Indiscriminate females and choosy males: within- and between-species variation in Drosophila.
The classic view of choosy, passive females and indiscriminate, competitive males gained theoretical foundations with parental investment theory. When females invest more in offspring than males, parental investment theory says that selection operates so that females discriminate among males for mates (i.e., females are choosy and passive) and males are indiscriminate (i.e., males are profligate and competitive). Here we report tests of predictions using Drosophila pseudoobscura and D. melanogaster, with typical asymmetry in gamete sizes (females > males), and in D. hydei with far less asymmetry in gamete size. Experimental observations revealed that the labels "choosy, passive females" and "profligate, indiscriminate males" did not capture the variation within and between species in premating behavior. In each of the species some females were as active in approaching males (or more so) than males in approaching females, and some males were as discriminating (or more so) than females. In pairs focal males and females responded differently to opposite-sex than to same-sex conspecifics. Drosophila hydei were less sex-role stereotyped than the other two species consistent with parental investment theory. However, D. pseudoobscura females approached males more often than did D. melanogaster females, and male D. hydei approached females as often as males of the other two species, both results inconsistent with parental investment theory. Male D. pseudoobscura and D. hydei were more likely to approach males in same-sex pairs than male D. melanogaster, inconsistent with parental investment theory.


Quantification of myocardial oxygen extraction fraction: A proof-of-concept study.
To demonstrate a proof of concept for the measurement of myocardial oxygen extraction fraction (mOEF) by a cardiovascular magnetic resonance technique. The mOEF measurement was performed using an electrocardiogram-triggered double-echo asymmetric spin-echo sequence with EPI readout. Seven healthy volunteers (22-37 years old, 5 females) were recruited and underwent the same imaging scans at rest on 2 different days for reproducibility assessment. Another 5 subjects (23-37 years old, 4 females) underwent cardiovascular magnetic resonance studies at rest and during a handgrip isometric exercise with a 25% of maximal voluntary contraction. Both mOEF and myocardial blood volume values were obtained in septal regions from respective maps. The reproducibility was excellent for the measurements of mOEF in septal myocardium (coefficient of variation: 3.37%) and moderate for myocardial blood volume (coefficient of variation: 19.7%). The average mOEF and myocardial blood volume of 7 subjects at rest were 0.61 ± 0.05 and 11.0 ± 4.3%, respectively. The mOEF agreed well with literature values that were measured by PET in healthy volunteers. In the exercise study, there was no significant change in mOEF (0.61 ± 0.06 vs 0.62 ± 0.07) or myocardial blood volume (12 ± 6% vs 13 ± 4%) from rest to exercise, as expected. The implemented cardiovascular magnetic resonance method shows potential for the quantitative assessment of mOEF in vivo. Future technical work is needed to improve image quality and to further validate mOEF measurements.


Leadership Views on the Barriers and Incentives to Clinical Preceptorship.
Clinical education often relies on a one-to-one student-preceptor model. Recruiting and retaining quality preceptors to sustain this model has become increasingly difficult at academic institutions across the nation. While ample literature describes preceptor barriers and incentives as viewed by physician educators, few studies explore the issue from institutional leadership perspectives. This study aimed to describe leadership perceptions across an academic institution to better understand knowledge gaps, system barriers, and proposed solutions to help institutions take action and address preceptor shortages. Between February and July 2019, the researchers conducted one-on-one semi-structured interviews with sampled representation of Medical College of Wisconsin leadership. The researchers reviewed transcriptions of each interview verbatim and used a qualitative grounded theory approach to generate content codes and themes. Researchers iteratively refined codes using the constant comparison method until all interviews were analyzed and final themes and subthemes were defined. Twelve institutional leaders participated, of whom 5 were clinical executives, 1 was an academic executive, 4 were academic deans, and 2 were educational directors. Analysis yielded 4 major themes: student impact, recognition, physician well-being, and leadership. Each content theme highlighted areas to consider when addressing preceptor issues within an institution: (1) leadership knowledge gaps regarding the scope of preceptor challenges, particularly time commitments and the number of preceptors required; (2) improving career advancement or promotion criteria to recognize teaching efforts; (3) enhanced physician well-being from teaching, while important, may no longer be sufficient for participation, especially without financial compensation; (4) distributed leadership may be needed to address issues at the course, clinic, and system levels.


A multiresponse parathyroid hormone assay: an inhibitor has agonist properties in vivo.
Vitamin D-deficient rats subjected to thyroparathyroidectomy (TPTX) were used to evaluate in vivo the biological properties of native bovine parathyroid hormone (bPTH) and chemically synthesized fragments and analogues of the hormone on several parameters of hormone action: calcium and phosphorus fluxes, generation of cyclic adenosine 3',5'-monophosphate (cAMP), and the metabolism of 25-hydroxyvitamin D3 [25(OH)D3]. Vitamin D-deficient rats, after TPTX or sham operation, were intravenously infused with a nutrient containing 7.5 mM CaCl2 for 30 h. During the last 7 h, PTH or one of its analogues was infused intravenously at rates between 0.04 and 20 nmol/h. One hour after the start of the peptide infusion, tritiated 25(OH)D3 was injected. Urine was collected hourly for phosphate and cAMP determinations and, at the end of the experiment, blood was obtained to determine the relative accumulation of tritiated 1,25-dihydroxyvitamin D3 ([3H]1,25(OH)2D3). Infusion of bPTH-(1--84), bPTH-(1--34), human (h)PTH-(1--34), or [Nle8, Nle18, Tyr34]bPTH-(1--34) amide was accompanied by a comparable dose-dependent decrease in plasma phosphate and a dose-dependent increase in plasma calcium and [3H]-1,25(OH)2D3, and urinary excretion of phosphate and cAMP. An evaluation of [Nle8, Nle18, Tyr34]bPTH-(3--34) amide, a potent inhibitor of PTH action in vitro in the renal adenylate cyclase assay, revealed that the analogue possessed weak agonist properties in vivo. The analogue increased excretion of both cAMP and phosphate in the urine, decreased plasma phosphate levels, and increased the accumulation of [3H]-1,25(OH)2D3 in the plasma. This multiparameter model system should aid in the elucidation of the in vivo biological effects of PTH and its analogues.


18F-FDG PET for evaluation of bone marrow infiltration in staging of lymphoma: a meta-analysis.
The ability of PET with (18)F-FDG to evaluate bone marrow infiltration in patients with lymphoma has been a matter of extensive investigation with controversial results. Therefore, we aimed to evaluate systematically, with a meta-analysis, the diagnostic performance of (18)F-FDG PET in this setting. Relevant studies were identified with MEDLINE and EMBASE searches (last update, August 2004). Data on the diagnostic performance of (18)F-FDG PET were combined quantitatively across eligible studies. We estimated weighted summary sensitivities and specificities, summary receiver-operating-characteristic (SROC) curves, and weighted summary likelihood ratios. We also conducted separate analyses according to various subgroups. Bone marrow biopsy (BMB) was used as the reference standard. Thirteen eligible nonoverlapping studies, which enrolled a total of 587 patients, were included in the meta-analysis. The independent random-effects weighted estimates of sensitivity and specificity against BMB were 51% (95% confidence interval [CI], 38%-64%) and 91% (95% CI, 85%-95%), respectively. Results were consistent in the SROC curve: a sensitivity of 51% corresponds to a specificity of 92%, whereas a specificity of 91% corresponds to a sensitivity of 55%. The weighted positive likelihood ratio (LR+) was 5.75 (95% CI, 348-9.48) and the negative likelihood ratio (LR-) was 0.67 (95% CI, 0.55-0.82). Six of 12 patients with positive (18)F-FDG PET and negative initial biopsy were found to have bone marrow involvement when biopsy was performed at the sites with positive imaging signals. Subgroup analyses showed better sensitivity in patients with Hodgkin's disease and in aggressive histologic types of non-Hodgkin's lymphoma than in patients with less aggressive histologic types and in studies using unilateral BMB compared with those using bilateral biopsy. This meta-analysis showed that (18)F-FDG PET has good, but not excellent, concordance with the results of BMB for the detection of bone marrow infiltration in the staging of patients with lymphoma. (18)F-FDG PET may complement the results of BMB and its performance may vary according to the type of lymphoma.


Plasma and urine neutrophil gelatinase-associated lipocalin in septic versus non-septic acute kidney injury in critical illness.
Sepsis is the most common trigger for acute kidney injury (AKI) in critically ill patients. We sought to determine whether there are unique patterns to plasma and urine neutrophil gelatinase-associated lipocalin (NGAL) in septic compared with non-septic AKI. Prospective observational study. Two adult ICUs in Melbourne, Australia. Critically ill patients with septic and non-septic AKI. None. Blood and urine specimens collected at enrollment, 12, 24 and 48 h to measure plasma and urine NGAL. Eighty-three patients were enrolled (septic n = 43). Septic AKI patients had more co-morbid disease (p = 0.005), emergency surgical admissions (p < 0.001), higher illness severity (p = 0.008), more organ dysfunction (p = 0.008) and higher white blood cell counts (p = 0.01). There were no differences at enrollment between groups in AKI severity. Septic AKI was associated with significantly higher plasma (293 vs. 166 ng/ml) and urine (204 vs. 39 ng/mg creatinine) NGAL at enrollment compared with non-septic AKI (p < 0.001). Urine NGAL remained higher in septic compared with non-septic AKI at 12 h (p < 0.001) and 24 h (p < 0.001). Plasma NGAL showed fair discrimination for AKI progression (area under receiver-operator characteristic curve 0.71) and renal replacement therapy (AuROC 0.78). Although urine NGAL performed less well (AuROC 0.70, 0.70), peak urine NGAL predicted AKI progression better in non-septic AKI (AuROC 0.82). Septic AKI patients have higher detectable plasma and urine NGAL compared with non-septic AKI patients. These differences in NGAL values in septic AKI may have diagnostic and clinical relevance as well as pathogenetic implications.


The systemic nature of CKD.
The accurate definition and staging of chronic kidney disease (CKD) is one of the major achievements of modern nephrology. Intensive research is now being undertaken to unravel the risk factors and pathophysiologic underpinnings of this disease. In particular, the relationships between the kidney and other organs have been comprehensively investigated in experimental and clinical studies in the last two decades. Owing to technological and analytical limitations, these links have been studied with a reductionist approach focusing on two organs at a time, such as the heart and the kidney or the bone and the kidney. Here, we discuss studies that highlight the complex and systemic nature of CKD. Energy balance, innate immunity and neuroendocrine signalling are highly integrated biological phenomena. The diseased kidney disrupts such integration and generates a high-risk phenotype with a clinical profile encompassing inflammation, protein-energy wasting, altered function of the autonomic and central nervous systems and cardiopulmonary, vascular and bone diseases. A systems biology approach to CKD using omics techniques will hopefully enable in-depth study of the pathophysiology of this systemic disease, and has the potential to unravel critical pathways that can be targeted for CKD prevention and therapy.


