Lineage replacement and evolution captured by 3 years of the United Kingdom Coronavirus (COVID-19) Infection Survey.
The Office for National Statistics Coronavirus (COVID-19) Infection Survey (ONS-CIS) is the largest surveillance study of SARS-CoV-2 positivity in the community, and collected data on the United Kingdom (UK) epidemic from April 2020 until March 2023 before being paused. Here, we report on the epidemiological and evolutionary dynamics of SARS-CoV-2 determined by analysing the sequenced samples collected by the ONS-CIS during this period. We observed a series of sweeps or partial sweeps, with each sweeping lineage having a distinct growth advantage compared to their predecessors, although this was also accompanied by a gradual fall in average viral burdens from June 2021 to March 2023. The sweeps also generated an alternating pattern in which most samples had either S-gene target failure (SGTF) or non-SGTF over time. Evolution was characterized by steadily increasing divergence and diversity within lineages, but with step increases in divergence associated with each sweeping major lineage. This led to a faster overall rate of evolution when measured at the between-lineage level compared to within lineages, and fluctuating levels of diversity. These observations highlight the value of viral sequencing integrated into community surveillance studies to monitor the viral epidemiology and evolution of SARS-CoV-2, and potentially other pathogens.


Effect of molecular crowding on DNA polymerase activity.
Live cells contain high concentrations of macromolecules, but almost all experimental biochemical data have been generated from dilute solutions that do not reflect conditions in vivo. To understand biomolecular behavior in vivo, properties studied in vitro are extrapolated to conditions in vivo; however, the molecular conditions within live cells are inherently crowded. The present study investigates the effect of molecular crowding on DNA polymerase activity using polyethylene glycol PEG of various molecular weights as a crowding agent. Polymerase activity assays under various conditions demonstrated that the activities of T7 and Taq DNA polymerases depend on the molecular weight and concentration of the crowding agent. Furthermore, equilibrium and kinetic analyses demonstrated that the binding affinity and catalytic activity of the polymerase increase and decrease, respectively, with increasing PEG concentrations. Based on quantitative parameters of the polymerase reactions, we improved the efficiency of PCR amplification under conditions of molecular crowding. These results suggest that quantitative measurements of biomolecular structure and function are useful for understanding the behavior of biomolecules in vivo and for biotechnology applications in vitro.


Frequency of headache and migraine in Qatar.
Headache is a worldwide problem that affects all ages. To date there are no epidemiological data regarding headaches in Qatar. The aim of this study was to use the International Headache Society (IHS) diagnostic criteria to estimate the prevalence of migraine and headache in an adult population. This is a cross-sectional population-based study. The study was carried out at 12 Primary Health Care Centers during the period from October to December 2004 in Qatar. Of the 1,200 subjects who were approached to participate in this study, 913 gave their consent, representing a response rate of 76.1%. Subjects were selected by a multistage stratified sampling procedure. We used the IHS criteria for migraine. Face-to-face interviews were based on a questionnaire that included variables such as age, sex, socioeconomic status, lifestyle habits, exacerbating factors, warning symptoms, associated features, family history and associated factors. Of the studied subjects, 54.9% were males and 45.1% females. The prevalence of headache and migraine among men was 67.9%, 7.8%, and among women 78.2%, 8.0%, respectively. Subjects aged 25-34 years had higher prevalence of headache comparing to other age groups (27.3%). Prevalence of headache was higher in women. Subjects with less than a high school education had higher frequency of headache (49.1%). The most common warning symptoms prior to headache were abnormal vision (53.0%) and weakness (30.4%). Stress (71.8%) and weather (49.5%) made headache worse in subjects. The majority of the headache sufferers looked for a dark (33.7%) and quiet place (74.5%) to rest. Headachedisorders constitute a public health problem in a newly developed Qatari society, due to fast urbanization. More women suffered from headache and migraine than men. The study revealed that there was a strong association between hemoglobin level and headache and its severity. Furthermore, the prevalence of headache and migraine in Qatar is higher than in Western and African countries.


Development and validation of a model for individualized prediction of cervical insufficiency risks in patients undergoing IVF/ICSI treatment.
Women who conceived with in vitro fertilization (IVF) or intracytoplasmic sperm injection (ICSI) are more likely to experience adverse pregnancy outcomes than women who conceived naturally. Cervical insufficiency (CI) is one of the important causes of miscarriage and premature birth, however there is no published data available focusing on the potential risk factors predicting CI occurrence in women who received IVF/ICSI treatment. This study aimed to identify the risk factors that could be integrated into a predictive model for CI, which could provide further personalized and clinically specific information related to the incidence of CI after IVF/ICSI treatment. This retrospective study included 4710 patients who conceived after IVF/ICSI treatment from Jan 2011 to Dec 2018 at a public university hospital. The patients were randomly divided into development (n = 3108) and validation (n = 1602) samples for the building and testing of the nomogram, respectively. Multivariate logistic regression was developed on the basis of pre-pregnancy clinical covariates assessed for their association with CI occurrence. A total of 109 patients (2.31%) experienced CI among all the enrolled patients. Body mass index (BMI), basal serum testosterone (T), gravidity and uterine length were associated with CI occurrence. The statistical nomogram was built based on BMI, serum T, gravidity and uterine length, with an area under the curve (AUC) of 0.84 (95% confidence interval: 0.76-0.90) for the developing cohort. The AUC for the validation cohort was 0.71 (95% confidence interval: 0.69-0.83), showing a satisfactory goodness-of-fit and discrimination ability in this nomogram. The user-friendly nomogram which graphically represents the risk factors and a pre-pregnancy predicted tool for the incidence of CI in patients undergoing IVF/ICSI treatment, provides a useful guide for medical staff on individualized decisions making, where preventive measures could be carried out during the IVF/ICSI procedure and subsequent pregnancy.


Picornavirus genome replication: roles of precursor proteins and rate-limiting steps in oriI-dependent VPg uridylylation.
The 5' ends of all picornaviral RNAs are linked covalently to the genome-encoded peptide, VPg (or 3B). VPg linkage is thought to occur in two steps. First, VPg serves as a primer for production of diuridylylated VPg (VPg-pUpU) in a reaction catalyzed by the viral polymerase that is templated by an RNA element (oriI). It is currently thought that the viral 3AB protein is the source of VPg in vivo. Second, VPg-pUpU is transferred to the 3' end of plus- and/or minus-strand RNA and serves as primer for production of full-length RNA. Nothing is known about the mechanism of transfer. We present biochemical and biological evidence refuting the use of 3AB as the donor for VPg uridylylation. Our data are consistent with precursors 3BC and/or 3BCD being employed for uridylylation. This conclusion is supported by in vitro uridylylation of these proteins, the ability of a mutant replicon incapable of producing processed VPg to replicate in HeLa cells and cell-free extracts and corresponding precursor processing profiles, and the demonstration of 3BC-linked RNA in mutant replicon-transfected cells. These data permit elaboration of our model for VPg uridylylation to include the use of precursor proteins and invoke a possible mechanism for location of the diuridylylated, VPg-containing precursor at the 3' end of plus- or minus-strand RNA for production of full-length RNA. Finally, determinants of VPg uridylylation efficiency suggest formation and/or collapse or release of the uridylylated product as the rate-limiting step in vitro depending upon the VPg donor employed.


Failure to report and provide commentary on research ethics board approval and informed consent in medical journals.
The Declaration of Helsinki prohibits the publication of articles that do not meet defined ethical standards for reporting of research ethics board (REB) approval and informed consent. Despite this prohibition and a call to highlight the deficiency for the reader, articles with potential ethical shortcomings continue to be published. To determine what proportion of articles in major medical journals lack statements confirming REB approval and informed consent, and whether accompanying commentary alerts readers to this deficiency. Retrospective, observational study. Online review of five major medical journals. All clinical research articles published online between 1 January 2005 and 31 December 2006 in the BMJ, Lancet, Annals of Internal Medicine, JAMA and the New England Journal of Medicine. Statement of REB approval and informed consent. Of 1780 articles reviewed, 1133 (63.7%) met inclusion criteria (manuscripts reporting human subjects, human tissue or identifiable personal data research), 36 (3.2%) articles lacked a statement of REB approval, 62 (5.5%) lacked disclosure of informed consent and 15 (1.3%) articles lacked both. Articles that did not state REB approval were associated with not stating informed consent (p<0.001). There were no editorial comments to alert readers to the lack of either REB approval or informed consent statements associated with any of the deficient articles. Articles that lack explicit statements of REB approval and informed consent are infrequent but continue to be published in major medical journals without editorial statements to alert the reader to this deficiency.


The relationship between physician empathy and disease complications: an empirical study of primary care physicians and their diabetic patients in Parma, Italy.
To test the hypothesis that scores of a validated measure of physician empathy are associated with clinical outcomes for patients with diabetes mellitus. This retrospective correlational study included 20,961 patients with type 1 or type 2 diabetes mellitus from a population of 284,298 adult patients in the Local Health Authority, Parma, Italy, enrolled with one of 242 primary care physicians for the entire year of 2009. Participating physicians' Jefferson Scale of Empathy scores were compared with occurrence of acute metabolic complications (hyperosmolar state, diabetic ketoacidosis, coma) in diabetes patients hospitalized in 2009. Patients of physicians with high empathy scores, compared with patients of physicians with moderate and low empathy scores, had a significantly lower rate of acute metabolic complications (4.0, 7.1, and 6.5 per 1,000 patients, respectively, P < .05). Logistic regression analysis showed physicians' empathy scores were associated with acute metabolic complications: odds ratio (OR) = 0.59 (95% confidence interval [CI], 0.37-0.95, contrasting physicians with high and low empathy scores). Patients' age (≥69 years) also contributed to the prediction of acute metabolic complications: OR = 1.7 (95% CI, 1.2-1.4). Physicians' gender and age, patients' gender, type of practice (solo, association), geographical location of practice (mountain, hills, plain), and length of time the patient had been enrolled with the physician were not associated with acute metabolic complications. These results suggest that physician empathy is significantly associated with clinical outcome for patients with diabetes mellitus and should be considered an important component of clinical competence.


