Ketone body metabolism in normal and diabetic human skeletal muscle.
Although the liver is considered the major source of ketone bodies (KB) in humans, these compounds may also be formed by nonhepatic tissues. To study this aspect further, 3-[14C]hydroxybutyrate (BOH) or [3-14C]acetoacetate (AcAc) were constantly infused after a priming dose and contemporaneous arterial and venous samples were taken at splanchnic, heart, kidney, and leg sites in eight normal subjects (N) undergoing diagnostic catheterization and at the forearm site in five normal and six ketotic diabetic (D) subjects. After 70 min of infusion, tracer and tracee levels of AcAc and BOH reached a steady state in the artery and vein in both normal and diabetic subjects. The venous-arterial (V-A) difference at the forearm step for cold KB was negligible both in normal and diabetic subjects, whereas for labeled KB it was approximately 10-fold higher in diabetic subjects (V-A AcAc, -31 +/- 7 and -270 +/- 34 dpm/ml in N and D, respectively; V-A BOH, -38 +/- 6 and -344 +/- 126 dpm/ml in N and D, respectively). We assumed that the V-A difference in tracer concentration was consistent with dilution of the tracer by newly synthesized tracee inside the muscle and calculated that the forearm muscle produces KB at a rate of 16.2 +/- 3.3 mumol/min in D and 0.9 +/- 0.9 mumol/min in N. These findings can be accounted for by the hypothesis that the disappearance flux of KB from circulation was replaced by an equivalent flux of KB entering the vein at the muscle step in D but not in N. Moreover, in N KB were not only produced but also utilized by the splanchnic area (39 +/- 9 mumol/min).(ABSTRACT TRUNCATED AT 250 WORDS)


Prospective Memory Is a Key Predictor of Functional Independence in Older Adults.
Prospective memory (PM), the ability to execute delayed intentions, has received increasing attention in neuropsychology and gerontology. Most of this research is motivated by the claim that PM is critical for maintaining functional independence; yet, there is a dearth of empirical evidence to back up the claims. Thus, the present study tested whether PM predicts functional independence in older adults using validated behavioral performance measures for both PM and functional independence. Fifty-eight healthy older adults performed a computerized PM paradigm, the Virtual Week task, as well as a timed version of an instrumental activities of daily living (TIADL) task. Furthermore, we assessed vocabulary, processing speed, and self-reported prospective remembering. TIADL scores correlated significantly with performance in the Virtual Week task, vocabulary, and processing speed. Hierarchical linear regressions revealed that vocabulary and Virtual Week performance were significant predictors for TIADL. However, self-reported PM scores did not predict everyday functioning. The findings indicate that PM is an important cognitive ability for successful and independent everyday life beyond vocabulary. Moreover, the results show a substantial incremental contribution of intact PM performance for the prediction of everyday functioning by using objective PM measures. (JINS, 2018, 24, 640-645).


Combinatorics of traces of Hecke operators.
We investigate the combinatorial properties of the traces of the nth Hecke operators on the spaces of weight 2k cusp forms of level N. We establish examples in which these traces are expressed in terms of classical objects in enumerative combinatorics (e.g., tilings and Motzkin paths). We establish in general that Hecke traces are explicit rational linear combinations of values of Gegenbauer (also known as ultraspherical) polynomials. These results arise from "packaging" the Hecke traces into power series in weight aspect. These generating functions are easily computed by using the Eichler-Selberg trace formula.


Bone evaluation in paediatric chronic kidney disease: clinical practice points from the European Society for Paediatric Nephrology CKD-MBD and Dialysis working groups and CKD-MBD working group of the ERA-EDTA.
Mineral and bone disorder (MBD) is widely prevalent in children with chronic kidney disease (CKD) and is associated with significant morbidity. CKD may cause disturbances in bone remodelling/modelling, which are more pronounced in the growing skeleton, manifesting as short stature, bone pain and deformities, fractures, slipped epiphyses and ectopic calcifications. Although assessment of bone health is a key element in the clinical care of children with CKD, it remains a major challenge for physicians. On the one hand, bone biopsy with histomorphometry is the gold standard for assessing bone health, but it is expensive, invasive and requires expertise in the interpretation of bone histology. On the other hand, currently available non-invasive measures, including dual-energy X-ray absorptiometry and biomarkers of bone formation/resorption, are affected by growth and pubertal status and have limited sensitivity and specificity in predicting changes in bone turnover and mineralization. In the absence of high-quality evidence, there are wide variations in clinical practice in the diagnosis and management of CKD-MBD in childhood. We present clinical practice points (CPPs) on the assessment of bone disease in children with CKD Stages 2-5 and on dialysis based on the best available evidence and consensus of experts from the CKD-MBD and Dialysis working groups of the European Society for Paediatric Nephrology and the CKD-MBD working group of the European Renal Association-European Dialysis and Transplant Association. These CPPs should be carefully considered by treating physicians and adapted to individual patients' needs as appropriate. Further areas for research are suggested.


Functional outcome measures for infantile Charcot-Marie-Tooth disease: a systematic review.
A functional outcome measure for infants (aged 0-3 years) with Charcot-Marie-Tooth (CMT) disease is needed for upcoming disease-modifying trials. A systematic review of outcome measures for infants with neuromuscular disorders was completed to determine if validated measures were available for the CMT infant population. We assessed 20,375 papers and identified seven functional outcome measures for infants with neuromuscular disorders. Six were developed and validated for spinal muscular atrophy (SMA). There were no CMT-specific outcome measures identified; however, one (motor function measure) assessed a range of neuromuscular disorders including 13 infants and children with CMT. The included studies exhibited "good" face, discriminant, convergent and concurrent validity, and reported excellent intra- and inter-rater reliability. No outcome measure was subjected to item response theory. Studies reported outcome measures comprising of 51 different items assessing six domains of function: reflexive movement, axial movement, limb movement, positioning, gross motor, and fine-motor skills. Scoring of items ranged from 2- to 7-point rating scales; and none were scaled to normative reference values to account for changes in growth and development. The SMA focus of most items is likely to produce ceiling effects and lack sensitivity and responsiveness for within and between types of CMT in infants. Nevertheless, several items across scales assessing distal strength, gross- and fine-motor function, could be included in the development of a composite functional outcome measure for infants with CMT to assess disease-modifying interventions.


