Influence of top-down and bottom-up manipulations on the R-BT065 subcluster of beta-proteobacteria, an abundant group in bacterioplankton of a freshwater reservoir.
We studied the effects of nutrient availability and protistan grazing on bacterial dynamics and community composition (BCC) in different parts of the canyon-shaped Rímov reservoir (Czech Republic). The effects of protistan grazing on BCC were examined using a size fractionation approach. Water from the dam area with only bacteria (<0.8 microm), bacteria and heterotrophic nanoflagellates (<5 microm), or whole water were incubated in situ inside dialysis bags. Top-down or predator manipulations (size fractionation) were also combined with bottom-up or resource manipulations, i.e., transplantation of samples to the middle and upper inflow parts of the reservoir with increased phosphorus availability. Significant genotypic shifts in BCC occurred with transplantation as indicated by denaturing gradient gel electrophoresis. Using different probes for fluorescence in situ hybridization, we found that 10 to 50% of total bacteria were members of the phylogenetically small cluster of beta-proteobacteria (targeted with the probe R-BT065). These rod-shaped cells of very uniform size were vulnerable to predation but very fast growing and responded markedly to the different experimental manipulations. In all the grazer-free treatments, the members of the R-BT065 cluster showed the highest net growth rates of all studied bacterial groups. Moreover, their relative abundance was highly correlated with bacterial bulk parameters and proportions of bacteria with high nucleic acid (HNA) content. In contrast, increasing protistan bacterivory yielded lower proportions of R-BT065-positive and HNA bacteria substituted by increasing proportions of the class Actinobacteria, which profited from the enhanced protistan bacterivory.


RIOK3 potentially regulates osteogenesis-related pathways in ankylosing spondylitis and the differentiation of bone marrow mesenchymal stem cells.
RNA-binding proteins (RBPs), which are key effectors of gene expression, play critical roles in inflammation and immune regulation. However, the potential biological function of RBPs in ankylosing spondylitis (AS) remains unclear. We identified differentially expressed genes (DEGs) in peripheral blood mononuclear cells (PBMCs) of five patients with AS and three healthy persons by RNA-seq, obtained differentially expressed RBPs by overlapping DEGs and RBPs summary table. RIOK3 was selected as a target RBP and knocked down in mouse bone marrow mesenchymal stem cells (mBMSCs), and transcriptomic studies of siRIOK3 mBMSCs were performed again using RNA-seq. Results showed that RIOK3 knockdown inhibited the expression of genes related to osteogenic differentiation, ribosome function, and β-interferon pathways in mBMSCs. In vitro experiments have shown that RIOK3 knockdown reduced the osteogenic differentiation ability of mBMSCs. Collectively, RIOK3 may affect the differentiation of mBMSCs and participate in the pathogenesis of AS, especially pathological bone formation.


Value of procalcitonin, C-reactive protein, and neopterin in exacerbations of chronic obstructive pulmonary disease.
The identification of biological markers in order to assess different aspects of COPD is an area of growing interest. The objective of this study was to investigate whether levels of procalcitonin (PCT), C-reactive protein (CRP), and neopterin in COPD patients could be useful in identifying the etiological origin of the exacerbation and assessing its prognosis. We included 318 consecutive COPD patients: 46 in a stable phase, 217 undergoing an exacerbation, and 55 with pneumonia. A serum sample was collected from each patient at the time of being included in the study. A second sample was also collected 1 month later from 23 patients in the exacerbation group. We compared the characteristics, biomarker levels, microbiological findings, and prognosis in each patient group. PCT and CRP were measured using an immunofluorescence assay. Neopterin levels were measured using a competitive immunoassay. PCT and CRP showed significant differences among the three patient groups, being higher in patients with pneumonia, followed by patients with exacerbation (P < 0.0001). For the 23 patients with paired samples, PCT and CRP levels decreased 1 month after the exacerbation episode, while neopterin increased. Neopterin showed significantly lower levels in exacerbations with isolation of pathogenic bacteria, but no differences were found for PCT and CRP. No significant differences were found when comparing biomarker levels according to the Gram result: PCT (P = 0.191), CRP (P = 0.080), and neopterin (P = 0.109). However, median values of PCT and CRP were high for Streptococcus pneumoniae, Staphylococcus aureus, and enterobacteria. All biomarkers were higher in patients who died within 1 month after the sample collection than in patients who died later on. According to our results, biomarker levels vary depending on the clinical status. However, the identification of the etiology of infectious exacerbation by means of circulating biomarkers is encouraging, but its main disadvantage is the absence of a microbiological gold standard, to definitively demonstrate their value. High biomarker levels during an exacerbation episode correlate with the short-term prognosis, and therefore their measurement can be useful for COPD management.


Changes in multidimensional pain inventory profile after a pain rehabilitation programme indicate the risk of receiving sick leave benefits one year later.
To determine whether coping profile changes after rehabilitation, assessed with the Multidimensional Pain Inventory (MPI), can predict which persons disabled by chronic musculoskeletal pain will be in receipt of sick leave benefits in the long term. Study of MPI data from 2,784 patients (709 men and 2,075 women) collected from the Swedish Quality Register for Pain Rehabilitation (SQRP) before and at the end of rehabilitation and compared with independent sick leave data for 1 year later. After rehabilitation there was a significantly decreased share of Dysfunctional profiles (DYS) among both men (44% before, 31% after) and women (39% before, 26% after), but an increased share of Adaptive Coper profiles (men 15% before, 24% after, women 14% before, 24% after). The number of patients on full-time sick leave decreased significantly among men (from 57% to 46%) and women (from 57% to 50%). Persons with a DYS profile after rehabilitation had a low probability of having no or part-time sick leave. The number of persons with DYS profiles decreased after rehabilitation. Those with other profiles had less full-time sick leave one year later than those with DYS profiles, indicating that leaving the DYS profile is a positive prognostic sign long-term. Furthermore, the gender differences observed suggest the need to tailor rehabilitative strategies differently for men and women.


Effects of shaded versus unshaded wallows on behavior, performance, and physiology of the outdoor lactating sow.
The objectives of this study were to determine the effects of shading wallows during the summer months on lactating sow behavior, performance, and physiology. A total of 128 sows were used during warm weather (May to August 2001) to determine the effects of unshaded (control; n = 8) vs. shaded (SH; n = 8) wallows. Sows ranged over 6 parities and were fed a completely balanced sorghum-based diet. Behavioral data were collected by 15-min scan samples over a 24-h period/wk for a total of 16 wk. All sows were observed twice when litter age was 5 and 15 d, respectively. Respiration rates (breaths/min) were collected on 50 sows (control, n = 25; SH, n = 25) over an 8-wk period when the maximum temperature exceeded 32 degrees C. Ten milliliters of clotted blood and 20 mL of whole blood were obtained by jugular puncture from each sow on the day of weaning to determine total white blood cells, acute phase proteins, packed-cell volume, and chemotaxis and chemokinesis. Descriptive water temperature profiles were measured by using data loggers positioned at 3 levels per wallow: surface water, shallow mud, and deep mud. Behavioral, postural, location, performance, and physiological measurements did not differ (P > 0.05) among wallow treatments. Regardless of treatment, sows spent approximately 82% of their total time budget inside the farrowing hut and only approximately 7% of their total time budget in the wallow. A total of 428 piglets died, 219 in the control treatment and 209 in the SH treatment. The majority of piglets in both treatments died of crushing within the first 72 h after parturition, and most of the piglets had suckled. Shade kept the shallow water profile cooler during the hotter afternoon temperatures compared with the control wallows. In SH for both the shallow and deep mud profiles, temperatures were consistent throughout the day. In conclusion, sows spent a large percentage of their daily time budget inside the farrowing hut and spent only brief episodes in the wallow. Shading the wallow did not result in increased wallow use time or improvements in sow physiology and overall performance.


Multiple stressor effects on marine infauna: responses of estuarine taxa and functional traits to sedimentation, nutrient and metal loading.
Sedimentation, nutrients and metal loading to coastal environments are increasing, associated with urbanization and global warming, hence there is a growing need to predict ecological responses to such change. Using a regression technique we predicted how maximum abundance of 20 macrobenthic taxa and 22 functional traits separately and interactively responded to these key stressors. The abundance of most taxa declined in response to sedimentation and metal loading while a unimodal response was often associated with nutrient loading. Optimum abundances for both taxa and traits occurred at relatively low stressor levels, highlighting the vulnerability of estuaries to increasing stressor loads. Individual taxa were more susceptible to stress than traits, suggesting that functional traits may be less sensitive for detecting changes in ecosystem health. Multiplicative effects were more common than additive interactions. The observed sensitivity of most taxa to increasing sedimentation and metal loading and the documented interaction effects between multiple stressors have important implications for understanding and managing the ecological consequences of eutrophication, sedimentation and contaminants on coastal ecosystems.


Theoretical analysis of single-frequency Raman fiber amplifier system operating at 1178 nm.
We analyze the scalability of amplifying the output from a single-frequency diode laser operating at 1178 nm through the utilization of a core pumped Raman fiber amplifier. A detailed model that accounts for stimulated Raman scattering (SRS) and stimulated Brillouin scattering (SBS) in relation to the fiber mode field diameter, length, seed power, and available pump power in both co-pumped and counter-pumped configurations is developed. The backward travelling Stokes light is initiated from both spontaneous Brillouin and spontaneous Raman processes. It is found that when fiber length is optimized, the amplifier output scales linearly with available pump power. Although higher amplifier efficiency is obtained with higher seed power, the output power diminishes. In order to mitigate the SBS process for further power scaling, we employ and optimize a multi-step temperature distribution. Finally, we consider the feasibility of generating the D(2a) and D(2b) lines in a sodium guide star beacon from a single Raman amplifier by examining four-wave mixing (FWM).


Quality and completeness of histopathology reports of rectal cancer resections. Results of an audit in Brittany.
Few data are available in France about implementation of guidelines for pathology reports of rectal cancer resections. The purpose of this study was to audit quality and completeness of histopathology reports of rectal cancer resections in Brittany by comparing results with French guidelines published in 1998. All inhabitants in Brittany who were beneficiaries of the general health insurance system and who underwent surgery for rectal cancer between February 1999 and January 2000 were included in the study. Twenty-one pathology laboratories, including 14 private laboratories, participated in this study. All pathology reports were examined by two physician-consultants of the health insurance system trained in the analysis of pathology reports (search for consigned or missing data). Results were compared with guidelines published in 1998. Of 234 pathology reports, 204 (84%) mentioned the number of examined lymph nodes and 217 (93%) the number of those found positive. The criterion of at least 8 examined lymph nodes was noted in 53.4% of reports. Longitudinal margin involvement was recorded in 92% of reports and circumferential margin involvement in 27% only. Venous and/or lymphatic and neural invasion were recorded in 34% and 18% of reports, respectively. Tumor staging was made by using UICC (pTNM) in 67% of reports. This study shows that the quality and completeness of histopathology reports of rectal cancer resections could be improved in Brittany. Despite its documented value as an important predictor of local recurrence, circumferential margin involvement is too frequently omitted. Standardisation of the examination procedures and exhaustive reporting would be useful to improve practice quality.