Influence of Cooking Method on the Nutritional Quality of Organic and Conventional Brazilian Vegetables: A Study on Sodium, Potassium, and Carotenoids.
Vegetable consumption is associated with increased health benefits, and vegetables are consumed both in cooked form and raw form in salads. All cooking techniques cause changes in a vegetable's the nutrient content. Consumers are increasingly health-conscious and have less time to prepare meals, and they do not know which cooking times and cooking methods are best suited to preserve the nutrients. This study aimed to determine the best method of cooking vegetables to maintain minerals (potassium and sodium) and carotenoids. The studied vegetables were broccoli (Brassica oleracea, var. Italica), carrots (Daucus carota), and zucchini (Cucurbita moschata). The cooking methods were: boiling, steaming, combined oven, microwave steaming, and microwave cooking. Samples of organic and conventionally grown vegetables were prepared in triplicate. Samples were analyzed to determine the availability of target minerals and carotenoids in the raw food and in each recommended cooking situation according to technical standards. Only the carrot showed a higher concentration in organic cultivation for carotenoids in raw vegetables, with both zucchini and broccoli having higher concentrations when grown by conventional cultivation. The zucchini from organic cultivation presented a reduction of potassium and sodium, almost consistently, in all cooking techniques. Regarding the conventionally cultivated zucchini, potassium remained stable in boiling. Broccoli from organic and conventional cultivation showed similar potassium levels for boiling and traditional steam cooking. Organic carrots showed easier sodium extraction compared with conventional cultivation. Heat treatment, in general, improves the accessibility of carotenoids.


[The effect of the noise factor on the course of pregnancy in women and on the saccharase activity of the small intestine in their newborns].
The course of pregnancy, labors and the status of neonates from 104 mothers working in the setting exposed to increased industrial noise were analyzed. The noise was found to have a negative impact on the health of women and their neonates. Small intestinal saccharose deficiency was detected in the newborn.


Maximum likelihood estimation of frequencies of known haplotypes from pooled sequence data.
DNA samples are often pooled, either by experimental design or because the sample itself is a mixture. For example, when population allele frequencies are of primary interest, individual samples may be pooled together to lower the cost of sequencing. Alternatively, the sample itself may be a mixture of multiple species or strains (e.g., bacterial species comprising a microbiome or pathogen strains in a blood sample). We present an expectation-maximization algorithm for estimating haplotype frequencies in a pooled sample directly from mapped sequence reads, in the case where the possible haplotypes are known. This method is relevant to the analysis of pooled sequencing data from selection experiments, as well as the calculation of proportions of different species within a metagenomics sample. Our method outperforms existing methods based on single-site allele frequencies, as well as simple approaches using sequence read data. We have implemented the method in a freely available open-source software tool.


Cleavage by RNase P of gene N mRNA reduces bacteriophage lambda burst size.
RNase P, an enzyme essential for tRNA biosynthesis, can be directed to cleave any RNA when the target RNA is in a complex with a short, complementary oligonucleotide called an external guide sequence (EGS). RNase P from Escherichia coli can cleave phage lambda N mRNA in vitro or in vivo when the mRNA is in a complex with an EGS. The EGS can either be separate from or covalently linked to M1 RNA, the catalytic RNA subunit of RNase P. The requirement for Mg2+ in the reaction in vitro is lower when the EGS is covalently linked to M1 RNA. Substrates made of DNA can also be cleaved by RNase P in vitro in complexes with RNA EGSs. When either kind of EGS construct is used in vivo, burst size of phage lambda is reduced by > or = 40%. Reduction in burst size depends on efficient expression of the EGS constructs. The product of phage lambda gene N appears to function in a stoichiometric fashion.


Neural reflexes in inflammation and immunity.
The mammalian immune system and the nervous system coevolved under the influence of infection and sterile injury. Knowledge of homeostatic mechanisms by which the nervous system controls organ function was originally applied to the cardiovascular, gastrointestinal, musculoskeletal, and other body systems. Development of advanced neurophysiological and immunological techniques recently enabled the study of reflex neural circuits that maintain immunological homeostasis, and are essential for health in mammals. Such reflexes are evolutionarily ancient, dating back to invertebrate nematode worms that possess primitive immune and nervous systems. Failure of these reflex mechanisms in mammals contributes to nonresolving inflammation and disease. It is also possible to target these neural pathways using electrical nerve stimulators and pharmacological agents to hasten the resolution of inflammation and provide therapeutic benefit.


Effects of one-year treatment with isoflavone extract from red clover on prostate, liver function, sexual function, and quality of life in men with elevated PSA levels and negative prostate biopsy findings.
To asses the effect of an isoflavonoid extract from red clover on the prostate, liver function, quality of life, and sexual function in men with an initial elevated prostate-specific antigen (PSA) level and negative prostate biopsy findings during a 1-year treatment period. A total of 20 men (mean age 65 years) were treated with a daily 60-mg dose of an isoflavone extract for 1 year. Liver function, sexual hormone levels (total testosterone, estrogen, luteotropic hormone, follicle-stimulating hormone, and dehydroepiandrosterone sulfate), transrectal ultrasonography volumetry of the prostate, PSA level, International Prostate Symptom Score, and International Index of Erectile Function score were recorded at the beginning of the study and every 3 months for 1 year. The average PSA level was 10.16 ng/mL at baseline versus 7.15 ng/mL after 12 months, for a statistically significant reduction of 33% (P <0.019). The mean prostate volume had decreased slightly from 49.3 cm3 to 44.3 cm3 after 12 months (P <0.097). The sexual hormone levels did not change throughout the study. We registered a significant increase in all three liver transaminases after 3 months (P <0.001). The International Prostate Symptom Score showed a mean value of 7.9 at baseline and 6.68 after 12 months (P <0.421). Sexual function was not influenced by the treatment. Daily oral administration of 60 mg of an isoflavone extract was well tolerated and caused no side effects. The significant increase in liver transaminases and the significant decrease in total PSA levels by >30% indicates that patients, general practitioners, and urologists should be informed about these potential effects of red clover extracts and possibly other phytoestrogens, on liver transaminases and elevated total PSA levels.


Hybrid charge division multiplexing method for silicon photomultiplier based PET detectors.
Silicon photomultiplier (SiPM) is widely utilized in various positron emission tomography (PET) detectors and systems. However, the individual recording of SiPM output signals is still challenging owing to the high granularity of the SiPM; thus, charge division multiplexing is commonly used in PET detectors. Resistive charge division method is well established for reducing the number of output channels in conventional multi-channel photosensors, but it degrades the timing performance of SiPM-based PET detectors by yielding a large resistor-capacitor (RC) constant. Capacitive charge division method, on the other hand, yields a small RC constant and provides a faster timing response than the resistive method, but it suffers from an output signal undershoot. Therefore, in this study, we propose a hybrid charge division method which can be implemented by cascading the parallel combination of a resistor and a capacitor throughout the multiplexing network. In order to compare the performance of the proposed method with the conventional methods, a 16-channel Hamamatsu SiPM (S11064-050P) was coupled with a 4 × 4 LGSO crystal block (3 × 3 × 20 mm3) and a 9 × 9 LYSO crystal block (1.2 × 1.2 × 10 mm3). In addition, we tested a time-over-threshold (TOT) readout using the digitized position signals to further demonstrate the feasibility of the time-based readout of multiplexed signals based on the proposed method. The results indicated that the proposed method exhibited good energy and timing performance, thus inheriting only the advantages of conventional resistive and capacitive methods. Moreover, the proposed method showed excellent pulse shape uniformity that does not depend on the position of the interacted crystal. Accordingly, we can conclude that the hybrid charge division method is useful for effectively reducing the number of output channels of the SiPM array.


Influence of low temperature on comparative arsenic accumulation and release by three Pteris hyperaccumulators.
Arsenic (As) hyperaccumulator Pteris ferns are renowned for their capacity to accumulate As and have been used to remediate As-contaminated environmental. However, there is less information on how they perform under low temperature though it is important for practical phytoremediation. The purpose of this study was to identify the effect of temperature on As accumulation by three As hyperaccumulators, Pteris multifida, Pteris cretica and Pteris vittata. Ferns were cultured with 5 mg/L As addition under 25 °C to 5 °C for 15 days. The results showed that dropping of temperatures reduced As accumulation by P. vittata moderately but not P. multifida and P. cretica until 10 °C. At 5 °C, all ferns discontinued As accumulation, and the morphology showed necrosis in P. vittata, wherein P. multifida and P. cretica kept healthy. The As distribution showed that As was mainly accumulated in fronds, while P. multifida stored partial As in its root. Both translocation factor and As efflux showed that temperate zone ferns manage As more strictly as compared to P. vittata. Our findings demonstrated that temperature should be considered when applying Pteris ferns for As phytoremediation, and P. multifida could be the most suitable fern for treating As-contaminated water in temperate zone area.


Degranulation and abnormal bactericidal function of granulocytes procured by reversible adhesion to nylon wool.
Granylocyte bactericidal capacity, chemotaxis, hexose monophosphate shung activity (before and after phagocytic stimulus), and quantitative nitroblue tetrazolium reduction and enzyme content were examined in cells obtained by filtration leukaphresis (FL) and continuous-flow centrifugation (CFC). A decrease in the bactericidal efficiency of FL-produced cells compared to that of both normal and CFC-procured granulocytes was found; the decrease was 17% with a cell-to-bacteria ratio of 5:1, and 55% with a 1:1 ratio. Moreover, FL-acquired cells were often vacuolated and consistently contained less acid phosphatase and beta-glucuronidase than did normal granulocytes. When normal cells were incubated for 1-2 hr with nylon wool, 30% of the total acid phosphatase and beta-glucuronidase was released, with no evidence of cell death, thus suggesting degranulation. Similar results were obtained with glass, cotton, or polysulfone plastic fibers. Electron microscopic and peroxidase cytochemical studies of the adherence of normal granulocytes to nylon fibers were also carried out. After 30 min of incubation, cell-to-fiber attachment and cellular aggregation had occurred, although the cells per se appeared normal. After 60 and 120 min, other changes became apparent: (1) a decrease in the amount of cytoplasmic granules; (2) large, intracytoplasmic vaculoles; and (3) extracellular peroxidase on fiber surfaces. We conclude that granulocytes obtained by adherence to nylon fibers show both morphological and biochemical evidence of degranulation and diminished bactericidal capacity, and that these abnormalities may be causally related to decreased granulocyte survival in transfusion recipients.