Emergence of tick-borne encephalitis (TBE) in the Netherlands.
Recently, tick-borne encephalitis virus (TBEV) was detected in the Netherlands for the first time, in ticks collected in 2015 in the National Park Sallandse heuvelrug in response to the detection of anti-TBEV antibodies in roe deer. Hereafter, two human cases of autochthonous TBE have been reported, occurring in 2016. One case was geographically linked to the area of the previously reported ticks, which harbored a genetically divergent TBEV-Eu strain variant (TBEV-NL). So far these are the few reported events that point to endemic transmission of TBEV in the Netherlands and the true prevalence of TBEV and TBE disease in the Netherlands and its impact on the human population remains to be determined. We describe the third human case, identified in 2017, which geographically clusters with the aforementioned case and TBEV-positive ticks. We also describe the identification of another TBEV-NL-positive tick in the Netherlands, collected 2 years after the initial find in that same region (in 2017). These observations support the concept of continued circulation of TBEV-NL and the presence of a possible TBEV hot spot in the Sallandse Heuvelrug region.


Visual and cognitive deficits predict stopping or restricting driving: the Salisbury Eye Evaluation Driving Study (SEEDS).
To determine the visual and other factors that predict stopping or restricting driving in older drivers. A group of 1425 licensed drivers aged 67 to 87 years, who were residents of greater Salisbury, participated. At 1 year after enrollment, this group was categorized into those who had stopped driving, drove only within their neighborhood, or continued to drive beyond their neighborhood. At baseline, a battery of structured questionnaires, vision, and cognitive tests were administered. Multivariate analysis determined the factors predictive of stopping or restricting driving 12 months later. Of the 1425 enrolled, 1237 (87%) were followed up at 1 year. Excluding those who were already limiting their driving at baseline (n = 35), 1.5% (18/1202) had stopped and 3.4% (41/1202) had restricted their driving. The women (odds ratio [OR], 4.01; 95% confidence interval [CI], 2.05-8.20) and those who prefer to be driven (OR, 3.91; 95% CI, 1.91-8.00) were more likely to stop or restrict driving. Depressive symptoms increased likelihood of restricting or stopping driving (OR, 1.08; 95% CI, 1.009-1.16 per point Geriatric Depression Scale). Slow visual scanning and psychomotor speed (Trail Making Test, Part A: OR, 1.02; 95% CI, 1.01-1.03), poor visuoconstructional skills (Beery-Buktenica Test of Visual Motor Integration: OR, 1.14; 95% CI, 1.05-1.25), and reduced contrast sensitivity (OR, 1.15; 95% CI, 1.03-1.28) predicted stopping or reducing driving. Visual field loss and visual attention were not associated. The effect of vision on changing driving behavior was partially mediated by cognition, depression, and baseline driving preferences. In this cohort, contrast sensitivity and cognitive function were independently associated with incident cessation or restriction of driving space. These data suggest drivers with functional deficits make difficult decisions to restrict or stop driving.


Preoperative maximal removal rate of technetium-99m-galactosyl-human serum albumin of the remnant liver is associated with postoperative tumor relapse in hepatitis C virus-related hepatocellular carcinoma.
Prognosis in patients with hepatocellular carcinoma (HCC) is not only influenced by tumor-related factors but also by the background liver functions. The maximal removal rate of technetium-99m-galactosyl human serum albumin (GSA-Rmax) of the remnant liver (rGSA-Rmax) is a useful candidate for predicting the liver function and clarifying the relationship between the remnant liver functional reserve and tumor-free survival in patients who have undergone hepatectomy. One hundred and sixty-five patients with HCC who underwent curative hepatectomy were divided into three groups of hepatitis B virus (B-HCC; n=42), hepatitis C virus (C-HCC, n=58), and non-B, non-C (NBNC-HCC, n=65). The relationship between rGSA-Rmax and survival was examined by univariate and multivariate analyses. In the C-HCC group, the albumin, or LHL15, level was significantly lower, and alanine aminotransferase, ICGR15, and the prevalence of grade B liver damage were significantly higher than other two groups (P<0.05). GSA-Rmax or rGSA-Rmax was not different between the three groups. Lower GSA-Rmax and rGSA-Rmax were only significantly associated with lower tumor-free survival in the C-HCC group by the univariate analysis (P<0.05) but not significantly by the multivariate analysis. GSA-Rmax and rGSA-Rmax reflect the severity of liver dysfunction and furthermore, the lower rGSA-Rmax is useful as a complementary factor to predict the early HCC recurrence after hepatectomy.


Deficiency of phosphofructo-1-kinase/muscle subtype in humans impairs insulin secretion and causes insulin resistance.
Non-insulin-dependent diabetes mellitus (NIDDM) is caused by peripheral insulin resistance and impaired beta cell function. Phosphofructo-1-kinase (PFK1) is a rate-limiting enzyme in glycolysis, and its muscle subtype (PFK1-M) deficiency leads to the autosomal recessively inherited glycogenosis type VII Tarui's disease. It was evaluated whether PFK1-M deficiency leads to alterations in insulin action or secretion in humans. A core family of four members was evaluated for PFK1-M deficiency by DNA and enzyme-activity analyses. All members underwent oral and intravenous glucose tolerance tests (oGTT and ivGTT) and an insulin-sensitivity test (IST) using octreotide. Enzyme activity determinations in red blood cells showed that the father (46 yr, body mass index [BMI] 22. 4 kg/m2) and older son (19 yr, BMI 17.8 kg/m2) had a homozygous, while the mother (47 yr, BMI 28.4 kg/m2) and younger son (13 yr, BMI 16.5 kg/m2) had a heterozygous PFK1-M deficiency. DNA analyses revealed an exon 5 missense mutation causing missplicing of one allele in all four family members, and an exon 22 frameshift mutation of the other allele of the two homozygously affected individuals. The father showed impaired glucose tolerance, and the mother showed NIDDM. By ivGTT, both parents and the older son had decreased first-phase insulin secretion and a diminished glucose disappearance rate. The IST showed marked insulin resistance in both parents and the older, homozygous son, and moderate resistance in the younger son. PFK1-M deficiency causes impaired insulin secretion in response to glucose, demonstrating its participation in islet glucose metabolism, and peripheral insulin resistance. These combined metabolic sequelae of PFK-1 deficiency identify it as a candidate gene predisposing to NIDDM.


