT allowed sufficient differentiation between the PWS patients from the healthy controls and, importantly, with individuals exhibiting conventional obesity. Further morphologic examinations revealed a significant deformity in erythrocytes and mild hemolysis in PWS patients. Comprehensive mechanistic investigations unveiled compromised membrane skeletal assembly and membrane lipid composition, and revealed a reduced F-actin/G-actin ratio in PWS patients. We ascribed these phenotypic changes in erythrocytes to the observed genetic defects, including DNA methylation abnormalities. Our collective data allowed us to uncover RBC deformation in children with PWS, and this may constitute an auxiliary indicator of PWS in early childhood.


Functionally distinct CD8+ memory T cell subsets in persistent EBV infection are differentiated by migratory receptor expression.
Human memory T lymphocytes have recently been re-defined as central or effector memory cells (Sallusto, F., Lenig, D., Forster, R., Lipp, M. and Lanzavecchia, A., Nature 1999. 401: 708-712). Effector memory cells (T(em)) are targeted to the peripheral tissues and show rapid effector function in response to antigenic stimulation. Central memory (T(cm)) cells are targeted to the lymph nodes and cannot be immediately activated. In this report HLA-A2-Epstein-Barr virus (EBV) peptide tetramers have been used to characterize the EBV-specific CD8+ T cell subsets in persistent EBV infection. In short-term activation studies two populations of tetramer-positive T cells were identified. One group resembled T(em) cells in that they rapidly produced IFN-gamma and lacked the lymph node homing receptor, CD62L, the second was similar to T(cm) cells since they were CD62L+ but could not be immediately induced to express IFN-gamma.


Thymine inhibits suppression by an Escherichia coli nonsense and frameshift suppressor.
A mutant strain of Escherichia coli suppressed a frameshift and some UAG, UAA, and UGA mutants of bacteriophage T4 at 37 degrees C but not at 31 degrees C. This suppression was inhibited by the addition of thymine or thymidine to the medium used to test phage growth. Furthermore, the suppressor strain required thymine or thymidine for growth on minimal medium at 43 degrees C and if this auxotrophy was removed by reversion or recombination the strain no longer suppressed. These results suggest a link between thymidine nucleotide biosynthesis and suppression.


σ-Electrons Responsible for Cooperativity and Ring Equalization in Hydrogen-Bonded Supramolecular Polymers.
Invited for this month's cover are collaborators from the TheoCheM group of the Vrije Universiteit Amsterdam and the University of Perugia. The cover picture shows a σ-electron traveling through a hydrogen-bonded squaramide linear chain. The charge transfer within the σ-electronic system is the cause for the cooperativity in the investigated urea, deltamide, and squaramide polymers. More information can be found in the Full Paper by Célia Fonseca Guerra, and co-workers.


Immune responses to bovine collagen implants. Significance of pretreatment serology.
Bovine collagen implants are in widespread use for the correction of soft tissue deficiencies. A small portion of the population responds adversely to these implants, evidenced by either positive skin tests or treatment site reactions. Antibodies against bovine collagen implants have been detected in the sera of virtually all patients with adverse treatment reactions and in small percentages of untreated individuals and treated individuals without adverse reactions. In this study we examined the levels of anti-bovine collagen implant antibodies in 150 individuals prior to bovine collagen implant therapy and then monitored these patients for adverse reactions to skin tests and bovine collagen implant treatments. Statistical analyses were performed to determine the value of pretreatment serology in predicting positive skin tests or adverse reactions. It was determined that individuals with greatly elevated levels (greater than 2 standard deviations [SDs] above the mean reactivity of pretreatment sera) of anti-bovine collagen implant antibodies prior to treatment were approximately sixfold more likely to suffer adverse reactions than patients with no elevation of antibodies prior to treatment.


Analysis of estrogen sulphate mixtures in pharmaceutical formulations by reversed-phase chromatography.
The relative retention times of estrogen sulphates for reversed-phase, ion-pair and argentous chromatography have been determined. Detector wavelengths of 200 nm, 200 nm and 270 nm were used, respectively. Samples are prepared by dissolution of pharmaceutical formulations in the high-performance liquid chromatography (HPLC) eluent. With reversed-phase chromatography, the estrone, equilin and 17 alpha-dihydroequilin estrogen sulphates are resolved from other estrogen sulphates in 20 min. Any traces of free estrogens which might be present are eluted too slowly for the formation of chromatographic peaks. With ion-pair chromatography, any hydrolyzed estrogens would be eluted before the sulphates with the formation of resolved chromatographic peaks. With argentous chromatography the retention times of the estrogen sulphates varied by nearly a factor of four. The utility of HPLC for the analysis of estrogen sulphates in pharmaceuticals is discussed.


Improving alcohol health literacy and reducing alcohol consumption: recommendations for Germany.
Although the detrimental health effects of alcohol are well established, consumption levels are high in many high-income countries such as Germany. Improving alcohol health literacy presents an integrated approach to alcohol prevention and an important complement to alcohol policy. Our aim was to identify and prioritize measures to enhance alcohol health literacy and hence to reduce alcohol consumption, using Germany as an example. A series of recommendations for improving alcohol health literacy were derived from a review of the literature and subsequently rated by five experts. Recommendations were rated according to their likely impact on enhancing (a) alcohol health literacy and (b) reducing alcohol consumption. Inter-rater agreement was assessed using a two-way intra-class correlation coefficient (ICC). Eleven recommendations were established for three areas of action: (1) education and information, (2) health care system, and (3) alcohol control policy. Education and information measures were rated high to increase alcohol health literacy but low to their impact on alcohol consumption, while this pattern was reversed for alcohol control policies. The ratings showed good agreement (ICC: 0.85-0.88). Improving alcohol health literacy and reducing alcohol consumption should be considered complementary and become part of a comprehensive alcohol strategy to curb the health, social, and economic burden of alcohol.


Split-bolus single-phase cardiac multidetector computed tomography for reliable detection of left atrial thrombus: comparison to transesophageal echocardiography.
Evaluation of a new cardiac MDCT protocol using a split-bolus contrast injection protocol and single MDCT scan for reliable diagnosis of LA/LAA thrombi in comparison to TEE, optimizing radiation exposure and use of contrast agent. A total of 182 consecutive patients with drug refractory AF scheduled for PVI (62.6 % male, mean age: 64.1 ± 10.2 years) underwent routine diagnostic work including TEE and cardiac MDCT for the evaluation of LA/LAA anatomy and thrombus formation between November 2010 and March 2012. Contrast media injection was split into a pre-bolus of 30 ml and main bolus of 70 ml iodinated contrast agent separated by a short time delay. In this study, split-bolus cardiac MDCT identified 14 of 182 patients with filling defects of the LA/LAA. In all of these 14 patients, abnormalities were found in TEE. All 5 of the 14 patients with thrombus formation in cardiac MDCT were confirmed by TEE. MDCT was 100 % accurate for thrombus, with strong but not perfect overall results for SEC equivalent on MDCT. • Patients with no filling defect or thrombus in MDCT in the LA/LAA region are unlikely to have thrombus and may undergo PVI without TEE.• Here, the role of an additional TEE in pre-procedural management prior to PVI in patients with AF has to be redefined.• Using a split-bolus injection protocol increases the diagnostic accuracy of thrombus in the LA/LAA region.


Increasing cardiac rate by measurement of right ventricular temperature.
A pacemaker that increases pacing rate in response to exercise enables the pacemaker patient to improve his state of well-being. Possible sensors include atrial rate, Q-T interval, pH, venous oxygen saturation, respiratory rate, cardiac output, body motion, and blood temperature. To determine the efficacy of right ventricular blood temperature as an indicator of exercise, an experimental temperature-controlled pacemaker was developed and was evaluated in dogs. A resulting algorithm was implemented in a microcomputer-based VVI pacemaker which reliably detected exercise and rest from blood temperature when totally implanted. The pacemaker generated two pacing rates to provide increased cardiac output. Data is being obtained to establish feasibility in humans.