Maternal Intrapartum Antibiotic Administration and Infantile Colic: Is there a Connection?
The aetiology of infantile colic remains unknown. However, altered gut microbiota composition has been reported in children with the disorder. The objective of this study was to determine the associations between perinatal factors potentially affecting gut colonization and infantile colic. Altogether 48 infants with colic and 29 controls were selected from 2 ongoing clinical studies. Infants with and without colic were comparable with regard to their background characteristics. A significant difference was detected in intrapartum antibiotic use and breastfeeding rates between infants with and without colic. The association between exposure to intrapartum antibiotics and infantile colic remained statistically significant after adjusting for potential confounding factors. Since intrapartum antibiotic exposure may have an effect on early gut colonization, our finding is consistent with the association between aberrant early gut microbiota composition and development of colic. Antibiotic-exposed neonates may represent a novel target group for preventive intervention studies.


[Spondylo-epiphyseal dysplasia associated with craniosynostosis, cleft palate and mental retardation. A case report].
Skeletal dysplasias are a heterogeneous group of disorders characterized by an alteration of the organization of osseous tissue causing a distortion on the growth and development pattern of bones. In 1998, four Japanese sibs were described by the first time, three males and one female who presented a previously undescribed spondylo-epiphyseal dysplasia associated with craniosynostosis, cataracts, cleft palate and different grades of mental retardation. A probable autosomic recessive inheritance was suggested, but a germinal mosaicism could not be discarded. This is a case report of a patient with clinical and radiological findings similar to the ones previously described, born to second degree consanguineous parents. This supports the postulated presumption of a mutation with an autosomic recessive inheritance. The present communication represents the fifth case reported in the literature and the second familiar group affected.


Herbaspirillum seropedicae signal transduction protein PII is structurally similar to the enteric GlnK.
PII-like proteins are signal transduction proteins found in bacteria, archaea and eukaryotes. They mediate a variety of cellular responses. A second PII-like protein, called GlnK, has been found in several organisms. In the diazotroph Herbaspirillum seropedicae, PII protein is involved in sensing nitrogen levels and controlling nitrogen fixation genes. In this work, the crystal structure of the unliganded H. seropedicae PII was solved by X-ray diffraction. H. seropedicae PII has a Gly residue, Gly108 preceding Pro109 and the main-chain forms a beta turn. The glycine at position 108 allows a bend in the C-terminal main-chain, thereby modifying the surface of the cleft between monomers and potentially changing function. The structure suggests that the C-terminal region of PII proteins may be involved in specificity of function, and nonenteric diazotrophs are found to have the C-terminal consensus XGXDAX(107-112). We are also proposing binding sites for ATP and 2-oxoglutarate based on the structural alignment of PII with PII-ATP/GlnK-ATP, 5-carboxymethyl-2-hydroxymuconate isomerase and 4-oxalocrotonate tautomerase bound to the inhibitor 2-oxo-3-pentynoate.


Neural and orofacial defects in Folp1 knockout mice [corrected].
Folic acid is essential for the development of the nervous system and other associated structures. Mice deficient in the folic acid-binding protein one (Folbp1) gene display multiple developmental abnormalities, including neural and craniofacial defects. To better understand potential interactions between Folbp1 gene and selected genes involved in neural and craniofacial morphogenesis, we evaluated the expression patterns of a panel of crucial differentiation markers (Pax-3, En-2, Hox-a1, Shh, Bmp-4, Wnt-1, and Pax-1). Folbp1 mice were supplemented with low dosages of folinic add to rescue nullizygotes from dying in utero before gestational day 10. The gene marker analyses were carried out by in situ hybridization. In nullizygote embryos with open cranial neural tube defects, the downregulation of Pax-3 and En-2 in the impaired midbrain, along with an observed upregulation of the ventralizing marker Shh in the expanded floor plate, suggested an important regulatory interaction among these three genes. Moreover, the nullizygotes also exhibit craniofacial abnormalities, such as cleft lip and palate. Pax-3 signals in the impaired medial nasal primordia were significantly increased, whereas Pax-1 showed no expression in the undeveloped lateral nasal processes. Although Shh was downregulated, Bmp-4 was strongly expressed in the medial and lateral nasal processes, highlighting the antagonistic activities of these molecules. Impairment of Folbp1 gene function adversely impacts the expression of several critical signaling molecules. Mis-expression of these molecules, perhaps mediated by Shh, may potentially contribute to the observed failure of neural tube closure and the development of craniofacial defects in the mutant mice.


The mechanism of monomer transfer between two structurally distinct PrP oligomers.
In mammals, Prion pathology refers to a class of infectious neuropathologies whose mechanism is based on the self-perpetuation of structural information stored in the pathological conformer. The characterisation of the PrP folding landscape has revealed the existence of a plethora of pathways conducing to the formation of structurally different assemblies with different biological properties. However, the biochemical interconnection between these diverse assemblies remains unclear. The PrP oligomerisation process leads to the formation of neurotoxic and soluble assemblies called O1 oligomers with a high size heterodispersity. By combining the measurements in time of size distribution and average size with kinetic models and data assimilation, we revealed the existence of at least two structurally distinct sets of assemblies, termed Oa and Ob, forming O1 assemblies. We propose a kinetic model representing the main processes in prion aggregation pathway: polymerisation, depolymerisation, and disintegration. The two groups interact by exchanging monomers through a disintegration process that increases the size of Oa. Our observations suggest that PrP oligomers constitute a highly dynamic population.


Epstein-Barr virus nuclear antigen 2 specifically induces expression of the B-cell activation antigen CD23.
Epstein-Barr virus (EBV) infection of EBV-negative Burkitt lymphoma (BL) cells induces some changes similar to those seen in normal B lymphocytes that have been growth transformed by EBV. The role of individual EBV genes in this process was evaluated by introducing each of the viral genes that are normally expressed in EBV growth-transformed and latently infected lymphoblasts into an EBV-negative BL cell line, using recombinant retrovirus-mediated transfer. Clones of cells were derived that stably express the EBV nuclear antigen 1 (EBNA-1), EBNA-2, EBNA-3, EBNA-leader protein, or EBV latent membrane protein (LMP). These were compared with control clones infected with the retrovirus vector. All 10 clones converted to EBNA-2 expression differed from control clones or clones expressing other EBV proteins by growth in tight clumps and by markedly increased expression of one particular surface marker of B-cell activation, CD23. Other activation antigens were unaffected by EBNA-2 expression, as were markers already expressed on the parent BL cell line, including BL markers (cALLA and BLA), proliferation markers (transferrin receptor and BK19.9), and cell adhesion-related molecules (LFA-1 and LFA-3). Increased CD23 expression in cells expressing EBNA-2 was apparent from monoclonal anti-CD23 antibody binding to the cell surface, from immunoprecipitation of the 45-kDa and 90-kDa CD23 proteins with monoclonal antibody, and from RNA blots probed with labeled CD23 DNA. The results indicate that EBNA-2 is a specific direct or indirect trans-activator of CD23. This establishes a link between an EBV gene and cell gene expression. Since CD23 has been implicated in the transduction of B-cell growth signals, its specific induction by EBNA-2 could be important in EBV induction of B-lymphocyte transformation.


The HNK-1 epitope and the elastic fiber system of the human ciliary body. An immunoelectron microscopic study.
To localize at the electron microscopic level the cell adhesion-related HNK-1 carbohydrate epitope in the inner connective tissue layer (ICTL) of the human ciliary body. Seven specimens representing the pars plicata (age range, 12 to 86 years) and three specimens representing the pars plana (age range, 29 to 71 years) of the ciliary body were sampled at death from eight normal human eyes. Three additional specimens from the pars plicata were taken from three eyes with exfoliation syndrome (age range, 78 to 81 years). All specimens were embedded in LR white resin and studied by postembedding immunogold labeling using two monoclonal antibodies (mAbs) HNK-1 and NC-1 in detecting the HNK-1 epitope. In the ICTL of the pars plicata and the pars plana, mAbs HNK-1 and NC-1 constantly bound to the surface of fibroblast-like cells present in the subepithelial connective tissue matrix. The immunoreaction was localized along the cell membrane, both around the cell body and around its long, slender cytoplasmic processes. Long microfibrillar bundles, which consisted of approximately 10-nm thick microfibrils in close association with these subepithelial matrix cells and elastic fibers, were also labeled. The periphery of elastic fibers was likewise immunolabeled for the HNK-1 epitope. The reaction pattern was essentially the same, regardless of age and presence or absence of exfoliation syndrome. At the ultrastructural level, the HNK-1 epitope in the ICTL is a common denominator to the subepithelial matrix cells, microfibrillar bundles, and elastic fibers. This suggests that the subepithelial matrix cells secrete this epitope, and that molecules hearing it may be involved in joining these connective tissue elements that structurally stabilize the ciliary body.


The evolutionary innovation of root suberin lamellae contributed to the rise of seed plants.
Seed plants overtook ferns to become the dominant plant group during the late Carboniferous, a period in which the climate became colder and dryer1,2. However, the specific innovations driving the success of seed plants are not clear. Here we report that the appearance of suberin lamellae (SL) contributed to the rise of seed plants. We show that the Casparian strip and SL vascular barriers evolved at different times, with the former originating in the most recent common ancestor (MRCA) of vascular plants and the latter in the MRCA of seed plants. Our results further suggest that most of the genes required for suberin formation arose through gene duplication in the MRCA of seed plants. We show that the appearance of the SL in the MRCA of seed plants enhanced drought tolerance through preventing water loss from the stele. We hypothesize that SL provide a decisive selective advantage over ferns in arid environments, resulting in the decline of ferns and the rise of gymnosperms. This study provides insights into the evolutionary success of seed plants and has implications for engineering drought-tolerant crops or fern varieties.


Pharmacokinetics of antofloxacin hydrochloride, a new fluoroquinolone antibiotic, after single oral dose administration in Chinese healthy male volunteers.
Antofloxacin hydrochloride is a newly developed fluoroquinolone antibacterial in China, which has comparable in vitro and animal pharmacological and toxicological properties to levofloxacin and is worthy of further clinical trial. Human tolerance of single escalating doses from 50 to 500 mg was shown to be safe. The purpose of the present study was to evaluate the pharmacokinetic characteristics of a single oral dose of antofloxacin hydrochloride in Chinese healthy male volunteers. Twelve subjects were randomized for administration of a single dose of 300, 400 and 500 mg antofloxacin hydrochloride in a 3-way crossover design. High pressure liquid chromatography (HPLC) was used to assay the serum and urine concentrations of antofloxacin in samples collected over a period of 72 h following drug administration. All three dosages were well tolerated. Antofloxacin hydrochloride demonstrated linear pharmacokinetic characteristics with mean pharmacokinetic values varying from 0.09 to 0.10 l/h/kg for CL/F and from 2.75-3.00 l/kg for V/F. The t(1/2beta) was around 20 h and the mean fraction of dose excreted in urine varied from 0.40 to 0.46. Based on these data, 300 mg of antofloxacin hydrochloride administered once daily was selected for further investigation in a multiple dose administration study.