Staurosporine inhibits neutrophil phagocytosis but not iC3b binding mediated by CR3 (CD11b/CD18).
C receptor CR3 (iC3b-receptor, CD11b/CD18) plays an essential role in several phagocytic and adhesive neutrophil functions. Recent evidence suggests that stimulus-induced phosphorylation of the CR3 beta-chain, CD18, may mediate certain neutrophil functions by transiently converting the molecule to an activated state. Staurosporine, a protein kinase C inhibitor that blocks PMA-induced CD18 phosphorylation, was used to study the functional relevance of this event. Neutrophils adhered to glass were assayed for binding and phagocytosis of iC3b-opsonized sheep E (EC3bi) in the presence or absence of PMA and/or staurosporine. Binding of EC3bi was markedly increased, not only by PMA, but also by staurosporine and by a combination of both agents (three- to sevenfold). The enhancement of rosetting by staurosporine was likely caused by increased surface expression of CR3 via exocytosis of specific granular contents. In contrast, staurosporine alone did not stimulate phagocytosis of EC3bi and markedly inhibited PMA-induced phagocytosis. Staurosporine also inhibited phagocytosis of yeast beta glucan particles, a CR3 ligand that, in contrast to EC3bi, is bound and ingested without additional prior treatment with PMA. beta glucan phagocytosis was associated with a low level of CD18 phosphorylation. Staurosporine did not block phagocytosis in general, because this agent had relatively little effect on FcR-mediated phagocytosis. These data demonstrate that phagocytosis mediated by CR3 requires activation of CR3 via a staurosporine-sensitive pathway. Increased binding of EC3bi, a function of increased surface expression of CR3, does not require activation of CR3 by such a pathway, confirming previous evidence for the independence of these two phenomena. A direct role for CD18 phosphorylation in the activation of CR3 for phagocytosis is consistent with these data.


[Activation of adenyl cyclase from bovine brain caudate nucleus by synthetic analogs of guanyl nucleotides].
In order to investigate the regulatory properties of dopamine-sensitive adenylate cyclase from bovine brain caudate nucleus some non-hydrolyzed guanyl nucleotides modified at the 8th position of the purine base and at phosphate site were synthesized. The activating effects of these compounds and that of GppCH2p on the membrane preparation of adenylate cyclase were studied. The activating effect of GppCH2p was decreased when the substituents were introduced at the 8th position of the purine base or when its terminal phosphate group was modified. The GDP analog ClCH2ppG-8-NH(CH2)6NHCOCH2Cl whose molecule contains simultaneously two substituents with alkylating chloromethyl groups produced an activating effect. This effect was abolished when the length of the substituent at the 8th position was reduced or one of the chloromethyl groups was lacking. The decrease of adenylate cyclase activation by ClCH2ppG-8-NH(CH2)NH(CH2)Cl in the presence of GDP is suggestive of a specific type of interaction of this compound with the enzyme. It is assumed that the components of the adenylate cyclase system may be covalently linked by this guanyl nucleotide.


Germinated brown rice extract shows a nutraceutical effect in the recovery of chronic alcohol-related symptoms.
Chronic ethanol abuse can cause liver damage and unfavorable lipid profiles in humans and rodents. Phytonutrients have the potential to partially reverse some of the adverse effects of alcoholism. In this study, a germinated brown rice grown under conditions that favor high concentrations of gamma-aminobutyric acid (GABA) was evaluated for protective effects against the toxic consequences of chronic ethanol use. Serum and hepatic lipid concentrations and enzymes indicative of liver damage were determined in mice chronically administered ethanol. Balb/c mice were fed with either AIN-76 diet (control), control diet plus ethanol, or control diet plus ethanol and supplemental brown rice extract for 30 days. The extract naturally contained 841 nmol GABA per milliliter and was prepared from germinated brown rice. Serum low-density lipoprotein cholesterol (LDL-C), liver aspartate aminotransferase, and liver alanine aminotransferase levels were increased in mice administered ethanol, but not in mice given ethanol and brown rice extract. The brown rice extract significantly increased serum and liver high-density lipoprotein cholesterol (HDL-C) concentrations. Furthermore, administration of the extract prevented ethanol-induced increases in liver triglyceride and total cholesterol concentrations. These findings raise the possibility that brown rice extracts containing a high level of GABA may have a nutraceutical role in the recovery from and prevention of chronic alcohol-related diseases.


The sway-density curve and the underlying postural stabilization process.
The sway-density curve (SDC) is computed by counting, for each time instant, the number of consecutive samples of the statokinesigram falling inside a circle of small radius R. The authors evaluated the sensitivity of the curve to the variation of R and found that in the range 3-5 mm the sensitivity was low, indicating that SDC is a robust descriptor of posturographic patterns. In addition, they investigated the relationship between SDC and the underlying postural stabilization process by decomposing the total ankle torque into three components: a tonic component (over 69 & of the total torque), an elastic torque caused by ankle stiffness (about 19 &), and an anticipatory active torque (about 12 &). The last component, although the smallest in size, is the most critical for the overall stability of the standing posture and appears to be correlated with the SDC curve.


Role of advanced glycation end products in diabetic neuropathy.
Diabetic neuropathy is the commonest form of peripheral neuropathy in the developed countries of the world. In diabetic patients, the presence of peripheral neuropathy increases their risks for developing foot ulceration and subsequent necrosis that necessitates lower limb amputation. Although the precise mechanisms underlying diabetic neuropathy remain unclear, there is evidence that hyperglycemia-induced formation of advanced glycation end products (AGEs) is related to diabetic neuropathy; AGE-modified peripheral nerve myelin is susceptible to phagocytosis by macrophages and contributes to segmental demyelination; modification of major axonal cytoskeletal proteins such as tubulin, neurofilament, and actin by AGEs results in axonal atrophy/degeneration and impaired axonal transport; and glycation of extracellular matrix protein laminin leads to impaired regenerative activity in diabetic neuropathy. Recently, the receptor for AGEs (RAGE) has been found to colocalize with AGEs in diabetic peripheral nerves. This suggests that, in diabetic neuropathy, AGEs and AGE/RAGE interactions induce oxidative stress, result in upregulation of nuclear factor (NF)-kappaB and various NF-kappaB-mediated proinflammatory genes, and exaggerate neurological dysfunction, including altered pain sensation. Additionally, AGE/RAGE-induced oxidative stress further accelerates formation of glycoxidation products such as Nepsilon-(carboxymethyl)lysine and pentosidine. Although new drugs that inhibit the formation of AGEs and block the AGE-RAGE interaction are being studied, no effective treatment modalities against AGE-induced nerve injury are currently available clinically. A therapeutic strategy to prevent and ameliorate diabetic neuropathy using anti-AGE agents needs to be established. In this review, the current issues involved in the role of the glycation process and the potential treatment options for diabetic neuropathy are explored.


Phosphorylation of the rev gene product of human immunodeficiency virus type 1.
Replication of human immunodeficiency virus type 1 requires the functional expression in trans of the virally encoded rev gene product (previously called art/trs). Here we demonstrate that this protein can be metabolically labeled with 32Pi. The phosphate receptor in the rev protein is shown to be exclusively serine. Treatment of rev-expressing cells with phorbol ester, a specific activator of protein kinase C, led to significant but transient enhancement of the level of rev phosphorylation. These results indicate that the rev protein is posttranslationally modified in vivo and suggest that the level of this modification is subject to modulation by extracellular stimuli.


Multiple aspects of sexual orientation: prevalence and sociodemographic correlates in a New Zealand national survey.
Sexual orientation consists of multiple components. This study investigated both sexual identity and same-sex sexual behavior. Data came from the New Zealand Mental Health Survey, a nationally representative community sample of New Zealanders aged 16 years or older, interviewed face-to-face (N = 12,992, 48% male). The response rate was 73.3%. Self-reported sexual identity was 98.0% heterosexual, 0.6% bisexual, 0.8% homosexual, 0.3% "Something else," and 0.1% "Not sure." Same-sex sexual behavior with a partner was more common: 3.2% reported same-sex sexual experience only and 1.9% reported both experience and a relationship. For analysis of childhood and lifecourse, five sexuality groups were investigated: homosexual, bisexual, and heterosexual divided into those with no same-sex sexual experience, experience only, and experience and relationship. The non-exclusively heterosexual groups were more likely to have experienced adverse events in childhood. Educational achievement and current equivalized household income did not differ systematically across the sexuality groups. Only 9.4% of the exclusively heterosexual lived alone, compared with 16.7% of bisexuals and 19.0% of homosexuals. Heterosexuals were more likely than bisexuals or homosexuals to have ever married or had biological children, with differences more marked for males than for females. Heterosexuals with no same-sex sexual experience were more likely to be currently married than the other two heterosexual groups. Restricting comparisons to heterosexual, bisexual, and homosexual identification ignores the diversity within heterosexuals. Differences between the bisexual and homosexual groups were small compared with the differences between these groups and the exclusively heterosexual group, except for sex (80.8% of bisexuals were female).


Characterization of two different five-coordinate soluble guanylate cyclase ferrous-nitrosyl complexes.
Soluble guanylate cyclase (sGC), a hemoprotein, is the primary nitric oxide (NO) receptor in higher eukaryotes. The binding of NO to sGC leads to the formation of a five-coordinate ferrous-nitrosyl complex and a several hundred-fold increase in cGMP synthesis. NO activation of sGC is influenced by GTP and the allosteric activators YC-1 and BAY 41-2272. Electron paramagnetic resonance (EPR) spectroscopy shows that the spectrum of the sGC ferrous-nitrosyl complex shifts in the presence of YC-1, BAY 41-2272, or GTP in the presence of excess NO relative to the heme. These molecules shift the EPR signal from one characterized by g 1 = 2.083, g 2 = 2.036, and g 3 = 2.012 to a signal characterized by g 1 = 2.106, g 2 = 2.029, and g 3 = 2.010. The truncated heme domain constructs beta1(1-194) and beta2(1-217) were compared to the full-length enzyme. The EPR spectrum of the beta2(1-217)-NO complex is characterized by g 1 = 2.106, g 2 = 2.025, and g 3 = 2.010, indicating the protein is a good model for the sGC-NO complex in the presence of the activators, while the spectrum of the beta1(1-194)-NO complex resembles the EPR spectrum of sGC in the absence of the activators. Low-temperature resonance Raman spectra of the beta1(1-194)-NO and beta2(1-217)-NO complexes show that the Fe-NO stretching vibration of the beta2(1-217)-NO complex (535 cm (-1)) is significantly different from that of the beta1(1-194)-NO complex (527 cm (-1)). This shows that sGC can adopt different five-coordinate ferrous nitrosyl conformations and suggests that the Fe-NO conformation characterized by this unique EPR signal and Fe-NO stretching vibration represents a highly active sGC state.