Antitumor effects of citrinin in an animal model of Sarcoma 180 via cytogenetic mechanisms.
Citrinin (CIT) is a cytotoxic, hepatotoxic, nephrotoxic and cardiotoxic metabolite obtained from Penicillium citrinum, that has been increasingly searched as an anticancer drug candidate. In this study, we assessed the antitumor effects of citrinin, using cytogenetic biomarkers for genotoxicity in Sarcoma 180 (S-180) ascitic fluid cells of mice. Citrinin, extracted from P. citrinum acetonitrile extract, was characterized by LC-MS. Cytotoxic assessment was done through using comet (alkaline version) and micronucleus assays. In S-180 cells, CI50 of CIT was 3.77 μg/mL, while at 12.5 and 100 μg/mL, CIT was as cytotoxic as doxorubicin (2 μg/mL). At 0.5, 1.0 and 2.0 μg/mL, it induced genotoxicity and mutagenicity in S-180 cells, especially at 2 μg/mL, triggering oxidative damage similar to hydrogen peroxide (10 mM). The antitumor effects were evidenced by a marked increase in S-180 cells apoptosis and necrosis due to clastogenic and/or aneugenic cytogenetic effects (micronucleus formation), as well as by induction of nucleoplasm bridges and nuclear buds, culminating in S-180 apoptosis and necrosis. CIT has potential as drug candidate for antitumor purposesbyinvolving cytogenetic mechanisms.


[Up-to-date methods of tomographic diagnostics for the evaluation of effectiveness of the treatment of laryngeal and laryngopharyngeal cancer].
Currently employed methods of tomographic diagnostics for the evaluation of effectiveness of the treatment of laryngeal and laryngopharyngeal cancer are described. The authors underscore that increasingly more attention tends to be given to magnetoresonance imaging (MRI) and especially X-ray computer tomography (CT) due to their wide application and high informative value whereas nuclear medicine techniques, such as single-photon emission CT using thallium-200, 11C-methionine and (18)F-positron emission tomography, are considered to be less important despite their great diagnostic potentialities. It is emphasized that X-ray CT with intravenous administration of contrast material should be regarded as the method of choice for the evaluation of larynx and pharynx conditions following treatment of their tumours. MRI should be reserved for patients in whom X-ray CT with contrast enhancement proves impracticable.


Direct oral anticoagulants versus warfarin for the treatment of inferior vena cava thrombus.
To evaluate the efficacy and safety of direct oral anticoagulants (DOACs) compared to warfarin in patients with inferior vena cava (IVC) thrombus. This was a single-system, retrospective cohort study of hospitalized adult patients with IVC thrombus treated with a DOAC or warfarin therapy. The primary efficacy endpoint was the thrombus resolution on imaging, and the primary safety endpoint was major bleeding, both assessed within 6 months of hospital discharge. Secondary endpoints included hospitalization for a bleeding-related event, pulmonary embolism, or death within 6 months of hospital discharge. A total of 33 patients were included in the study. Twenty-three (70%) patients received a DOAC, and 10 (30%) received warfarin. Of the 10 patients with repeat imaging available, complete resolution was noted in two (33%) DOAC patients and no warfarin patients (p = .5). Major bleeding occurred in two (8.7%) DOAC patients and one (10%) warfarin patient (p = .9). No significant differences in secondary endpoints were observed between groups. There were no differences in efficacy and safety between patients receiving DOACs or warfarin for the treatment of IVC thrombus, although results are limited by the small patient population and number of patients with repeat imaging available.


Malaria in the Limbé River valley of northern Haiti: a hospital-based retrospective study, 1975-1997.
In the Limbé River valley of northern Haiti a retrospective study at the Bon Samaritain Hospital (BSH) determined the total number of cases and the cyclical nature of malaria from 1975 through 1997, examined the relationship between rainfall and malaria from 1975 through 1985, and compared the incidence of malaria at that hospital with general trends for Haiti for 1975 through 1996 as reported by the World Health Organization (WHO). During 1975-1997, 27,078 positive cases of malaria were diagnosed at BSH; 50% of these cases occurred during 16 weeks out of the year, during a summer peak in June and July and a winter peak in December and January. For 1975-1985, there was no significant correlation between the incidence of malaria and annual rainfall. The strongest correlation was observed between weekly rainfall and weekly incidence of malaria when the data was staggered to allow a lag of 9-11 weeks between rainfall and new malaria cases. The lag period is explained by the time required for the creation of breeding sites after rain, the life cycles of the Anopheles albimanus mosquito and the Plasmodium falciparum parasite, and the incubation period for falciparum malaria. The incidence of malaria in the Limbé River valley loosely followed the trends in all of Haiti and also supported WHO reports indicating that malaria in Haiti has been in a general decline since the mid-1980s. By showing the seasonal trends for malaria in the Limbé valley and the relationship between rainfall and malaria over an extended time period, this study provides a means to measure the effectiveness of malaria control efforts in the region.


Two component mie scattering models of sargasso sea particles.
The volume scattering function is calculated for particle suspensions consisting of two components systematically distributed in a manner consistent with Coulter Counter observations in the Sargasso Sea. The components are assigned refractive indices 1.01-0.01i and 1.15 to represent organic and inorganic particles, respectively. The only models found that reproduce observed scattering functions require a considerable fraction of the suspended particle volume to be organic in nature. This fraction, however, contributes less than 10% to the total scattering function. The model finally chosen indicates that the inorganic particles smaller than 2.5 micro do not occur in large enough concentrations to have a significant effect on the volume scattering function.


Chronic rhinosinusitis: an enhanced immune response to ubiquitous airborne fungi.
Chronic rhinosinusitis (CRS) is one of the most common long-term illnesses in the United States. The etiology of CRS is unknown, and no effective treatment has been established. We investigated the hypothesis that abnormal immunologic responses to ubiquitous airborne fungi contribute to the pathogenesis of this disease. The proliferative and cytokine responses of PBMCs to extracts from 4 common airborne fungi-including Alternaria , Aspergillus , Cladosporium , and Penicillium -were examined by in vitro culture. Serum specimens were tested for specific IgE and IgG to these fungi. PBMCs from approximately 90% of the patients with CRS, but not those from normal individuals, produced both IL-5 and IL-13 when exposed to Alternaria. Furthermore, PBMCs from patients with CRS produced significantly more IFN-gamma than PBMCs from normal individuals in response to Alternaria (median, 553 pg/mL vs 98 pg/mL; P < .01). Levels of serum IgG antibodies to Alternaria and Cladosporium were clearly increased in patients with CRS compared with normal individuals ( P < .01). Less than 30% of the patients with CRS had specific IgE antibodies to Alternaria or Cladosporium. The increased humoral (serum IgG) response strongly correlated with the increased cellular (IL-5 production) response to Alternaria ( r = 0.619; P < .01). Patients with CRS show exaggerated humoral and cellular responses, both T(H)1 and T(H)2 types, to common airborne fungi, particularly Alternaria. The anomalous immune and inflammatory responses to ubiquitous fungi may explain the chronicity of airway inflammation in CRS.