Xeno-free generation of human induced pluripotent stem cells from donor-matched fibroblasts isolated from dermal and oral tissues.
Induced pluripotent stem cells (iPS) can be generated from various somatic cells and can subsequently be differentiated to multiple cell types of the body. This makes them highly promising for cellular therapy in regenerative medicine. However, to facilitate their clinical use and to ensure safety, iPS culturing protocols must be compliant with good manufacturing practice guidelines and devoid of xenogenic products. Therefore, we aimed to compare the efficiency of using humanized culture conditions, specifically human platelet lysate to fetal bovine serum, for iPS generation from different sources, and to evaluate their stemness. iPS were generated via a platelet lysate or fetal bovine serum-based culturing protocol from matched dermal, buccal and gingival human fibroblasts, isolated from healthy donors (n = 2) after informed consent, via episomal plasmid transfection. Pluripotency, genotype and phenotype of iPS, generated by both protocols, were then assessed by various methods. More attempts were generally required to successfully reprogram xeno-free fibroblasts to iPS, as compared to xenogenic cultured fibroblasts. Furthermore, oral fibroblasts generally required more attempts for successful iPS generation as opposed to dermal fibroblasts. Morphologically, all iPS generated from fibroblasts formed tight colonies surrounded by a reflective "whitish" outer rim, typical for iPS. They also expressed pluripotency markers at both gene (SOX2, OCT4, NANOG) and protein level (SOX2, OCT4). Upon stimulation, all iPS showed ability to differentiate into the three primary germ layers via expression of lineage-specific markers for mesoderm (MESP1, OSR1, HOPX), endoderm (GATA4) and ectoderm (PAX6, RAX). Genome analysis revealed several amplifications and deletions within the chromosomes of each iPS type. The xeno-free protocol had a lower reprogramming efficiency compared to the standard xenogenic protocol. The oral fibroblasts generally proved to be more difficult to reprogram than dermal fibroblasts. Xeno-free dermal, buccal and gingival fibroblasts can successfully generate iPS with a comparable genotype/phenotype to their xenogenic counterparts.


An endoplasmic reticulum transmembrane prolyl 4-hydroxylase is induced by hypoxia and acts on hypoxia-inducible factor alpha.
Prolyl 4-hydroxylases (P4Hs) act on collagens (C-P4Hs) and the oxygen-dependent degradation domains (ODDDs) of hypoxia-inducible factor alpha subunits (HIF-P4Hs) leading to degradation of the latter. We report data on a human P4H possessing a transmembrane domain (P4H-TM). Its gene is also found in zebrafish but not in flies and nematodes. Its sequence more closely resembles those of the C-P4Hs than the HIF-P4Hs, but it lacks the peptide substrate-binding domain of the C-P4Hs. P4H-TM levels in cultured cells are increased by hypoxia, and P4H-TM is N-glycosylated and is located in endoplasmic reticulum membranes with its catalytic site inside the lumen, a location differing from those of the HIF-P4Hs. Despite this, P4H-TM overexpression in cultured neuroblastoma cells reduced HIF-alpha ODDD reporter construct levels, and its small interfering RNA increased HIF-1alpha protein level, in the same way as those of HIF-P4Hs. Furthermore, recombinant P4H-TM hydroxylated the two critical prolines in HIF-1alpha ODDD in vitro, with a preference for the C-terminal proline, whereas it did not hydroxylate any prolines in recombinant type I procollagen chains.


Regurgitant bile acids and mucosal injury of the gastric remnant after partial gastrectomy.
Three groups, each consisting of seven patients who had undergone either Billroth I, Billroth II, or pylorus-preserving gastrectomies, were evaluated more than 18 months postoperatively in terms of concentration and amount of bile acids in the gastric aspirate and histologic changes in the gastric remnant mucosa. Concentrations of bile acids were determined by gas chromatography and mucosal specimens were obtained by endoscopic biopsy. The total bile acid concentration and all of the individual fractional bile acid levels, whether free or conjugated, were significantly higher in the Billroth II group than in the other two groups. The amount of gastric aspirate was also highest in the Billroth II group. Endoscopic biopsy revealed glandular dysplasia to be predominantly in the Billroth II group. The presence of bile acids in the gastric remnant may contribute to mucosal injury, possibly leading to cancer in the gastric remnant, especially after the Billroth II operation.


Controlling protected designation of origin of wine by Raman spectroscopy.
In this paper, a Fourier Transform Raman spectroscopy method, to authenticate the provenience of wine, for food traceability applications was developed. In particular, due to the specific chemical fingerprint of the Raman spectrum, it was possible to discriminate different wines produced in the Piedmont area (North West Italy) in accordance with i) grape varieties, ii) production area and iii) ageing time. In order to create a consistent training set, more than 300 samples from tens of different producers were analyzed, and a chemometric treatment of raw spectra was applied. A discriminant analysis method was employed in the classification procedures, providing a classification capability (percentage of correct answers) of 90% for validation of grape analysis and geographical area provenance, and a classification capability of 84% for ageing time classification. The present methodology was applied successfully to raw materials without any preliminary treatment of the sample, providing a response in a very short time.


Effect of simultaneously replacing putative TM6 and TM12 of human NBCe1-A with those from NBCn1 on surface abundance in Xenopus oocytes.
HCO₃⁻ translocation across the plasma membrane via the electrogenic Na/HCO₃⁻ cotransporter NBCe1 plays an important role in intracellular pH regulation and transepithelial HCO₃⁻ transport. However, the structural determinants of transporter function remain largely unknown. A previous study showed that the putative fourth extracellular loop (EL4) plays an essential role in determining the electrogenicity of NBCe1. In the present study, we generated eight new chimeras of human NBCe1-A and NBCn1-A. All possess the putative NBCe1 EL4 and are electrogenic. Chimera O, in which the putative sixth transmembrane segment (TM6) and EL5 through the C terminus (Ct) of NBCe1 was replaced by corresponding NBCn1 sequence, produces the smallest hyperpolarization (1-2 mV) when CO₂/HCO₃⁻ is added to the extracellular solution. Biotinylation experiments show that O has a very low abundance at the plasma membrane. However, chimeras in which we simultaneously replaced the putative TM6 and smaller subdomains of the EL5-Ct region for the NBCn1 sequence were strongly electrogenic except for chimera T, in which we replaced TM6 and TM12 of NBCe1 with the corresponding regions of NBCn1. T exhibited greatly reduced transporter surface expression compared to wild-type NBCe1-A, while retaining at least some electrogenic character. We hypothesize that putative TM6 and TM12 are part of a functional unit and that if the two TMs are replaced by those of the same transporter type, high surface expression would require that the surrounding TMs are also from the same transporter type.


Health care IT collaboration in Massachusetts: the experience of creating regional connectivity.
The state of Massachusetts has significant early experience in planning for and implementing interoperability networks for exchange of clinical and financial data. Members of our evolving data-sharing organizations gained valuable experience that is of potential benefit to others regarding the governance, policies, and technologies underpinning regional health information organizations. We describe the history, roles, and evolution of organizations and their plans for and success with pilot projects.