[Vertical distribution of acanthocephalans of the order Echinorhynchida in Lake Baikal].
Vertical distribution of acanthocephalans of the order Echinorhynchida is studied in Lake Baikal. Four species and subspecies from cottid fishes (Perciformes: Cottoidei) were examined, namely Pseudoechinorhynchus borealis (Linstow, 1901), Metechinorhynchus salmonis salmonis (Muller, 1780), M.s. baicalensis Bogolepova, 1957, M. truttae (Schrank, 1788). In the littoral (0-5 m) and sublittoral (5-100 m) areas all these species and subspecies were occurred, white in the profundal (100-300 m) and abyssal (900-1600 m) areas only Metechinorhynchus salmonis baisalensis has been found.


Diagnostic, pathogenetic, and therapeutic aspects of diabetic neuropathy.
Since nearly all patients with diabetes mellitus will manifest evidence of dysfunction of their peripheral and autonomic nerves, neuropathy can be considered to be the most common of the so-called "late complications of diabetes." Mononeuropathies are uncommon, but can affect virtually any cranial or nerve and are thought to be due to vascular occlusion. Distal symmetrical polyneuropathy usually begins insidiously with the development of sensory symptoms involving the feet. These symptoms progressively ascend and are accompanied by progressive motor dysfunction as well as cardiac, gastrointestinal, and genitourinary manifestations of autonomic neuropathy. Numerous biochemical abnormalities including impaired neural protein synthesis, abnormal axoplasmic transport, disturbed polyol pathway activity, and abnormal myo-inositol metabolism have been described and have been postulated to underlie the development of diabetic peripheral and autonomic neuropathies. At the present time, the available treatments for diabetic neuropathies are largely palliative and are not curative. In the future, a better understanding of the pathophysiologic basis of diabetic neuropathy will undoubtedly lead to the development of more rational therapeutic approaches.


Thermoregulatory value of cracking-clay soil shelters for small vertebrates during extreme desert conditions.
Deserts exhibit extreme climatic conditions. Small desert-dwelling vertebrates have physiological and behavioral adaptations to cope with these conditions, including the ability to seek shelter. We investigated the temperature (T) and relative humidity (RH) regulating properties of the soil cracks that characterize the extensive cracking-clay landscapes of arid Australia, and the extent of their use by 2 small marsupial species: fat-tailed and stripe-faced dunnarts (Sminthopsis crassicaudata and Sminthopsis macroura). We measured hourly (over 24-h periods) the T and RH of randomly-selected soil cracks compared to outside conditions, during 2 summers and 2 winters. We tracked 17 dunnarts (8 Sminthopsis crassicaudata and 9 Sminthopsis macroura) to quantify their use of cracks. Cracks consistently moderated microclimate, providing more stable conditions than available from non-crack points, which often displayed comparatively dramatic fluctuations in T and RH. Both dunnart species used crack shelters extensively. Cracks constitute important shelter for small animals during extreme conditions by providing a stable microclimate, which is typically cooler than outside conditions in summer and warmer in winter. Cracks likely play a fundamental sheltering role by sustaining the physiological needs of small mammal populations. Globally, cracking-clay areas are dominated by agricultural land uses, including livestock grazing. Management of these systems should focus not only on vegetation condition, but also on soil integrity, to maintain shelter resources for ground-dwelling fauna.


Recent developments in patent anti-cancer agents targeting the matrix metalloproteinases (MMPs).
Matrix metalloproteinases (MMPs) belong to a family of closely related calcium- and zinc-dependent endopeptidases involved in the degradation and remodeling of extracellular matrix (ECM) proteins that are associated with the tumorigenic processes. MMPs promote tumor invasion and metastasis, regulating host defense mechanisms and normal cell function. Thus, MMP inhibitors (MMPIs) are expected to be useful chemotherapeutic agents for the treatment of malignant cancer, osteoarthritis, and rheumatoid arthritis. A vast number of small molecular MMPIs have been developed in recent years. Although there have been considerable preclinical and clinical studies on these inhibitors, most of the effective candidates in clinical trials, however, have yielded unsatisfactory results, thus they are as yet unavailable for use as therapeutic drugs. Currently, more efforts have been directed to the design of specific inhibitors towards certain MMP family members for selective usage. This review will focus primarily on an analysis of recent developed MMPIs that have entered preclinical or clinical trials, and recently registered patents with regard to new highly selective MMPIs in USA or patent applications related to the specific inhibitors of MMPs. We also analyze the clinical failure and discuss the possible strategies to best optimize the development of these novel agents.


Effectiveness of resident-prepared conferences in teaching imaging utilization guidelines to radiology residents.
The authors evaluated the effectiveness of a resident-prepared conference series for teaching imaging utilization guidelines to radiology residents. Brown University radiology residents (n = 17) gave 61 presentations on imaging utilization to their colleagues during 16 1-hour conferences. The residents were later examined on the topics presented and surveyed about their familiarity with the American College of Radiology appropriateness criteria, their exposure to issues of cost-effectiveness, and their degree of confidence in providing imaging consultation. The same examination and survey were administered to control residents from the University of Wisconsin (n = 14) and the Oregon Health Sciences University (n = 14). Scores were compared by using linear regression and Wilcoxon rank sum tests. Controlling for years in radiology residency, residents at Brown scored on average 16.0% (standard error = 2.2%) higher than residents at the other universities (P < .001). Controlling for institution, 3rd- and 4th-year residents scored on average 7.4% (standard error = 2.1%) higher than 1st- and 2nd-year residents (P = .001). Brown residents expressed more familiarity with American College of Radiology appropriateness criteria and appeared to have more exposure to cost-effectiveness issues in conferences than residents at Wisconsin or Oregon Health Sciences University (P < .005). Residents from the three universities did not differ in their level of confidence in providing imaging consultation. Resident-prepared conferences are an effective means of teaching imaging utilization guidelines to residents, but they do not affect the residents' perception of their ability to provide imaging consultation.


The adolescent with an inborn error of metabolism: medical issues and transition to adulthood.
As patients with inborn errors of metabolism survive longer, understanding of potential medical and psychiatric complications adolescence and adulthood has increased. In general, detailed therapeutic guidelines for specific metabolic disorders are not available, and medical management must be tailored to the individual patient. Close interaction between the biochemical genetics clinic staff, primary care physician, mental health professional, and other specialists is necessary to formulate an integrated care plan. The education of the patient and family is a critical function of the biochemical genetics clinic, and transition from dependence on parents or other care providers to full independence is gradual. The ultimate goal is for the patient to have the essential knowledge and motivation required to cope responsibly with dietary and medical therapeutic regimens by adolescence or early adulthood. Specific illustrative inborn errors of metabolism are discussed (aminoacidemias, urea cycle defects, organic acidemias, fatty acid oxidation defects, disorders of carbohydrate metabolism, lysosomal storage disorders) in light of potential problems encountered in adolescence and adulthood, including issues involving pregnancy and long-term medical, psychosocial, and psychiatric complications.


A meta-analysis of imitation abilities in individuals with autism spectrum disorders.
Although imitation impairments are often reported in individuals with autism spectrum disorders (ASD), previous work has not yet determined whether these impairments are significant, specific to ASD, and present across the entire spectrum. This report of 53 studies on imitation in ASD seeks to determine whether individuals with ASD show significant imitation deficits, the magnitude of these deficits, and whether they are specific to ASD. Using standard meta-analytic techniques in a random-effects model, the data reviewed suggest that individuals with ASD show deficits in imitation, performing on average 0.81 SDs below individuals without ASD on imitation tasks. This deficit was specific to the condition of having ASD. Moderator analyses revealed that the average Autism Diagnostic Observation Schedule (ADOS) scores of groups of ASD participants were significantly and strongly negatively associated with the imitation abilities of these subjects, but average participant IQ was not associated with imitation abilities. Study setting, novelty of actions, format of imitation tasks (live vs. not), number of actions to imitate, or verbal prompts were not found to significantly affect the sizes of the imitation differences between individuals with and without ASD. The manner in which imitation was operationalized, however, had significant effects on whether imitation deficits were found between individuals with and without ASD. In tests that measured imitation of both form and end points, participants with ASD showed significant deficits compared with those without ASD; on tests of end point emulation only, individuals with ASD showed no deficits.


Hemodynamic and regional circulatory alterations in dog during anaphylactic challenge.
Microspheres were employed to measure the cardiac output (CO), percent distribution of CO (%CO), and blood flow (BF) to various tissues in the dog during anaphylactic shock. Five minutes after challenge there was a 66 and 83% drop from control for mean arterial pressure and CO, respectively. Arterial glucose and lactic acid rose 62 and 537%, respectively. Furthermore, there was a narrowing of the systemic arteriovenous difference for PO2 and a widening of the venoarterial difference for PCO2. Pressure compensation toward control values began 5 min postchallenge and continued through 180 min. Circulatory perturbations during severe hypotension (less than mmHg) included depressed %CO and BF to the pancreas and spleen and elevated %CO to the brain and adrenals. Cerebral and adrenal BF remained unchanged at this period. BF to the kidneys, liver (hepatic artery), and gastrointestinal tract dropped markedly as a function of increased peripheral resistance and decreased CO. As cardiac performance improved, blood flow to the pancreas, spleen, liver, diaphragm, and gastrointestinal tissues returned toward control but was elevated to the adrenals and decreased to the brain.