Genetics and bitter taste responses to goitrin, a plant toxin found in vegetables.
The perceived bitterness of cruciferous vegetables such as broccoli varies from person to person, but the functional underpinnings of this variation are not known. Some evidence suggests that it arises, in part, from variation in ability to perceive goitrin (5-vinyloxazolidine-2-thione), a potent antithyroid compound found naturally in crucifers. Individuals vary in ability to perceive synthetic compounds similar to goitrin, such as 6-propyl-2-thiouracil (PROP) and phenylthiocarbamide (PTC), as the result of mutations in the TAS2R38 gene, which encodes a bitter taste receptor. This suggests that taste responses to goitrin itself may be mediated by TAS2R38. To test this hypothesis, we examined the relationships between genetic variation in TAS2R38, functional variation in the encoded receptor, and threshold taste responses to goitrin, PROP, and PTC in 50 subjects. We found that threshold responses to goitrin were associated with responses to both PROP (P = 8.9 x 10(-4); r(s) = 0.46) and PTC (P = 7.5 x 10(-4); r(s) = 0.46). However, functional assays revealed that goitrin elicits a weaker response from the sensitive (PAV) allele of TAS2R38 (EC(50) = 65.0 μM) than do either PROP (EC(50) = 2.1 μM) or PTC (EC(50) = 1.1 μM) and no response at all from the insensitive (AVI) allele. Furthermore, goitrin responses were significantly associated with mutations in TAS2R38 (P = 9.3 × 10(-3)), but the same mutations accounted for a smaller proportion of variance in goitrin response (r(2) = 0.16) than for PROP (r(2) = 0.50) and PTC (r(2) = 0.57). These findings suggest that mutations in TAS2R38 play a role in shaping goitrin perception, but the majority of variance must be explained by other factors.


[Coronary calcifications in a subgroup of post-menopausal women with metabolic syndrome].
The aim of this study was to evaluate the burden of coronary calcifications in a subgroup of post-menopausal women with metabolic syndrome (MS) in agreement with the National Cholesterol Educational Program-Third Report of the Expert Panel on Detection, Evaluation, and Treatment of High Blood Cholesterol in Adults (NCEP-ATP III) definition. We studied 81 women (43 control subjects and 38 women with MS) in agreement with the NCEP-ATP III definition undergoing multislice computed tomography for evaluation of coronary calcifications. The patients were similar for Framingham risk score. The severity and extent of coronary artery calcifications were higher in individuals with MS (10.8 +/- 15.8 vs 3.02 +/- 5.6; p = 0.006). In all patients total cholesterol, low-density lipoproteins and triglycerides were correlated with calcium score (p < 0.05) while high-density lipoproteins were inversely correlated with coronary calcifications. In women with MS total cholesterol and low-density lipoprotein cholesterol were correlated with calcium score. Women with MS have a higher burden of subclinical coronary atherosclerosis. The correlation between MS and calcium score concerned more the presence rather than the severity of coronary calcifications. Moreover, no correlation was observed among single components of MS in agreement with the NCEP-ATP III definition.


Understanding cadherin EGF LAG seven-pass G-type receptors.
The cadherin epidermal growth factor (EGF) laminin G (LAG) seven-pass G-type receptors (CELSRs) are a special subgroup of adhesion G protein-coupled receptors, which are pivotal regulators of many biologic processes such as neuronal/endocrine cell differentiation, vessel valve formation, and the control of planar cell polarity during embryonic development. All three members of the CELSR family (CELSR1-3) have large ecto-domains that form homophilic interactions and encompass more than 2000 amino acids. Mutations in the ecto-domain or other gene locations of CELSRs are associated with neural tube defects and other diseases in humans. Celsr knockout (KO) animals have many developmental defects. Therefore, specific agonists or antagonists of CELSR members may have therapeutic potential. Although significant progress has been made regarding the functions and biochemical properties of CELSRs, our knowledge of these receptors is still lacking, especially considering that they are broadly distributed but have few characterized functions in a limited number of tissues. The dynamic activation and inactivation of CELSRs and the presence of endogenous ligands beyond homophilic interactions remain elusive, as do the regulatory mechanisms and downstream signaling of these receptors. Given this motivation, future studies with more advanced cell biology or biochemical tools, such as conditional KO mice, may provide further insights into the mechanisms underlying CELSR function, laying the foundation for the design of new CELSR-targeted therapeutic reagents. The cadherin EGF LAG seven-pass G-type receptors (CELSRs) are a special subgroup of adhesion G protein-coupled receptors (GPCRs), which have large ecto-domains that form homophilic interactions and encompass more than 2000 amino acids. Recent studies have revealed that CELSRs are pivotal regulators of many biological processes, such as neuronal/endocrine cell differentiation, vessel valve formation and the control of planar cell polarity during embryonic development.


Reconstructing patterns of reticulate evolution in plants.
Until recently, rigorously reconstructing the many hybrid speciation events in plants has not been practical because of the limited number of molecular markers available for plant phylogenetic reconstruction and the lack of good, biologically based methods for inferring reticulation (network) events. This situation should change rapidly with the development of multiple nuclear markers for phylogenetic reconstruction and new methods for reconstructing reticulate evolution. These developments will necessitate a much greater incorporation of population genetics into phylogenetic reconstruction than has been common. Population genetic events such as gene duplication coupled with lineage sorting and meiotic and sexual recombination have always had the potential to affect phylogenetic inference. For tree reconstruction, these problems are usually minimized by using uniparental markers and nuclear markers that undergo rapid concerted evolution. Because reconstruction of reticulate speciation events will require nuclear markers that lack these characteristics, effects of population genetics on phylogenetic inference will need to be addressed directly. Current models and methods that allow hybrid speciation to be detected and reconstructed are discussed, with a focus on how lineage sorting and meiotic and sexual recombination affect network reconstruction. Approaches that would allow inference of phylogenetic networks in their presence are suggested.


Plasma protein binding and metabolic clearance of phenytoin in the rat.
The purpose [corrected] of this investigation was to determine the effects of certain changes in plasma protein binding on the disposition of phenytoin after i.v. administration in the rat. Treatment of rats with sulfisoxazole and oleic acid significantly reduced plasma protein binding of phenytoin. The displacement of phenytoin from plasma proteins by sulfisoxazole had no significant effect on the elimination of phenytoin whereas comparable displacement by oleic acid produced an increase in the apparent volume of distribution and a marked decrease in the metabolic clearance of the drug. A similar difference in metabolic clearance was noted when phenytoin elimination was determined as a function of the intrinsic ability of the rat to bind phenytoin in the plasma. Rats showing relatively high plasma protein binding of phenytoin cleared the drug much more rapidly than rats showing relatively low plasma protein binding of phenytoin. Assuming that an endogenous inhibitor is responsible for both the decreased plasma protein binding and decreased metabllic clearance of phenytoin in rats with an intrinsically reduced ability to bind phenytoin in plasma, this inhibitor is evidently similar to oleic acid in its effects.


A first-in-human study of Brozopentyl Sodium, following single and multiple ascending intravenous infusion in Chinese healthy volunteers.
Brozopentyl Sodium (BZP), a novel agent for ischemic stroke, has shown promising results in preclinical pharmacological studies, prompting the initiation of the first-in-human investigation. This study aimed to assess the safety, tolerability, and pharmacokinetic (PK) characteristics of BZP in Chinese healthy volunteers. The study consisted of two parts. Part I was a single-center, randomized, single-blinded, placebo-controlled, single-ascending study with six BZP dose cohorts (SAD: 25, 50, 100, 200, 300, and 400 mg). Part II was a single-center, randomized, single-blinded, placebo-controlled, multi-dose- and dose-elevated study with three BZP dose cohorts (MAD: 50, 100, and 200 mg). Doses were administered once daily on days 1 and 7 and twice daily on days 2-6. The PK properties of BZP and its bioactive metabolites, BNBP, were assessed. Safety and tolerability evaluations were also conducted. In the SAD study, BZP reached peak plasma concentrations (Tmax) at the end of administration, with median Tmax values ranging from 1 to 1.03 h, while BNBP reached Tmax between 1.25 to 1.38 h. The terminal half-lives (T1/2) were approximately 8 h for BZP and 15 h for BNBP. In the MAD study, steady-state plasma concentrations of BZP were reached by day 5. There was minimal accumulation of both BZP and BNBP after 7 days of administration. The area under the plasma concentration-time curve from 0 to time of the last measurable concentration (AUC0-t) and maximum plasma drug concentration (Cmax) showed dose-proportional increases for BZP but not for BNBP in both study parts. Single and multiple doses of BZP demonstrated a good safety profile and were well-tolerated. BZP displayed safety, good tolerability and predictable PK characteristics following both single and multiple ascending intravenous administrations. These findings provide a basis for further clinical development of BZP for ischemic stroke patients.


Species Distribution and Comparison between EUCAST and Gradient Concentration Strips Methods for Antifungal Susceptibility Testing of 112 Aspergillus Section Nigri Isolates.
Aspergillus niger, the third species responsible for invasive aspergillosis, has been considered as a homogeneous species until DNA-based identification uncovered many cryptic species. These species have been recently reclassified into the Aspergillus section Nigri However, little is yet known among the section Nigri about the species distribution and the antifungal susceptibility pattern of each cryptic species. A total of 112 clinical isolates collected from 5 teaching hospitals in France and phenotypically identified as A. niger were analyzed. Identification to the species level was carried out by nucleotide sequence analysis. The MICs of itraconazole, voriconazole, posaconazole, isavuconazole, and amphotericin B were determined by both the EUCAST and gradient concentration strip methods. Aspergillus tubingensis (n = 51, 45.5%) and Aspergillus welwitschiae (n = 50, 44.6%) were the most common species while A. niger accounted for only 6.3% (n = 7). The MICs of azole drugs were higher for A. tubingensis than for A. welwitschiae The MIC of amphotericin B was 2 mg/liter or less for all isolates. Importantly, MICs determined by EUCAST showed no correlation with those determined by the gradient concentration strip method, with the latter being lower than the former (Spearman's rank correlation tests ranging from 0.01 to 0.25 depending on the antifungal agent; P > 0.4). In conclusion, A. niger should be considered as a minority species in the section Nigri The differences in MICs between species for different azoles underline the importance of accurate identification. Significant divergences in the determination of MIC between EUCAST and the gradient concentration strip methods require further investigation.