Assessing Volumetric Absorptive Microsampling Coupled with Stable Isotope Dilution Assay and Liquid Chromatography-Tandem Mass Spectrometry as Potential Diagnostic Tool for Whole Blood 5-Methyltetrahydrofolic Acid.
Volumetric absorptive microsamplers (VAMS) have been developed recently as a promising tool for clinical blood sampling. Compared to dried blood spot samples analyzed by accurate stable isotope dilution assays (SIDAs), the new technique could provide further substantial miniaturizing of folate assays by eliminating hematocrit effects and uneven analyte distribution within the sample. Herein, we present a miniaturized SIDA coupled with LC-MS/MS measurement of 5-methyltetrahydrofolic acid as main folate vitamer in whole blood (WB) using [13C5]-5-methyltetrahydrofolic acid as internal standard. Elution and extraction of only 10.8 μL-dried WB were carried out by centrifugation followed by enzymatic treatment for polyglutamate deconjugation. Matrix separation was achieved by heating and centrifugation. To verify applicability, WB folate status of 11 volunteers was screened. Limits of detection and limits of quantitation were 9 and 26 nmol·L-1, respectively, which is sufficiently low for screening folate status. Recoveries were 97 (±5.8), 99 (±2.8), and 96 (±6.1)% for 800, 400, and 200 nmol L-1 5-methyltetrahydrofolic acid, respectively. Precision of the LC-MS/MS instrument and inter-assay precision trials revealed CVs of 8.1 and 3.5% (294 nmol L-1), respectively, thus confirming reproducible and precise quantitation. Compared to fresh WB, no significant degradation of 5-methyltetrahydrofolate was observed after 2.5 h of drying at room temperature. VAMS 5-CH3-H4folate was stable for at least 3 weeks at -20°C. In our pilot study, accurate and diagnostically conclusive determination of folate status was verified. Nevertheless, blood sampling should be performed by trained individuals to avoid substantial errors concerning the absorbed volume. Endogenous folate in rat serum and chicken pancreas caused a significant background especially at low blood 5-CH3-H4folate levels and, thus, for polyglutamate deconjugation, these background folates or alternative mixtures need to be removed. The superior feasibility of a minimized blood collection with VAMS allows further progress regarding time- and cost-effective methodologies in newborn or population screenings for 5-methyltetrahydrofolate status. Further steps toward minimization could include an automated assay coupled with UPLC-MS/MS.


Mitigation Effects of Selenium Nanoparticles on Depression-Like Behavior Induced by Fluoride in Mice via the JAK2-STAT3 Pathway.
Depression is a mental health problem with typically high levels of distress and dysfunction, and 150 mg/L fluoride (F) can induce depression-like behavior. The development of depression is correlated with neuronal atrophy, insufficient secretion of monoamine neurotransmitters, extreme deviations from the normal microglial activation status, and immune-inflammatory response. Studies found that Se supplementation was related to the improvement of depression. In this study, we applied selenium nanoparticles (SeNPs) for F-induced depression disease mitigation by regulating the histopathology, metabolic index, genes, and protein expression related to the JAK2-STAT3 signaling pathway in vivo. Results showed that F and 2 mg Se/kg BW/day SeNPs lowered the dopamine (DA) content (P < 0.05), altered the microglial morphology, ramification index as well as solidity, and triggered the microglial neuroinflammatory response by increasing the p-STAT3 nuclear translocation (P < 0.01). Furthermore, F reduced the cortical Se content and the number of surviving neurons (P < 0.05), increasing the protein expressions of p-JAK2/JAK2 and p-STAT3/STAT3 of the cortex (P < 0.01), accompanied by the depression-like behavior. Importantly, 1 mg Se/kg BW/day SeNPs alleviated the microglial ramification index as well as solidity changes and decreased the interleukin-1β secretion induced by F by suppressing the p-STAT3 nuclear translocation (P < 0.01). Likewise, 1 mg Se/kg BW/day SeNPs restored the F-disturbed dopamine and noradrenaline secretion, increased the number of cortical surviving neurons, and reduced the vacuolation area, ultimately suppressing the occurrence of depression-like behavior through inhibiting the JAK2-STAT3 pathway activation. In conclusion, 1 mg Se/kg BW/day SeNPs have mitigation effects on the F-induced depression-like behavior. The mechanism of how SeNPs repair neural functions will benefit depression mitigation. This study also indicates that inhibiting the JAK/STAT pathway can be a promising novel treatment for depressive disorders.


Establishment of a mouse- and egg-adapted strain for the evaluation of vaccine potency against H3N2 variant influenza virus in mice.
Sporadic spreads of swine-origin influenza H3N2 variant (H3N2v) viruses were reported in humans, resulting in 437 human infections between 2011 and 2021 in the USA. Thus, an effective vaccine is needed to better control a potential pandemic for these antigenically distinct viruses from seasonal influenza. In this study, a candidate vaccine strain with efficient growth capacity in chicken embryos was established through serial blind passaging of A/Indiana/08/2011 (H3N2)v in mice and chicken embryos. Seven amino acid substitutions (M21I in PA; A138T, N165K, and V226A in HA; S312L in NP; T167I in M1; G62A in NS1 proteins) were found in the passaged viruses without a major change in the antigenicity. This mouse- and egg-adapted virus was used as a vaccine and challenge strain in mice to evaluate the efficacy of the H3N2v vaccine in different doses. Antibodies with high neutralizing titers were induced in mice immunized with 100 µg of inactivated whole-virus particles, and those mice were significantly protected from the challenge of homologous strain. The findings indicated that the established strain in the study was useful for vaccine study in mouse models.


Changes in Respiratory Muscle Thickness during Mechanical Ventilation: Focus on Expiratory Muscles.
The lateral abdominal wall muscles are recruited with active expiration, as may occur with high breathing effort, inspiratory muscle weakness, or pulmonary hyperinflation. The effects of critical illness and mechanical ventilation on these muscles are unknown. This study aimed to assess the reproducibility of expiratory muscle (i.e., lateral abdominal wall muscles and rectus abdominis muscle) ultrasound and the impact of tidal volume on expiratory muscle thickness, to evaluate changes in expiratory muscle thickness during mechanical ventilation, and to compare this to changes in diaphragm thickness. Two raters assessed the interrater and intrarater reproducibility of expiratory muscle ultrasound (n = 30) and the effect of delivered tidal volume on expiratory muscle thickness (n = 10). Changes in the thickness of the expiratory muscles and the diaphragm were assessed in 77 patients with at least two serial ultrasound measurements in the first week of mechanical ventilation. The reproducibility of the measurements was excellent (interrater intraclass correlation coefficient: 0.994 [95% CI, 0.987 to 0.997]; intrarater intraclass correlation coefficient: 0.992 [95% CI, 0.957 to 0.998]). Expiratory muscle thickness decreased by 3.0 ± 1.7% (mean ± SD) with tidal volumes of 481 ± 64 ml (P < 0.001). The thickness of the expiratory muscles remained stable in 51 of 77 (66%), decreased in 17 of 77 (22%), and increased in 9 of 77 (12%) patients. Reduced thickness resulted from loss of muscular tissue, whereas increased thickness mainly resulted from increased interparietal fasciae thickness. Changes in thickness of the expiratory muscles were not associated with changes in the thickness of the diaphragm (R2 = 0.013; P = 0.332). Thickness measurement of the expiratory muscles by ultrasound has excellent reproducibility. Changes in the thickness of the expiratory muscles occurred in 34% of patients and were unrelated to changes in diaphragm thickness. Increased expiratory muscle thickness resulted from increased thickness of the fasciae.