Comparison of Bone Preserving and Radical Surgical Treatment in 32 Cases of Calcaneal Osteomyelitis.
Introduction. Radical procedures like calcanectomy and amputation performed for calcaneal osteomyelitis are regarded as effective in eradication of infection even though potentially functionally disabling. Bone sparing procedures offer better functional result at the expense of potentially worse infection control. The aim of the study has been to assess the influence of the surgical radicalism as much as the extent of bone infection on the final outcome in the surgical therapy of chronic calcaneal osteomyelitis (CO). Material and method. 32 patients with chronic CO have comprised the group under study: 8 with superficial type, 12 localised type and 12 with diffuse type according to Cierny-Mader classification. The aim of the treatment was to heal infection, preserve the heel shape and achieve good skin coverage over the calcaneus. The therapy consisted of 9 debridement surgeries with or without flaps, 8 drilling-operations of the calcaneus with application of collagen-gentamicin-sponge in bore holes, 15 partial and 2 total calcanectomies, and 4 below-the knee amputations. Results. The healing of infection and wound has been achieved after 7 of 9 debridements, 6 of 8 drilling-operations, 13 of 15 partial and all total calcanectomies. Conclusion. Bone preserving operations in chronic calcaneal osteomyelitis provided inferior infection control (76,47% vs 88,24%) and worse patient satisfaction (88,24% vs 100%) and almost camparable ambulation (100% vs 93,33%). Drilling of the calcaneus with application of collagen sponge containing gentamicin performed in chronic diffuse calcaneal osteomyelitis seems to offer a viable alternative to partial or radical calcanectomy. V.


Modulation of Antimicrobial Peptide Potency in Stressed Lipid Bilayers.
It is shown that the tendency of an archetypal antimicrobial peptide to insert into and perforate a simple lipid bilayer is strongly modulated by tensile stress in the membrane. The results, obtained through molecular dynamics simulations, have been demonstrated with several lipid compositions and appear to be general, although quantitative details differ. The findings imply that the potency of antimicrobial peptides may not be a purely intrinsic chemical property and, instead, depends on the mechanical state of the target membrane.


Light dosimetry in vivo.
This paper starts with definitions of radiance, fluence (rate) and other quantities that are important with regard to in vivo light dosimetry. The light distribution in mammalian tissues can be estimated from model calculations using measured optical properties or from direct measurements of fluence rate using a suitable detector. A historical introduction is therefore followed by a brief discussion of tissue optical properties and of calculations using diffusion theory, the P3-approximation or Monte Carlo simulations. In particular the form of the scattering function is considered in relation to the fluence rate close to the tissue boundary, where light is incident. Non-invasive measurements of optical properties yield the absorption coefficient mu a and mu s(1 - g), where mu s is the scattering coefficient and g is the mean cosine of the scattering angle. An important question is whether this combination is sufficient, or whether g itself must be known. It appears that for strongly forward scattering, as in mammalian tissues, rather detailed knowledge of the scattering function is needed to reliably calculate the fluence rate close to the surface. Deeper in the tissue mu s (1 - g) is sufficient. The construction, calibration and use of fibre-optic probes for measurements of fluence rate in tissues or optical phantoms is discussed. At present, minimally invasive absolute fluence (rate) measurements seem to be possible with an accuracy of 10-20%. Examples are given of in vivo measurements in animal experiments and in humans during clinical treatments. Measurements in mammalian tissues, plant leaves and marine sediments are compared and similarities and differences pointed out. Most in vivo light fluence rate measurements have been concerned with photodynamic therapy (PDT): Optical properties of the same normal tissue may differ between patients. Tumours of the same histological type may even show different optical properties in a single patient. Treatment-induced changes of optical properties may also occur. Scattered light appears to contribute substantially to the light dose. All these phenomena emphasize the importance of in situ light measurements. Another important dosimetric parameter in PDT is the concentration and distribution of the photosensitizer. Apart from in vivo fluorescence monitoring, the photosensitizer part of in vivo PDT dosimetry is still in its infancy.


Synthesis, crystal structures and magnetic properties of mer-cyanideiron(III)-based 1D heterobimetallic cyanide-bridged chiral coordination polymers.
Two pairs of cyanide-bridged Fe(III)–Mn(III)/Cu(II) chiral enantiomer coordination polymers {[Mn(S,S/R,R-Salcy)(CH3OH)2]{[Mn(S,S/R,R-Salcy)][Fe(bbp)(CN)3]}}2n (1,2) (bbp = bis(2-benzimidazolyl)pyridine dianion) and {[Cu(S,S/R,R-Chxn)2]2[Fe2(tbbp)(CN)6]}n (3,4) (tbbp = tetra(3-benzimidazolyl)-4,4′-bipyridine tetraanion) have been successfully prepared by employing mer-tricyanometallate [PPh4]2[Fe(bbp)(CN)3] or the newly bimetallic mer-cyanideiron(III) precursor K4[Fe2(tbbp)(CN)6] as building blocks and with chiral manganese(III)/copper(II) compounds as assemble segments. The four complexes have been characterized by elemental analysis, IR spectroscopy, circular dichroism (CD) and magnetic circular dichroism (MCD) spectra. Single X-ray diffraction reveals that complexes 1 and 2 possess a single anionic chain structure consisting of the asymmetric chiral {[Mn(S,S/R,R-Salcy)][Fe(bbp)(CN)3]}2(2−) unit with free [Mn(S,S/R,R-Salcy)](+) as balanced cations. The cyanide-bridged Fe(III)–Cu(II) complexes 3 and 4 can be structurally characterized as neutral ladder-like double chains composed of the alternating cyanide-bridged Fe–Cu units. Our investigation of magnetic susceptibilities reveals the antiferromagnetic coupling between the cyanide-bridged Fe(III) and Mn(III)/Cu(II) ions for complexes 1–4. These results have been further confirmed by theoretical simulation through numerical matrix diagonalization techniques using a Fortran program or a uniform chain model, leading to the coupling constants J = −7.36 cm(−1), D = −1.52 cm(−1) (1) and J = −4.35 cm(−1) (3), respectively.


Alterations in mRNA stability during rat liver regeneration.
We examined the in vivo transcriptional and posttranscriptional regulation of various genes involved in hepatocyte growth and replication that exhibited changes in steady-state mRNA levels after 70% partial hepatectomy (PH). Of the 19 genes examined by nuclear run-on assay, 17 demonstrated no change in transcriptional activity through the first 96 h of regeneration. However, results from in vivo half-life determinations indicated that changes in mRNA stability played a critical role in regulating transcript levels during liver regeneration. For many of the genes, alterations in transcript abundance correlated with similar changes in mRNA half-lives. Inhibition of protein synthesis by cycloheximide was generally associated with increased levels of mRNA expression, but no detectable changes in transcriptional rates in both control and regenerating rat liver. Finally, genomic methylation status was investigated by Southern analysis for several genes that displayed changes in mRNA stability. Interestingly, increases in mRNA half-lives for the genes p53, c-myc, H-ras, and ornithine decarboxylase were associated with decreased genomic methylation. In conclusion, regulation of gene expression beyond the immediate early phase of te cell cycle during rat liver regeneration after PH occurs predominantly at the posttranscriptional level. mRNA stability appears to be a significant factor in this control, and may itself be modulated by the methylation status of the corresponding genomic DNA.


The pedicled lateral arm flap for oncologic reconstruction near the shoulder.
Oncologic resections near the shoulder create challenging reconstructive problems. In addition to severe contour defects and the risk for functional deficits resulting from contractures, there may be exposed bone and peripheral nerves rendering simple skin graft closures suboptimal long term. The lateral arm flap is a versatile septofasciocutaneous flap based on the posterior branch of the radial collateral artery in the lateral intermuscular septum of the upper arm. This paper details our experience with 3 patients who underwent wide local excision or radical resection of soft tissue tumors of the upper arm and closure with pedicled lateral arm fasciocutaneous flaps. Flap size ranged from 6 × 10 cm to 5 × 15 cm. Two flaps were transferred as 180-degree pivotal flaps; 1 flap was transferred as an advancement flap. There were no immediate postoperative complications or long-term functional deficits. The pedicled lateral arm flap is a safe and reliable option for soft tissue coverage of anterolateral shoulder defects. The vascular pedicle lies deep within the mid-upper arm and is rarely at risk during extensive tumor extirpation. The adjacent donor site allows for a single operative field without the need for repositioning and furthermore results in a skin paddle requirement that is one half the width of the original defect.


Impaired metabolic control and socio-demographic status in immigrant children at onset of Type 1 diabetes.
The aim of the present study was to compare clinical and socio-demographic conditions at the onset of Type 1 diabetes in children born to immigrant families and children born to Swedish families, and to assess whether those conditions had an impact on metabolic status. This was an observational nationwide population-based matched cohort study on prospectively recorded registry data of all children with diabetes in Sweden and their families during 2000-2010. Out of a total of 13 415 children from the Swedish Childhood Diabetes Registry (SWEDIABKIDS), 879 children born to immigrant parents were collected. To these we added 2627 children with Swedish-born parents, matched for gender, age and year of onset of Type 1 diabetes. The proportion of low capillary pH (< 7.30) at onset was higher in the immigrant cohort [25.8% vs. 16.4% in the Swedish cohort (P < 0.001)]. HbA1c was also higher [95 mmol/mol (10.8%) vs. 88 mmol/mol (10.2%), respectively (P < 0.001)]. In a logistic regression model with low pH as the dependent variable, we were unable to reveal any significant association to socio-demographic factors, but the odds ratio for HbA1c was 0.983 (95% CI 0.976-0.991) and for plasma glucose was 0.953 (95% CI 0.933-0.973). Children born to immigrant parents have lower capillary pH and higher HbA1c at diabetes onset. Immigrant families harbour lower socio-demographic living conditions, but this fact does not seem to influence the inferior metabolic condition at diabetes onset.


Increased Serum Matrix Metalloproteinase-9 Levels are Associated with Anti-Jo1 but not Anti-MDA5 in Myositis Patients.
Matrix metalloproteinases 9 (MMP9) is a member of the zinc-ion-dependent proteinases family and plays a pathogenic role in chronic inflammatory autoimmune diseases. However, its roles in the pathogenesis of myositis have not been elucidated. In this study, we aimed to determine the gene expression and serum level of MMP9 and their relationship with clinical features and serological parameters in myositis. Our results showed that MMP9 mRNA in peripheral blood mononuclear cells (PBMC) was upregulated in myositis patients compared to that in healthy controls. Myositis patients positive for anti-Jo1 antibodies exhibited significantly higher serum MMP9 than anti-MDA5 positive or antibody-negative patients and healthy controls. However, the presence of interstitial lung disease (ILD) did not affect MMP9 levels. We further identified that anti-Jo1-positive myositis patients showed higher numbers of white blood cells (WBC), lymphocytes and neutrophils; increased levels of creatine kinase (CK), lactate dehydrogenase (LDH), and C-reactive protein (CRP); and higher erythrocyte sedimentation rate (ESR) than anti-MDA5 positive patients. In addition, serum MMP-9 levels were positively correlated with WBCs, neutrophils, CK, CRP, ESR, and LDH in myositis patients. In vitro experiments showed that purified serum IgG from Jo-1-positive patients could stimulate PBMCs to release more MMP9 than the IgG from MDA-5-positive sera. These results indicated that increased MMP9 in anti-Jo1-positive myositis patients was associated with the extent of muscle involvement, but not pulmonary damage. The distinct pattern of serum MMP9 perhaps clarifies the differences in pathophysiology between anti-Jo1 and anti-MDA5 in patients with myositis.