IL-16 represses HIV-1 promoter activity.
IL-16 is produced by CD8+ lymphocytes and has been reported to inhibit HIV-1 and SIV replication in infected PBMCs. CD4 serves as a receptor for the secreted form of IL-16, and IL-16 binding to CD4 induces signal transduction, which affects the activation state of the cell. We hypothesized, therefore, that the effect of IL-16 on HIV-1 replication might occur at the level of virus expression. In transient transfection studies with HIV-1 LTR-reporter gene constructs we found that pretreatment of CD4+ lymphoid cells with recombinant IL-16 repressed HIV-1 promoter activity up to 60-fold, preventing both PMA and Tat activation. This effect of IL-16 required sequences contained within the core enhancer, but was not simply due to the down-regulation of transcription factors binding to this element.


Transcriptome 3'end organization by PCF11 links alternative polyadenylation to formation and neuronal differentiation of neuroblastoma.
Diversification at the transcriptome 3'end is an important and evolutionarily conserved layer of gene regulation associated with differentiation and dedifferentiation processes. Here, we identify extensive transcriptome 3'end-alterations in neuroblastoma, a tumour entity with a paucity of recurrent somatic mutations and an unusually high frequency of spontaneous regression. Utilising extensive RNAi-screening we reveal the landscape and drivers of transcriptome 3'end-diversification, discovering PCF11 as critical regulator, directing alternative polyadenylation (APA) of hundreds of transcripts including a differentiation RNA-operon. PCF11 shapes inputs converging on WNT-signalling, and governs cell cycle, proliferation, apoptosis and neurodifferentiation. Postnatal PCF11 down-regulation induces a neurodifferentiation program, and low-level PCF11 in neuroblastoma associates with favourable outcome and spontaneous tumour regression. Our findings document a critical role for APA in tumorigenesis and describe a novel mechanism for cell fate reprogramming in neuroblastoma with potentially important clinical implications. We provide an interactive data repository of transcriptome-wide APA covering > 170 RNAis, and an APA-network map with regulatory hubs.


A Randomized Controlled Trial of Green Tea Beverages on the in vivo Radical Scavenging Activity in Human Skin.
Oxidative stress plays an important role in the pathogenesis of various skin diseases. Thus, the antioxidant network of the skin relies on the uptake of exogenous antioxidants to ensure cell protection against radical formation. Green tea is one of the main sources of polyphenolic antioxidant compounds, but only few data are available on its cutaneous antioxidant effects. The radical scavenging properties of Benifuuki and Yabukita green tea were investigated in the human skin. Thirty-two participants who met the inclusion criteria were randomized to consume 3 cups per day of either Benifuuki tea, Yabukita tea, or water (control group) for 2 weeks. Electron paramagnetic resonance (EPR) spectroscopy was applied to measure the radical scavenging capacity of the skin in vivo before and after the intervention. Both Yabukita and Benifuuki tea led to an increase in the radical scavenging activity of the skin by 28 and 29%, respectively, and the difference was significant when compared to the control group. Benifuuki tea, previously reported to be superior in bioavailability due to a highly absorbable methylated catechin, did not prove to be more effective than the common Yabukita tea. The results show that green tea enhances the radical scavenging capacity of the skin and support the hypothesis that green tea may offer protection against cutaneous oxidative stress.


Impact of mesophilic anaerobic digestion and post-treatment of digestates on the transfer of conjugative antimicrobial resistance plasmids.
Manure is a major source of antimicrobial-resistant bacteria and resistance genes carried by mobile genetic elements such as plasmids. In France, the number of on-farm biogas plants has increased significantly in recent years. Our study investigated the impact of mesophilic anaerobic digestion (AD) and the post-treatment of digestates on the fate of conjugative plasmids, along with their potential transfer of antimicrobial resistance. Samples of raw manure, digestates and post-treated digestates were collected from three on-farm biogas plants. Conjugative plasmids were captured using the Escherichia coli CV601 recipient strain and media supplemented with rifampicin and kanamycin - to which the recipient strain is resistant - and tetracycline, sulfamethoxazole, gentamicin, trimethoprim, amoxicillin, cefotaxime, ciprofloxacin or colistin. Putative transconjugants were identified and characterised by disc diffusion and whole genome sequencing. The results showed that the antimicrobial resistance genes transferred from the different matrices conferred resistance to tetracyclines, sulphonamides, trimethoprim, and/or streptomycin. Transconjugants were obtained from raw manure samples but not from digestates or post-digestates, suggesting that mesophilic AD processes may produce fewer conjugative plasmids potentially able to be transferred to Enterobacterales.


Antifungal activity, yield, and composition of Ocimum gratissimum essential oil.
Ocimum gratissimum L. or clove basil, belongs to the Lamiaceae family, has various desirable uses and applications. Beyond its aromatic, seasoning, and medicinal applications, this plant also has antimicrobial activity. This study was aimed at assessing the antifungal activity, yield, and composition of the essential oil (EO) of O. gratissimum. The species was cultivated in garden beds with dystrophic red latosol soil type containing high organic-matter content. The EO was obtained by hydrodistillation of dried leaves in a modified Clevenger apparatus, followed by determination of its content. Chemical characterization was carried out by gas chromatography-mass spectrometry (GC-MS). Microbial activity was assessed using the broth microdilution method, by determining the minimum inhibitory concentration (MIC), in order to compare the antimicrobial effect of EO in 10 isolates-Fusarium oxysporum f. sp tracheiphilum (CMM-0033), F. oxysporum f. sp. cubense (CMM-0813 and CMM-2819), F. oxysporum f. sp lycopersici (CMM-1104), F. solani (CMM-3828), Rhizoctonia solani (CMM-3274), and Macrophomina phaseolina (CMM-2715, CMM-3875, CMM-3615, and CMM-3650). The EO was a highly effective inhibitor of the studied phytopathogenic fungi, with MICs varying from 31.25 to 125 µg/mL. F. oxysporum f. sp lycopersici and R. solani were the most sensitive; both were inhibited at an MIC of 31.25 µg/mL. The EO content in the plant extract was 0.18%. Thirty chemical compounds were detected via GC-MS, with linalool (32.9%) being the major compound followed by 1,8-cineole (21.9%), both oxygenated monoterpenes. It can be concluded that clove basil EO is a highly effective antifungal agent, and therefore, a potential alternative for the control of plant pathogenic diseases.


Hydrodynamic performance and heat generation by centrifugal pumps.
For over a century, centrifugal pumps (CP) have been used in various applications, from large industrial pumps to flow pumps for aquariums. However, the use of CP as blood pumps has a rather short history. Consequently, the hydraulic performance data for a blood CP are limited. The aim of our investigation was to study the hydraulic performance and the heat generation of three commercially available CP: Bio-Medicus Bio-Pump BP80 (Medtronic), Rotaflow (Jostra Medizintechnik), and DeltaStream DP2 (MEDOS Medizintechnik AQ). The study was performed using a circuit primed with a water-glycerin mixture with a dynamic viscosity of 0.00272 pa/s. Pressure-flow curves were obtained by a stepwise stagnation of the pump outlet or inlet. The temperature changes were observed using ThermaCAM SC2000 (Flir Systems). The pumps' performance in close to clinical conditions ('operating region') was analysed in this report. The 'operating region' in the case of the BP80 is positioned around the pressure-flow curve at a pump speed of 3000 rpm. In the case of the Rotaflow, the 'operating region' was between the pump pressure-flow curves at a speed of 3000 and 4000 rpm, and the DP2 was found between 7000 and 8000 rpm. The standard deviation of mean pressure through the pump was used to characterise the stability of the pump. In experiments with outlet stagnation, the BP80 demonstrated high negative association between flow and pressure variability (r = -0.68, p < 0.001). In experiments with the DP2, this association was positive (r = 0.68, p < 0.001). All pumps demonstrated significantly higher variability of pressure in experiments with inlet stagnation in comparison to the experiments with outlet stagnation. The rise of relative temperature in the inlet of a pump was closely related to the flow rate. The heating of fluid was more pronounced in the 'zero-flow' mode, especially in experiments with inlet stagnation. In summary, (1) the 'zero-flow' regime, which is described in the manuals of some commercially-available pumps, is the use of the pump outside the allowable operating region. It is potentially dangerous and should, therefore, never be used in clinical settings. (2) Using centrifugal pumps for kinetic-assisted venous return can only be performed safely when the negative pressure at the inlet of the pump is monitored continuously. The maximum allowable negative pressure has to be defined for each type of pump, and must be based on pump performance.


Limb length restoration during total hip arthroplasty: use of a caliper to control femoral component insertion and accurate acetabular placement relative to the transverse acetabular ligament.
Current methods for restoring or preserving limb length following total hip arthroplasty largely depend on restoring the distance between a fixed point on the pelvis and femur. Each of these techniques allows length correction to be made by combining the effects of both acetabular and femoral height into a single measurement. These methods help to minimise inequality but are anatomically flawed, as they do not allow independent control of placement of the femoral and acetabular components which both contribute to leg length. To address this we present and evaluate a technique that uses a caliper to control the vertical placement of the femoral component and the transverse acetabular ligament to control the vertical height of the acetabular component. Limb lengths were measured in 200 patients who had undergone primary total hip arthroplasty using this technique. Using this method 94% had a postoperative limb length inequality that was 6mm or less (average, +0.38 mm). The maximum measured limb length inequality was +/-8 mm.


Impact of gamification on green consumption behavior integrating technological awareness, motivation, enjoyment and virtual CSR.
Gamification entails integrating game design elements, including rewards, points, competition, and interactive challenges, into non-game contexts to engage and motivate individuals. In the context of green consumption, gamification can encourage individuals to acquire more sustainable consumption behaviors. The proposed study aims to examine the influence of gamification on green consumption behavior among Chinese university students. However, university students are considered an important target group for such interventions due to their technological savvy and high interest in environmental issues. A self-determination theory (SDT) was used to measure the motivating factors of gamification for adopting green consumption behavior-a convenience sampling technique in which survey-based research designs were used to collect the data. A survey was conducted on a sample of 332 university students in China, using a questionnaire with structural equation modeling (SEM) to test the hypotheses and assess the proposed relationships between the variables. The finding of this study reveals that gamification has a significant negative relation with green consumption behavior. Further, technological awareness, hedonic motivation, and perceived enjoyment significantly mediate the relationship between gamification and green consumption behavior. Additionally, virtual CSR significantly moderates the relationship between gamification and technological awareness, hedonic motivation, and perceived enjoyment. The findings of this study could have implications for the development of more effective interventions for policy makers and industrialists aimed at promoting sustainable consumption behaviors in China.