Thermodynamics of interaction of phthalocyanine-oligonucleotide conjugates with single- and double-stranded DNA.
Thermodynamics of interaction of phthalocyanine-oligonucleotide conjugates with single- and double-stranded DNA resulting in formation of duplexes and triplexes was measured by UV melting method. It was shown that a phthalocyanine moiety of conjugates stabilized the formation of duplexes and triplexes.


Intestinal angioedema induced by angiotensin-converting enzyme inhibitors: an underrecognized cause of abdominal pain?
Intestinal angioedema caused by angiotensin-converting enzyme inhibitors such as lisinopril is rare but well documented in the literature. Patients with this condition typically present with common symptoms such as diffuse abdominal pain, cramping, nausea, and emesis. Imaging is needed to reveal segmental edema of the small intestine, often associated with free fluid in the abdomen. The authors report 2 cases of intestinal angioedema caused by angiotensin-converting enzyme inhibitors. Awareness of this allergic reaction and careful history taking--noting temporal relationship to occurrence of symptoms--are essential to diagnose this condition; laboratory and radiologic findings are needed to confirm the diagnosis. An accurate diagnosis helps the patient recover quickly and avoid complications from unnecessary tests and invasive procedures.


Atomic force microscopy as a tool to study the proteasome assemblies.
Proteasome is an exceptional enzyme because of its essential physiological role, multiple activities, and structural complexity. It is, in fact, a family of enzymes sharing a common catalytic core and equipped with distinct protein attachments regulating the core and adding to its new functional capabilities. As a drug target and a major regulator of cellular processes, proteasome is extensively studied with tools of structural, biochemical, and molecular biology. Atomic force microscopy (AFM) besides X-ray crystallography and electron microscopy is one of the most attractive methods to study proteasome. The noninvasive nature of this method is particularly well suited for investigating the structure-function relationship within the core particle (CP) as well as in higher-order assemblies. Here we review, from the methodological point of view, AFM-based studies on the proteasome. First, we will present the application of height distribution analysis of proteasome complexes to dissect the subunit organization in the base of the regulatory particle (RP). The RP is considered the most physiologically important among all the attachments of the CP; however, its structure remains enigmatic. Then, we will outline the use of AFM imaging to research on structural dynamics of the proteasome, a phenomenon which is starting to gain a broad interest. We will finish with a brief presentation of nanotechnological studies performed using ordered proteasomes and nanolithography carried out with the particles. The presented AFM research offers a unique and often unexpected insight into the structure and function of the proteasome.


Heart rates in hospitalized children by age and body temperature.
Heart rate (HR) is frequently used by clinicians in the hospital to assess a patient's severity of illness and make treatment decisions. We sought to develop percentiles that characterize the relationship of expected HR by age and body temperature in hospitalized children and to compare these percentiles with published references in both primary care and emergency department (ED) settings. Vital sign data were extracted from electronic health records of inpatients <18 years of age at 2 large freestanding children's hospitals from July 2011 to June 2012. We selected up to 10 HR-temperature measurement pairs from each admission. Measurements from 60% of patients were used to derive the percentile curves, with the remainder used for validation. We compared our upper percentiles with published references in primary care and ED settings. We used 60,863 observations to derive the percentiles. Overall, an increase in body temperature of 1°C was associated with an increase of ∼ 10 beats per minute in HR, although there were variations across age and temperature ranges. For infants and young children, our upper percentiles were lower than in primary care and ED settings. For school-age children, our upper percentiles were higher. We characterized expected HR by age and body temperature in hospitalized children. These percentiles differed from references in primary care and ED settings. Additional research is needed to evaluate the performance of these percentiles for the identification of children who would benefit from further evaluation or intervention for tachycardia.


Evolutionary dynamics of cancer in response to targeted combination therapy.
In solid tumors, targeted treatments can lead to dramatic regressions, but responses are often short-lived because resistant cancer cells arise. The major strategy proposed for overcoming resistance is combination therapy. We present a mathematical model describing the evolutionary dynamics of lesions in response to treatment. We first studied 20 melanoma patients receiving vemurafenib. We then applied our model to an independent set of pancreatic, colorectal, and melanoma cancer patients with metastatic disease. We find that dual therapy results in long-term disease control for most patients, if there are no single mutations that cause cross-resistance to both drugs; in patients with large disease burden, triple therapy is needed. We also find that simultaneous therapy with two drugs is much more effective than sequential therapy. Our results provide realistic expectations for the efficacy of new drug combinations and inform the design of trials for new cancer therapeutics. DOI:http://dx.doi.org/10.7554/eLife.00747.001.


Smoking initiation among young adults in the United States and Canada, 1998-2010: a systematic review.
Young adults have the highest smoking rate of any age group in the United States and Canada, and recent data indicate that they often initiate smoking as young adults. The objective of this study was to systematically review peer-reviewed articles on cigarette smoking initiation and effective prevention efforts among young adults. We searched 5 databases for research articles published in English between 1998 and 2010 on smoking initiation among young adults (aged 18-25) living in the United States or Canada. We extracted the following data from each study selected: the measure of initiation used, age range of initiation, age range of study population, data source, target population, sampling method, and sample size. We summarized the primary findings of each study according to 3 research questions and categories of data (eg, sociodemographic) that emerged during the data extraction process. Of 1,072 identified studies, we found 27 articles that met our search criteria, but several included a larger age range of initiation (eg, 18-30, 18-36) than we initially intended to include. Disparities in young adult smoking initiation existed according to sex, race, and educational attainment. The use of alcohol and illegal drugs was associated with smoking initiation. The risk of smoking initiation among young adults increased under the following circumstances: exposure to smoking, boredom or stress while serving in the military, attending tobacco-sponsored social events while in college, and exposure to social norms and perceptions that encourage smoking. Effective prevention efforts include exposure to counter-marketing, denormalization campaigns, taxation, and the presence of smoke-free policies. Much remains to be learned about young adult smoking initiation, particularly among young adults in the straight-to-work population. Dissimilar measures of smoking initiation limit our knowledge about smoking initiation among young adults. We recommend developing a standardized measure of initiation that indicates progression to regular established smoking.