The role of glial cells in epilepsy.
Brain research in the last century was mainly directed at neurons, with the role of glia assumed to be limited to repair, supplying nutrients and above all acting as a packing material between neurons. In recent years, the importance of glial cells for normal brain function has been recognised. This article summarizes knowledge of glial cells of relevance to epilepsy. The article is based on a literature search in PubMed as well as the authors' clinical and research experience. Astrocytes are the largest subgroup of glial cells and, in common with neurons, have diverse membrane transporters, ion channels and receptors. Among the most important roles of astrocytes are the uptake and redistribution of ions and water, glucose metabolism and communication with nerve cells. Disturbances in all of these functions have been associated with epilepsy. Epilepsy has previously been regarded as exclusively a disturbance in the functioning of neurons and especially of their contact points, the synapses. The mechanisms of action of today's anti-epileptic drugs are therefore primarily directed at neuronal channels and receptors. New knowledge of the role played by glial cells could increase our understanding of how epilepsy arises and could lead to new treatment strategies.


Catheter interventional treatment of congenital portosystemic venous shunts in childhood.
The study aimed on the catheter interventional treatment of congenital portosystemic venous shunt (CPSVS) in childhood and the impact on vascular growth of hypoplastic portal veins. Clinical course and follow up of partial or complete closure of CPSVS are described. CPSVS is a very rare vascular malformation of the portal venous drainage resulting in potentially life threatening abdominal, pulmonary, cerebral, and cardiac complications. The anatomic type, clinical course, and diagnosis must be determined for optimal management. Single centre case series. Eight (6 female) children with extrahepatic (n = 5) and intrahepatic (n = 3) CPSVS were diagnosed invasively by catheterization, including test balloon occlusion of the shunt and simultaneous retrograde angiography, and treated by catheter interventions (n = 5) with partial (n = 2) and complete (n = 3) occlusion of CPSVS at a median age of 3.9 years (range 0.7-21). Congenital heart disease (CHD) was the most frequent associated organ manifestation (n = 5) followed by mild to severe pulmonary arterial hypertension (n = 4), hepatopulmonary syndrome (n = 2), and portosystemic encephalopathy (n = 1). CHD was simple (n = 3) or complex type (n = 2). Three patients were untreated so far, because they were in excellent clinical condition at an age <1 year, refused treatment, or planned for later treatment. Accurate invasive diagnosis of CPSVS with test balloon occlusion of the shunt is mandatory to depict the anatomic situation. Catheter interventional treatment of CPSVS offers a feasible and safe approach with complete or partial closure of the vascular malformation inducing potentially significant vascular growth of a former hypoplastic portal venous system. © 2015 Wiley Periodicals, Inc.


De Novo Damaging Variants, Clinical Phenotypes, and Post-Operative Outcomes in Congenital Heart Disease.
De novo genic and copy number variants are enriched in patients with congenital heart disease, particularly those with extra-cardiac anomalies. The impact of de novo damaging variants on outcomes following cardiac repair is unknown. We studied 2517 patients with congenital heart disease who had undergone whole-exome sequencing as part of the CHD GENES study (Congenital Heart Disease Genetic Network). Two hundred ninety-four patients (11.7%) had clinically significant de novo variants. Patients with de novo damaging variants were 2.4 times more likely to have extra-cardiac anomalies (P=5.63×10-12). In 1268 patients (50.4%) who had surgical data available and underwent open-heart surgery exclusive of heart transplantation as their first operation, we analyzed transplant-free survival following the first operation. Median follow-up was 2.65 years. De novo variants were associated with worse transplant-free survival (hazard ratio, 3.51; P=5.33×10-04) and longer times to final extubation (hazard ratio, 0.74; P=0.005). As de novo variants had a significant interaction with extra-cardiac anomalies for transplant-free survival (P=0.003), de novo variants conveyed no additional risk for transplant-free survival for patients with these anomalies (adjusted hazard ratio, 1.96; P=0.06). By contrast, de novo variants in patients without extra-cardiac anomalies were associated with worse transplant-free survival during follow-up (hazard ratio, 11.21; P=1.61×10-05) than that of patients with no de novo variants. Using agnostic machine-learning algorithms, we identified de novo copy number variants at 15q25.2 and 15q11.2 as being associated with worse transplant-free survival and 15q25.2, 22q11.21, and 3p25.2 as being associated with prolonged time to final extubation. In patients with congenital heart disease undergoing open-heart surgery, de novo variants were associated with worse transplant-free survival and longer times on the ventilator. De novo variants were most strongly associated with adverse outcomes among patients without extra-cardiac anomalies, suggesting a benefit for preoperative genetic testing even when genetic abnormalities are not suspected during routine clinical practice. Registration: URL: https://www.clinicaltrials.gov. Unique identifier: NCT01196182.


A comparison of three indicators for identifying Mexican Americans in epidemiologic research. Methodological findings from the San Antonio Heart Study.
Because the issue of how to empirically identify Mexican Americans in health-related research is still unresolved, the authors compared the performance of three indicators for identifying Mexican Americans across five distinct population subgroups: men and women in two age strata, and residents in low, middle, and high socioeconomic neighborhoods. Individual surname had the lowest sensitivity, specificity, and predictive values in the pooled population sample and varied the most widely on these parameters across population subgroups. Parental surnames, which are available on vital statistics and could easily be added to other health records used in secondary analyses, offered a significant improvement over individual surname in classifying persons as Mexican American. The San Antonio Heart Study (SAHS) algorithm, a nine-item indicator which uses parental surnames, birthplace of both parents, self-declared ethnic identity, and ethnic background of grandparents, had the highest sensitivity, specificity, and predictive values and varied the least on these parameters across different sex, age, and socioeconomic status population subgroups. The performance of all indicators was lower at the higher socioeconomic status levels. The findings suggest that it may be useful to use parental surnames as an indicator for Mexican-American ethnicity in research involving vital statistics and to add parental surnames to other health records frequently used in secondary analyses. Since the SAHS algorithm can be adapted for use with non-Mexican origin Hispanic subgroups, it may be a useful indicator for Mexican-American (or other Hispanic) ethnicity in survey research.


[Methodological aspects of pharmacological regulation of emotional stress in persons with dangerous occupations].
The significance of the emotional stress problem for persons of dangerous professions makes it necessary to search for the optimal procedure for emotional status correction. However, taking into account the specific features of military specialists' professional activity, it is highly difficult to choose a single drug. The authors apply a differential approach to solving the problem by considering the efficacy of tranquilizers, stimulants, adrenergic blocking agents, and adaptogens. The use of anxiolytics corrects an emotional response to the level which has marked adaptational sense. A combination of tranquilizers with stimulants is beneficial for the prevention or abortion of fatigue states. beta-adrenergic blockers act as stress protective agents in the cases when the implementation of highly coordinated skills is of crucial importance. By evoking physiological and biochemical changes in the body, adaptogens are involved in the elaboration of an adequate behavioral pattern and in adaptive rearrangement under the influence of unfavourable factors of the environment and activities.


Novel tau fragments in cerebrospinal fluid: relation to tangle pathology and cognitive decline in Alzheimer's disease.
Tau is an axonal microtubule-binding protein. Tau pathology in brain and increased tau concentration in the cerebrospinal fluid (CSF) are hallmarks of Alzheimer's disease (AD). Most of tau in CSF is present as fragments. We immunoprecipitated tau from CSF and identified several endogenous peptides ending at amino acid (aa) 123 or 224 using high-resolution mass spectrometry. We raised neo-epitope-specific antibodies against tau fragments specifically ending at aa 123 and 224, respectively. With these antibodies, we performed immunohistochemistry on brain tissue and designed immunoassays measuring N-123, N-224, and x-224 tau. Immunoassays were applied to soluble brain fractions from pathologically confirmed subjects (81 AD patients, 33 controls), CSF from three cross-sectional and two longitudinal cohorts (a total of 133 AD, 38 MCI, 20 MCI-AD, 31 PSP, 15 CBS patients, and 91 controls), and neuronally- and peripherally-derived extracellular vesicles (NDEVs and PDEVs, respectively) in serum from four AD patients and four controls. Anti-tau 224 antibody stained neurofibrillary tangles and neuropil threads, while anti-tau 123 only showed weak cytoplasmic staining in AD. N-224 tau was lower in the AD soluble brain fraction compared to controls, while N-123 tau showed similar levels. N-224 tau was higher in AD compared to controls in all CSF cohorts (p < 0.001), but not N-123 tau. Decrease in cognitive performance and conversion from MCI to AD were associated with increased baseline CSF levels of N-224 tau (p < 0.0001). N-224 tau concentrations in PSP and CBS were significantly lower than in AD (p < 0.0001) and did not correlate to t-tau and p-tau. In a longitudinal cohort, CSF N-224 tau levels were stable over 6 months, with no significant effect of treatment with AChE inhibitors. N-224 tau was present in NDEVs, while N-123 tau showed comparable concentrations in both vesicle types. We suggest that N-123 tau is produced both in CNS and PNS and represents a general marker of tau metabolism, while N-224 tau is neuron-specific, present in the tangles, secreted in CSF, and upregulated in AD, suggesting a link between tau cleavage and propagation, tangle pathology, and cognitive decline.


Semen parameter improvements after microsurgical subinguinal varicocele repair are durable for more than 12 months.
Varicoceles account for the most common correctable cause of male infertility, with varicocele repair leading to improvements in semen quality. However, there is little evidence to establish the durability of varicocele repair. We analyzed the durability of improvements in postoperative semen parameters following microsurgical subinguinal varicocele repair. We evaluated all men who underwent microscopic subinguinal varicocelectomy from 2015-2019. Patients were included if they desired fertility and had a followup of at least 12 months. We assessed the baseline characteristics of these patients, as well as semen volume, total motile sperm count (TMSC), concentration, percent motility, and morphology. Semen parameters were analyzed at baseline (preoperative) and at approximately three months and ≥12 months postoperatively. Of 105 men who underwent varicocelectomy, 18 men had a followup of at least 12 months. These men presented with median age of 34.5 (27-38) years for a median followup duration of 14.5 (13-22.5) months. TMSC levels increased from 6.4 (1.1-24.5) million at baseline to 11.1 (2.4-38.4) million at approximately three months and remained similar at 12.5 (1.6-31.5) million at ≥12 months. The study is limited by its retrospective nature and limited sample size. Microscopic subinguinal varicocele repairs can result in durable improvements of semen quality beyond one year, as demonstrated by upgrade in median TMSC. Further studies should be performed to confirm our findings.