Stimulus size selectivity and receptive field organization of ectostriatal neurons in the pigeon.
The avian ectostriatum is the telencephalic recipient zone of the tectofugal pathway, which may be homologous to the colliculo-pulvinar-cortical pathway in mammals. The present paper studies the visual response properties and receptive field organization of ectostriatal cells in pigeons with extracellular recording and computer mapping techniques. The results indicate that 90% of ectostriatal cells prefer forward, downward and upward motion of visual stimuli at velocities in the range of 16-128 degrees s(-1). They respond optimally to small stimuli (1-4 degrees visual angle), mostly preferring a target of 2 degrees. Most cells (78.8%) have one excitatory receptive field that usually possesses one or two hotspots, some cells (13.5%) have two excitatory receptive fields each having one or two hotspots, and a few cells (7.7%) have one excitatory receptive field with an unresponsive region in the center. An inhibitory receptive field is not found surrounding the excitatory receptive field in the ectostriatal cells examined. These response properties and receptive field organization may reflect the possible roles of the ectostriatum in stimulus discrimination and visual cognition.


Scale matters in understanding the complexity of Amazon fires: A response to the Editor.
The Amazon Basin is experiencing climate change, altered hydrological cycles, and forest loss. The processes causing increased fires are complex, and therefore cannot be attributed to climate change or human-induced deforestation alone. Here, we show why the Amazon fires must be understood across spatial scales within the regional coupled system.


Activated T lymphocytes produce a matrix-degrading heparan sulphate endoglycosidase.
We have previously found that lines of activated T lymphocytes specifically autosensitized to the basic protein of myelin (BP), on intravenous inoculation into syngeneic rats, were able to penetrate blood vessels, accumulate in the nervous system and cause experimental autoimmune encephalomyelitis (EAE). An important question is how effector T cells reach such targets outside the walls of blood vessels. To investigate this we have studied in vitro the interaction of anti-BP effector T lymphocytes with the basement membrane-like extracellular matrix produced by vascular endothelial cells. We now report that activated but not resting T lymphocytes produce an endoglycosidase capable of degrading heparan sulphate side chains of the proteoglycan scaffold of the extracellular matrix. Moreover, the anti-BP T lymphocytes respond to BP presented by extracellular matrix by markedly enhanced elaboration of the endoglycosidase. These results suggest that tissue-specific antigens on blood vessel walls could direct lymphocyte homing by activating enzymes that facilitate penetration of the subendothelial basal lamina. They also suggest that effector T lymphocytes can recognize antigen which is not associated with a major histocompatibility complex signal.


Androgen receptor signaling regulates the transcriptome of prostate cancer cells by modulating global alternative splicing.
Androgen receptor (AR), is a transcription factor and a member of a hormone receptor superfamily. AR plays a vital role in the progression of prostate cancer and is a crucial target for therapeutic interventions. While the majority of advanced-stage prostate cancer patients will initially respond to the androgen deprivation, the disease often progresses to castrate-resistant prostate cancer (CRPC). Interestingly, CRPC tumors continue to depend on hyperactive AR signaling and will respond to potent second-line antiandrogen therapies, including bicalutamide (CASODEX®) and enzalutamide (XTANDI®). However, the progression-free survival rate for the CRPC patients on antiandrogen therapies is only 8-19 months. Hence, there is a need to understand the mechanisms underlying CRPC progression and eventual treatment resistance. Here, we have leveraged next-generation sequencing and newly developed analytical methodologies to evaluate the role of AR signaling in regulating the transcriptome of prostate cancer cells. The genomic and pharmacologic stimulation and inhibition of AR activity demonstrates that AR regulates alternative splicing within cancer-relevant genes. Furthermore, by integrating transcriptomic data from in vitro experiments and in prostate cancer patients, we found that a significant number of AR-regulated splicing events are associated with tumor progression. For example, we found evidence for an inadvertent AR-antagonist-mediated switch in IDH1 and PL2G2A isoform expression, which is associated with a decrease in overall survival of patients. Mechanistically, we discovered that the epithelial-specific splicing regulators (ESRP1 and ESRP2), flank many AR-regulated alternatively spliced exons. And, using 2D invasion assays, we show that the inhibition of ESRPs can suppress AR-antagonist-driven tumor invasion. Our work provides evidence for a new mechanism by which AR alters the transcriptome of prostate cancer cells by modulating alternative splicing. As such, our work has important implications for CRPC progression and development of resistance to treatment with bicalutamide and enzalutamide.


Cardiovascular effects and molecular targets of resveratrol.
Resveratrol (3,5,4'-trihydroxy-trans-stilbene) is a polyphenol phytoalexin present in a variety of plant species and has been implicated to explain the health benefits of red wine. A wide range of health beneficial effects have been demonstrated for resveratrol in animal studies. In this review, we summarize the cardiovascular effects of resveratrol with emphasis on the molecular targets of the compound. In this regard, resveratrol stimulates endothelial production of nitric oxide, reduces oxidative stress, inhibits vascular inflammation and prevents platelet aggregation. In animal models of cardiovascular disease, resveratrol protects the heart from ischemia-reperfusion injury, reduces blood pressure and cardiac hypertrophy in hypertensive animals, and slows the progression of atherosclerosis. A number of direct and indirect target molecules mediating the aforementioned cardiovascular effects of resveratrol have been identified. These include, among others, the estrogen receptor α, the adenosine receptors, the cyclooxygenase 1, the histone/protein deacetylase sirtuin 1, the AMP-activated protein kinase, the Akt kinase, the nuclear factor-E2-related factor-2, and NF-κB. Molecular mechanisms involved in the signal cascades are discussed.


Affinity labeling of catalytic subunit of bovine heart muscle cyclic AMP-dependent protein kinase by 5'-p-fluorosulfonylbenzoyladenosine.
Incubation of 5'-p-fluorosulfonylbenzoyladenosine with the catalytic subunit of bovine cardiac muscle cyclic AMP-dependent protein kinase led to the formation of an inactive enzyme irreversibly modified with approximately one mol of reagent per mol of subunit. The inactivation reaction followed pseudofirst order kinetics. The rate of inactivation at various reagent concentrations exhibited saturation kinetics implying that the reagent reversibly binds to the enzyme prior to inactivation. The addition of MgATP, MgADP, or MgAMP-PNP to the reaction mixture fully protected the enzyme from inactivation by 5'-p-fluorosulfonylbenzoyladenosine. The reagent was demonstrated to be a competitive inhibitor of MgATP with a Ki of 0.235 mM. Metal-free nucleotides were without effect upon the reaction rate while metal ions alone accelerated the inactivation rate up to 7-fold. The inclusion of casein or synthetic peptide substrate in the incubation mixture did not affect the reaction kinetics. Reaction of 5'-p-fluorosulfonylbenzoyladenosine with the kinase subunit exhibits all of the characteristics of affinity labeling of the MgATP-binding site.


Alveolar macrophages: antigen presentation activity in vivo.
Previous studies on the antigen-presenting activity of alveolar macrophages (AM) have been restricted to the use of in vitro assay systems, and have generally shown suppression of T cell activation by these macrophages. The present study exploits the rapid adaptability of AM to changes in oxygen tension, which facilitates their transfer to other tissues in a functional state. Employing this rationale, antigen-pulsed AM were shown to trigger T cell-dependent immune responses following subcutaneous or intraperitoneal inoculation, while suppressing similar responses in tissue culture.