Quantitative phosphoproteomic analysis reveals reciprocal activation of receptor tyrosine kinases between cancer epithelial cells and stromal fibroblasts.
Cancer-associated fibroblasts (CAFs) are one of the most important components of tumor stroma and play a key role in modulating tumor growth. However, a mechanistic understanding of how CAFs communicate with tumor cells to promote their proliferation and invasion is far from complete. A major reason for this is that most current techniques and model systems do not capture the complexity of signal transduction that occurs between CAFs and tumor cells. In this study, we employed a stable isotope labeling with amino acids in cell culture (SILAC) strategy to label invasive breast cancer cells, MDA-MB-231, and breast cancer patient-derived CAF this has already been defined above cells. We used an antibody-based phosphotyrosine peptide enrichment method coupled to LC-MS/MS to catalog and quantify tyrosine phosphorylation-mediated signal transduction events induced by the bidirectional communication between patient-derived CAFs and tumor cells. We discovered that distinct signaling events were activated in CAFs and in tumor epithelial cells during the crosstalk between these two cell types. We identified reciprocal activation of a number of receptor tyrosine kinases including EGFR, FGFR1 and EPHA2 induced by this bidirectional communication. Our study not only provides insights into the mechanisms of the interaction between CAFs and tumor cells, but the model system described here could be used as a prototype for analysis of intercellular communication in many different tumor microenvironments.


Gender differences in physician-patient communication. Evidence from pediatric visits.
To determine whether physician gender and patient gender influence the process of communication and parent and child satisfaction during pediatric office visits. Content analysis of videotaped pediatric office visits. University-based pediatric primary care practice. Videotaped communication between 212 children, ages 4 to 14 years, parents, and physicians. Thirty-eight percent were child health supervision visits, and 62% were for the management of minor or chronic illnesses. An established coding system of physician-patient communication and measures of parent and child satisfaction with medical care. Female physician visits were 29% longer than those of male physicians (P < .001). Compared with male physicians, female physicians engaged in more social exchange (P < .01), more encouragement and reassurance (P < .01), more communication during the physical examination (P < .05), and more information gathering (P < .01) with children. Male and female physicians engaged in similar amounts of discussions regarding illness management. Children were more satisfied with physicians of the same gender (P < .05), while parents were more satisfied with female physicians (P < .05). Children communicate more with female than with male physicians and show preferences for physicians of the same gender. These findings are consistent with communication patterns in adult patients and may have a significant influence on gender disparities in health care. Efforts at improving the process and outcome of medical care should address gender differences.


What stops children with a chronic illness accessing health care: a mixed methods study in children with Chronic Fatigue Syndrome/Myalgic Encephalomyelitis (CFS/ME).
Paediatric Chronic Fatigue Syndrome/Myalgic Encephalomyelitis (CFS/ME) is relatively common and disabling with a mean time out of school of more than one academic year. NICE guidelines recommend referral to specialist services immediately if severely affected, within 3 months if moderately affected and within 6 months if mildly affected. However, the median time-to-assessment by a specialist service in the UK is 18 months. This study used a mixed-methods approach to examine factors associated with time taken to access specialist services. Time-to-assessment was analysed as a continuous "survival-time" variable in Cox regression models using data from self-completed assessment forms for children attending a regional specialist CFS/ME service between January 2006 and December 2009. Semi-structured interviews about barriers experienced in accessing healthcare for their child were conducted with nine parents of children aged < 17 years (8 individual and one parent couple). Interviews were digitally recorded and analysed using "thematic analysis". 405 children were assessed between 2006 and 2009 and information on school attendance was available on 388. Only 1/125 with severe CFS/ME and 49/263 (19%) with mild to moderate CFS/ME were seen within NICE recommended timeframe. Increased fatigue was associated with shorter time to assessment (HR = 1.15; 95% CI 1.03, 1.29 per unit increase in Chalder fatigue score; P = 0.01). Time-to-assessment was not associated with disability, mood, age or gender. Parents described difficulties accessing specialist services because of their own as well as their GP's and Paediatrician's lack of knowledge. They experienced negative attitudes and beliefs towards the child's condition when they consulted GPs, Paediatricians and Child Psychiatrists. Parents struggled to communicate an invisible illness that their child and not themselves were experiencing. GPs, Child Psychiatrists and Paediatricians need more knowledge about CFS/ME and the appropriate referral pathways to ensure timeliness in referral to specialist services.


The Fc region of an antibody impacts the neutralization of West Nile viruses in different maturation states.
Flavivirus-infected cells secrete a structurally heterogeneous population of viruses because of an inefficient virion maturation process. Flaviviruses assemble as noninfectious, immature virions composed of trimers of envelope (E) and precursor membrane (prM) protein heterodimers. Cleavage of prM is a required process during virion maturation, although this often remains incomplete for infectious virus particles. Previous work demonstrated that the efficiency of virion maturation could impact antibody neutralization through changes in the accessibility of otherwise cryptic epitopes on the virion. In this study, we show that the neutralization potency of monoclonal antibody (MAb) E33 is sensitive to the maturation state of West Nile virus (WNV), despite its recognition of an accessible epitope, the domain III lateral ridge (DIII-LR). Comprehensive epitope mapping studies with 166 E protein DIII-LR variants revealed that the functional footprint of MAb E33 on the E protein differs subtly from that of the well-characterized DIII-LR MAb E16. Remarkably, aromatic substitutions at E protein residue 306 ablated the maturation state sensitivity of E33 IgG, and the neutralization efficacy of E33 Fab fragments was not affected by changes in the virion maturation state. We propose that E33 IgG binding on mature virions orients the Fc region in a manner that impacts subsequent antibody binding to nearby sites. This Fc-mediated steric constraint is a novel mechanism by which the maturation state of a virion modulates the efficacy of the humoral immune response to flavivirus infection.


Immunoexpression of N-cadherin, Twist and Vimentin in Bladder Urothelial Carcinomas.
Epithelial mesenchymal transition consists in the acquisition of neoplastic epithelial cells of a mesenchymal phenotype the process being involved in cancers invasion and metastasis. In this study were analyzed the expression of N-cadherin, Twist and Vimentin in bladder urothelial carcinomas according to the main prognostic parameters. The study included 20 bladder urothelial carcinomas which were analyzed histopathological, immunohistochemical and statistical. N-cadherin was identified in 45% of cases, which belonged to high-grade carcinomas with deep invasion and lymph node metastases. Twist immunoreaction was identified in all cases and was significantly increased in advanced stages carcinomas. Vimentin was present only in the advancing edge in 25% of cases, which belonged to highly invasive carcinomas. Urothelial carcinoma metastases were N-cadherin and Twist and Vimentin negative. We found a linear positive distribution of N-cadherin and Twist values. the used markers are useful for identifying aggressive urothelial carcinomas in the context of reciprocal stimulation mechanisms inside of urothelial epithelial-mesenchymal transition process.