Giant cell tumor of bone arising in long bones possibly originates from the metaphyseal region.
Giant cell tumor of bone (GCTB) is a primary benign bone tumor with a locally aggressive character. Definitive descriptions of the site of origin for this type of tumor are not available. The aim of the present study was to evaluate the site of origin of GCTB of long bones with regards to epiphyseal lines by means of radiographic examination. For that purpose, plain X-ray scans of 71 GCTBs arising in long bones were retrospectively reviewed. The tumor locations were the distal femur in 31 cases, proximal femur in 11 cases, proximal tibia in 13 cases, distal radius in 6 cases, proximal humerus in 5 cases and proximal fibula in 5 cases. The vertical center (VC) of the tumor was determined with X-ray anteroposterior view, and the correlation between the VC and the epiphyseal line, and between the distance from the epiphyseal line to the VC and tumor area or volume were analyzed using a regression model equation based on scatter plot diagrams. The VC of the tumor was located in the metaphyseal region in 57 cases, in the epiphyseal line in 11 cases and in the epiphyseal region in 3 cases. In cases of GCTB located in the distal femur or proximal tibia, significant correlations between the distance from the VC to the epiphyseal line and tumor area or volume were identified. The site of origin of GCTB was estimated to be located in the metaphyseal region. GCTB often occurs in mature patients, which renders it challenging to estimate the true site of origin of this lesion, since the metaphyseal line has disappeared in mature patients. The results of the present study suggest that GCTB possibly originates in the metaphyseal region.


Beyond Self-Protection: Self-Affirmation Benefits Hedonic and Eudaimonic Well-Being.
Limited work has examined how self-affirmation might lead to positive outcomes beyond the maintenance of a favorable self-image. To address this gap in the literature, we conducted two studies in two cultures to establish the benefits of self-affirmation for psychological well-being. In Study 1, South Korean participants who affirmed their values for 2 weeks showed increased eudaimonic well-being (need satisfaction, meaning, and flow) relative to control participants. In Study 2, U.S. participants performed a self-affirmation activity for 4 weeks. Extending Study 1, after 2 weeks, self-affirmation led both to increased eudaimonic well-being and hedonic well-being (affect balance). By 4 weeks, however, these effects were non-linear, and the increases in affect balance were only present for vulnerable participants-those initially low in eudaimonic well-being. In sum, the benefits of self-affirmation appear to extend beyond self-protection to include two types of well-being.


Breast cancer awareness of rural women in Malaysia: is it the same as in the cities?
Breast cancer is the most common cancer among women globally. This study was conducted to compare the awareness of breast cancer and the practice of breast self-examination (BSE), clinical breast examination (CBE) and mammography screening among rural females in Pahang and Perak. A cross-sectional study was carried out in five selected rural districts of Pahang and Perak. Two hundred and fifty households were randomly selected and interviewed face to face using a semi-structured questionnaire. The majority of residents from both states were Malay, aged between 50 and 60 years and had a secondary level of education. Malay women aged 40-49 years and women with a higher level of education were significantly more aware of breast cancer (p<0.05). About half of these women practiced BSE (60.7%) and CBE (56.1%), and 7% had underwent mammography screening. The results of this study suggest that women in Pahang and Perak have good awareness of breast cancer and that more than half practice BSE and CBE. The women's level of education appears to contribute to their level of knowledge and health behaviour. However, more effort is needed to encourage all women in rural areas to acquire further knowledge on breast cancer.


[Stereochemical enhancement of specificity in biological recognition systems].
Specificity of the reaction of equilibrium association as a model of recognition process has been analysed. It has been shown that subdividing the centres of recognition into two uncoupled groups may lead to the effect of stereochemical enhancement of specificity, i. e. an increase of the fraction of the correct complex. If the second group of centers is unspecific, then a compensation of mistakes on a specific group of centers by unspecific ones takes place. Stereochemical enhancement of specificity requires the presence in the recognition protein molecule of a mechanism of switching on - off interaction on a group of centers. The most universal method of such switching on - off is a conformational transition in the protein molecule and (or) the "substrate".


One-year outcome of hip fracture patients admitted to a Singapore hospital: quality of life post-treatment.
A prospective and consecutive documentation of hip fracture care was performed. Outcomes, including quality of life, mortality, complication rates, were documented; and mobility, ambulatory status, freedom from pain and activities of daily living one year before and after treatment, were compared. We prospectively reviewed the medical records of 70 consecutive patients admitted to the Singapore General Hospital, following either a cervical or intertrochanteric femoral fracture from February 2004 to May 2004. Patients' progress was reviewed at one year post-treatment, and the EuroQOL was used to quantify the patients' quality of life. Description of any problems encountered was also recorded. The follow-up rate at one year for the 70 patients described in this report was 98.6 percent. Mortality rate was 27.1 percent. Early complication rate was 5.7 percent. Outcome was satisfactory in all but two patients. Mortality for surgically-operated patients was 25.4 percent. About a quarter of the patients had excellent ambulatory status and 40.0 percent were able to walk independently. Eight percent suffered from falls after discharge, but no recurrence of hip fracture was recorded. None was re-admitted under suspicion of hip fracture. The average self-scoring system (EuroQOL) yielded an average of 66.6 out of 100. Hip fractures can be treated surgically with good results and low early complication rates, without drastically affecting patients' quality of life.


Conditional knockout of ITGB4 in bronchial epithelial cells directs bronchopulmonary dysplasia.
Neonatal respiratory system disease is closely associated with embryonic lung development. Our group found that integrin β4 (ITGB4) is downregulated in the airway epithelium of asthma patients. Asthma is the most common chronic respiratory illness in childhood. Therefore, we suspect whether the deletion of ITGB4 would affect fetal lung development. In this study, we characterized the role of ITGB4 deficiency in bronchopulmonary dysplasia (BPD). ITGB4 was conditionally knocked out in CCSP-rtTA, Tet-O-Cre and ITGB4f/f triple transgenic mice. Lung tissues at different developmental stages were collected for experimental detection and transcriptome sequencing. The effects of ITGB4 deficiency on lung branching morphogenesis were observed by fetal mouse lung explant culture. Deleting ITGB4 from the airway epithelial cells results in enlargement of alveolar airspaces, inhibition of branching, the abnormal structure of epithelium cells and the impairment of cilia growth during lung development. Scanning electron microscopy showed that the airway epithelial cilia of the β4ccsp.cre group appear to be sparse, shortened and lodging. Lung-development-relevant factors such as SftpC and SOX2 significantly decreased both mRNA and protein levels. KEGG pathway analysis indicated that multiple ontogenesis-regulating-relevant pathways converge to FAK. Accordingly, ITGB4 deletion decreased phospho-FAK, phospho-GSK3β and SOX2 levels, and the correspondingly contrary consequence was detected after treatment with GSK3β agonist (wortmannin). Airway branching defect of β4ccsp.cre mice lung explants was also partly recovered after wortmannin treatment. Airway epithelial-specific deletion of ITGB4 contributes to lung developmental defect, which could be achieved through the FAK/GSK3β/SOX2 signal pathway.


Pharmacology of sensory stimulation-evoked increases in frontal cortical acetylcholine release.
Recent research has demonstrated that a variety of sensory stimuli can increase acetylcholine release in the frontal cortex of rats. The aim of the present experiments was to investigate the pharmacological regulation of sensory stimulation-induced increases in the activity of basal forebrain cholinergic neurons. To this end, the effects of agonists and antagonists at a variety of neurotransmitter receptors on basal and tactile stimulation-evoked increases in frontal cortical acetylcholine release were studied using in vivo brain microdialysis. Tactile stimulation, produced by gently stroking the rat's neck with a nylon brush for 20 min, significantly increased frontal cortical acetylcholine release by more than 100% above baseline. The noradrenergic alpha2 agonist clonidine (0.1 or 0.2 mg/kg) and alpha1 antagonist prazosin (1 mg/kg) failed to affect basal cortical acetylcholine release; however, both compounds significantly reduced the increases evoked by sensory stimulation. In contrast, the alpha2 antagonist yohimbine (3 mg/kg) increased basal cortical acetylcholine release, thereby preventing meaningful investigation of its effects on tactile stimulation-evoked increases. The benzodiazepine agonist diazepam (5 mg/kg) reduced, and the GABA(A) receptor antagonist picrotoxin (2 mg/kg) increased basal cortical acetylcholine release; in addition, diazepam attenuated the increases in cortical acetylcholine release evoked by tactile stimulation. While dopaminergic D1 (SCH 23390, 0.15 mg/kg) and D2 (raclopride, 1 mg/kg) receptor antagonists did not by themselves significantly influence the increases evoked by tactile stimulation, their co-administration produced a significant reduction. The opioid receptor antagonist naltrexone (1.5 mg/kg) failed to affect either basal or tactile stimulation-evoked increases in acetylcholine overflow. Finally, the non-competitive N-methyl-D-aspartate receptor antagonist, dizocilpine maleate (MK-801; 0.025 and 0.05 mg/kg) increased basal cortical acetylcholine release. These results confirm that cortically projecting cholinergic neurons are activated by sensory stimuli, and indicate that the increases in cortical acetylcholine release produced by tactile stimulation are inhibited by stimulation of alpha2 or blockade of alpha1 noradrenergic receptors, and by enhanced GABAergic transmission. In addition, simultaneous blockade of dopamine D1 and D2 receptors appears necessary to achieve a significant reduction of sensory stimulation-evoked acetylcholine release in the frontal cortex. The results are consistent with the hypothesis that cortical acetylcholine release is a component of the neurochemistry of arousal and/or attention and indicate that this is modulated by GABAergic, noradrenergic and dopaminergic systems. In contrast, endogenous opioid actions do not appear to be involved.


Interactions of CD80 and CD86 with CD28 and CTLA4.
CD80 and CD86 are cell surface glycoproteins expressed on a variety of professional APCs. They have attracted much attention due to their function as potent costimulators of T lymphocyte function through their interaction with CD28 and possibly CTLA4. Because inhibitors of this interaction may have therapeutic relevance in human autoimmune disease, we investigated the properties of linear peptides derived from conserved regions of CTLA4 and CD80 known to be essential for binding. None of these peptides were sufficient to bind ligand, nor did they act as potent competitive inhibitors. Conformationally constrained versions of the CTLA4 motif were also inactive. These results suggested that other parts of the proteins are important in determining binding, so a series of modified CD80 and CD86 molecules were constructed in an attempt to identify other binding determinants. Insertion of two residues between the two Ig domains of CD80 resulted in decreased affinity for CTLA4, but a similar mutation in CD86 was without effect. We also identified another asymmetry between CD80 and CD86 in that the V domain of CD86 but not that of CD80 is sufficient for CTLA4 binding. The CD86-V domain appears to have CTLA4 binding properties equivalent to that of intact CD86. These data illustrate a fundamental difference between these costimulatory molecules and suggest a mechanism by which they may be differentially recognized by receptors on the T cell surface.