Application of sensory evaluation, GC-ToF-MS, and E-nose to discriminate the flavor differences among five distinct parts of the Chinese blanched chicken.
Blanched chicken, boiling the whole chicken without removing head and butt parts during process, is a traditional poultry product in China. In this manuscript, the flavor differences of five parts (skin, breast, thigh, head and butt) of the Chinese blanched chicken (CBC) were studied using sensory evaluation and instrumental analysis. Check-all-that-apply (CATA), rate-all-that-apply (RATA), and 9-point hedonic preference were used to collect consumers' sensory results. Gas chromatograph-time of flight mass spectrometer (GC-ToF-MS) and electronic nose (E-nose) were used to discriminate flavor variances. CATA perceptions showed different parts of CBC stimulated consumers' odor and emotion feelings differently. GC-ToF-MS and E-nose data elaborated that the flavor of breast with chicken-like flavor was better than the other parts. Consumers preferred the chicken breast part the most. This analysis of different parts of CBC could provide advice to cater consumers' preference and predict the quality change might induce.


Partial in vitro analysis of toxic and antigenic activities of eleven Peruvian pitviper snake venoms.
This work used eleven Peruvian snake venoms (Bothrops andianus, Bothrops atrox, Bothrops barnetti, Bothrops castelnaudi, Bothriopsis chloromelas, Bothrocophias microphthalmus, Bothrops neuwiedi, Bothriopsis oligolepis, Bothriopsis peruviana, Bothrops pictus and Bothriopsis taeniata) to perform in vitro experimentation and determine its main characteristics. Hyaluronidase (HYAL), phospholipase A2 (PLA2), snake venom metalloproteinase (SVMP), snake venom serine protease (SVSP) and L-amino acid oxidase (LAAO) activities; toxicity by cell viability assays using MGSO3, VERO and HeLa cell lineages; and crossed immunoreactivity with Peruvian (PAV) and Brazilian (BAV) antibothropic polyvalent antivenoms, through ELISA and Western Blotting assays, were determined. Results show that the activities tested in this study were not similar amongst the venoms and each species present their own peculiarities, highlighting the diversity within Bothrops complex. All venoms were capable of reducing cell viability of all tested lineages. It was also demonstrated the crossed recognition of all tested venoms by both antivenoms.


Characteristics of a community-based distribution of home blood pressure in Ohasama in northern Japan.
To evaluate the distribution, reference values and day-to-day variation of blood pressure of untreated subjects measured at home. Cross-sectional study of a cohort. General community in northern Japan. Blood pressure was measured in 871 subjects (mean +/- SD age 46.0 +/- 19.5 years, range 7-98, constituting 38.7% of the local population of Uchikawama region, Ohasama) who were not receiving antihypertensive medication. Subjects measured their own blood pressure at home at least three times (mean +/- SD 19.7 +/- 8.4) each morning using a semi-automatic oscillometric blood pressure measuring device. Screening blood pressure was measured once. Distribution of home blood pressure in the study population as a whole and with respect to age and sex, and the distribution of day-to-day variation of home blood pressure were determined. Mean home blood pressure was 117.3 +/- 13.4/69.3 +/- 9.7 mmHg (95% confidence interval 116.4-118.2/68.7-70.0). The 95th centile value was 143/85 mmHg, mean+SD 131/79 mmHg and mean + 2SD 144/89 mmHg. Mean screening blood pressure was 126.2 +/- 18.9/72.1 +/- 11.7 mmHg (95th centile 159/92 mmHg). Age- and sex-specific 95th centile values as well as mean +/- SD were obtained. Mean+SD, mean + 2SD and the 95th centile values obtained as reference upper limits of home blood pressure from subjects identified as normotensive by screening blood pressure (n = 707) were 125/77, 137/86 and 134/83 mmHg, respectively. Home blood pressure increased gradually with increasing age in both men and women, although blood pressure was significantly higher in men until 50 years of age. Day-to-day variation of home systolic blood pressure also increased with age. Since the distribution of home blood pressure values was affected by age and sex, age- and sex-matched reference values for home blood pressure should be established. Home blood pressure values in elderly subjects should be evaluated carefully, since these exhibit greater day-to-day variation.


Is the androgen-dependent increase in preoptic estradiol-17 beta formation due to aromatase induction?
Aromatization of testosterone in the preoptic area of the male ring dove is required for the expression of specific estrogen-sensitive sexual behavior. Estradiol metabolism and/or binding of the aromatization product(s) to the preoptic estrogen receptors (ER) could interfere with the apparent increase in estradiol formation that occurs after androgen administration to castrated males. Therefore, we have re-examined the induction mechanism using a direct in vitro assay of aromatase activity. Since levels of aromatase activity and enzyme kinetics were similar irrespective of the assay used, we can conclude that increased estradiol formation after androgen stimulation is mainly due to enzyme induction and that estrogen binding and/or metabolism do not interfere significantly with the induction process.


DDX23 of black carp negatively regulates MAVS-mediated antiviral signaling in innate immune activation.
Mammalian DDX23 is involved in multiple biological processes, such as RNA processing and antiviral responses. However, the function of teleost DDX23 still remains unclear. In this paper, we have cloned the DDX23 homologue of black carp (Mylopharyngodon piceus) (bcDDX23) and elucidated its role in the antiviral innate immunity. The coding region of bcDDX23 comprises 2427 nucleotides and encodes 809 amino acids. The transcription of bcDDX23 was promoted by the stimulation of LPS, poly(I:C), and SVCV; and immunoblotting (IB) assay showed that bcDDX23 migrated aground 94.5 kDa. Immunofluorescence (IF) assay revealed that bcDDX23 was mainly distributed in the nucleus, and the amount of cytosolic bcDDX23 was significantly increased after SVCV infection. The reporter assay showed that bcDDX23 inhibited bcMAVS-mediated transcription of the IFN promoter. And the co-immunoprecipitation (co-IP) assays identified the interaction between bcDDX23 and bcMAVS. Furthermore, co-expressed bcDDX23 significantly inhibited bcMAVS-mediated antiviral ability against SVCV in EPC cells, and knockdown of bcDDX23 enhanced the resistance of host cells against SVCV. Overall, our results conclude that bcDDX23 targets bcMAVS and suppresses MAVS-mediated IFN signaling, which sheds light on the regulation of IFN signaling in teleost fish.


[Pancreatic pseudocyst in children: what is the best therapeutic approach?].
Pancreatic pseudocyst is an uncommon disorder in children and the majority of reported cases are secondary to trauma. Treatment options range from medical management to different forms of drainage procedure. The aim of this study was to discuss therapeutic strategies. The authors report herein pancreatic pseudocyst in four children aged 7, 9, 12, and 13 years with non-resolving pancreatic pseudocyst over a 2-year period from January 2006 to July 2008. The etiology of pancreatic pseudocyst was abdominal trauma in two cases and acute pancreatitis in two cases. Ultrasound and computed tomography scans confirmed the diagnosis. Two patients had endoscopic drainage. There were no procedure-related complications, nor was there a recurrence of the cyst. In one case, the pancreatic pseudocyst resolved spontaneously. This report suggests that children with non-spontaneously resolving pancreatic pseudocyst can be treated successfully and safely with endoscopic drainage. Surgical treatment remains an important alternative in the therapeutic armamentarium of this affection.


A retrospective cost-effectiveness analysis of the treatment of onychomycosis in general practice.
Analysis of the computer records of 100 general practices from the CompuFile Doctors Independent Network revealed 1492 patients receiving treatment for onychomycosis in the first 6 months of 1994 with terbinafine, tioconazole, amorolfine or griseofulvin. These records indicated the average treatment time for each agent, number of general practitioner consultations and incidence of hospital referrals and minor surgery. Applying standard costs to this resource consumption gave the direct costs for each of these four agents. Published clinical and mycological cure rates allowed a cost per success to be calculated. These were as follows: terbinafine (n = 511) pound258, amorolfine (n = 315) pound312, griseofulvin (n = 196) pound356 and tioconazole (n = 470) pound520. Sensitivity analysis showed that terbinafine remained the most cost-effective option despite variations in resource costs. The cost impact on a typical practice for switching from a less to a more cost-effective treatment is discussed.