Leprosy Associated with Atypical Cutaneous Leishmaniasis in Nicaragua and Honduras.
In Central America, few cases of leprosy have been reported, but the disease may be unrecognized. Diagnosis is based on clinical criteria and histology. Preliminary field work in Nicaragua and Honduras found patients, including many children, with skin lesions clinically suggestive of atypical cutaneous leishmaniasis or indeterminate leprosy. Histology could not distinguish these diseases although acid-fast organisms were visible in a few biopsies. Lesions healed after standard antimicrobial therapy for leprosy. In the present study, patients, family members, and other community members were skin-tested and provided nasal swabs and blood samples. Biopsies were taken from a subgroup of patients with clinical signs of infection. Two laboratories analyzed samples, using local in-house techniques. Mycobacterium leprae, Leishmania spp. and Leishmania infantum were detected using polymerase chain reactions. Mycobacterium leprae DNA was detected in blood samples and nasal swabs, including some cases where leprosy was not clinically suspected. Leishmania spp. were also detected in blood and nasal swabs. Most biopsies contained Leishmania DNA and coinfection of Leishmania spp. with M. leprae occurred in 33% of cases. Mycobacterium leprae DNA was also detected and sequenced from Nicaraguan and Honduran environmental samples. In conclusion, leprosy and leishmaniasis are present in both regions, and leprosy appears to be widespread. The nature of any relationship between these two pathogens and the epidemiology of these infections need to be elucidated.


Introduction of HIV-1 in a rural city of Zaire.
In the rural city of Inongo (Bandundu region, Zaire), the prevalence of HIV-1 infection has been investigated during a survey on HTLV-I infection and associated neurological diseases, using a cluster sampling technique. Thirteen individuals were infected by HIV-1, 11 adults and 2 children, giving a crude prevalence rate in the cluster sample of 1.1%. More women were infected than men. After direct standardization on the age and sex structure given by the 1984 census, the prevalence of HIV-1 in the city may be estimated at 1.0% for the whole population and 1.6% for the population above 15 years of age. The importance is stressed of protecting rural populations, particularly women of childbearing age from the HIV epidemic, by information campaigns focused on the male migrant population, including both seasonal and travelling workers.


Sample-size calculations for short-term proof-of-concept studies of tissue protection and repair in multiple sclerosis lesions via conventional clinical imaging.
New multiple sclerosis (MS) lesion activity on magnetic resonance imaging (MRI) can test immunomodulatory therapies in proof-of-concept trials. Comparably powerful endpoints to assess tissue protection or repair are lacking. The objective of this paper is to report sample-size calculations for assessment of new lesion recovery. In two sets of six active MS cases, new lesions were observed by monthly MRI for approximately 12 months. Averages and quartiles of normalized (proton density/T1/T2 weighted) and quantitative (T1/T2 and mean diffusivity maps for dataset 1, T2 and magnetization transfer ratio maps for dataset 2) measures were used to compare the lesion area before lesion appearance to afterward. A linear mixed-effects model incorporating lesion- and participant-specific random effects estimated average levels and variance components for sample-size calculations. In both datasets, greatest statistical sensitivity was observed for the 25th percentile of normalized proton density-weighted signal. At 3T, using new lesions ⩾15 mm(3), as few as nine participants/arm may be required for a six-month placebo-controlled add-on trial postulating a therapeutic effect size of 20% and statistical power of 90%. Lesion recovery is a powerful outcome measure for proof-of-concept clinical trials of tissue protection and repair in MS. The trial design requires active cases and is therefore best implemented near disease onset.


The Investigations into What Happened at the Gosport War Memorial Hospital - Did the Coroner's Process Help?
The Gosport Independent Panel was established to review the care of older patients at the Gosport War Memorial Hospital in England over some 20 years. There had been a number of internal and external investigations that included police investigations, clinical care audits, GMC investigations and inquests. The Panel provided a means of public disclosure of much of the contents of the prior investigations and resulted in the creation of a catalogue of all relevant information. The report indicated that many of the investigative processes had failed to address the concerns of family and staff. In part this appears to have been the result of some investigations being limited in their ability to deal with social and community concerns and focusing on whether criminal prosecutions should be brought. Legislative restrictions regarding the nature and outcomes of the inquest process in the United Kingdom compounded these concerns. It is interesting to speculate whether a more proactive inquest system brought into play earlier might have alleviated many of the community and professional concerns regarding patient care.


Collapse of growth cone structure on contact with specific neurites in culture.
We have studied the morphology of embryonic chick retinal and sympathetic growth cones as they meet retinal and sympathetic neurites grown in culture. Growth cones preserve their normal morphology and ability to locomote when retinal growth cones contact retinal neurites or when sympathetic growth cones contact sympathetic neurites. Growth cones collapse and their motility ceases when retinal growth cones contact sympathetic neurites or when sympathetic growth cones contact retinal neurites. Collapse was never observed before a growth cone touched a neurite. As a growth cone collapses, the neurite it leads retracts. After a brief pause, a new growth cone is organized and extension recommences. These results suggest that contact-mediated inhibition of locomotion could play a role in growth cone guidance.


Conceptual synthesis in community ecology.
Community ecology is often perceived as a "mess, "given the seemingly vast number of processes that can underlie the many patterns of interest, and the apparent uniqueness of each study system. However, at the most general level, patterns in the composition and diversity of species--the subject matter of community ecology--are influenced by only four classes of process: selection, drift, speciation, and dispersal. Selection represents deterministic fitness differences among species, drift represents stochastic changes in species abundance, speciation creates new species, and dispersal is the movement of organisms across space. All theoretical and conceptual models in community ecology can be understood with respect to their emphasis on these four processes. Empirical evidence exists for all of these processes and many of their interactions, with a predominance of studies on selection. Organizing the material of community ecology according to this framework can clarify the essential similarities and differences among the many conceptual and theoretical approaches to the discipline, and it can also allow for the articulation of a very general theory of community dynamics: species are added to communities via speciation and dispersal, and the relative abundances of these species are then shaped by drift and selection, as well as ongoing dispersal to drive community dynamics.


Virtual Disassembling of Historical Edifices: Experiments and Assessments of an Automatic Approach for Classifying Multi-Scalar Point Clouds into Architectural Elements.
3D heritage documentation has seen a surge in the past decade due to developments in reality-based 3D recording techniques. Several methods such as photogrammetry and laser scanning are becoming ubiquitous amongst architects, archaeologists, surveyors, and conservators. The main result of these methods is a 3D representation of the object in the form of point clouds. However, a solely geometric point cloud is often insufficient for further analysis, monitoring, and model predicting of the heritage object. The semantic annotation of point clouds remains an interesting research topic since traditionally it requires manual labeling and therefore a lot of time and resources. This paper proposes an automated pipeline to segment and classify multi-scalar point clouds in the case of heritage object. This is done in order to perform multi-level segmentation from the scale of a historical neighborhood up until that of architectural elements, specifically pillars and beams. The proposed workflow involves an algorithmic approach in the form of a toolbox which includes various functions covering the semantic segmentation of large point clouds into smaller, more manageable and semantically labeled clusters. The first part of the workflow will explain the segmentation and semantic labeling of heritage complexes into individual buildings, while a second part will discuss the use of the same toolbox to segment the resulting buildings further into architectural elements. The toolbox was tested on several historical buildings and showed promising results. The ultimate intention of the project is to help the manual point cloud labeling, especially when confronted with the large training data requirements of machine learning-based algorithms.


Robust Blind Learning Algorithm for Nonlinear Equalization Using Input Decision Information.
In this paper, we propose a new blind learning algorithm, namely, the Benveniste-Goursat input-output decision (BG-IOD), to enhance the convergence performance of neural network-based equalizers for nonlinear channel equalization. In contrast to conventional blind learning algorithms, where only the output of the equalizer is employed for updating system parameters, the BG-IOD exploits a new type of extra information, the input decision information obtained from the input of the equalizer, to mitigate the influence of the nonlinear equalizer structure on parameters learning, thereby leading to improved convergence performance. We prove that, with the input decision information, a desirable convergence capability that the output symbol error rate (SER) is always less than the input SER if the input SER is below a threshold, can be achieved. Then, the BG soft-switching technique is employed to combine the merits of both input and output decision information, where the former is used to guarantee SER convergence and the latter is to improve SER performance. Simulation results show that the proposed algorithm outperforms conventional blind learning algorithms, such as stochastic quadratic distance and dual mode constant modulus algorithm, in terms of both convergence performance and SER performance, for nonlinear equalization.


Thermohalobacter berrensis gen. nov., sp. nov., a thermophilic, strictly halophilic bacterium from a solar saltern.
A new thermophilic, strictly halophilic, anaerobic, non-sporulating rod-shaped bacterium, measuring 0.5 x 3.0-8.0 microns and designated strain CTT3T, was isolated from a solar saltern. Strain CTT3T stained Gram-negative, was motile by means of laterally inserted flagella, had a genome G + C content of 33 mol% and grew optimally at 65 degrees C and pH 7.0 with 5% NaCl. The strain also grew readily at 70 degrees C in the presence of 15% NaCl. Strain CTT3T fermented cellobiose, fructose, glucose, maltose, mannitol, mannose, sucrose, glycerol, N-acetylglucosamine, starch, pyruvate and bio-Trypticase. It produced acetate, ethanol, H2 and presumably CO2 from glucose. 16S rRNA gene sequence analysis indicated that it is a member of cluster XII of the Clostridiales and related genera of the subphylum of the Gram-positive bacteria containing genomes of low G + C content. Its phenotypic and phylogenetic characteristics clearly differentiated it from all other members of this cluster. Based on the findings it is proposed that strain CTT3T be designated as a new species of a new genus, Thermohalobacter berrensis gen. nov., sp. nov. The type strain is CTT3T (= CNCM 105955T).