Cost effectiveness of screening for tuberculosis in a general medical clinic.
Over a 3-year period, only 22 new active cases of tuberculosis were discovered as a result of taking 48,000 70-mm X-ray films of generally elderly and chronically ill patients. The cases were among the patients of the general medical outpatient clinic of Grady Memorial Hospital in Atlanta, Ga. Costs for the X-ray screening program exceeded $90,000. Ten of the 22 patients with new cases were alcohol abusers. Further analysis of the program revealed a yield of 1 new active case per 240 alcoholics screened, but only 1 new active case per 3,806 nonalcoholic patients screened. It is argued that screening of alcoholics for tuberculosis should be widely practiced, but routine yearly X-rays for detection of tuberculosis--even in a chronically ill population--is unwarranted.


Bacterial community structures are unique and resilient in full-scale bioenergy systems.
Anaerobic digestion is the most successful bioenergy technology worldwide with, at its core, undefined microbial communities that have poorly understood dynamics. Here, we investigated the relationships of bacterial community structure (>400,000 16S rRNA gene sequences for 112 samples) with function (i.e., bioreactor performance) and environment (i.e., operating conditions) in a yearlong monthly time series of nine full-scale bioreactor facilities treating brewery wastewater (>20,000 measurements). Each of the nine facilities had a unique community structure with an unprecedented level of stability. Using machine learning, we identified a small subset of operational taxonomic units (OTUs; 145 out of 4,962), which predicted the location of the facility of origin for almost every sample (96.4% accuracy). Of these 145 OTUs, syntrophic bacteria were systematically overrepresented, demonstrating that syntrophs rebounded following disturbances. This indicates that resilience, rather than dynamic competition, played an important role in maintaining the necessary syntrophic populations. In addition, we explained the observed phylogenetic differences between all samples on the basis of a subset of environmental gradients (using constrained ordination) and found stronger relationships between community structure and its function rather than its environment. These relationships were strongest for two performance variables--methanogenic activity and substrate removal efficiency--both of which were also affected by microbial ecology because these variables were correlated with community evenness (at any given time) and variability in phylogenetic structure (over time), respectively. Thus, we quantified relationships between community structure and function, which opens the door to engineer communities with superior functions.


Immunomodulatory properties of stem cells from human exfoliated deciduous teeth.
Stem cells from human exfoliated deciduous teeth (SHED) have been identified as a population of postnatal stem cells capable of differentiating into osteogenic and odontogenic cells, adipogenic cells, and neural cells. Herein we have characterized mesenchymal stem cell properties of SHED in comparison to human bone marrow mesenchymal stem cells (BMMSCs). We used in vitro stem cell analysis approaches, including flow cytometry, inductive differentiation, telomerase activity, and Western blot analysis to assess multipotent differentiation of SHED and in vivo implantation to assess tissue regeneration of SHED. In addition, we utilized systemic SHED transplantation to treat systemic lupus erythematosus (SLE)-like MRL/lpr mice. We found that SHED are capable of differentiating into osteogenic and adipogenic cells, expressing mesenchymal surface molecules (STRO-1, CD146, SSEA4, CD73, CD105, and CD166), and activating multiple signaling pathways, including TGFbeta, ERK, Akt, Wnt, and PDGF. Recently, BMMSCs were shown to possess an immunomodulatory function that leads to successful therapies for immune diseases. We examined the immunomodulatory properties of SHED in comparison to BMMSCs and found that SHED had significant effects on inhibiting T helper 17 (Th17) cells in vitro. Moreover, we found that SHED transplantation is capable of effectively reversing SLE-associated disorders in MRL/lpr mice. At the cellular level, SHED transplantation elevated the ratio of regulatory T cells (Tregs) via Th17 cells. These data suggest that SHED are an accessible and feasible mesenchymal stem cell source for treating immune disorders like SLE.


An indolent case of isolated cerebral mucormycosis: an uncommon presentation.
This case is a presentation of isolated central nervous system (CNS) Mucormycosis in an immunocompetent patient. This case is unique in its demonstration of isolated CNS involvement while lacking clear evidence elucidating an entry point. The patient is a 36-year-old man without a pertinent past medical history, who initially presented with altered mental status and a 5-day history of progressively slurred speech. His social history is significant for intravenous drug use and outdoor pest control work. The patient's head computed tomography (CT) scan without contrast demonstrated the presence of possible bilateral infarcts or masses involving the basal ganglia and periventricular white matter. The patient then progressed to facial diplegia with new onset hemiplegia. High-dose steroids were initiated due to concern for neurosarcoidosis. A lumbar puncture was ordered due to minimal improvement and suggested an inflammatory process. A stereotactic brain biopsy was then performed, demonstrating non-caseating granulomatous inflammation with giant cells. Liposomal amphotericin B was added to cover possible fungal etiology. The pathology report was consistent with an isolated cerebral mucormycosis infection. The etiology remained elusive with clear paranasal sinuses and no cutaneous manifestations. Due to extensive gray matter involvement, the patient was not a candidate for surgery. This is a report of mucormycosis in a seemingly immunocompetent patient with either isolated CNS involvement or disseminated mucormycosis without an identifiable source. Although this patient did have two risk factors including intravenous drug use and outdoor working history, his lack of peripheral involvement demonstrates an uncommon presentation.


Effect of a competitive angiotensin antagonist on the renal haemodynamic changes induced by inhibition of prostaglandin synthesis in rats.
1. Inhibition of prostaglandin synthesis by indomethacin induced an increase in blood pressure which did not occur when rats were bilaterally nephrectomized. 2. The blood pressure effect was related to the state of sodium balance and thus to the activity of the renin--angiotensin system. 3. Indomethacin induced a decrease in renal blood flow. 4. Angiotensin receptor blockade with Sar1-Ala8-angiotensin II blunted the blood pressure effect and prevented the renal haemodynamic changes induced by indomethacin.


Evolution of basal deuterostome nervous systems.
Understanding the evolution of deuterostome nervous systems has been complicated by the by the ambiguous phylogenetic position of the Xenocoelomorpha (Xenoturbellids, acoel flat worms, nemertodermatids), which has been placed either as basal bilaterians, basal deuterostomes or as a sister group to the hemichordate/echinoderm clade (Ambulacraria), which is a sister group of the Chordata. None of these groups has a single longitudinal nerve cord and a brain. A further complication is that echinoderm nerve cords are not likely to be evolutionarily related to the chordate central nervous system. For hemichordates, opinion is divided as to whether either one or none of the two nerve cords is homologous to the chordate nerve cord. In chordates, opposition by two secreted signaling proteins, bone morphogenetic protein (BMP) and Nodal, regulates partitioning of the ectoderm into central and peripheral nervous systems. Similarly, in echinoderm larvae, opposition between BMP and Nodal positions the ciliary band and regulates its extent. The apparent loss of this opposition in hemichordates is, therefore, compatible with the scenario, suggested by Dawydoff over 65 years ago, that a true centralized nervous system was lost in hemichordates.


Construction of a ceRNA regulatory network to explore potential pathogenesis mechanisms involved in human hepatocellular carcinoma.
Worldwide, primary liver cancer is the third leading cause of cancer-related death. Hepatocellular carcinoma (HCC) accounts for the majority of primary liver cancers. Recent studies have shown that circular RNAs (circRNAs) that interact with microRNAs (miRNAs) are involved in the occurrence and development of various tumours. Transcriptional profile analysis was used to analyse expression of circRNAs in HCC in this study. The top ten upregulated circRNAs were selected and validated by quantitative reverse transcription polymerase chain reaction (qRT-PCR) in another 34 HCC patients. MiRNAs and mRNAs downstream of these circRNAs were explored through database analysis, and finally, the competitive endogenous RNA (ceRNA) networks were constructed for 5 selected circRNAs. We identified 9658 differentially expressed circRNAs by transcriptional profile analysis. QRT-PCR was performed to validate the top ten upregulated circRNAs, and five circRNAs were selected for further analysis. The miRNAs and mRNAs downstream of these five circRNAs were predicted to construct ceRNA network diagrams. Further analysis revealed five circRNA-miRNA-mRNA axes that correlate negatively with HCC prognosis. Numerous differentially expressed circRNAs exist in HCC, and they can regulate the biological behaviour of HCC through ceRNA networks. Bioinformatics analysis showed that ceRNA regulatory axes involved in HCC have high diagnostic and prognostic value and deserve further exploration.


Serological profile of asymptomatic HCV positive patients with low level of cryoglobulins.
Clinical spectrum of hepatitis C virus (HCV)-related cryoglobulinemia varies from an asymptomatic presentation to severe vasculitis and lymphoma. A recent study in HCV-negative patients suggests that low cryoglobulins (CGs) levels are responsible for severe renal and neurological complications. The aim of this study was to identify a panel of serological biomarkers associated with low levels of CGs in HCV-positive patients. We studied a population of 79 untreated patients with chronic HCV infection: 13 naïve patients without CGs; 28 patients with asymptomatic mixed cryoglobulinemia (MC) and low levels of CGs (16/28 with polyclonal type III and 12/28 with microheterogeneous type III CGs); 38 patients with symptomatic MC and high levels of type II CGs. Serum samples were collected and examined for rheumatoid factor (RF) IgG and IgM, free light chains (FLCs) and C3 and C4 complement components. We found that RF-IgG and IgM, free k chains and k+λ were increased while C4 component was reduced, both in symptomatic and asymptomatic patients. Our results suggest that, even in absence of MC symptoms, the low levels of CGs may represent a trigger of activation for immune system in course of HCV infection. The identification of a correlated biomarkers panel could improve the clinical management of these patients and pave the way for target treatment strategies. © 2018 BioFactors, 45(3):318-325, 2019.