Improving packed red blood cell storage with a high-viscosity buffered storage solution.
Massive transfusion with older packed red blood cells is associated with increased morbidity and mortality. As packed red blood cells age, they undergo biochemical and structural changes known as the storage lesion. We developed a novel solution to increase viscosity in stored packed red blood cells. We hypothesized that packed red blood cell storage in this solution would blunt storage lesion formation and mitigate the inflammatory response after resuscitation. Blood was obtained from 8- to 10-week-old C57BL/6 male donor mice or human volunteers and stored as packed red blood cell units for 14 days for mice or 42 days for humans in either standard AS-3 storage solution or EAS-1587, the novel packed red blood cell storage solution. Packed red blood cells were analyzed for microvesicles, cell-free hemoglobin, phosphatidylserine, band-3 protein, glucose utilization, and osmotic fragility. Additional mice underwent hemorrhage and resuscitation with packed red blood cells stored in either AS-3 or EAS-1587. Serum was analyzed for inflammatory markers. Murine packed red blood cells stored in EAS-1587 demonstrated reductions in microvesicle and cell-free hemoglobin accumulation as well as preserved band-3 expression, increase glucose utilization, reductions in phosphatidylserine expression, and susceptibility to osmotic stress. Serum from mice resuscitated with packed red blood cells stored in EAS-1587 demonstrated reduced proinflammatory cytokines. Human packed red blood cells demonstrated a reduction in microvesicle and cell-free hemoglobin as well as an increase in glucose utilization. Storage of packed red blood cells in a novel storage solution mitigated many aspects of the red blood cell storage lesion as well as the inflammatory response to resuscitation after hemorrhage. This modified storage solution may lead to improvement of packed red blood cell storage and reduce harm after massive transfusion.


Hepatic heterogeneity on CT in Budd-Chiari syndrome: correlation with regional disturbances in portal flow.
A comparative study of the imaging findings of computed tomography (CT), selective arteriography, CT arteriography, and/or CT portography is presented in 4 patients with Budd-Chiari syndrome. Hepatic differences in attenuation and morphologic changes were generally found to be closely related with regional disturbances in portal flow. Areas with complete hepatic vein obstruction were hypodense on pre- and postcontrast scans, probably due to portal flow inversion. In 2 of 4 cases, these were subsequently atrophied, while areas receiving the remaining venous outflow appeared hypertrophied. They were markedly enhanced on postcontrast scans. Enhancement may be patchy due to portal and sinusoidal stasis.


Vasopressin neuropeptides and acquisition of heroin and cocaine self-administration in rats.
The effect of the vasopressin neuropeptide des-glycinamide (Arg8)-vasopressin (DGAVP) on reducing the acquisition of intravenous heroin self-administration in rats was analyzed. When rats reduced in body weight were allowed to self-administer heroin for 1 h per day in the presence of a fixed time, non contingent food delivery schedule, it appeared that heroin intake was related in an orderly way to the unit dose of heroin delivered. DGAVP decreased heroin intake during days 4 and 5 of acquisition, especially when a high dose of heroin was delivered. DGAVP decreased heroin intake more effectively when rats were tested without the food delivery schedule and for 6 h instead of 1 h sessions per day. Structure activity relationship studies revealed that the peptide (pGlu4, Cyt6)AVP-(4-8) was the shortest active sequence mimicking the effect of DGAVP and that this peptide was somewhat more potent than DGAVP in this respect. The peptide (pGlu4,Cyt6)AVP-(4-9) increased the heroin intake of the rats. DGAVP and (pGlu4,Cyt6)-AVP-(4-8) also decreased cocaine intake of body weight reduced rats given the opportunity to self-administer cocaine intravenously in daily 6 h sessions. It is concluded that vasopressin neuropeptides may decrease the reinforcing efficacy of heroin and cocaine during acquisition of drug self-administration rather than interact with nutritional and environmental factors influencing drug taking behavior.


The myristoylation site of meiotic lamin C2 promotes local nuclear membrane growth and the formation of intranuclear membranes in somatic cultured cells.
Lamin C2 is a splice product of the mammalian lamin A gene and expressed in primary spermatocytes where it is distributed in the form of discontinuous plaques at the nuclear envelope. We have previously shown that the aminoterminal hexapetide GNAEGR of lamin C2 following the start methionine is essential for its association with the nuclear envelope and that the aminoterminal glycine of the hexapeptide is myristoylated. Here we have analyzed the ultrastructural changes induced in COS-7 and Xenopus A6 cells by overexpressing rat lamin C2 or a human lamin C mutant possessing the lamin C2-specific hexapeptide at its aminoterminus. Both lamins were targeted to the nuclear envelope of mammalian and amphibian cells and induced the formation of intranuclear membranes, whereas wild-type human lamin C and a lamin C2 mutant, that both lack this lipid moiety, did not. Our data indicate that the myristoyl group of lamin C2 has besides its demonstrated role in nuclear envelope association additional functions during spermatogenesis. Our present study complements previously published results where we have shown that the CxxM motif of lamins promotes nuclear membrane growth (Prüfert et al., 2004. J. Cell Sci. 117, 6105-6116).


Endocarditis of the forgotten valve: vacuum-assisted aspiration (AngioVac) of the eustachian valve: a case report.
Eustachian valve endocarditis (EVE) is a rare entity that traditionally has been treated with antibiotics or surgery, if refractory to antibiotic treatment. A 64-year-old man presented with right shoulder pain and new-onset hypoxia. His blood cultures were positive for methicillin-sensitive staphylococcal aureus (MSSA) 1 month ago and he was treated with antibiotics at that time. Blood cultures during this admission were again positive for MSSA. Trans-oesophageal echocardiogram showed a large independently mobile echogenic density consistent with vegetation (3.0 × 1.6 cm) on the eustachian valve (EV). The patient was a poor surgical candidate due to his multiple co-morbidities, and therefore, a non-invasive procedure called AngioVac® was selected. In the setting of infective endocarditis refractory to antibiotics, the large-bore percutaneous mechanical aspiration (AngioVac®, AngioDynamics, Latham, NY, USA) system is gaining increasing momentum as the treatment of choice over standard surgical intervention for debulking large vegetations. AngioVac® has provided a minimally invasive and effective measure especially in those unable to tolerate surgery. The novel percutaneous technique is linked to great success in right-sided endocarditis, with the tricuspid valve accounting for a majority of the cases. However, in rare instances, the EV may be involved. To our knowledge, we report the first case of EVE treated with AngioVac®.


Immunoglobulin allotypes in Southwest Asia: populations at the crossroads.
Southwest Asia has a long history of contact with Central Asian and with Sub-Saharan African populations. Is the genetic structure of these populations reflective of these historical facts? To study this, data was generated on the immunoglobulin heavy chain (GM) and light chain (KM) allotypes from seven Arab and three non-Arab populations in SW Asia to examine the relationship of these populations to SE European, NW Indian, Sub-Saharan African, and Central Asian populations. Like mtDNA and Y chromosome markers, the GM haplotypes are largely continent specific making them an excellent tool for the detection of gene flow whereas the KM markers are less informative. Six of the nine GM haplotypes detected in SW Asians are Indo-European, Sub-Saharan African, or East Asian specific. The allotype results indicate variable but significantly higher Sub-Saharan African gene flow in Arab populations (average 26.9%; 15.0-61.6%) vs. the non-Arab populations (average 7.3%; 9.0-13.4%), but higher levels of Central Asian gene flow in the non-Arab populations (average 28.8%; 10.5-48.8%) vs. the Arab populations (average 9.0%; 0.0-26.4%). Principal components analysis and hierarchical cluster analysis based on the immunoglobulin allotypes are consistent with the historical population contacts of this part of the world and reflect the power of the GM haplotypes in dissecting population relationships. However, the KM*1 frequencies were only correlated with the degree of African gene flow (Pearson r = 0.69, P = 0.026) in SW Asian populations.