Mesenchymal Stem Cells for Coronavirus (COVID-19)-Induced Pneumonia: Revisiting the Paracrine Hypothesis with New Hopes?
Mesenchymal stem cells (MSCs) bear a promising potential for regenerative medicine therapies and they repair damaged tissue through secretion of immune modulatory and anti-inflammatory molecules acting in a paracrine fashion. Coronavirus disease 2019 (COVID-19) has spread all over the world with high morbidity and mortality rates and there is no specific treatment for this infection. A recent study published in the journal reports that MSC infusion is safe and effective in patients suffering from COVID-19 induced pneumonia. In the light of this study and previous reports, we make additional comments about possible therapeutic effects of MSCs in COVID-19 infection.


The natural compound chebulagic acid inhibits vascular endothelial growth factor A mediated regulation of endothelial cell functions.
Vascular endothelial growth factor A (VEGFA) plays an important role in tumour angiogenesis and its angiogenic action is mainly mediated through its VEGF receptor 2 (VEGFR-2). Therefore drugs targeting VEGFA/VEGFR-2 are being presently used in the clinics for treatment of several types of solid malignant tumours. We here in report that low dose of chebulagic acid (CA), a hydrolysable tannin found in myrobalan fruits can inhibit VEGFA induced vascular permeability, endothelial cell proliferation, migration, tube formation and thereby, angiogenesis by suppressing VEGFR-2 phosphorylation. CA may thus be an effective and useful natural inhibitor of VEGFA mediated angiogenesis.


Defined contribution defined: health insurance for the next century.
The consumerism movement will dramatically affect the current payer model and present a new series of challenges for managed care organizations. Employers will fuel the changes, as they create health benefit programs that are modeled after retirement programs. In these cases, employers will shift a major portion of financial responsibility to employees, who will be asked to make buying decisions often previously determined by managed care organizations. New business entities known as health navigators will be formed to aid consumers. However, many structural and policy obstacles may slow or transform the consumerism movement.


D-cycloserine to enhance extinction of cue-elicited craving for alcohol: a translational approach.
Cue-elicited craving for alcohol is well established but extinction-based treatment to extinguish this response has generated only modest positive outcomes in clinical trials. Basic and clinical research suggests that D-cycloserine (DCS) enhances extinction to fear cues under certain conditions. However, it remains unclear whether DCS would also accelerate extinction of cue-elicited craving for alcohol. The goal of the current study was to examine whether, compared with placebo (PBO), DCS enhanced extinction of cue-elicited craving among treatment-seeking individuals with alcohol use disorders (AUDs). Participants were administered DCS (50 mg) or PBO 1 h before an alcohol extinction paradigm in a simulated bar environment on two occasions. The extinction procedures occurred 1 week apart and were fully integrated into outpatient treatment. Subjective craving for alcohol was the primary variable of interest. Follow-up cue reactivity sessions were conducted 1 week and 3 weeks later to ascertain persisting DCS effects. Drinking outcomes and tolerability were also examined. DCS was associated with augmented reductions in alcohol craving to alcohol cues during the first extinction session and these effects persisted through all subsequent sessions, suggesting facilitation of extinction. Participants in the DCS condition reported significant short-term reductions in drinking, although these did not persist to follow-up, and found the medication highly tolerable. These findings provide evidence that DCS enhances extinction of cue-elicited craving for alcohol in individuals with AUDs in the context of outpatient treatment. The potential clinical utility of DCS is discussed, including methodological considerations and context-dependent learning.


Simultaneous measurement of magnetic vortex polarity and chirality using scanning electron microscopy with polarization analysis (SEMPA).
The magnetic vortex structure is an equilibrium configuration frequently found in patterned magnetic nanostructures. It is characterized by an in-plane curling of the magnetization with clockwise or anticlockwise chirality and by an out-of-plane vortex core that can have a positive or negative polarity. The small size of the vortex core, on the order of 10nm, makes it technologically interesting due to potential data storage, but also difficult to measure or image directly. In this work, we used Scanning Electron Microscopy with Polarization Analysis (SEMPA) to directly image magnetic vortex cores in patterned NiFe/Ta bilayer structures. With SEMPA we can simultaneously measure the in-plane and the out-of-plane component of the surface magnetization and thereby determine both the vortex chirality and the vortex core polarity in a single measurement. Our magnetic simulation of the vortex core, considering only the exchange and magnetostatic energy, is in good agreement with the SEMPA measurement of the magnetization when other experimental factors are taken into account.


Dilemmas of Adopting Goal-Directed Perfusion in Extracorporeal Circulation: A Narrative Review.
Extracorporeal circulation (ECC) is generally based on standards established in the last decade. In recent years, a concept of perfusion management during ECC, goal-directed perfusion (GDP), has emerged to create optimal conditions for oxygen delivery and extraction, initiated by Rannuci et al. The aim of the present work was to determine whether the ECC procedure can truly be optimized with the current state of knowledge and understanding of human physiology. Discussed articles from 2017 to 2022 were selected from the MEDLINE (PubMed) database using the keywords "cardiopulmonary bypass" AND "cardiac surgery" AND "oxygen delivery" with the conditions of "clinical trial" OR "randomized controlled trial." The concept of GDP is an attempt to reproduce the physiological conditions of tissue respiration during ECC. Published articles, also due to their retrospective nature, are based on standards and recommendations that do not fully fit the field of physiological circulation. There are still insufficient tools to assess the relationship between volemia, perfusion pressure, and pump performance. Limitations include indications for vasoactive drugs. Methodology has rarely taken into account the period of starting and stopping the heart-lung machine, the most pronounced periods of circulatory destabilization with reduced oxygen delivery. Problems associated with ECC such as acute kidney injury, liver failure, vasoplegic syndrome, and others must await its resolution. The use of advanced monitoring technology and data engineering may allow the development of baseline hemodynamic models, which may make the ECC procedure more physiologic and thus improve the safety of the procedure.


Role of oxidative stress in rabies virus infection of adult mouse dorsal root ganglion neurons.
Rabies virus infection of dorsal root ganglia (DRG) was studied in vitro with cultured adult mouse DRG neurons. Recent in vivo studies of transgenic mice that express the yellow fluorescent protein indicate that neuronal process degeneration, involving both dendrites and axons, occurs in mice infected with the challenge virus standard (CVS) strain of rabies virus by footpad inoculation. Because of the similarities of the morphological changes in experimental rabies and in diabetic neuropathy and other diseases, we hypothesize that neuronal process degeneration occurs as a result of oxidative stress. DRG neurons were cultured from adult ICR mice. Two days after plating, they were infected with CVS. Immunostaining was evaluated with CVS- and mock-infected cultures for neuron specific beta-tubulin, rabies virus antigen, and amino acid adducts of 4-hydroxy-2-nonenal (4-HNE) (marker of lipid peroxidation and hence oxidative stress). Neuronal viability (by trypan blue exclusion), terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling (TUNEL) staining, and axonal growth were also assessed with the cultures. CVS infected 33 to 54% of cultured DRG neurons. Levels of neuronal viability and TUNEL staining were similar in CVS- and mock-infected DRG neurons. There were significantly more 4-HNE-labeled puncta at 2 and 3 days postinfection in CVS-infected cultures than in mock-infected cultures, and axonal outgrowth was reduced at these time points in CVS infection. Axonal swellings with 4-HNE-labeled puncta were also associated with aggregations of actively respiring mitochondria. We have found evidence that rabies virus infection in vitro causes axonal injury of DRG neurons through oxidative stress. Oxidative stress may be important in vivo in rabies and may explain previous observations of the degeneration of neuronal processes.


Dual-functional gold nanoparticles with antimicrobial and proangiogenic activities improve the healing of multidrug-resistant bacteria-infected wounds in diabetic mice.
Gold nanoparticles (Au NPs) are conjugated with the vascular endothelial growth factor-A165 (VEGF-A165) and (11-mercaptoundecyl)-N,N,N-trimethylammonium (11-MTA) cation to form dual-functional gold nanoparticles (11-MTA/VEGF-Au NPs) that possess antimicrobial and proangiogenic activities for wound healing in diabetic (db/db) mice. VEGF-A165 is a popular proangiogenic growth factor that stimulates multiple components in the wound-healing cascade. On the other hand, 11-MTA possesses antibacterial activity and can be bound to Au NPs easily through Au-S bonding. We have found that the surface density of VEGF-A165 plays a vital role in promoting the proliferation, migration, and tube formation of human umbilical vein endothelial cells. 11-MTA tethered on the VEGF-modified Au NPs enables the nanocomposites (i.e., 11-MTA/VEGF-Au NPs) to exhibit a strong antimicrobial activity against multidrug-resistant bacteria [methicillin-resistant S. aureus (MRSA)]. The minimal inhibition concentration of 11-MTA/VEGF-Au NPs is ∼450-fold lower than that of 11-MTA, revealing their high antibacterial efficiency. 11-MTA/VEGF-Au NPs exhibit high biocompatibility. 11-MTA/VEGF-Au NPs as dressing materials to treat MRSA-infected wounds in diabetic mice not only show strong in vivo bactericidal activities but also enhance the healing process of the formation of collagen fibers and epithelialization. Our results show that dual-functional 11-MTA/VEGF-Au NPs are promising agents for clinical applications like treating chronic wound infections.


Wide-band tracheids in genera of Portulacaceae: novel, non-xylary tracheids possibly evolved as an adaptation to water stress.
Wide-band tracheids (WBTs) are novel tracheids with wide, lignified secondary walls that intrude deeply into the cell lumen when viewed in transverse sections. These tracheids are found in a few genera in related families in the order Caryophyllales: Aizoaceae, Cactaceae, and Portulacaceae. WBTs in these three families vary in (1) systematic occurrence (found in more highly derived genera in each family), (2) location in plant organs, and (3) structure and dimensions. In addition, an analysis was conducted of WBT cell walls to test the hypothesis that WBTs evolved as an adaptation to water stress (i.e., the wide secondary walls should prevent collapse of the primary wall during water stress). The cell wall data show that primary cell walls in WBTs cannot inwardly collapse to occlusion, thus providing support for the water stress hypothesis of WBT evolution. With consideration of their systematic occurrence, the molecular phylogenetic data, and data here showing support for a water stress adaptive origin, it is logical to assume that WBTs evolved in genera that were adapting to environments undergoing a rapid trend toward aridification.