Inhibition of human neuroblastoma by dopamine antagonists.
The effects of dopamine agonists and antagonists were investigated in human neuroblastoma (HNB) tissue culture cell lines and correlated with the presence of specific membrane-bound dopamine-binding activity ("receptor"). In four HNB cell lines the dopamine antagonists domperidone, pimozide, and spiroperidol inhibited macromolecular synthesis in vitro as indicated by decreased 3H-TdR and 14C-leu incorporation in a dose-response fashion with at least 50% inhibition noted at 10(-6)M concentration of each drug. Dopamine agonists showed no significant inhibition. Scatchard analysis of competitive dopamine-binding assays in all four HNB cell lines and in five of eight solid tumors obtained at surgery demonstrated high affinity, limited-capacity binding consistent with a single class of receptor sites with receptor concentrations (Rc) ranging from 8.8 to 26.7 pmol/gm wet weight of tissue with dissociation constants (KD) from 0.40 to 6.6 nmol/L, compared with a mean Rc of 28.1 +/- 5.2 pmol/gm wet weight of tissue and KD = 0.38 +/- 0.09 nmol/L in receptor-rich dog caudate nucleus, the normal dopamine-sensitive control. Survival was prolonged after inoculation of the SK-N-AS cell line into nude mice and subsequent domperidone administration by 50% (24 days after drug initiation versus 16 days in control mice). These data demonstrate inhibition of macromolecular synthesis in HNB by dopamine antagonists and suggest that dopamine receptor is associated with this inhibition. The determination of dopamine receptors may prove useful in the selection of dopamine antagonists as specific chemotherapy for patients with neuroblastoma.


Noise, sign problems, and statistics.
We show how sign problems in simulations of many-body systems can manifest themselves in the form of heavy-tailed correlator distributions, similar to what is seen in electron propagation through disordered media. We propose an alternative statistical approach for extracting ground state energies in such systems, illustrating the method with a toy model and with lattice data for unitary fermions.


Diltiazem attenuates oxidative stress in diabetic rats.
Diabetic nephropathy is the main cause of end stage renal damage. Oxidative stress is involved in the etiology of diabetic nephropathy and intracellular calcium is reported to play a considerable role in the development of renal damage in the diabetic kidney. Calcium antagonism can slow the progression of renal impairment in diabetes. The present study was thus designed to examine the effect of a nondihydropyridine calcium channel blocker, diltiazem, on renal function, oxidative stress, and nitric oxide (NO) release in streptozotocin (STZ)-induced diabetic rats. Diabetes was induced by a single intraperitoneal injection of STZ (65 mg/kg) in rats. After 4 weeks of STZ injection, the rats were divided in to four groups: control rats, diabetic rats treated with saline, and two groups of diabetic rats treated with diltiazem (5 and 10 mg/kg, i.p, respectively) for 8 weeks starting from 4 weeks after STZ injection. Renal function was assessed by creatinine, blood urea nitrogen, creatinine clearance, and urea clearance. Oxidative stress was measured by renal malondialdehyde (MDA), reduced glutathione (GSH), superoxide dismutase (SOD), and catalase. We also measured renal nitrite levels. At the end of the 8 weeks, diabetic rats exhibited renal dysfunction as evidenced by reduced creatinine and urea clearance along with enhanced albumin excretion rate as compared with control rats. Biochemical analysis of kidneys revealed a marked increase in oxidative stress demonstrated by increased lipid peroxidation and decreased activities of key antioxidant enzymes, GSH, SOD, and catalase in diabetic rats. Release of NO also significantly higher in diabetic rats than controls. Chronic treatment with diltiazem in diabetic rats significantly attenuated both renal dysfunction and oxidative stress along with increased NO levels as compared with untreated diabetic rats. The kidneys of diabetic rats showed morphological changes such as hyaline casts, glomerular thickening, and moderate interstitial fibrosis and arteriolopathy, whereas diltiazem administration markedly prevented diabetic-induced renal morphological alterations. The present study suggests that oxidative stress/nitrosative stress is increased in the diabetic kidney and calcium channel blockage can prevent these changes. The results also suggest that in STZ-induced diabetic rats, the protective action of diltiazem might be mediated, at least in part, by its effect on tissue oxidant/antioxidant status.


Antiepileptic drug interactions - principles and clinical implications.
Antiepileptic drugs (AEDs) are widely used as long-term adjunctive therapy or as monotherapy in epilepsy and other indications and consist of a group of drugs that are highly susceptible to drug interactions. The purpose of the present review is to focus upon clinically relevant interactions where AEDs are involved and especially on pharmacokinetic interactions. The older AEDs are susceptible to cause induction (carbamazepine, phenobarbital, phenytoin, primidone) or inhibition (valproic acid), resulting in a decrease or increase, respectively, in the serum concentration of other AEDs, as well as other drug classes (anticoagulants, oral contraceptives, antidepressants, antipsychotics, antimicrobal drugs, antineoplastic drugs, and immunosupressants). Conversely, the serum concentrations of AEDs may be increased by enzyme inhibitors among antidepressants and antipsychotics, antimicrobal drugs (as macrolides or isoniazid) and decreased by other mechanisms as induction, reduced absorption or excretion (as oral contraceptives, cimetidine, probenicid and antacides). Pharmacokinetic interactions involving newer AEDs include the enzyme inhibitors felbamate, rufinamide, and stiripentol and the inducers oxcarbazepine and topiramate. Lamotrigine is affected by these drugs, older AEDs and other drug classes as oral contraceptives. Individual AED interactions may be divided into three levels depending on the clinical consequences of alterations in serum concentrations. This approach may point to interactions of specific importance, although it should be implemented with caution, as it is not meant to oversimplify fact matters. Level 1 involves serious clinical consequences, and the combination should be avoided. Level 2 usually implies cautiousness and possible dosage adjustments, as the combination may not be possible to avoid. Level 3 refers to interactions where dosage adjustments are usually not necessary. Updated knowledge regarding drug interactions is important to predict the potential for harmful or lacking effects involving AEDs.


Calculating pH in pig manure taking into account ionic strength.
Models such as the Anaerobic Digestion Model No. 1 (ADM1) assume that pH can be calculated directly from the concentration of hydrogen ions. However because pH is, by definition, the negative logarithm of the hydrogen ion activity, and thus pH measurements represent hydrogen ion activities, this approach may lead to a bias between measured and predicted pH values. Implementing ionic strength effects into the charge balance equation and the calculation of pH is a theoretical improvement to this. In this study a model, implementing a procedure for calculating pH, was developed to analyse the effect of ionic strength on pH in a pig manure. By adding KCl to samples of pig manure, experimental results could be analysed with help from the model. A modified form of the Davies equation was found to give the most accurate prediction of pH in the pig manure studied in this paper with changing ionic strength.


Identification and characterization of the karrikins signaling gene SsSMAX1 in Sapium sebiferum.
SUPPRESSOR OF MAX2 LIKE 1 (SMAX1) is a member of the SUPPRESSOR of MAX2 1‑LIKE family of genes and is known as a target protein of KARRIKIN INSENSITIVE2 (KAI2)-MORE AXILLARY BRANCHES2 (MAX2), which mediates karrikin signaling in Arabidopsis. SMAX1 plays a significant role in seed germination, hypocotyl elongation, and root hair development in Arabidopsis. SMAX1 has not yet been identified and characterized in woody plants. This study identified and characterized SsSMAX1 in Sapium sebiferum and found that SsSMAX1 was highly expressed in the seed, hypocotyl, and root tips of S. sebiferum. SsSMAX1 was functionally characterized by ectopic expression in Arabidopsis. SsSMAX1 overexpression lines of Arabidopsis showed significantly delayed seed germination and produced seedlings with longer hypocotyl and roots than wild-type and Atsmax1 functional mutants. SsSMAX1 overexpression lines of Arabidopsis also had broader and longer leaves and petioles than wild-type and Atsmax1, suggesting that SsSMAX1 is functionally conserved. This study characterizes the SMAX1 gene in a woody and commercially valuable bioenergy plant, Sapium sebiferum. The results of this study are beneficial to future research on the molecular biology of woody plants.


VISTA is highly expressed on MDSCs and mediates an inhibition of T cell response in patients with AML.
Treatment of acute myeloid leukemia (AML) remains challenging. Enhancement of anti-tumor responses by blocking negative immune regulators is a promising strategy for novel effective leukemia therapeutics. V-domain Ig suppressor of T-cell activation (VISTA) is a recently defined negative regulator mediating immune evasion in cancer. To investigate the effect of VISTA on anti-leukemia immune response in AML, we initiated a study using clinical samples collected from AML patients. Here we report that VISTA is highly expressed on myeloid-derived suppressor cells (MDSCs) in the peripheral blood of AML patients. Both the frequency and intensity of VISTA expression on MDSCs are significantly higher in newly diagnosed AML than in healthy controls. Importantly knockdown of VISTA by specific siRNA potently reduced the MDSCs-mediated inhibition of CD8 T cell activity in AML, suggesting a suppressive effect of VISTA on anti-leukemia T cell response. Furthermore, we observed a strong positive association between MDSC expression of VISTA and T cell expression of PD-1 in AML. These results support the strategy of VISTA-targeted treatment for AML and underscore the strong potential for combined blockade of VISTA and PD-1 pathways in effective leukemia control.


Using genome wide association studies to identify common QTL regions in three different genetic backgrounds based on Iberian pig breed.
One of the major limitation for the application of QTL results in pig breeding and QTN identification has been the limited number of QTL effects validated in different animal material. The aim of the current work was to validate QTL regions through joint and specific genome wide association and haplotype analyses for growth, fatness and premier cut weights in three different genetic backgrounds, backcrosses based on Iberian pigs, which has a major role in the analysis due to its high productive relevance. The results revealed nine common QTL regions, three segregating in all three backcrosses on SSC1, 0-3 Mb, for body weight, on SSC2, 3-9 Mb, for loin bone-in weight, and on SSC7, 3 Mb, for shoulder weight, and six segregating in two of the three backcrosses, on SSC2, SSC4, SSC6 and SSC10 for backfat thickness, shoulder and ham weights. Besides, 18 QTL regions were specifically identified in one of the three backcrosses, five identified only in BC_LD, seven in BC_DU and six in BC_PI. Beyond identifying and validating QTL, candidate genes and gene variants within the most interesting regions have been explored using functional annotation, gene expression data and SNP identification from RNA-Seq data. The results allowed us to propose a promising list of candidate mutations, those identified in PDE10A, DHCR7, MFN2 and CCNY genes located within the common QTL regions and those identified near ssc-mir-103-1 considered PANK3 regulators to be further analysed.


Near-infrared fluorescein dyes containing a tricoordinate boron atom.
Bora-fluoresceins (BFs), fluorescein analogues containing a tricoordinate boron atom instead of an oxygen atom at the 10-position of the fluorescein skeleton, were synthesized as a new family of fluorescein analogues. The deprotonated BFs exhibited absorption and fluorescence in the near-infrared region, which were significantly red-shifted relative to those of hitherto-known heteroatom-substituted fluorescein analogues on account of the orbital interaction between the tricoordinate boron atom and the fluorescein skeleton. BFs also showed multi-stage changes resulting from a Lewis acid-base equilibrium at the boron center in combination with a Brønsted acid-base equilibrium at the phenolic hydroxy group.