Percutaneous closure of patent ductus arteriosus in small infants with significant lung disease may offer faster recovery of respiratory function when compared to surgical ligation.
To describe our experience with percutaneous closure of patent ductus arteriosus (PDA) in small infants and compare outcomes to matched surgical patients. Ligation via thoracotomy has been used to close PDAs in small infants, but has been associated with respiratory and hemodynamic compromise. We hypothesized that percutaneous closure would offer faster recovery of respiratory function. Patients <4 kg requiring positive pressure ventilation who underwent percutaneous PDA closure between January 2000 and April 2012 were reviewed and matched to contemporary surgical patients on gestational age (GA), birth weight (BW), procedure weight (WT), and ventilation mode. Patients returned to baseline respiratory status when the product of mean airway pressure and FiO2 returned to pre-procedural levels. Eight matched pairs were included. Median BW, GA, and WT were 1.43 kg (0.52-2.97), 29.8 weeks (24-39), and 2.8 kg (2.2-3.9) for catheter patients and 1.55 kg (0.48-3.04), 29 weeks (23-37), and 2.75 kg (2.3-4.2) for surgical patients. Complete PDA closure occurred in all. The median time to return to baseline respiratory status was significantly shorter in the percutaneous group (17 hr (range 0-113) vs. 53 hr (range 13-219), P < 0.05). In the percutaneous group, two patients developed mild aortic coarctation, one mild left pulmonary artery stenosis, and four femoral vascular thromboses which all resolved with medical therapy. Surgical complications included significant respiratory and cardiac compromise, rib fractures and urinary retention. Percutaneous closure of PDA in small infants on respiratory support is equivalent in safety and efficacy and may offer shorter recovery time than surgical ligation.


Reduction of intrahepatic vascular space in the pathogenesis of portal hypertension. In vitro and in vivo studies in the rat.
To elucidate a possible role for reduction of intrahepatic vascular space in the pathogenesis of portal hypertension, we studied a partially hepatectomized rat model in vitro and in vivo. The in vitro study used livers from normal, 1/3-hepatectomized, and 2/3-hepatectomized rats, and rats with cirrhosis caused by chronic bile duct ligation, for isolated, perfused flow-pressure plotting. Resistance to perfusion increased such that significant differences were found between all groups except the last two, which showed similar resistances. The in vivo study measured splanchnic blood flow by radioactive microspheres and portal pressure in anesthetized sham-operated and 1/3- and 2/3-hepatectomized rats. Although absolute portal tributary blood flows did not change, portal flow per gram of remnant liver showed significant increases: 1.57 +/- 0.32 ml/min X g liver, 2.52 +/- 0.60 ml/min X g liver, p less than 0.01; 3.48 +/- 1.04 ml/min X g liver, p less than 0.01, respectively. Although intrahepatic resistance increased significantly only in the 2/3-hepatectomized group, portal pressures increased significantly in both groups of hepatectomized rats: normal, 7.5 +/- 1.1 mmHg; 1/3-hepatectomized, 9.4 +/- 1.1 mmHg; and 2/3-hepatectomized, 11.1 +/- 1.2 mmHg. Thus, decreased intrahepatic vascular space caused by resection and hepatocellular hypertrophy leads to portal hypertension, thereby suggesting that this reduction in space may be the pathogenic factor common to a number of different theories of portal hypertension.


Fiberglass dermatitis: clinical presentations, prevention, and treatment - a review of literatures.
Fiberglass dermatitis is a common form of occupational irritant contact dermatitis. Individuals experience a negative reaction to fiberglass fragments because of mechanical irritation. This review comprehensively analyzes the information in the existing literature on fiberglass dermatitis. Recognizing the different environmental risks factors for acquiring fiberglass dermatitis is necessary for better diagnosis and prevention. In addition to the identification of high-risk populations exposed to fiberglass particles, the common clinical presentation and preventive measures that can reduce the risk of acquiring fiberglass dermatitis are thoroughly discussed.


The identification of polyoxyethylene glycols and related compounds by capillary gas chromatography.
A sample work-up method for gas chromatographic profiling of polyoxyethylene glycol (PEG)-related compounds in pharmaceutical matrices is described. After a short sample clean-up, carbon-oxygen linkages are partially cleaved with 0.07 M boron tribromide in dichloromethane at room temperature. The reaction is stopped after 1 min by addition of 0.01 M HCl. The products are trimethylsilylated and injected onto a WCOT 50 m x 0.25 mm CP-SIL 5 CB fused silica column. Eleven model compounds, representing four common types of PEG-derivatives, have been evaluated by this method. The results show that characteristic profiles can be obtained from PEG-derivatives carrying different functional groups. Minimum detectable amounts are in the range of 200 micrograms.


Review of current research at Boston Children's Hospital.
Both clinical and laboratory studies are being undertaken to investigate the deleterious neurologic and developmental effects associated with cardiopulmonary bypass, hypothermia, and circulatory arrest in the neonate and infant. A prospective, randomized clinical study of 171 neonates and young infants compared circulatory arrest with low-flow bypass (50 mL.kg-1.min-1). Circulatory arrest was associated with a higher incidence of early postoperative seizures as well as greater release of creatine kinase-BB. There was a strong correlation between duration of circulatory arrest and seizures (p = 0.004). The late consequences of these findings will be known at the completion of developmental assessment of all patients at 1 and 4 years of age. Laboratory studies have used a miniature piglet model that closely replicates clinical circulatory arrest. High-energy phosphate stores determined by magnetic resonance spectroscopy were maintained in animals undergoing 1 hour of low-flow bypass but became undetectable after 32 minutes of a 1-hour period of circulatory arrest. However, they returned to baseline within 3 hours of reperfusion as did cerebral blood flow and metabolism determined by microsphere studies. Piglets undergoing 1 hour of circulatory arrest showed more rapid recovery of cerebral adenosine triphosphate content and intracellular pH when managed with the pH-stat strategy during hypothermic bypass than with the more alkaline alpha-stat strategy. Other laboratory studies have examined pharmacologic methods of reducing cerebral injury associated with circulatory arrest including aprotinin, anti-CD18, neuronal receptor antagonists (MK801, NBQX), and blockade of glutamate release with adenosine in a cerebroplegia solution. These studies have suggested a number of promising approaches to improving the technique of circulatory arrest.


Recent developments and emerging directions in ion chromatography.
This review provides a selected overview of some of the significant developments that have occurred in ion chromatography (IC) over recent years, together with some views on the future directions that IC will follow. The topics covered in the review include new stationary phases (especially monolithic phases), miniaturized IC systems (capillary, microchip and low-pressure formats), enhanced peak capacity through the use of complex eluent profiles and associated computer tools for simulation and prediction of retention, hyphenated IC systems and their use for speciation studies and metallomics, and the rapidly increasing use of IC in bioanalysis.


Minimally invasive renal auto-transplantation: the first report.
Regarding transplant surgery, the minimally invasive revolution was started in 1995 by laparoscopic living donor nephrectomy (L-LDN). In 2006, we made the first report on a minimally invasive technique for kidney transplantation; restricted to a 7-9 cm transverse incision targeted on the anastomotic area of the iliac vessels, and with the meticulously prepared kidney placed in a fitting, retroperitoneal pouch lateral to the skin incision. By combining "hand-assisted laparoscopic nephrectomy" and "minimally invasive kidney transplantation" - using the same incision (7-8 cm) for hand-assistance, kidney harvesting, and transplantation - we have during 2009 conducted "minimally invasive renal auto-transplantation" in two patients. In both cases, the postoperative course was uneventful. When examined 3 mo postoperatively, both auto-transplants were shown to have excellent function by renal scintigraphy. Renal auto-transplantation, a traditionally major surgical procedure, can be made minimally invasive by a similar incision as that used for L-LDN. Taking into regard the highly traumatic conventional incisions, we expect the generally proven minimally invasive benefits to be considerable.