The Adoption of AI in Mental Health Care-Perspectives From Mental Health Professionals: Qualitative Descriptive Study.
Artificial intelligence (AI) is transforming the mental health care environment. AI tools are increasingly accessed by clients and service users. Mental health professionals must be prepared not only to use AI but also to have conversations about it when delivering care. Despite the potential for AI to enable more efficient and reliable and higher-quality care delivery, there is a persistent gap among mental health professionals in the adoption of AI. A needs assessment was conducted among mental health professionals to (1) understand the learning needs of the workforce and their attitudes toward AI and (2) inform the development of AI education curricula and knowledge translation products. A qualitative descriptive approach was taken to explore the needs of mental health professionals regarding their adoption of AI through semistructured interviews. To reach maximum variation sampling, mental health professionals (eg, psychiatrists, mental health nurses, educators, scientists, and social workers) in various settings across Ontario (eg, urban and rural, public and private sector, and clinical and research) were recruited. A total of 20 individuals were recruited. Participants included practitioners (9/20, 45% social workers and 1/20, 5% mental health nurses), educator scientists (5/20, 25% with dual roles as professors/lecturers and researchers), and practitioner scientists (3/20, 15% with dual roles as researchers and psychiatrists and 2/20, 10% with dual roles as researchers and mental health nurses). Four major themes emerged: (1) fostering practice change and building self-efficacy to integrate AI into patient care; (2) promoting system-level change to accelerate the adoption of AI in mental health; (3) addressing the importance of organizational readiness as a catalyst for AI adoption; and (4) ensuring that mental health professionals have the education, knowledge, and skills to harness AI in optimizing patient care. AI technologies are starting to emerge in mental health care. Although many digital tools, web-based services, and mobile apps are designed using AI algorithms, mental health professionals have generally been slower in the adoption of AI. As indicated by this study's findings, the implications are 3-fold. At the individual level, digital professionals must see the value in digitally compassionate tools that retain a humanistic approach to care. For mental health professionals, resistance toward AI adoption must be acknowledged through educational initiatives to raise awareness about the relevance, practicality, and benefits of AI. At the organizational level, digital professionals and leaders must collaborate on governance and funding structures to promote employee buy-in. At the societal level, digital and mental health professionals should collaborate in the creation of formal AI training programs specific to mental health to address knowledge gaps. This study promotes the design of relevant and sustainable education programs to support the adoption of AI within the mental health care sphere.


Primary spinal epidural extraosseous Ewing's sarcoma.
The case of a 7-year-old boy with a spinal epidural extraosseous Ewing's sarcoma (EES) is presented. He is in complete remission without neurologic deficit 40 months after diagnosis. Another 15 cases were found in the literature and are discussed together with this patient. Twelve of them were male patients. The mean age of the patients was 17.5 years (range, 4 to 47). Symptoms included back pain and/or radicular pain (100%), paresis of one or both legs (83%), sensory disturbances, and bladder and bowel dysfunction. The mean diagnostic delay was 5.8 months. Each patient underwent laminectomy; complete resection of the tumor was impossible in more than 50% of the cases. Most patients received radiation therapy and/or chemotherapy. Four patients suffered from local recurrence, eight from metastases. Ten (63%) patients died, 1 to 48 months (mean, 16) after diagnosis. The differential diagnosis is discussed, including disk herniation and several benign and malignant tumors.


[Content of antibodies to heat shock proteins in uncomplicated and complicated course of acute appendicitis].
The article presents information on the content and dynamics of changes in heat shock protein (HSP-70) in plasma and serum of patients with acute appendicitis before and after treatment. It was shown that higher content was noted in the group of patients with a complicated course. The use of traditional methods resulted in the decreased level of HSP-70 but in the group of patients with a complicated course the concentrations of shaperons remains elevated.


Electrophysiological evidence of atypical processing underlying mental set shifting in ecstasy polydrug and polydrug users.
Executive functioning deficits are reported in ecstasy users. However research into mental set switching has been equivocal, with behavioral studies suggesting the function is preserved. The current study sought to address the issue of switching deficits in ecstasy users by combining behavioral performance with electrophysiological correlates (electroencephalography; EEG). Twenty ecstasy polydrug users, 20 nonecstasy polydrug users, and 20 drug naive controls were recruited. Participants completed questionnaires about their drug use, sleep quality, fluid intelligence, and current mood state. Each participant completed a mental set switching task (the number-letter task) while EEG measures were recorded. Analysis of variance (ANOVA) revealed no between-group differences on performance of the task; however a regression suggested that ecstasy use was a significant predictor for performance, after controlling for cannabis use. Mixed ANOVA revealed a significant effect of group on the P3, with significant differences between both drug groups and naives. There was also an interaction between electrode and group on the P2 component, with ecstasy users differing from both other groups. On the P3 component the results suggest a reduction in positivity at parieto-occipital electrodes for drug users compared to controls. Furthermore a significant increase in negativity in ecstasy users compared to control groups could be observed in several occipito-parietal electrodes at an N2 component as well as observable atypicalities in early processing (P2) displayed by ecstasy users and polydrug controls. The present study provides evidence of atypical processing of attentional shifting in ecstasy and polydrug users. Deficits in this executive function could reflect cognitive inflexibility and paucity of rapid behavioral adjustment, which may be problematic in real world situations.


Investigation of occupational asthma: sputum cell counts or exhaled nitric oxide?
The measure of sputum eosinophil counts is a useful tool in the investigation of occupational asthma (OA), but processing sputum is time consuming. Measuring the fractional concentration of exhaled nitric oxide (FENO) may be an alternative in clinical practice. The aim of this study was to assess the respective changes of sputum eosinophil counts and FENO following exposure to occupational agents in the routine practice of two tertiary centers in North America and Europe. Workers undergoing specific inhalation challenges (SICs) for possible OA in tertiary clinics in both Canada and Belgium were enrolled. Sputum cell counts and FENO were collected at the end of the control day and at 7 and 24 h after exposure to the offending agent. Forty-one subjects had a negative SIC; 26 subjects had OA proven by a positive SIC. In subjects with positive SIC, there was a significant increase in sputum eosinophils at 7 h (9.0 [9.9]%) and 24 h (11.9 [14.9]%) after exposure compared with the baseline (2.8 [4.2]%), whereas there was a significant increase in FENO only 24 h after exposure (26.0 [30.5] ppb) compared with the baseline (16.6 [18.5] ppb). A 2.2% change in sputum eosinophil counts achieved a much higher sensitivity and positive predictive value than a 10-ppb change in FENO with similar specificity and negative predictive value for predicting a 20% decrease in FEV(1) during SICs. Sputum eosinophil counts constitute a more reliable tool than FENO to discriminate positive and negative SICs.


Effects of Different Doses of Eucalyptus Oil From Eucalyptus globulus Labill on Respiratory Tract Immunity and Immune Function in Healthy Rats.
Eucalyptol (1,8-cineole), the major constituent of eucalyptus oil (EO), was used in traditional medicine as a remedy for colds and bronchitis. This study aimed at clarifying the effect of eucalyptol on respiratory immune function of CD8 and CD4 cells, and alveolar macrophages (AM). Thirty male Sprague-Dawley rats were divided into experimental and control groups. The drug was given once a day for 3 weeks and the experimental group was divided according to the eucalyptol dose into: 30, 100, and 300 mg·kg-1 groups. Flow cytometry was used to detect the phagocytic function of CD4, CD8 cells, and AM in the bronchopulmonary lavage fluid. The 30 and 100 mg·kg-1 groups had an up-regulation effect on CD8 (p < 0.05), with no significant effect on macrophage phagocytosis. The 300 mg·kg-1 group had an inhibitory effect on CD8 and macrophage phagocytosis (p < 0.05), with no significant difference in CD4 between groups. Further investigation was conducted to evaluate the effect of EO on immune function in rats by detecting blood T, B, and NK cells using flow cytometry, and blood IgA, IgG, IgM, and IFN-γ levels by ELISA. High dosage of eucalyptol significantly reduced the proportion of blood B and NK cells (p < 0.05). IgA was decreased in the 100 and 300 mg·kg-1 groups (p < 0.05). There are no significant differences between the number of T cells and the IgG, IgM, and IFN-γ levels between experimental and control groups. Rational use of EO containing eucalyptol can improve the immune function of the respiratory tract and the body immunity, while high dose could have damaging effects, through modifying the phagocytic function of CD8 cells and reducing the proportion of blood B cells, NK cells, and IgA.


The association of having a monitoring or blunting coping style with psychological distress, health-related quality of life and satisfaction with healthcare in gastrointestinal stromal tumour (GIST) patients.
There are two main coping styles regarding information seeking under medical threat; monitoring (information-seeking) and blunting (information-avoiding). The aim of this study is to (1) determine factors associated with a monitoring or blunting coping style in gastro-intestinal stromal tumour (GIST) patients and (2) investigate its association with psychological distress, cancer-related concerns, health-related quality of life and satisfaction with healthcare. In a cross-sectional study, Dutch GIST patients completed the shortened version of the Threatening Medical Situations Inventory to determine their coping style, the Hospital Anxiety and Depression Scale, Cancer Worry Scale, EORTC QLQ-C30 and part of the EORTC QLQ-INFO25. A total of 307 patients were classified as blunters (n = 175, 57%) or monitors (n = 132, 43%). Coping style was not associated with tumour or treatment variables, but being a female (OR 2.5; 95%CI 1.5-4.1; p= <.001) and higher educated (OR 5.5; 95%CI 2.5-11.9, p= <.001) were associated with higher odds of being a monitor. Monitors scored significantly lower on emotional functioning (mean = 86.8 vs mean = 90.9, p=.044), which is considered a trivial difference, more often experienced severe fear of cancer recurrence or progression (53.0% vs 37.7%, p=.007), and had more concerns about dying from GIST in the future (60.6% vs 47.4%, p=.025). Compared to blunters, monitors were less satisfied with the received healthcare and information, and would have liked to receive more information. GIST patients with a monitoring coping style experience a higher emotional burden. Additionally, monitors exhibit a greater need for information. Although this need for information could potentially result in fears and concerns, recognising it may also create an opening for tailored communication and information.