Association of newly replicated DNA with the nuclear matrix of Physarum polycephalum.
We have studied the role of the nuclear matrix in DNA replication in a naturally synchronized eucaryote, Physarum polycephalum. When P. polycephalum. When P. polycephalum macroplasmodia were pulse labeled with 3H-thymidine, the DNA remaining tightly associated with the matrix was highly enriched in newly synthesized DNA. This enrichment was found both in nuclei that had just initiated DNA replication as well as in nuclei isolated later during S phase. Pulse chase experiments showed that the association of newly replicated DNA with the matrix is transient, since most of the newly replicated DNA could be chased from the matrix by incubating pulse labeled macroplasmodia in media containing unlabeled thymidine. Studies measuring the size distribution of the matrix DNA supported the hypothesis that replication forks are attached to the nuclear matrix. Reconstitution controls indicated that these results were unlikely to be due to preferential, nonspecific binding of nascent DNA to the matrix during the extraction procedures. These results with P. polycephalum in combination with previous studies in non-synchronized rodent cells, suggest that the association of newly replicated DNA with the nuclear matrix may be a general feature of eucaryotic DNA replication.


L-type cardiac calcium channels in doxorubicin cardiomyopathy in rats morphological, biochemical, and functional correlations.
Doxorubicin (DXR) is an effective antitumor agent in a wide spectrum of neoplasms. Chronic treatment is associated with cardiomyopathy and characteristic myocardial ultrastructural changes, which include swelling of the t tubules. Accordingly, we investigated excitation-contraction coupling in cardiomyopathic rat heart resulting from chronic DXR treatment. Using the whole-cell patch clamp technique, we studied the L-type calcium channel in single cells enzymatically isolated from normal (CTRL) and DXR rat hearts. Despite similar cell dimensions, the total membrane capacitance was significantly smaller in the DXR cells (138 +/- 9 pF) than in the CTRL cells (169 +/- 11 pF) (mean +/- SEM, n = 9, P less than 0.05). The mean current and the current density-voltage relationships of the CTRL and the DXR cells were significantly different (n = 9, P less than 0.001) with the maximal peak L-type calcium current (ICa) density increased from 6.4 +/- 0.9 in CTRL cells to 10.5 +/- 2.4 microA/cm2 in the DXR cells (P less than 0.05). There was no shift either in the current-voltage relationship or the steady-state inactivation curve in the two cell groups. However, the fast time constant of inactivation was increased at a membrane voltage of -10 to 10 mV. Calcium channel antagonist equilibrium binding assays using [3H]-PN200-110 revealed no difference in the maximal receptor binding capacity (CTRL, 194 +/- 27 and DXR 211 +/- 24 fmol/mg protein; P greater than 0.05, n = 6) and in receptor affinity (CTRL, 0.15 +/- 0.05 and DXR 0.13 +/- 0.03 nM; P less than 0.05). These data suggest that a decrease in effective capacitance might be associated with t-tubular damage. Despite this decrease, ICa was increased in the DXR cells. Such an increase may result from an alteration in the properties of the calcium channels and/or recruitment of "hibernating" channels in the remaining surface and t-tubular membranes.


Fuel utilization in colonocytes of the rat.
In incubated colonocytes isolated from rat colons, the rates of utilization O2, glucose or glutamine were linear with respect to time for over 30 min, and the concentrations of adenine nucleotides plus the ATP/ADP or ATP/AMP concentration ratios remained approximately constant for 30 min. Glutamine, n-butyrate or ketone bodies were the only substrates that caused increases in O2 consumption by isolated incubated colonocytes. The maximum activity of hexokinase in colonic mucosa is similar to that of 6-phosphofructokinase. Starvation of the donor animal decreased the activities of hexokinase and 6-phosphofructokinase, whereas it increased those of glucose-6-phosphatase and fructose-bisphosphatase. Isolated incubated colonocytes utilized glucose at about 6.8 mumol/min per g dry wt., with lactate accounting for 83% of glucose removed. These rates were not affected by the addition of glutamine, acetoacetate or n-butyrate, and starvation of the donor animal. Isolated incubated colonocytes utilized glutamine at about 5.5 mumol/min per g dry wt., which is about 21% of the maximum activity of glutaminase. The major end-products of glutamine metabolism were glutamate, aspartate, alanine and ammonia. Starvation of the donor animal decreased the rate of glutamine utilization by colonocytes, which is accompanied by a decrease in glutamate formation and in the maximum activity of glutaminase. Isolated incubated colonocytes utilized acetoacetate at about 3.5 mumol/min per g dry wt. This rate was not markedly affected by addition of glucose or by starvation of the donor animal. When colonocytes were incubated with n-butyrate, both acetoacetate and 3-hydroxybutyrate were formed, with the latter accounting for only about 19% of total ketones produced.


Effect of submental sensitive transcutaneous electrical stimulation on virtual lesions of the oropharyngeal cortex.
The aim of this study was to assess the effect of submental sensitive transcutaneous electrical stimulation (SSTES) on pharyngeal cortical representation after the creation of an oropharyngeal cortical virtual lesion in healthy subjects. Motor-evoked potential amplitude of the mylohyoid muscles was measured with transcranial magnetic stimulation (TMS), the oropharyngeal cortex was mapped by cartography, and videofluoroscopic parameters of swallowing function were measured before and after SSTES (at the end of SSTES [0 min] and at 30 and 60 min), after the creation of a cortical virtual lesion (repetitive TMS, 1 Hz, 20 min on the dominant swallowing hemisphere). Nine subjects completed the study. After 20 min of SSTES, motor-evoked potential amplitude increased (P<0.05), as did swallow reaction time after repetitive TMS, as seen on videofluoroscopy, which was reversed after electrical stimulation. On cortical mapping, the number of points with a cortical response increased in the dominant lesioned hemisphere (P<0.05), remaining constant at 60 min (P<0.05). SSTES may be effective for producing cortical plasticity for mylohyoid muscles and reverses oropharyngeal cortical inhibition in healthy subjects. It could be a simple non-invasive way to treat post-stroke dysphagia.


Heterogeneity of CD4+ T cells involved in anti-allo-class I H-2 immune responses. Functional discrimination between the major proliferating cells and helper cells assisting cytotoxic T cell responses.
MLR in various combinations with class I H-2 disparity revealed that there are three patterns of MLR in the aspect of responding T subset (CD4 vs CD8) dominance. Irrespective of the CD8 vs CD4 dominance, a single i.v. administration of class I-disparate allogeneic spleen cells resulted in almost complete abrogation of anti-class I proliferative capacity of both CD4+ and CD8+ T cells in six combinations. The suppression of proliferative responses was correlated with the striking reduction in the ability to produce IL-2 upon stimulation with the relevant class I alloantigens. In contrast, i.v. presensitized recipient mice exhibiting only marginal MLR/Il-2 production could generate comparable magnitudes of anti-allo class I CTL as well as graft rejection responses to those induced by normal unpresensitized mice. The administration in vivo of anti-CD4 antibody along with the i.v. presensitization not only suppressed the generation of CTL responses by spleen cells but also induced appreciable prolongation of allo-class I-disparate skin grafts under conditions in which neither alone did it. These results demonstrate that 1) the suppression of graft rejection responses is not necessarily reflected on the reduction of MLR; 2) CD8+ CTL precursors responsible for graft rejection can be activated by either allo-class I-reactive CD8+ or CD4+ Th cells; 3) i.v. presensitization induces functional elimination of CD8+ and CD4+ proliferative/IL-2-producing T cells but not of CD8+ CTL precursors and CD4+ Th whose capacity is expressed by assistance of CTL induction but not by their own proliferation. Thus, this study illustrates the heterogeneity of class I alloantigen-reactive CD4+ T cells in the aspect of their capacity to proliferate themselves vs contribute to CTL induction as well as graft rejection.


Delivery of dangerous goods: type III secretion in enteric pathogens.
Type III secretion systems (TTSSs) of Gram-negative pathogens are molecular syringes that inject bacterial virulence factors directly into host cells. These virulence factors manipulate host cell pathways to aid bacterial survival within the host. Four important enteric pathogens use TTSSs to colonize and persist within the intestinal environment. The following is a brief review of the way in which TTSSs and their effectors contribute to the pathogenic nature of the prototypic diarrheal pathogens Salmonella, Shigella, Yersinia and enteropathogenic Escherichia coli (EPEC).


Sex differences in behavioral decision-making and the modulation of shared neural circuits.
Animals prioritize behaviors according to their physiological needs and reproductive goals, selecting a single behavioral strategy from a repertoire of possible responses to any given stimulus. Biological sex influences this decision-making process in significant ways, differentiating the responses animals choose when faced with stimuli ranging from food to conspecifics. We review here recent work in invertebrate models, including C. elegans, Drosophila, and a variety of insects, mollusks and crustaceans, that has begun to offer intriguing insights into the neural mechanisms underlying the sexual modulation of behavioral decision-making. These findings show that an animal's sex can modulate neural function in surprisingly diverse ways, much like internal physiological variables such as hunger or thirst. In the context of homeostatic behaviors such as feeding, an animal's sex and nutritional status may converge on a common physiological mechanism, the functional modulation of shared sensory circuitry, to influence decision-making. Similarly, considerable evidence suggests that decisions on whether to mate or fight with conspecifics are also mediated through sex-specific neuromodulatory control of nominally shared neural circuits. This work offers a new perspective on how sex differences in behavior emerge, in which the regulated function of shared neural circuitry plays a crucial role. Emerging evidence from vertebrates indicates that this paradigm is likely to extend to more complex nervous systems as well. As men and women differ in their susceptibility to a variety of neuropsychiatric disorders affecting shared behaviors, these findings may ultimately have important implications for human health.


Investigating the oxidation mechanism of facet-dependent pyrite: implications for the environment and sulfur evolution.
The oxidation of pyrite (FeS2) not only adversely affects the environment, but also plays a critical role in the geochemical evolution of Fe and S elements. However, the oxidation rate of FeS2 is often controlled by its exposed crystal facets. Herein, the oxidation behaviors and mechanisms of naturally existing FeS2(100) and FeS2(210) crystals are investigated. The adsorption models of O2 on FeS2(100) and FeS2(210) facets are established, additionally, their corresponding surface energies, O2 adsorption sites and energies are also obtained using Density Functional Theory (DFT) calculations. These results suggest that the FeS2(210) facet more readily reacts with O2 because it has more unsaturated coordination of Fe atoms compared with the FeS2(100) facet. Moreover, electrochemical results such as EIS, Tafel and CV curves further prove that FeS2(210) possesses a higher oxidation rate than that of FeS2(100). The results of chemical oxidation experiments and XPS analyses show that FeS2(210) can produce more total Fe, SO42- and H+ than FeS2(100). Furthermore, various intermediate S species such as SO32-, S2O32-, S3O62-, S4O62- and S5O62- are also detected. This work can provide a basis for understanding the oxidation mechanism of facet-dependent FeS2 and the geochemical evolution of Fe and S elements.