Liver glycogen synthesis and glucose tolerance in rats adapted to diets with a high proportion of fructose or glucose.
Glucose tolerance and in vivo incorporation of glucose into liver glycogen were investigated in rats fed high carbohydrate diets containing glucose or fructose as the sole carbohydrate source. As compared with control glucose-adapted rats, a slight deterioration of the glucose tolerance was observed in fructose-adapted rats. The possivle cause of the deteriorated glucose tolerance in fructose-adapted rats seems to be among others reduced glucose incorporation into liver glycogen and a smaller depression of endogenous glucose production by exogenous glucose.


Facioscapulohumeral muscular dystrophy type 2: an update on the clinical, genetic, and molecular findings.
Facioscapulohumeral muscular dystrophy (FSHD) is a common genetic disease of the skeletal muscle with a characteristic pattern of weakness. Facioscapulohumeral muscular dystrophy type 2 (FSHD2) accounts for approximately 5% of all cases of FSHD and describes patients without a D4Z4 repeat contraction on chromosome 4. Phenotypically FSHD2 shows virtually no difference from FSHD1 and both forms of FSHD arise via a common downstream mechanism of epigenetic derepression of the transcription factor DUX4 in skeletal muscle cells. This results in expression of DUX4 and target genes leading to skeletal muscle toxicity. Over the past decade, major progress has been made in our understanding of the genetic and epigenetic architecture that underlies FSHD2 pathogenesis, as well as the clinical manifestations and disease progression. These include the finding that FSHD2 is a digenic disease and that mutations in the genes SMCHD1, DNMT3B, and more recently LRIF1, can cause FSHD2. FSHD2 is complex and it is important that clinicians keep abreast of recent developments; this review aims to serve as an update of the clinical, genetic, and molecular research into this condition.


CDK-mediated Yku80 Phosphorylation Regulates the Balance Between Non-homologous End Joining (NHEJ) and Homologous Directed Recombination (HDR).
There are two major pathways for repairing DNA double-strand breaks (DSBs): homologous directed recombination (HDR) and non-homologous end-joining (NHEJ). While NHEJ functions throughout the cell cycle, HDR is only possible during S/G2 phases, suggesting that there are cell cycle-specific mechanisms regulating the balance between the two repair systems. The regulation exerted by CDKs on HDR has been extensively demonstrated, and here we present evidence that the CDK Pho85, in association with the G1 cyclin Pcl1, phosphorylates Yku80 on Ser 623 to regulate NHEJ activity. Cells bearing a non-phosphorylatable version of Yku80 show increased NHEJ and reduced HDR activity. Accordingly, yku80S623A cells present diminished viability upon treatment with the DSB-producer bleomycin, specifically in the G2 phase of the cell cycle. Interestingly, the mutation of the equivalent residue in human Ku80 increases sensitivity to bleomycin in several cancer cell lines, suggesting that this mechanism is conserved in humans. Altogether, our results reveal a new mechanism whereby G1-CDKs mediate the choice between HDR and NHEJ repair pathways, putting the error prone NHEJ on a leash and enabling error free HDR in G2 when homologous sequences are available.


The effect of paraspinal muscle on functional status and recovery in patients with lumbar spinal stenosis.
To investigate the association of paraspinal muscle quantity and quality with functional status in patients with lumbar spinal stenosis (LSS) and explore whether degeneration of paraspinal muscle could predict patients' functional recovery. The data of 69 patients (26 males, 43 females; mean age 60.6 ± 9.4 years) with LSS was reviewed. The total cross-sectional area (tCSA), functional cross-sectional area (fCSA), and fatty infiltration (FI) of paraspinal muscle were measured. The Oswestry Disability Index (ODI) scores were used to reflect patients' functional status. Correlations between measurements of paraspinal muscle and ODI scores were investigated by the Pearson correlation analysis. The multiple linear regression analysis was used to explore the correlation between change of ODI and other potential influence factors. Receiver operating characteristic curve was used to find out the most optimum cut-off value for predicting functional recovery. The pre-operation ODI was significantly associated with multifidus muscle (MF) fCSA (r = - 0.304, p = 0.012), while the post-operation ODI was significantly correlated to MF FI (r = 0.407, p < 0.01). Preoperative MF FI was an independent influence factor for change of ODI. The best cut-off value of preoperative MF FI to predict improvement of functional status (change of ODI > 50%) was 33%. The preoperative degeneration of MF was significantly associated with patients' functional status. Preoperative MF FI was a good predictor for assessing improvement of patients' functional status. Evaluation of paraspinal muscle before operation could be helpful for surgeons to predict patients' functional status and recovery.


Reduced Mitochondrial Apoptotic Priming Drives Resistance to BH3 Mimetics in Acute Myeloid Leukemia.
Acquired resistance to BH3 mimetic antagonists of BCL-2 and MCL-1 is an important clinical problem. Using acute myelogenous leukemia (AML) patient-derived xenograft (PDX) models of acquired resistance to BCL-2 (venetoclax) and MCL-1 (S63845) antagonists, we identify common principles of resistance and persistent vulnerabilities to overcome resistance. BH3 mimetic resistance is characterized by decreased mitochondrial apoptotic priming as measured by BH3 profiling, both in PDX models and human clinical samples, due to alterations in BCL-2 family proteins that vary among cases, but not to acquired mutations in leukemia genes. BCL-2 inhibition drives sequestered pro-apoptotic proteins to MCL-1 and vice versa, explaining why in vivo combinations of BCL-2 and MCL-1 antagonists are more effective when concurrent rather than sequential. Finally, drug-induced mitochondrial priming measured by dynamic BH3 profiling (DBP) identifies drugs that are persistently active in BH3 mimetic-resistant myeloblasts, including FLT-3 inhibitors and SMAC mimetics.


Efficacy of stereotactic radiosurgery as a salvage treatment for recurrent malignant gliomas.
The objective of this prospective cohort study was to determine the efficacy of stereotactic radiosurgery (SRS) as a salvage treatment in patients with recurrent malignant gliomas. Between January 2000 and December 2006, 114 consecutive patients were treated with SRS as a salvage treatment for recurrent malignant gliomas at a single institution. Clinical outcome and its prognostic factors were analyzed and compared with the historical control group who were treated at the same institution between 1995 and 1999. The median overall survival from the time of diagnosis was 37.5 months (95% confidence interval [95% CI], 11.7-63.2 months) for patients with grade 3 gliomas (according to World Health Organization criteria) and was 23 months (95% CI, 16.2-29.3 months) for patients with glioblastomas. The median progression-free survival after SRS was 8.6 months (95% CI, 1.1-16.2 months) for patients with grade 3 gliomas and 4.6 months for patients with glioblastomas (95% CI, 4.0-5.2 months). With regard to treatment-related complications, radiation-induced necrosis was observed in 22 of 114 patients (24.4%). Compared with this historic control group, SRS significantly prolonged survival as a salvage treatment in patients with recurrent glioblastomas (23 months vs 12 months; P < .0001), but it was not found to provide a significant surgical benefit in patients with recurrent grade 3 gliomas (37.5 months vs 26 months; P = .789). On univariate analysis of prognostic factors, tumor volume (<10 mL) and low histologic grade were found to significantly influence better survival (P = .009 and P = .041, respectively). SRS is a safe and effective modality in selected patients with recurrent small-sized glioblastomas. However, the efficacy of SRS for recurrent grade 3 gliomas needs to be further evaluated in well-designed clinical studies.