[The personality of the doctor as a corner stone in the temple of gerontology].
The fundamentals of gerontology are not fundamental science and not dialectical materialism, but, in our opinion, particularly respect for elders. The very same attitude toward the elderly in the society has changed to the best party not so long ago, when the material ability had appeared to adequately support the elderly and implement the monotheistic values, such as the value to God of every life. Thanks to the presence in St. Petersburg at 1889 year well-organized places for medical care of oldster professor S. P. Botkin was able to implement the world's first large-scale complex research of old age, surveyed more than two thousand old people. Today's liberal model of providing education without upbringing is fraught with the most terrible consequences. We must not forget a number of artifacts in medicine, when the doctor in different civilizations ignored the Hippocratic Oath. These are Josef Mengele and Karl Brandt, the physicians of "Unit 731", Leiba Shatunovskii, the doctors of the clinic "Medicus", and many others. These facts force to carefully scrutinize the current time. So in the British newspapers the information appear about the dead from hunger and thirst old people in expensive nursing homes. It seems necessary to go back to the moral roots of medicine. The pride of the Russian medicine is a life-physician Eugene S. Botkin, who ascended to Calvary with his crowned patient, Emperor Nicholas II, because he could not change his word of honor. Thus, in the Russian tradition, glorious Hippocratic writings oath strengthened by noble honor. Knight code of the Russian nobleman, described us as "4D" (in Russian): duty, virtue, goodness and leisure, is the best formula for education and self-improvement noble doctor. Thus, it is proposed to pay attention to the unity of education and upbringing in preparing the next generation of doctors, which should strengthen the guarantees of the preservation of high moral relations to the old people and gains of fundamental gerontology.


Bronchial hyperreactivity to histamine in aspirin sensitive asthmatics: relationship to aspirin threshold and effect of aspirin desensitisation.
Twenty seven aspirin sensitive asthmatic patients were studied to determine the relationship between non-specific bronchial responsiveness to inhaled histamine and the degree of sensitivity to aspirin (aspirin threshold dose). No correlation was found between provocative concentration of histamine (PC20H) and aspirin threshold dose. In 11 patients the influence of aspirin desensitisation on bronchial reactivity to inhaled histamine was examined. Mean PC20H measured the day after the patients were desensitised to 600 mg of aspirin did not change significantly from the values before desensitisation. These observations suggest that sensitivity to aspirin and non-specific bronchial hyperreactivity in asthmatic patients are independent phenomena.


What, not just salad and veg? Consumer testing of the eatwell week.
To test the appeal of the eatwell week, a nutritionally balanced 7 d menu which satisfies nutritional guidelines of the Food Standards Agency in Scotland; determine the clarity and understanding of the main messages; and gather views on the usability and acceptability of the eatwell week resource format. Focus group discussions with consumers and health professionals. Four locations across the UK. The eatwell week was considered realistic by consumers as it contained foods they recognised and already ate. A preconceived idea had been that there would be more fruit and vegetables and fewer 'treats'. Consumers found the recipes simple and lack of cooking skills was not an apparent barrier. However, the message of 'balance' was poorly understood. Consumers often lacked the knowledge to make informed substitutions in the week. Both the general public and some health professionals felt the menu contained too much carbohydrate. Health professionals felt it was unclear who the eatwell week was intended for and what purpose it served. Use of familiar foods and the provision of simple, easy-to-follow recipes have the potential to overcome some barriers to healthy eating encountered by the general public and encourage improvements in dietary intakes. The eatwell week shows promise as a resource to facilitate implementation of the principles of the eatwell plate and supports government priorities and policies for health.


Molecular basis of P450 inhibition and activation: implications for drug development and drug therapy.
Three-dimensional homology models of cytochromes P450 (P450) 2B1 and P450 3A4 have been utilized along with site-directed mutagenesis to elucidate the molecular determinants of substrate specificity. Most of the key residues identified in 2B enzymes fall within five substrate recognition sites (SRSs) and have counterparts in bacterial P450 residues that regulate substrate binding or access. Docking of inhibitors into 2B models has provided a plausible explanation for changes in susceptibility to mechanism-based inactivation that accompany particular amino acid side-chain replacements. These studies provide a basis for predicting drug interactions due to P450 inhibition and for rational inhibitor design. In addition, the location of P450 3A4 residues capable of influencing homotropic stimulation by substrates and heterotropic stimulation by flavonoids has been identified. Steroid hydroxylation by the wild-type enzyme exhibits sigmoidal kinetics, indicative of positive cooperativity. Based on the 3A4 model and single-site mutants, a double mutant in SRS-2 has been constructed that exhibits normal Michaelis-Menten kinetics. Results of modeling and mutagenesis studies suggest that the substrate and effector bind at adjacent sites within a single large cavity in P450 3A4. A thorough understanding of the location and structural requirements of the substrate-binding and effector sites in cytochrome P450 3A4 should prove valuable in rationalizing and predicting interactions among the multitude of drugs and other compounds that bind to the enzyme.


Cholesterol-β1 AR interaction versus cholesterol-β2 AR interaction.
Two 8-µs all-atom molecular dynamics simulations have been performed on the two highly homologous G protein-coupled receptor (GPCR) subtypes, β1 - and β2 -adrenergic receptors, which were embedded in a lipid bilayer with randomly dispersed cholesterol molecules. During the simulations, cholesterol molecules accumulate to different surface regions of the two receptors, suggesting the subtype specificity of cholesterol-β-adrenergic receptor interaction and providing some clues to the physiological difference of the two subtypes. Meanwhile, comparison between the two receptors in interacting with cholesterols shed some new light on general determinants of cholesterol binding to GPCRs. Our results indicate that although the concave surface, charged residues and aromatic residues are important, neither of these stabilizing factors is indispensable for a cholesterol interaction site. Different combinations of these factors lead to the diversified binding modes of cholesterol binding to the receptors. Our long-time simulations, for the first time, revealed the pathway of a cholesterol molecule entering the consensus cholesterol motif (CCM) site, and the binding process of cholesterol to CCM is accompanied by a side chain flipping of the conserved Trp4.50. Moreover, the simulation results suggest that the I-/V-/L-rich region on the extracellular parts of helix 6 might be an alternatively conserved cholesterol-binding site for the class-A GPCRs.


Discovery of β-arrestin-biased β2-adrenoceptor agonists from 2-amino-2-phenylethanol derivatives.
β-Arrestins are a small family of proteins important for signal transduction at G protein-coupled receptors (GPCRs). β-Arrestins are involved in the desensitization of GPCRs. Recently, biased ligands possessing different efficacies in activating the G protein- versus the β-arrestin-dependent signals downstream of a single GPCR have emerged, which can be used to selectively modulate GPCR signal transduction in such a way that desirable signals are enhanced to produce therapeutic effects while undesirable signals of the same GPCR are suppressed to avoid side effects. In the present study, we evaluated agonist bias for compounds developed along a drug discovery project of β2-adrenoceptor agonists. About 150 compounds, including derivatives of fenoterol, 2-amino-1-phenylethanol and 2-amino-2-phenylethanol, were obtained or synthesized, and initially screened for their β-adrenoceptor-mediated activities in the guinea pig tracheal smooth muscle relaxation assay or the cardiomyocyte contractility assay. Nineteen bioactive compounds were further assessed using both the HTRF cAMP assay and the PathHunter β-arrestin assay. Their concentration-response data in stimulating cAMP synthesis and β-arrestin recruitment were applied to the Black-Leff operational model for ligand bias quantitation. As a result, three compounds (L-2, L-4, and L-12) with the core structure of 5-(1-amino-2-hydroxyethyl)-8-hydroxyquinolin-2(1H)-one were identified as a new series of β-arrestin-biased β2-adrenoceptor agonists, whereas salmeterol was found to be Gs-biased. These findings would facilitate the development of novel drugs for the treatment of both heart failure and asthma.


Postprandial lipid abnormalities in type 2 diabetes mellitus.
To study the postprandial lipid abnormalities in patients with type 2 diabetes mellitus. Postprandial lipids were studied in 20 male type 2 diabetic subjects (age 49.75 +/- 4.82 years) and 20 age and sex matched healthy controls (age 49.55 +/- 4.82 years) after an oral fat challenge which consisted of a meal providing 729 keal/m2 body surface area with 68 gm fat. Average duration of diabetes among diabetic was 2.32 +/- 3.03 years. The body mass index (cases 25.84 +/- 4.52; controls 25.74 +/- 5.0; p > 0.05) and waist-hip ratio (cases 1.06 +/- 0.13; controls 1.14 +/- 0.2; p > 0.05) were similar in both groups. While fasting serum lipids were not significantly different between the two groups, a number of serum lipid abnormalities were noted in type 2 diabetic subjects in the postprandial state. These included a higher triglyceride-area under curve (AUC) (cases 1298.08 +/- 485.2 vs. controls 922.15 +/- 390.47 mg/dl/8h; p=0.01), a higher triglyceride-area under incremental curve (AUIC) (cases 549.68 +/- 382.24; control 294.75 +/- 172.6 mg/dl/8h; p=0.01), a higher peak triglyceride level (cases 425.2 +/- 204.47 mg%, controls 283.9 +/- 11.6.94 mg%, p=0.01), a lower HDL-AUC (cases 130.35 +/- 33.55 vs. controls 168.48 +/- 56.01 mg/dl/8h, p=0.013) and a lower HDL nadir (Cases 28.05 +/- 10.94 mg%, controls 37.13 +/- 13.52 mg%, p < 0.02). Triglyceride AUC correlated significantly with fasting serum triglyceride (r=0.62) and BMI (r=0.7), but not with waist hip ratio or fasting serum insulin levels. Postprandial lipaemia did not correlate with age, duration of diabetes, fasting blood glucose or glycosylated hemoglobin. In conclusion, make type 2 diabetics demonstrate significant postprandial lipid abnormalities, particularly of triglycerides, which appear to be independent of glycaemic control.


Tembusu-like flavivirus (Perak virus) as the cause of neurological disease outbreaks in young Pekin ducks.
A neurological disease of young Pekin ducks characterized by ataxia, lameness, and paralysis was observed at several duck farms in Malaysia in 2012. Gross pathological lesions were absent or inconsistent in most of the cases, but severe and consistent microscopic lesions were found in the brain and spinal cord, characterized by non-purulent panencephalomyelitis. Several virus isolates were obtained in embryonated duck eggs and in cell cultures (Vero and DF-1) inoculated with the brain homogenates of affected ducks. After exclusion of other viruses, the isolates were identified as a flavivirus by flavivirus-specific reverse transcription-polymerase chain reaction (RT-PCR) assays. Inoculation of 2-week-old Pekin ducks with a flavivirus isolate by the subcutaneous or intramuscular route resulted in typical clinical signs and histological lesions in the brain and spinal cord. The inoculated virus was detected by RT-PCR from organ samples of ducks with clinical signs and histological lesions. With a few days delay, the disease was also observed among co-mingled contact control birds. Phylogenetic analysis of NS5 and E gene sequences proved that the isolates were representatives of a novel phylogenetic group within clade XI (Ntaya virus group) of the Flavivirus genus. This Malaysian Duck Tembusu Virus (DTMUV), named Perak virus, has moderate genomic RNA sequence similarity to a related DTMUV identified in China. In our experiment the Malaysian strain of DTMUV could be transmitted in the absence of mosquito vectors. These findings may have implications for the control and prevention of this emerging group of flaviviruses.