Differentiation, Evaluation, and Application of Human Induced Pluripotent Stem Cell-Derived Endothelial Cells.
The emergence of induced pluripotent stem cell (iPSC) technology paves the way to generate large numbers of patient-specific endothelial cells (ECs) that can be potentially delivered for regenerative medicine in patients with cardiovascular disease. In the last decade, numerous protocols that differentiate EC from iPSC have been developed by many groups. In this review, we will discuss several common strategies that have been optimized for human iPSC-EC differentiation and subsequent studies that have evaluated the potential of human iPSC-EC as a cell therapy or as a tool in disease modeling. In addition, we will emphasize the importance of using in vivo vessel-forming ability and in vitro clonogenic colony-forming potential as a gold standard with which to evaluate the quality of human iPSC-EC derived from various protocols.


VACTERL/VATER Association.
VACTERL/VATER association is typically defined by the presence of at least three of the following congenital malformations: vertebral defects, anal atresia, cardiac defects, tracheo-esophageal fistula, renal anomalies, and limb abnormalities. In addition to these core component features, patients may also have other congenital anomalies. Although diagnostic criteria vary, the incidence is estimated at approximately 1 in 10,000 to 1 in 40,000 live-born infants. The condition is ascertained clinically by the presence of the above-mentioned malformations; importantly, there should be no clinical or laboratory-based evidence for the presence of one of the many similar conditions, as the differential diagnosis is relatively large. This differential diagnosis includes (but is not limited to) Baller-Gerold syndrome, CHARGE syndrome, Currarino syndrome, deletion 22q11.2 syndrome, Fanconi anemia, Feingold syndrome, Fryns syndrome, MURCS association, oculo-auriculo-vertebral syndrome, Opitz G/BBB syndrome, Pallister-Hall syndrome, Townes-Brocks syndrome, and VACTERL with hydrocephalus. Though there are hints regarding causation, the aetiology has been identified only in a small fraction of patients to date, likely due to factors such as a high degree of clinical and causal heterogeneity, the largely sporadic nature of the disorder, and the presence of many similar conditions. New genetic research methods offer promise that the causes of VACTERL association will be better defined in the relatively near future. Antenatal diagnosis can be challenging, as certain component features can be difficult to ascertain prior to birth. The management of patients with VACTERL/VATER association typically centers around surgical correction of the specific congenital anomalies (typically anal atresia, certain types of cardiac malformations, and/or tracheo-esophageal fistula) in the immediate postnatal period, followed by long-term medical management of sequelae of the congenital malformations. If optimal surgical correction is achievable, the prognosis can be relatively positive, though some patients will continue to be affected by their congenital malformations throughout life. Importantly, patients with VACTERL association do not tend to have neurocognitive impairment.


In vitro and in vivo activity of amphotericin B-lipid formulations against experimental Trypanosoma cruzi infections.
The activities of four amphotericin B formulations, Fungizone, AmBisome, Amphocil, and Abelcet, were compared in vitro and in vivo against Trypanosoma cruzi infections. In vitro, Fungizone and Amphocil were highly active against T. cruzi Y strain amastigotes in macrophages with 50% effective dose (ED50) values of 0.027-0.028 microg/ml, which were 7-fold and 42-fold more active than AmBisome and Abelcet, respectively. In vitro activities of all formulations against T. cruzi amastigotes in Vero cells were similar, with ED50 values in the range of 2.0-4.2 microg/ml. Acute infections of the T. cruzi Y strain in BALB/c mice were suppressed in all animals by a single 25 mg/kg dose of the liposomal formulation AmBisome. At the same dose, the two other lipid formulations, Amphocil and Abelcet, increased the survival rate but did not suppress infection in all animals.


[Tobacco and pregnancy in Denmark].
The risks of smoking during pregnancy are serious and well-documented. It is assumed that we do not identify all pregnant women with tobacco and nicotine consumption. Therefore, a new national Danish guideline presents a new screening and referral model for use from early pregnancy. All tobacco and nicotine consumption must be covered, including previous smoking and exposure to tobacco smoke. All pregnant women should be offered help to stop smoking or use of smokeless tobacco and help to avoid any exposure to tobacco smoke. Reference can be made to the municipality, Stoplinien and/or the hospital.


Hematologic toxicity of zidovudine in HIV-infected patients.
Zidovudine is now used extensively in an effort to control infection with the human immunodeficiency virus (HIV). The drug is associated with major hematologic toxicity, especially anemia and granulocytopenia. Conservative management of hematologic toxicity includes dosage reduction or cessation of therapy, diagnosis and treatment of chronic debilitating diseases, and supportive care, such as blood transfusions. New investigational agents, including hematopoietic growth factors, are being studied to combat the toxicities associated with zidovudine. The efficacy of these agents has yet to be established. Recent advances in drug efficacy at reduced dosage and in combination therapy promise to permit use of zidovudine with markedly reduced toxicity.


Immediate retrograde amnesia induced by midazolam: a prospective, nonrandomized cohort study.
Midazolam, a short-acting benzodiazepine, has sedative, anxiolytic, amnestic, and anticonvulsant effect. Given its advantages of rapid onset, short duration, and low toxicity, midazolam is optimal for any procedural sedation. Midazolam is known to cause anterograde amnesia; however, the possibility of retrograde amnesia has also been raised. This prospective cohort, nonrandomized study evaluated the presence and extent of retrograde amnesia induced by midazolam during cesarean delivery. One-hundred parturients scheduled for elective cesarean delivery under spinal anesthesia were enrolled. As soon as giving birth, 6 picture cards were shown to the patients in one min intervals, and then midazolam (0.1 mg/kg) was given or not according to the patients' preference. This overall retrograde recall rate of six cards was the primary outcome of our study, which was asked by a blinded investigator. The overall retrograde card recall rate was lower in the midazolam group compared to the control group (77.0 ± 13.4 vs. 87.7 ± 3.9%, p <0.001), especially at one min before midazolam administration (58 vs. 88%, p <0.001). Decreased memory trend was observed as time progressed toward midazolam administration in the midazolam group (p = 0.035). More patients answered 'yes' to the factitious event in the midazolam group than in the control group (26% vs. 4%, p = 0.004). Intravenous midazolam could cause a brief-period retrograde amnesia in visual and event memory. Moreover, there were more spurious reports of intraoperative factitious events in the midazolam group, implying that episodic memories were also affected by midazolam.


Does Route of Full Feeding Affect Outcome among Ventilated Critically Ill COVID-19 Patients: A Prospective Observational Study.
The outbreak of the new coronavirus strain SARS-CoV-2 (COVID-19) highlighted the need for appropriate feeding practices among critically ill patients admitted to the intensive care unit (ICU). This study aimed to describe feeding practices of intubated COVID-19 patients during their second week of hospitalization in the First Department of Critical Care Medicine, Evaggelismos General Hospital, and evaluate potential associations with all cause 30-day mortality, length of hospital stay, and duration of mechanical ventilation. We enrolled adult intubated COVID-19 patients admitted to the ICU between September 2020 and July 2021 and prospectively monitored until their hospital discharge. Of the 162 patients analyzed (52.8% men, 51.6% overweight/obese, mean age 63.2 ± 11.9 years), 27.2% of patients used parenteral nutrition, while the rest were fed enterally. By 30 days, 34.2% of the patients in the parenteral group had died compared to 32.7% of the patients in the enteral group (relative risk (RR) for the group receiving enteral nutrition = 0.97, 95% confidence interval = 0.88-1.06, p = 0.120). Those in the enteral group demonstrated a lower duration of hospital stay (RR = 0.91, 95% CI = 0.85-0.97, p = 0.036) as well as mechanical ventilation support (RR = 0.94, 95% CI = 0.89-0.99, p = 0.043). Enteral feeding during second week of ICU hospitalization may be associated with a shorter duration of hospitalization and stay in mechanical ventilation support among critically ill intubated patients with COVID-19.


Machine Learning Enhances the Performance of Bioreceptor-Free Biosensors.
Since their inception, biosensors have frequently employed simple regression models to calculate analyte composition based on the biosensor's signal magnitude. Traditionally, bioreceptors provide excellent sensitivity and specificity to the biosensor. Increasingly, however, bioreceptor-free biosensors have been developed for a wide range of applications. Without a bioreceptor, maintaining strong specificity and a low limit of detection have become the major challenge. Machine learning (ML) has been introduced to improve the performance of these biosensors, effectively replacing the bioreceptor with modeling to gain specificity. Here, we present how ML has been used to enhance the performance of these bioreceptor-free biosensors. Particularly, we discuss how ML has been used for imaging, Enose and Etongue, and surface-enhanced Raman spectroscopy (SERS) biosensors. Notably, principal component analysis (PCA) combined with support vector machine (SVM) and various artificial neural network (ANN) algorithms have shown outstanding performance in a variety of tasks. We anticipate that ML will continue to improve the performance of bioreceptor-free biosensors, especially with the prospects of sharing trained models and cloud computing for mobile computation. To facilitate this, the biosensing community would benefit from increased contributions to open-access data repositories for biosensor data.


Tumor overgrowth after expandable metallic stent placement: experience in 583 patients with malignant gastroduodenal obstruction.
The objective of our study was to assess the incidence, predictive factors, and treatment of tumor overgrowth after placement of expandable metallic dual stents in patients with malignant gastroduodenal obstruction. Expandable metallic dual stents were inserted under fluoroscopic guidance in 583 patients with symptomatic malignant gastroduodenal obstruction. We retrospectively reviewed prospectively collected patient records to determine the incidence and treatment of tumor overgrowth after stent placement and used multivariate analysis to determine factors predicting tumor overgrowth. Tumor overgrowth occurred after stent placement in 22 of 583 patients (3.8%) (range, 41-634 days; mean, 179.0 days). Duodenal lesions (odds ratio [OR], 4.505; p = 0.002), longer survival time (OR, 1.003; p = 0.001), and length of obstruction (OR, 0.783; p = 0.035) were independent predictors of tumor overgrowth. Twenty of the 22 patients were successfully treated by placement of a second dual stent, whereas the other two patients refused placement of a second stent or other further treatment. Overall, 19 of 20 patients (95%) showed improvement in symptoms after second stent placement. Duodenal perforation occurred in one of the 20 patients 125 days after placement of a second stent and was treated surgically. Tumor overgrowth seems to be an uncommon complication of expandable metallic dual stent placement in patients with malignant gastroduodenal obstruction. Tumor overgrowth is associated with duodenal lesions, longer survival time, and shorter stricture length. Tumor overgrowth can be successfully managed by coaxial insertion of a second dual-expandable metallic stent into the obstructed first stent.