In Situ Electrochemical Tip-Enhanced Raman Spectroscopy with a Chemically Modified Tip.
Chemically modified tips in scanning tunneling microscopy (STM) and atomic force microscopy (AFM) have been used to improve the imaging resolution or provide richer chemical information, mostly in ultrahigh vacuum (UHV) environments. Tip-enhanced Raman spectroscopy (TERS) is a nanoscale spectroscopic technique that already provides chemical information and can provide subnanometer spatial resolution. Chemical modification of TERS tips has mainly been focused on increasing their lifetimes for ambient and in situ experiments. Under UHV conditions, chemical functionalization has recently been carried out to increase the amount of chemical information provided by TERS. However, this strategy has not yet been extended to in situ electrochemical (EC)-TERS studies. The independent control of the tip and sample potentials offered by EC-STM allows us to prove the in situ functionalization of a tip in EC-STM-TERS. Additionally, the Raman response of chemically modified TERS tips can be switched on and off at will, which makes EC-STM-TERS an ideal platform for the development of in situ chemical probes on the nanoscale.


Intracellular cleavage of glycosylphosphatidylinositol by phospholipase D induces activation of protein kinase Calpha.
Many proteins are anchored to the cell membrane by glycosylphosphatidylinositol (GPI). One of the functions proposed for the GPI anchor is as a possible mediator in signal transduction through its hydrolysis. GPI-specific phospholipase D (GPI-PLD) is a secretory protein that is suggested to be involved in the release of GPI-anchored protein from the membrane. In the present study we examined how GPI-PLD is involved in signal transduction. When introduced exogenously and overexpressed in cells, GPI-PLD cleaved the GPI anchors in the early secretory pathway, possibly in the endoplasmic reticulum, resulting in an increased production of diacylglycerol. Experiments in vitro and in vivo showed that the association of protein kinase Calpha (PKCalpha) with membranes was increased markedly by expression of GPI-PLD in cells. Furthermore, sucrose-density-gradient centrifugation and immunofluorescence microscopy demonstrated that PKCalpha was translocated to the endoplasmic reticulum membrane in cells expressing GPI-PLD, in contrast with its association with the plasma membrane in cells treated with PMA. We also confirmed that the phosphorylation of c-Fos as well as PKCalpha itself was greatly enhanced by the expression of GPI-PLD. Taken together, these results suggest that GPI-PLD is involved in intracellular cleavage of the GPI anchor, which is a new potential source of diacylglycerol production to activate PKCalpha.


Patient Experiences of Life Years After Severe Civilian Lower Extremity Trauma With Vascular Injury.
Severe lower limb trauma with arterial injury is often devastating for the individual. Many studies describe how to manage these injuries when they occur. Short-term functional outcome is quite well described, but the patients are often young, and their suffering is physical, mental, and social from a lifelong perspective. The aim of this study was to report patient experiences of their lives several years after their accidents, and to explore mechanisms of how to improve management. The Swedvasc registry was searched for participants from 1987 to 2011, living in the region of Uppsala, Sweden. Some amputated participants were added from the Walking Rehabilitation Center. There were five reconstructed patients with an intact limb, and three with amputations. In depth interviews were conducted and systematically analyzed, using A Giorgi's descriptive phenomenological method. Eight patients participated, five with reconstructed and three with amputated limbs. Life affecting functional impairments were described by all patients. The patients undergoing amputation had received more structured follow up and support through the Walking Rehabilitation Center. The satisfaction with the cosmetic result was poorer than expected. All patients had developed strategies of how to cope with their impairments and stated they now lived "normal lives." Despite substantial physical, psychological, and cosmetic impairments years after severe lower limb trauma, the participants described life as "normal" and mainly satisfactory. Transition to the new situation could have been facilitated by more frequent and continuous follow up after discharge from hospital, in particular among the non-amputated patients who tend to be lost to follow up. Findings also indicate that family members have to be acknowledged, strengthened, and supported.


Circadian pattern of continuous FUDR infusion reduces toxicities.
The toxicity and efficacy of at least a dozen of the most commonly used anticancer drugs vary predictably depending on when in the day they are given. In a series of murine experiments involving CD2F1 mice, we have demonstrated that the safest time for FUDR administration is the mid- to late activity span. In a clinical pilot study, an implantable programmable drug-administration device (Medtronic, Inc), allowing automatic dose modifications of continuous long-term infusion, was employed to investigate whether this is also true in cancer patients. Toxicity of FUDR given as continuous flat infusion was significantly greater than that of time-modified infusion regardless of the route of drug administration. Responses have occurred with at least equal frequency, and patients are currently being followed to see if the pattern of drug delivery has an effect on patient survival.


Short term efficacy of ibuprofen phonophoresis versus continuous ultrasound therapy in knee osteoarthritis.
To compare the effectiveness of ibuprofen phonophoresis (PH) with conventional ultrasound (US) therapy in knee osteoarthritis. Sixty patients with a mean age of 59.8 +/- 9.0 years were randomly assigned to PH or US groups. Continuous ultrasonic waves of 1 MHz frequency and 1 watt/cm2 power were applied for 5 minutes to the target knee joint. Acoustic gel without any active pharmacological agent was applied in the US group, whereas cream containing 5% ibuprofen was applied in the PH group for a total treatment period of 10 sessions. The Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC) scores, pain on passive and active motion, 20 metres walking time, knee range of motion (ROM), and global assessments of disease activity and treatment efficacy by the investigator and by the patients were evaluated before and after therapy. Primary outcome measure of the study was 30% improvement in total WOMAC scores at the end of the study with respective scores at baseline. At the end of two weeks, 30% improvement in total WOMAC score was observed in 12 (40%) and 14 (46.6%) of patients in the PH and US groups respectively, indicating no significant difference in improvement rates. Pain scores, knee ROM degrees, 20 metres walking time measurements and all global assessment scores also improved significantly in both groups, yet these variables showed no significant differences between the two groups. When treatment efficacy was assessed as satisfaction rates, investigator satisfaction rates were 96.7% and 90%, while patient satisfaction rates were 93.3% and 83.3% in the PH and US groups respectively, suggesting similar satisfaction rates for both treatment methods. Both therapeutic modalities were found to be effective and generally well tolerated after 10 therapy sessions. Ibuprofen PH was not superior to conventional ultrasound in patients with knee osteoarthritis.