Effects of magnetic shear on toroidal rotation in tokamak plasmas with lower hybrid current drive.
Application of lower hybrid (LH) current drive in tokamak plasmas can induce both co- and countercurrent directed changes in toroidal rotation, depending on the core q profile. For discharges with q(0) <1, rotation increments in the countercurrent direction are observed. If the LH-driven current is sufficient to suppress sawteeth and increase q(0) above unity, the core toroidal rotation change is in the cocurrent direction. This change in sign of the rotation increment is consistent with a change in sign of the residual stress (the divergence of which constitutes an intrinsic torque that drives the flow) through its dependence on magnetic shear.


Identification and functional analysis of antifungal immune response genes in Drosophila.
Essential aspects of the innate immune response to microbial infection appear to be conserved between insects and mammals. Although signaling pathways that activate NF-kappaB during innate immune responses to various microorganisms have been studied in detail, regulatory mechanisms that control other immune responses to fungal infection require further investigation. To identify new Drosophila genes involved in antifungal immune responses, we selected genes known to be differentially regulated in SL2 cells by microbial cell wall components and tested their roles in antifungal defense using mutant flies. From 130 mutant lines, sixteen mutants exhibited increased sensitivity to fungal infection. Examination of their effects on defense against various types of bacteria and fungi revealed nine genes that are involved specifically in defense against fungal infection. All of these mutants displayed defects in phagocytosis or activation of antimicrobial peptide genes following infection. In some mutants, these immune deficiencies were attributed to defects in hemocyte development and differentiation, while other mutants showed specific defects in immune signaling required for humoral or cellular immune responses. Our results identify a new class of genes involved in antifungal immune responses in Drosophila.


Characterization of Entamoeba histolytica adenosine 5'-phosphosulfate (APS) kinase; validation as a target and provision of leads for the development of new drugs against amoebiasis.
Amoebiasis, caused by Entamoeba histolytica infection, is a global public health problem. However, available drugs to treat amoebiasis are currently limited, and no effective vaccine exists. Therefore, development of new preventive measures against amoebiasis is urgently needed. Here, to develop new drugs against amoebiasis, we focused on E. histolytica adenosine 5'-phosphosulfate kinase (EhAPSK), an essential enzyme in Entamoeba sulfolipid metabolism. Fatty alcohol disulfates and cholesteryl sulfate, sulfolipids synthesized in Entamoeba, play important roles in trophozoite proliferation and cyst formation. These processes are closely associated with clinical manifestation and severe pathogenesis of amoebiasis and with disease transmission, respectively. We validated a combination approach of in silico molecular docking analysis and an in vitro enzyme activity assay for large scale screening. Docking simulation ranked the binding free energy between a homology modeling structure of EhAPSK and 400 compounds. The 400 compounds were also screened by a 96-well plate-based in vitro APSK activity assay. Among fifteen compounds identified as EhAPSK inhibitors by the in vitro system, six were ranked by the in silico analysis as having high affinity toward EhAPSK. Furthermore, 2-(3-fluorophenoxy)-N-[4-(2-pyridyl)thiazol-2-yl]-acetamide, 3-phenyl-N-[4-(2-pyridyl)thiazol-2-yl]-imidazole-4-carboxamide, and auranofin, which were identified as EhAPSK inhibitors by both in silico and in vitro analyses, halted not only Entamoeba trophozoite proliferation but also cyst formation. These three compounds also dose-dependently impaired the synthesis of sulfolipids in E. histolytica. Hence, the combined approach of in silico and in vitro-based EhAPSK analyses identified compounds that can be evaluated for their effects on Entamoeba. This can provide leads for the development of new anti-amoebic and amoebiasis transmission-blocking drugs. This strategy can also be applied to identify specific APSK inhibitors, which will benefit research into sulfur metabolism and the ubiquitous pathway terminally synthesizing essential sulfur-containing biomolecules.


Tissue lipids in leprosy.
A total of 30 cases of leprosy (15 lepromatous and 15 tuberculoid) were studied by histochemical procedures for lipids in the morbid skin. The possible origin and relation of lipids to the presence of lepra bacilli in the lepra cells is discussed.


The use of positive B cell flow cytometry crossmatch in predicting rejection among renal transplant recipients.
We performed retrospective flow cytometry crossmatch (FCXM) on 106 renal graft recipients who were transplanted based on current T cell negative serologic crossmatch. T and B cell FCXMs were performed on current and historical peak reactive post-transplant sera using 1024-channel flow cytometer and the shift in median channel fluorescence (SMCF) over the negative control was calculated. Cut-off values for a positive T and B crossmatch, > 40 and > 80 SMCF, respectively, were determined based on previous retrospective analysis of the data in the context of clinical outcome in our center, and were 1.5 times the standard deviation (SD) above the mean median channel fluorescence (MCF) of normal sera controls. The 1-yr graft survival was 95% for the total group of patients studied, and 87% for the recipients who had a positive T cell FCXM. To focus on the influence of a positive B cell FCXM on the incidence of rejection, primary transplant recipients who had a negative T cell FCXM (n = 81) were studied. Fifteen of 30 (50%) recipients with a positive B cell FCXM experienced at least one rejection episode within the first year. By contrast, only 15 of 51 (29.4%) of patients with a negative B cell FCXM experienced rejection (p = 0.05). The mean B cell SMCF in the group of patients who had no rejections was 45 +/- 59, while that of the group of patients who experienced at least one rejection was 97 +/- 97 (p = 0.012). By comparison, the rejection rate among the retransplant patients was 44.4%, and the mean B cell SMCF in the group with rejection was 94 +/- 75 while it was 5 +/- 7 among retransplant patients who did not have rejection (p = 0.031). Eighty-six percent of sensitized (panel reactivity antibodies (PRA) > 10%) patients who had a B positive/T negative FCXM experienced rejection, compared to 33% (n = 6 out of 16) of the B negative/T negative sensitized patients (p = 0.03). Furthermore, 62% (n = 13 out of 21) of donor-recipient mismatched patients with a B positive/T negative FCXM experienced rejection, compared to 38% (n = 13 out of 35) of patients with T negative/B negative FCXM who were similarly mismatched (p = 0.064). These data demonstrate the value of a positive B cell FCXM for predicting post-transplant rejections particularly when evaluated in the context of prior sensitization and/or DR mismatching. Our results suggest that B cell FCXM may have significant clinical implications, justifying its use in post-transplant management of recipients who have other risk factors of rejection.


Deciphering specificity and cross-reactivity in tachykinin NK1 and NK2 receptors.
The tachykinin receptors neurokinin 1 (NK1R) and neurokinin 2 (NK2R) are G protein-coupled receptors that bind preferentially to the natural peptide ligands substance P and neurokinin A, respectively, and have been targets for drug development. Despite sharing a common C-terminal sequence of Phe-X-Gly-Leu-Met-NH2 that helps direct biological function, the peptide ligands exhibit some degree of cross-reactivity toward each other's non-natural receptor. Here, we investigate the detailed structure-activity relationships of the ligand-bound receptor complexes that underlie both potent activation by the natural ligand and cross-reactivity. We find that the specificity and cross-reactivity of the peptide ligands can be explained by the interactions between the amino acids preceding the FxGLM consensus motif of the bound peptide ligand and two regions of the receptor: the β-hairpin of the extracellular loop 2 (ECL2) and a N-terminal segment leading into transmembrane helix 1. Positively charged sidechains of the ECL2 (R177 of NK1R and K180 of NK2R) are seen to play a vital role in the interaction. The N-terminal positions 1 to 3 of the peptide ligand are entirely dispensable. Mutated and chimeric receptor and ligand constructs neatly swap around ligand specificity as expected, validating the structure-activity hypotheses presented. These findings will help in developing improved agonists or antagonists for NK1R and NK2R.


Understanding high rates of stillbirth and neonatal death in a disadvantaged, high-migrant district in France: A perinatal audit.
The objective of this study is to investigate factors associated with risks of perinatal death in a disadvantaged, high-migrant French district with mortality rates above the national average. The study design is a perinatal audit in 2014 in all 11 maternity units in the Seine-Saint-Denis district (25 037 births). The data come from medical chart abstraction, maternal interviews and peer assessor confidential review of deaths. A representative sample of live births in the same district, from the 2010 French Perinatal Survey, was used for comparisons (n = 429). The main outcome measures were stillbirth and neonatal death (0-27 days) at ≥22 weeks of gestation. The audit included 218 women and 227 deaths (156 stillbirths, 71 neonatal deaths); 75 women were interviewed. In addition to primiparity and multiple pregnancy, overweight and obesity increased mortality risks (50% of cases, adjusted odds ratios [aOR] 1.7, 95% confidence interval [CI] 1.1-2.8, and aOR 1.9 [95% CI 1.1-3.2], respectively) as did the presence of preexisting medical/obstetric conditions (28.6% of cases, aOR 3.2, 95% CI 2.0-5.3). Problems accessing or complying with care were noted in 25% of medical records and recounted in 50% of interviews. Assessors identified suboptimal factors in 73.2% of deaths and judged 33.9% to be possibly or probably preventable. Care not adapted to risk factors and poor healthcare coordination were frequent suboptimal factors. Possibly preventable deaths were higher (P < .05) for women with gestational diabetes or hypertension (44.6%) than women without (29.0%). Preventive actions to improve healthcare referral and coordination, especially for overweight and obese women and women with medical and obstetrical risk factors, could reduce perinatal mortality in disadvantaged areas.


Modification of the Textural Properties of Chitosan to Obtain Biochars for CO2-Capture Processes.
Three chitosans with different morphologies have been used (commercial chitosan powder, chitosan in film form and chitosan in globular form synthesized by the freeze-dried method) for the synthesis of biochars. The pyrolytic treatment has revealed that the biochar synthesized from the chitosan formed by the freeze-dried method reaches the highest CO2-adsorption capacity (4.11 mmol/g at 0 °C and a pressure of 1 bar) due to this adsorbent is highly microporous. Moreover, this biochar is more resistant to the pyrolytic treatment in comparison to the biochars obtained from the commercial chitosan and chitosan in the form of film. CO2-adsorption studies at different temperatures have also shown that the adsorption capacity diminishes as the adsorption temperature increases, thus suggesting that the adsorption takes place by a physical process.