NEI-Supported Age-Related Macular Degeneration Research: Past, Present, and Future.
To review past and current National Eye Institute (NEI)-supported age-related macular degeneration (AMD) activities and initiatives and preview upcoming coordinated efforts for studying AMD. We conducted and summarized a portfolio analysis and literature review of NEI intramural and extramural AMD activities. The NEI supports a broad range of AMD research, both by individual independent investigators as well as through networks and consortia. The International AMD Genomics Consortium, Age-Related Eye Disease Study, Age-Related Eye Disease Study 2 (AREDS2), and Comparison of AMD Treatments Trial legacy work probed the complex genetics, clinical presentation, and standards of patient care, respectively. The NEI AMD Pathobiology Working Group identified gaps and opportunities for future research efforts. The AMD Ryan Initiative Study and clinical trials testing the efficacies of minocycline to modulate retinal microglia activity and induced pluripotent stem cells-derived retinal pigmented epithelium (RPE) patch implants to rescue photoreceptor cell death are among the future directions for NEI-supported AMD research. Finally, NEI commissioned the creation of AREDS2 participant-derived induced pluripotent stem cell (iPSC) lines linked to their associated genomic and phenotypic datasets. These datasets will also be linked to the data obtained using their associated iPSC-derived cells (RPE, retina, choroid) and made publicly available. Investments by NEI for AMD research will continue to provide invaluable resources to investigators committed to addressing this complex blinding disease and other retinal degenerative diseases. NEI now stands poised to expand the resources available to clinical investigators to uncover disease mechanisms and move experimental therapies into clinical trials.


Isocitrate dehydrogenase is important for nitrosative stress resistance in Cryptococcus neoformans, but oxidative stress resistance is not dependent on glucose-6-phosphate dehydrogenase.
The opportunistic intracellular fungal pathogen Cryptococcus neoformans depends on many antioxidant and denitrosylating proteins and pathways for virulence in the immunocompromised host. These include the glutathione and thioredoxin pathways, thiol peroxidase, cytochrome c peroxidase, and flavohemoglobin denitrosylase. All of these ultimately depend on NADPH for either catalytic activity or maintenance of a reduced, functional form. The need for NADPH during oxidative stress is well established in many systems, but a role in resistance to nitrosative stress has not been as well characterized. In this study we investigated the roles of two sources of NADPH, glucose-6-phosphate dehydrogenase (Zwf1) and NADP(+)-dependent isocitrate dehydrogenase (Idp1), in production of NADPH and resistance to oxidative and nitrosative stress. Deletion of ZWF1 in C. neoformans did not result in an oxidative stress sensitivity phenotype or changes in the amount of NADPH produced during oxidative stress compared to those for the wild type. Deletion of IDP1 resulted in greater sensitivity to nitrosative stress than to oxidative stress. The amount of NADPH increased 2-fold over that in the wild type during nitrosative stress, and yet the idp1Delta strain accumulated more mitochondrial damage than the wild type during nitrosative stress. This is the first report of the importance of Idp1 and NADPH for nitrosative stress resistance.


Nurturing care among adolescent mothers.
Many studies show the impact of adolescent mothers on child development. To prevent the impact of adolescent pregnancy, nurturing care is needed to support optimal children's development. This study aims to identify comprehensive nurturing care among adolescent mothers. This study used secondary data in the integration of the 2018 National Socioeconomic Survey and Basic Health Survey. To measure nurturing care, 5 components were used: health, adequate nutrition, security and safety, responsive caregiving, and opportunities for early learning with latent class analysis. The results of the analysis showed that 62% of adolescent mothers provided uncompleted nurturing care. About 21% of adolescent mothers need assistance in improving nurturing care related to adequate nutrition and learning opportunities, and 17% need assistance to improve health, nutrition, responsive caregiving, and opportunities for early learning. Adolescent mothers are in need of comprehensive nurturing care for their children. Practice is needed in order to enhance nurturing care initiatives, particularly for teenage mothers.


Effects of alcohol intake on plasma fatty acids assessed independently of diet and smoking habits.
1. This study was designed to determine prospectively whether changing alcohol consumption influenced the proportion of plasma linoleic acid independently of diet or smoking habits, and to evaluate changes in the plasma linoleic acid concentration as a potential marker of changes in alcohol consumption. 2. Fasting plasma fatty acid profiles were investigated in 72 male drinkers who were randomly assigned to drink low-alcohol beer or to maintain their usual drinking habits for a period of 4 weeks. 3. At entry to the study, a higher alcohol intake was associated with lower proportions of plasma linoleic acid, arachidonic acid and dihomolinolenic acid and higher proportions of plasma palmitoleic acid, independently of changes in body mass index. Smoking habits were unchanged and there were no major changes in diet during the period of the intervention. 4. Because the plasma palmitoleic acid concentration has been suggested as a possible marker of 'at risk' drinking, we investigated plasma fatty acid concentrations as indicators of alcohol intake. The plasma palmitoleic acid concentration was not a useful discriminator. Indices determined using logistic regression and combining plasma apolipoprotein A-II and linoleic acid concentrations gave better discrimination than either variable alone.


The SR Splicing Factors: Providing Perspectives on Their Evolution, Expression, Alternative Splicing, and Function in Populus trichocarpa.
Serine/arginine-rich (SR) proteins are important splicing factors in plant development and abiotic/hormone-related stresses. However, evidence that SR proteins contribute to the process in woody plants has been lacking. Using phylogenetics, gene synteny, transgenic experiments, and RNA-seq analysis, we identified 24 PtSR genes and explored their evolution, expression, and function in Popolus trichocarpa. The PtSR genes were divided into six subfamilies, generated by at least two events of genome triplication and duplication. Notably, they were constitutively expressed in roots, stems, and leaves, demonstrating their fundamental role in P. trichocarpa. Additionally, most PtSR genes (~83%) responded to at least one stress (cold, drought, salt, SA, MeJA, or ABA), and, especially, cold stress induced a dramatic perturbation in the expression and/or alternative splicing (AS) of 18 PtSR genes (~75%). Evidentially, the overexpression of PtSCL30 in Arabidopsis decreased freezing tolerance, which probably resulted from AS changes of the genes (e.g., ICE2 and COR15A) critical for cold tolerance. Moreover, the transgenic plants were salt-hypersensitive at the germination stage. These indicate that PtSCL30 may act as a negative regulator under cold and salt stress. Altogether, this study sheds light on the evolution, expression, and AS of PtSR genes, and the functional mechanisms of PtSCL30 in woody plants.


Comparative study of vitrectomy versus intravitreous triamcinolone for diabetic macular edema on randomized paired-eyes.
The present study was performed to compare the effects of pars plana vitrectomy (PPV) and single intravitreaous triamcinolone acetonide (IVTA) on diabetic macular edema (DME) in paired eyes. Prospective comparative study on randomized paired-eyes was carried out at two hospitals. Forty eyes of 20 patients with bilateral DME were included. One randomly-selected eye was treated with PPV (PPV group), and the other eye was treated with IVTA (4 mg, IVTA group). The central macular thickness (CMT) measured by optical coherence tomography (OCT) and best-corrected visual acuity (BCVA) were monitored for 12 months after treatment. Changes from baseline and differences between groups were analyzed using a mixed model. At 1 and 3 months, CMT decreased significantly in the IVTA group compared to baseline (p < 0.0001 both), but CMT then increased gradually and no significant difference was found at 12 months (p = 0.90). In the PPV group, CMT decreased continuously and reached a significant level at 12 months (p < 0.0001). CMT of the IVTA group was significantly less than that of the PPV group at 1 month (p = 0.009); however, there was no significant difference at 3 months. Conversely, CMT was significantly less in the PPV group than in the IVTA group at 12 months (p = 0.0003). The changes of BCVA paralleled those of CMT, but no significant difference was detected between baseline BCVA and any time point. Despite the short-term improvement, DME recurred 6 months after IVTA, while it remained resolved after PPV. Although this study did not reveal a significant change of BCVA with either treatment, PPV resolved DME more effectively than IVTA at 1 year.


[Protective effect of caffeine on chromosomal structures of mammalian cells irradiated with UV-rays].
The effect of caffeine (2mM) on the frequency of structural mutations induced by UV light (lambda = 265 nm at an incident dose of 40 erg/mm2) in the primary culture of mouse embryonic fibroblasts is studied. A half-hour treatment with caffeine of cells at the time of the first mitosis metaphase decreased approximately by 2 times the frequency of chromosome aberrations induced by UV light at the S stage and observed at the metaphase of this or the next C-mitosis. The frequency of both breaks and exchanges decreased as the result of caffeine treatment. The persistence of the protective effect of caffeine at the time of the second C-mitosis suggests that the observed decrease of the aberration rate is accompanied by the true reparation of pre-mutational lesions in chromosomes; the nature of the reparative process and the time when it takes place is as yet not clear. Caffeine did not decrease the frequency of spontaneous structural mutations.


The Benefits of MRI in the Prenatal Diagnosis of Congenital Diaphragmatic Hernia.
Aims: We searched for correlations between ultrasound findings in pregnancies with congenital diaphragmatic hernia (CDH) and magnetic resonance imaging (MRI) follow-up examinations; MRI was used to confirm and complete the investigation in these difficult cases. In some of them, new elements that ultrasound was not able to fully describe have been also brought. We were especially interested when MRI was superior to ultrasound. Material and methods: This is a retrospective study of 12 pregnancies with congenital diaphragmatic hernia that were diagnosed in two major university clinics of Bucharest, Romania. Ultrasounds and MRI examinations were performed to evaluate pulmonary hypoplasia and correctly asses the herniated organs. We used standard international protocols and guidelines for calculating different parameters. All patients signed an informed consent before being enrolled in the study. Results: We described the herniated organs, dimensions of the hernia and the remaining lung capacity, so that we could correctly evaluate the prognosis. We have also used the lung to head ratio (LHR) in an attempt to better determine the degree of lung hypoplasia. Conclusion: High quality ultrasound followed by an MRI examination helped correctly assess the prognostic, treatment possibilities and total affected lung volume. It not only confirmed the diagnosis, but also offered new information that ultrasound was not able to provide.