Streptococcus agalactiae and Prototheca spp. induce different mammary gland leukocyte responses in Holstein cows.
In this study, we investigated the association between natural subclinical intramammary infection (IMI) caused by Streptococcus agalactiae and Prototheca spp. and milk differential cell counts assessed by cytofluorimetric analysis in Holstein cows. After an initial bacteriological screening on 188 animals and a second assessment carried out 2 wk later aimed at confirming the bacteriological status, we collected milk samples from 47 animals and performed (1) milk composition analyses; (2) somatic cell counts and differential somatic cell counts (DSCC); and (3) cytofluorimetric analyses. Before statistical analyses, animals with co-infections were filtered out. Bacteriological status (negative, positive for Strep. agalactiae, or positive for Prototheca spp.) significantly affected the investigated traits. Compared with culture-negative samples, those that were positive for Strep. agalactiae and Prototheca spp. had higher SCS (+61% and +49%, respectively), DSCC (+4% and +19%, respectively), log polymorphonuclear neutrophil (PMN)-lymphocyte (LYM) counts (+59% and +71%, respectively), and log macrophage (MAC) counts (+63% and +72%, respectively). The individual leukocyte populations determined by cytofluorimetric analysis confirmed that mastitis infection increased the proportion of PMN in the milk samples compared with culture-negative samples, particularly when caused by Strep. agalactiae (+51%). In the case of MAC, the 2 pathogens behaved in opposite ways: Strep. agalactiae increased MAC by 41%, whereas Prototheca decreased MAC by 25%. Prototheca infection strongly increased the proportion of total T lymphocytes (TL; +87%) and T-helper lymphocytes (+83%). Accordingly, the (PMN+MAC):TL ratio increased with Strep. agalactiae infection (+95%) and decreased with Prototheca infection (-43%) compared with culture-negative samples. These results suggest the prevalence of an adaptive immune response and chronic inflammation in Prototheca infection, and an innate immune response to Strep. agalactiae. This knowledge might provide an important contribution to the development of novel and effective diagnostics and therapeutics.


Adherence to treatment guidelines in primary health care facilities in Botswana.
To assess the quality of nurses' prescribing through an assessment of their prescription in relation to diagnosis, and to investigate trends in drug use in Botswana primary health care. Key data regarding nurses' adherence to national prescription and treatment guidelines were obtained through participatory observation using a questionnaire, related to each consultation. Adherence was categorized into (i) Full adherence, (ii) acceptable adherence, (iii) acceptable adherence, but one or more useless, but not dangerous, drugs and (iv) insufficient or dangerous treatment. The study comprises data on nurses' prescriptions, diagnoses and quality of dispensing in 2994 consecutive consultations in 30 primary health care facilities in three districts of Botswana: Ngami East, Gaborone and Kgalagadi North. The average number of drugs prescribed per patient was 2.3. Antibiotics were prescribed in 27% of all encounters. Full adherence was found in 44%, acceptable compliance in 20%, 'acceptable, but one or more useless, but not dangerous, drugs' in 33% and 'insufficient or dangerous treatment' in 3% of the consultations. Four factors were found to be independently associated with full adherence: patient age 16-31 years, specified diagnosis, type of health facility and nurses' years of practice (4-11 years best). Although Botswana's health workers perform relatively well in terms of drug use indicators, there is a clear potential for improving health workers' adherence to national treatment guidelines.


Main genome characteristics of butanol-producing Clostridium sp. UCM В-7570 strain.
The rapid development of new molecular methods and approaches, sequencing technologies, has provided new insights into genetic and structural features of bacterial genomes. Information about the genetic organization of metabolic pathways and their regulatory elements has greatly contributed to the increase in the number of studies related to the construction of new bacterial strains with improved characteristics. In this study, the entire genome of the producing strain Clostridium sp. UCM В-7570 from the "Collection of producing strains of microorganisms and plant lines for food and agricultural biotechnology" of Institute of Food Biotechnology and Genomics of the National Academy of Sciences of Ukraine was sequenced and characterized. The genome was assembled into the scaffold with a total size of 4,470,321 bp and a GC content of 29.7%. The total number of genes identified was 4262, of which 4057 encoded proteins, 10 were rRNA operons, and 80 were tRNA genes. The genes of the sequenced genome encoding enzymes involved in butanol fermentation were found and analyzed. They were organized into cluster structures, and their protein sequences were found to be similar to the corresponding strains of C. acetobutylicum, C. beijerinckii, and C. pasteurianum type strains with the highest similarity to the latter. Thus, Clostridium sp. UCM В-7570 producing strain was identified as C. pasteurianum and suggested for metabolic engineering purposes.


(1)H NMR metabolomics to study the effects of diazepam on anisatin induced convulsive seizures.
The anticonvulsive properties of diazepam have been extensively studied, mainly focusing on the γ-amino butyrate (GABA) system. The aim of this investigation was to integrally analyze the metabolic events related to neuroprotection of diazepam on anisatin-induced convulsive seizures by a NMR-based metabolomic approach combined with histopathological examination and behavior examination. Multivariate analysis on metabolic profiles of the piriform cortex and cerebellum of mice revealed that diazepam could relieve mice suffering from the convulsive seizures by recovering destructed neurotransmitter and neuromodulator metabolism, ameliorating oxidative stress, alleviating the disturbance in energy, amino acid and nucleic acid metabolism in anisatin intoxicated mice. This integrated metabolomics study provided a powerful and highly effective approach to elucidate therapeutic effects and assessed the safety of diazepam. This study should be helpful for our understanding of convulsive seizures, and provide a holistic view of the treatment effects of benzodiazepine on convulsive seizures.


anaklasis: a compact software package for model-based analysis of specular neutron and X-ray reflectometry data sets.
anaklasis constitutes a set of open-source Python scripts that facilitate a range of specular neutron and X-ray reflectivity calculations, involving the generation of theoretical curves and the comparison/fitting of interfacial model reflectivity against experimental data sets. The primary focus of the software is twofold: on one hand to offer a more natural framework for model definition, requiring minimum coding literacy, and on the other hand to include advanced analysis methods that have been proposed in recent work. Particular attention is given to the ability to co-refine reflectivity data and to the estimation of model-parameter uncertainty and covariance using bootstrap analysis and Markov chain Monte Carlo sampling. The compactness and simplicity of model definition together with the streamlined analysis do not present a steep learning curve for the user, an aspect that may accelerate the generation of reproducible, easily readable and statistically accurate reports in future neutron and X-ray reflectivity related literature.


Methods to analyze novel liaisons between endosomes and centrosomes.
For some time, it has been known that recycling endosomes (REs) are organized in a nebulous "pericentrosomal" region in interphase cells. However, the collective use of previously developed methods, including centrosome isolation, live cell imaging, and electron microscopy, suggested that there is much more going on between the centrosome and the RE than previously imagined. By exploiting these approaches, we uncovered novel roles of the centrosome in RE function and, conversely, novel roles for REs in centrosome function. We first found that REs dynamically localized to the centrosome throughout the cell cycle. More specifically, we found that REs interacted with appendages of the older centriole in interphase cells to control endosome recycling, and this interaction was governed by RE-machinery including the small GTPase Rab11. We next determined that REs carry centrosome proteins to spindle poles as part of the "centrosome maturation" process. Here we discuss the methods used and materials needed to complete these types of studies.