Emerging novel sequence types of Staphylococcus aureus in Pakistan.
Despite an increasing incidence of Staphylococcus aureus infection and dissemination in Pakistan, the epidemiology of different Staphylococcus aureus research clones has been the subject of only a small number of investigations. By analyzing the collected data sequence, this study was designed to study the epidemiology of Staphylococcus aureus in the area using multilocus sequence typing (MLST). A total of 1015 staphylococcus strains collected from the city's tertiary care facilities were biochemically screened, followed by antimicrobial susceptibility testing against a panel of 13 antibiotics. Analyzed methicillin-resistant Staphylococcus aureus (MRSA) was subjected to molecular characterization using multilocus sequence typing (MLST), clonal complex analysis, recombination testing, and phylogenetic analysis. Approximately 421 bacteria were verified as Staphylococcus aureus by biochemical analysis. 57% of the isolates exhibited multidrug resistance, of which 89% were found to be methicillin-resistant Staphylococcus aureus (MRSA). MLST results in a total of 39 sequence types (ST) and 5 clonal complexes (CC), out of which twenty-two STs were newly documented worldwide. The most common CC identified was CC8. The direct sequencing data also revealed significant shifts at MLST loci, with point mutations resulting in the aroE-343 and tpi-278 alleles. This study concludes that there is high diversity in the locally circulating clones of Staphylococcus aureus present in nature and that they are defined by their geographic epidemiology. These findings have practical implications for public health, including the need for tailored infection control strategies, antibiotic stewardship, global surveillance, and a deeper understanding of bacterial evolution.


Improving the strength of sutureless laser-assisted vessel repair using preloaded longitudinal compression on tissue edge.
Little is known about the approximation of coapted edges in sutureless laser-assisted vessel welding. Tissue shrinkage by laser irradiation may cause coapted edges to separate, reducing strength of welding. This may be avoided by preloaded longitudinal compression on the tissue edges to be welded. This study compared welding strength with and without preloaded compression in ex vivo animal experiments. This study evaluated 24 samples of harvested porcine carotid arteries, each having a length of 3 cm and an inner diameter of 1.0-2.0 mm. A half circumferential incision was made at the center of each sample. A steel shaft 2.0 mm in diameter was inserted into each sample to approximate the incised edges. The samples were longitudinally compressed to 6 mm. Incision sites were repaired by irradiation with a 970-nm diode laser. No glue or die was used. The repair strength was evaluated by measuring the bursting point (BP) of all samples. In a pilot study, the welding conditions, including power, duration, and interval of the laser spots, were tested by trial and error in 18 samples, including six treated under optimum conditions. As a control group, six samples were welded under optimum conditions, but without compression. Optimum conditions, consisting of 2.4 W power, 30-second duration, and 1-mm intervals of laser spots, yielded the highest BP (623 ± 236 mmHg), which was significantly higher than in the control group without compression (204 ± 208 mmHg, P = 0.009). Defining BP > 400 mmHg as successful repair, the success rates in the compression and control groups were 83% and 17%, respectively. Preloaded longitudinal compression on the coapted edges may be important for sutureless laser-assisted vessel repair and anastomosis and may affect the strength of welding. Lasers Surg. Med. 49:533-538, 2017. © 2017 Wiley Periodicals, Inc.


Macromolecular organization and fine structure of the human basilar membrane - RELEVANCE for cochlear implantation.
Cochlear micromechanics and frequency tuning depend on the macromolecular organization of the basilar membrane (BM), which is still unclear in man. Novel techniques in cochlear implantation (CI) motivate further analyses of the BM. Normal cochleae from patients undergoing removal of life-threatening petro-clival meningioma and an autopsy specimen from a normal human were used. Laser-confocal microscopy, high resolution scanning (SEM) and transmission electron microscopy (TEM) were carried out in combination. In addition, one human temporal bone was decellularized and investigated by SEM. The human BM consisted in four separate layers: (1) epithelial basement membrane positive for laminin-β2 and collagen IV, (2) BM "proper" composed of radial fibers expressing collagen II and XI, (3) layer of collagen IV and (4) tympanic covering layer (TCL) expressing collagen IV, fibronectin and integrin. BM thickness varied both radially and longitudinally (mean 0.55-1.16 μm). BM was thinnest near the OHC region and laterally. There are several important similarities and differences between the morphology of the BM in humans and animals. Unlike in animals, it does not contain a distinct pars tecta (arcuate) and pectinata. Its width increases and thickness decreases as it travels apically in the cochlea. Findings show that the human BM is thinnest and probably most vibration-sensitive at the outer pillar feet/Deiter cells at the OHCs. The inner pillar and IHCs seem situated on a fairly rigid part of the BM. The gradient design of the BM suggests that its vulnerability increases apical wards when performing hearing preservation CI surgery.


International patterns and trends in the incidence of melanoma and cutaneous squamous cell carcinoma, 1989-2020.
Cutaneous squamous cell carcinoma (cSCC) and melanoma have different associations with sun exposure, so comparing the trends in incidence rates of the two different cancers may provide insights into changing patterns of exposure to ultraviolet radiation. We compared trends in the incidence of cSCC and melanoma in seven susceptible populations residing at mid-to-high latitudes: Finland, Norway, Sweden, Denmark, Scotland, the Netherlands and Tasmania (Australia). We fitted Joinpoint models to describe trends in age-standardized incidence rates for melanoma and cSCC and calculated the average annual percentage rate of change for the period 1989-2020 (1989-2018 for Tasmania). We calculated the incident rate ratio as the ratio of the age-standardized rates (European Standard Population) for cSCC to melanoma and conducted age-period-cohort modelling to compare age, period, and cohort effects. The ratio of cSCC-to-melanoma incidence increased with proximity to the equator and over time. In the most recent time period, the incidence of cSCC was higher than the incidence of melanoma for men and women in all seven populations. While the ratio of cSCC-to-melanoma incidence was higher for men compared with women, in most countries the cSCC-to-melanoma IRR increased over time to a greater extent in women compared with men. Melanoma incidence was higher among younger people and cSCC incidence was higher among older people; the age at which the incidence of cSCC overtook incidence of melanoma was progressively younger with proximity to the equator. Despite concerted international efforts to preserve the ozone layer over the past four decades resulting in significant reductions in surface UV-B at mid-latitudes, the incidence of skin cancer, particularly cSCC, continues to rise in those regions. Our findings are consistent with a stronger association with age-associated cumulative sun exposure for cSCC compared with melanoma, and suggest that women are receiving greater UV radiation exposure in recent decades than in the past.


Isolated necrotizing epiglottitis: report of a case in a neutropenic patient and review of the literature.
Isolated necrotizing epiglottitis (INE) is an unusual condition that may develop in immunocompromised patients. Only 3 cases of INE have been reported in the English-language literature; this is the fourth case, one in a 27-year-old woman who had neutropenia due to cytomegalovirus infection. Stenotrophomonas maltophilia and Candida albicans were isolated from the culture of necrotic material. The features of INE are discussed here by reviewing the literature and by examining the characteristics of this case.


Recreational concentrations of alcohol enhance synaptic inhibition of cerebellar unipolar brush cells via pre- and postsynaptic mechanisms.
Variation in cerebellar sensitivity to alcohol/ethanol (EtOH) is a heritable trait associated with alcohol use disorder in humans and high EtOH consumption in rodents, but the underlying mechanisms are poorly understood. A recently identified cellular substrate of cerebellar sensitivity to EtOH, the GABAergic system of cerebellar granule cells (GCs), shows divergent responses to EtOH paralleling EtOH consumption and motor impairment phenotype. Although GCs are the dominant afferent integrator in the cerebellum, such integration is shared by unipolar brush cells (UBCs) in vestibulocerebellar lobes. UBCs receive both GABAergic and glycinergic inhibition, both of which may mediate diverse neurological effects of EtOH. Therefore, the impact of recreational concentrations of EtOH (~10-50 mM) on GABAA receptor (GABAAR)- and glycine receptor (GlyR)-mediated spontaneous inhibitory postsynaptic currents (sIPSCs) of UBCs in cerebellar slices was characterized. Sprague-Dawley rat (SDR) UBCs exhibited sIPSCs mediated by GABAARs, GlyRs, or both, and EtOH dose-dependently (10, 26, 52 mM) increased their frequency and amplitude. EtOH increased the frequency of glycinergic and GABAergic sIPSCs and selectively enhanced the amplitude of glycinergic sIPSCs. This GlyR-specific enhancement of sIPSC amplitude resulted from EtOH actions at presynaptic Golgi cells and via protein kinase C-dependent direct actions on postsynaptic GlyRs. The magnitude of EtOH-induced increases in UBC sIPSC activity varied across SDRs and two lines of mice, in parallel with their respective alcohol consumption/motor impairment phenotypes. These data indicate that Golgi cell-to-UBC inhibitory synapses are targets of EtOH, which acts at pre- and postsynaptic sites, via Golgi cell excitation and direct GlyR enhancement.NEW & NOTEWORTHY Genetic variability in cerebellar alcohol/ethanol sensitivity (ethanol-induced ataxia) predicts ethanol consumption phenotype in rodents and humans, but the cellular and molecular mechanisms underlying genetic differences are largely unknown. Here it is demonstrated that recreational concentrations of alcohol (10-30 mM) enhance glycinergic and GABAergic inhibition of unipolar brush cells through increases in glycine/GABA release and postsynaptic enhancement of glycine receptor-mediated responses. Ethanol effects varied across rodent genotypes parallel to ethanol consumption and motor sensitivity phenotype.


Primary scalp cancer in Singapore.
Cancers of the scalp are uncommon. Owing to their unique site they pose problems in surgical treatment. A retrospective study of all scalp cancers notified to the Singapore Cancer Registry over 17 years has shown that squamous cell carcinoma is the most common cancer followed by basal cell carcinoma. Other cancers are sweat gland carcinoma, angiosarcoma, dermatofibrosarcoma protruberans, liposarcoma, fibrosarcoma and malignant lymphoma. Scalp cancers are more common in Chinese and very rare in Indians. There is no difference in the incidence between the sexes. The surgical treatment and reconstructive techniques employed in 52 patients are described. When localised, scalp cancers are managed like other skin cancers but once they invade bone, they become aggressive and are difficult to eradicate. Wide radical excision provides cure and the skull bone if involved should be removed.