Co-administration with Men-B vaccine increases Rotavirus vaccination coverage: A 5-year regionwide retrospective cohort study (STORM study).
In Italy Rotavirus vaccination (RVV) is provided free of charge from 2018, however, the coverage is scattered and suboptimal. The narrow time frame to complete the schedule is a barrier to uptake, and co-administration with other vaccines may potentially increase the coverage. Although the co-administration of RV vaccine and Meningococcal Group B vaccine (MenB) is not included in the product labels, we aimed at studying its impact on RVV coverage. This Surveillance study on Timing and cOverage of Rotavirus and MenB vaccine co-administration (STORM study) used the Regional Vaccination Registry to collect data about children born in Campania Region between January 2016 and December 2020, and receiving vaccines scheduled in the first year of life. Among the 224,110 children enrolled, 60,614 (27.0%) completed the RVV schedule, with a vaccination rate that increased over time from 1.15% in 2016 to 56.92% in 2020. The first and last dose of RVV schedule were administered beyond the recommended time in 6% of the study population, respectively. Co-administration of RV vaccine with MenB vaccine increased from 0.7 % in 2016 to 46.85 % in 2020. Children receiving RV/MenB vaccines concomitantly had a significantly higher chance of completing the RV schedule compared to those receiving RVV alone during a specific appointment (94.78 % vs 72.26 %, Prevalence Ratio -PR- 1.275, 95 %CI 1.245-1.295p < 0.00001). The positive driving effect of RV/MenB co-administration was more evident for children receiving pentavalent (PR 1.288) than monovalent RVV (PR 1.115) which was confirmed when adjusted for confounding variables (i.e., year of vaccination, local district, gender). Although still far from the target, RVV coverage has increased in recent years in Campania Region. Co-administration with MenB vaccine may aid in increasing RVV coverage, especially for pentavalent RVV. Further safety data are needed to support co-administration as a key tool to increase coverage.


Wnt Signaling Inhibits Osteoclast Differentiation by Activating Canonical and Noncanonical cAMP/PKA Pathways.
Although there has been extensive characterization of the Wnt signaling pathway in the osteoblast lineage, the effects of Wnt proteins on the osteoclast lineage are less well studied. We found that osteoclast lineage cells express canonical Wnt receptors. Wnt3a reduced osteoclast formation when applied to early bone-marrow macrophage (BMM) osteoclast differentiation cultures, whereas late addition did not suppress osteoclast formation. Early Wnt3a treatment inactivated the crucial transcription factor NFATc1 in osteoclast progenitors. Wnt3a led to the accumulation of nuclear β-catenin, confirming activation of canonical Wnt signaling. Reducing low-density lipoprotein receptor-related proteins (Lrp) 5 and Lrp6 protein expression prevented Wnt3a-induced inactivation of NFATc1; however, deletion of β-catenin did not block Wnt3a inactivation of NFATc1, suggesting that this effect was mediated by a noncanonical pathway. Wnt3a rapidly activated the cyclic adenosine monophosphate (cAMP)/protein kinase A (PKA) pathway and pharmacological stimulation of cAMP/PKA signaling suppressed osteoclast differentiation; Wnt3a-induced NFATc1 phosphorylation was blocked by inhibiting interactions between PKA and A-kinase anchoring proteins (AKAPs). These data indicate that Wnt3a directly suppresses osteoclast differentiation through both canonical (β-catenin) and noncanonical (cAMP/PKA) pathways in osteoclast precursors. In vivo reduction of Lrp5 and Lrp6 expressions in the early osteoclast lineage via Rank promoter Cre recombination reduced trabecular bone mass, whereas disruption of Lrp5/6 expression in late osteoclast precursors via cathepsin K (Ctsk) promoter Cre recombination did not alter the skeletal phenotype. Surprisingly, reduction of Lrp5/6 in the early osteoclast lineage decreased osteoclast numbers, as well as osteoblast numbers. Published studies have previously noted that β-catenin signaling is required for osteoclast progenitor proliferation. Our in vivo data suggest that Rank promoter Cre-mediated deletion of Lrp5/6 may similarly impair osteoclast progenitor proliferation.


PERMANENT GENETIC RESOURCES: Isolation of microsatellite loci from the kelp, Saccorhiza polyschides (Heterokontophyta, incertae sedis).
Kelps are ecologically important seaweeds that dominate the subtidal zones of rocky coasts. In Northern Europe, Saccorhiza polyschides is a pioneer species suspected of outcompeting the harvested kelp, Laminaria digitata. To examine how the process of species competition affects species distribution and genetic diversity in coastal environments, we developed 10 polymorphic microsatellite markers for S. polyschides using an enriched library (microsatellites are already available for L. digitata). These loci showed from three to 24 alleles with heterozygosities ranging from 0.36 to 0.92. This polymorphism is high enough for fine-scale population analyses including assignment tests to determine the origin of recruits.


Social Interaction Affects Neural Outcomes of Sign Language Learning As a Foreign Language in Adults.
Children naturally acquire a language in social contexts where they interact with their caregivers. Indeed, research shows that social interaction facilitates lexical and phonological development at the early stages of child language acquisition. It is not clear, however, whether the relationship between social interaction and learning applies to adult second language acquisition of syntactic rules. Does learning second language syntactic rules through social interactions with a native speaker or without such interactions impact behavior and the brain? The current study aims to answer this question. Adult Japanese participants learned a new foreign language, Japanese sign language (JSL), either through a native deaf signer or via DVDs. Neural correlates of acquiring new linguistic knowledge were investigated using functional magnetic resonance imaging (fMRI). The participants in each group were indistinguishable in terms of their behavioral data after the instruction. The fMRI data, however, revealed significant differences in the neural activities between two groups. Significant activations in the left inferior frontal gyrus (IFG) were found for the participants who learned JSL through interactions with the native signer. In contrast, no cortical activation change in the left IFG was found for the group who experienced the same visual input for the same duration via the DVD presentation. Given that the left IFG is involved in the syntactic processing of language, spoken or signed, learning through social interactions resulted in an fMRI signature typical of native speakers: activation of the left IFG. Thus, broadly speaking, availability of communicative interaction is necessary for second language acquisition and this results in observed changes in the brain.


Functional near-infrared-spectroscopy-based measurement of changes in cortical activity in macaques during post-infarct recovery of manual dexterity.
Because compensatory changes in brain activity underlie functional recovery after brain damage, monitoring of these changes will help to improve rehabilitation effectiveness. Functional near-infrared spectroscopy (fNIRS) has the potential to measure brain activity in freely moving subjects. We recently established a macaque model of internal capsule infarcts and an fNIRS system for use in the monkey brain. Here, we used these systems to study motor recovery in two macaques, for which focal infarcts of different sizes were induced in the posterior limb of the internal capsule. Immediately after the injection, flaccid paralysis was observed in the hand contralateral to the injected hemisphere. Thereafter, dexterous hand movements gradually recovered over months. After movement recovery, task-evoked hemodynamic responses increased in the ventral premotor cortex (PMv). The response in the PMv of the infarcted (i.e., ipsilesional) hemisphere increased in the monkey that had received less damage. In contrast, the PMv of the non-infarcted (contralesional) hemisphere was recruited in the monkey with more damage. A pharmacological inactivation experiment with muscimol suggested the involvement of these areas in dexterous hand movements during recovery. These results indicate that fNIRS can be used to evaluate brain activity changes crucial for functional recovery after brain damage.


Genomics and peptidomics of neuropeptides and protein hormones present in the parasitic wasp Nasonia vitripennis.
Neuropeptides and protein hormones constitute a very important group of signaling molecules, regulating central physiological processes such as reproduction, development, and behavior. Using a bioinformatics approach, we screened the recently sequenced genome of the parasitic wasp, Nasonia vitripennis, for the presence of these signaling molecules and annotated 30 precursor genes encoding 51 different mature neuropeptides or protein hormones. Twenty-four of the predicted mature Nasonia neuropeptides could be experimentally confirmed by mass spectrometry. We also discovered a completely novel neuropeptide gene in Nasonia, coding for peptides containing the C-terminal sequence RYamide. This gene has orthologs in nearly all arthropods with a sequenced genome, and its expression in mosquitoes was confirmed by mass spectrometry. No precursor could be identified for N-terminally extended FMRFamides, even though their putative G protein coupled receptor (GPCR) is present in the Nasonia genome. Neither the precursor nor the putative receptor could be identified for allatostatin-B, capa, the glycoprotein hormones GPA2/GPB5, kinin, proctolin, sex peptide, and sulfakinin, arguing that these signaling systems are truly absent in the wasp. Also, antidiuretic factors, allatotropin, and NPLP-like precursors are missing in Nasonia, but here the receptors have not been identified in any insect, so far. Nasonia (Hymenoptera) has the lowest number of neuropeptide precursor genes compared to Drosophila melanogaster, Aedes aegypti (both Diptera), Bombyx mori (Lepidoptera), Tribolium castaneum (Coleoptera), Apis mellifera (Hymenoptera), and Acyrthosiphon pisum (Hemiptera). This lower number of neuropeptide genes might be related to Nasonia's parasitic life.


Polarus nail fixation for proximal humeral fractures.
PURPOSE. To report the outcomes of Polarus nail fixation for displaced proximal humeral fractures. METHODS. Medical records of 19 men and 27 women aged 26 to 84 (mean, 63) years who underwent fixation using a standard Polarus nail (n=34) or Polarus plus nail with shaft extension (n=12) for proximal humeral fractures of 2 and 3 parts (n=34) and 2-part with extension to the shaft (n=12) were reviewed. Fracture union, neck shaft angle, and hardware complications were evaluated using radiographs. Outcomes were assessed using the Constant score, the visual analogue scale for pain, and subjective satisfaction. The range of motion, grip strength, and ability to return to work were also recorded. RESULTS. The mean follow-up was 20 (range, 15-28) months. The mean humeral neck shaft angle was 137.2º. 44 (96%) patients achieved fracture union after a mean of 14 (range, 11-16) weeks. Two patients with 3-part fractures did not unite; one of them underwent hemiarthroplasty and the other consented for nail removal only. Complications included temporary radial nerve palsy (n=2), impingement symptoms caused by the prominent metal work (n=10), and missing of the proximal screws in the nail (n=4). Respectively in patients aged <60 (n=15) and ≥ 60 years (n=31), the mean Constant scores were 79 and 67 (p=0.04). The mean pain score improved from 93 to 15.5 at 12 months. 16 patients were very satisfied, 22 were satisfied, and 8 were not satisfied with the outcome. Of the latter, 3 were aged <60 years and 5 were aged ≥ 60 years. CONCLUSION. Polarus nail fixation for displaced proximal humeral fractures achieved good fracture union and satisfactory functional outcome, particularly in younger patients and those in active employment, but elderly dependent patients tended to have poor functional results. The rates of minor hardware problems and non-union in 3-part fractures were relatively high.