Salmonella susceptibility patterns in hospitals from 1975 through 1984.
The susceptibility of salmonella to commonly used antibiotics was analyzed for the 10-year period 1975 through 1984. The susceptibilities of 19,200 strains isolated from a yearly average of 211 hospitals nationwide and 1,508 strains isolated at the Kings County Medical Center in Brooklyn, N.Y., were described. Nationally, the susceptibility to nine antibacterial agents, including tetracycline, did not change markedly for the decade. The slight increase in susceptibility to tetracycline observed nationally may reflect the decreased clinical use of this antibiotic during the decade studied.


Performance characteristics of an instrument-free point-of-care CD4 test (VISITECT®CD4) for use in resource-limited settings.
CD4+ T lymphocyte count remains the most common biomarker of immune status and disease progression in human immunodeficiency virus (HIV)-positive individuals. VISITECT®CD4 is an instrument-free, low-cost point-of-care CD4 test with a cut-off of 350 CD4 cells/μL. This study aimed to evaluate VISITECT®CD4 test's diagnostic accuracy. Two hundred HIV-positive patients attending a tertiary HIV centre in South India were recruited. Patients provided venous blood for reference and VISITECT®CD4 tests. An additional finger-prick blood sample was obtained for VISITECT®CD4. VISITECT®CD4's diagnostic performance in identifying individuals with CD4 counts ≤350 cells/μL was assessed by calculating sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV) taking flow cytometry as the reference. The overall agreement between VISITECT®CD4 and flow cytometry was 89.5% using venous blood and 81.5% using finger-prick blood. VISITECT®CD4 showed better performance using venous blood [sensitivity: 96.6% (95% confidence interval: 92.1%-98.9%), specificity: 70.9% (57.1%-82.4%), PPV: 89.7% (83.9%-94.0%) and NPV: 88.6% (75.4%-96.2%)] than using finger-prick blood [sensitivity: 84.8% (77.9%-90.2%), specificity: 72.7% (59.0%-83.9%), PPV: 89.1% (82.7%-93.8%) and NPV: 64.5% (51.3%-76.3%)]. VISITECT®CD4 performed well using venous blood, demonstrating its potential utility in decentralization of CD4 testing services in resource-constrained settings.


When and how to include vulnerable subjects in clinical trials.
There has been a good deal of discussion in the literature regarding which subjects are vulnerable in the context of clinical trials. There has been significantly less discussion regarding when and how to include vulnerable subjects in clinical trials. This lack of guidance is a particular problem for trials covered by the US regulations, which mandate strict requirements on the inclusion of three groups: pregnant women/fetuses, prisoners, and children. For the past 30 years, funders, investigators, and institutional review boards have frequently responded to these regulations by excluding pregnant women/fetuses, prisoners, and children from clinical trials. More recent work has emphasized the extent to which a default of exclusion can undermine the value of clinical trials, especially pragmatic trials. A default of exclusion also has the potential to undermine the interests of vulnerable groups, in both the short and the long term. These concerns raise the need for guidance on how to satisfy existing US regulations, while minimizing their negative impact on the value of clinical trials and the interests of vulnerable groups. The present manuscript thus describes a six-step decision procedure that institutional review boards can use to determine when and how to include vulnerable subjects in clinical trials, including pragmatic trials, that are covered by US regulations.


CD137 ligand activated microglia induces oligodendrocyte apoptosis via reactive oxygen species.
CD137 (4-1BB, TNFRSF9), a member of the tumor necrosis factor (TNF) receptor family, is a potent T cell co-stimulatory molecule. CD137 ligand (CD137L) is expressed by antigen presenting cells (APC) as a transmembrane protein and transmits activating signals into APC. In this study we investigated the effects of CD137L signaling in microglia, the resident APC in the central nervous system. In vitro, the murine microglia cell lines BV-2 and N9, as well as primary murine microglia responded with activation as evidenced by adherence and secretion of proinflammatory cytokines, MMP-9, and soluble intercellular adhesion molecule (ICAM). CD137L signaling is also important for microglia activation in vivo, since CD137L-deficient mice exhibited profoundly less microglia activation during experimental autoimmune encephalomyelitis (EAE) which is a well-established murine model for neuroinflammation and human multiple sclerosis (MS). Also CD137 is expressed in the CNS of mice during EAE. Activated microglia has been reported to mediate the destruction of axonal myelin sheaths and cause the death of oligodendrocytes, the main pathogenic mechanisms in EAE and MS. Corresponding to the lower microglia activation there were also fewer apoptotic oligodendrocytes in the CNS of CD137L-deficient mice. In vitro co-culture confirmed that CD137L-activated microglia induces apoptosis in oligodendrocytes, and identified reactive oxygen species as the mechanism of apoptosis induction. These data demonstrate activating effects of CD137L signaling to microglia, and show for the first time that the CD137 receptor/ligand system may be a mediator of neuroinflammatory and neurodegenerative disease, by activating microglia which in turn kill oligodendrocytes.


Quality of life in patients with minimal hepatic encephalopathy.
Minimal hepatic encephalopathy (MHE) represents the mildest type of hepatic encephalopathy (HE). This condition alters the performance of psychometric tests by impairing attention, working memory, psychomotor speed, and visuospatial ability, as well as electrophysiological and other functional brain measures. MHE is a frequent complication of liver disease, affecting up to 80% of tested patients, depending of the diagnostic tools used for the diagnosis. MHE is related to falls, to an impairment in fitness to drive and the development of overt HE, MHE severely affects the lives of patients and caregivers by altering their quality of life (QoL) and their socioeconomic status. MHE is detected in clinically asymptomatic patients through appropriate psychometric tests and neurophysiological methods which highlight neuropsychological alterations such as video-spatial orientation deficits, attention disorders, memory, reaction times, electroencephalogram slowing, prolongation of latency evoked cognitive potentials and reduction in the critical flicker frequency. Several treatments have been proposed for MHE treatment such as non-absorbable disaccharides, poorly absorbable antibiotics such rifaximin, probiotics and branched chain amino acids. However, because of the multiple diagnosis methods, the various endpoints of treatment trials and the variety of agents used in trials, to date the treatment of MHE is not routinely recommended apart from on a case-by-case basis. Aim of this review is analyze the burden of MHE on QoL of patients and provide a brief summary of therapeutic approaches.


Location choices of face-to-face interactions in academic buildings: an experience sampling approach.
Although it is recognised that face-to-face interactions are important for sharing interests and (new) knowledge, it remains unknown how and where students and university employees interact in academic buildings. Therefore, the aim of this study is to analyse the location choice for face-to-face interactions in an academic building, including several personal- and interaction characteristics. An Experience Sampling Method (ESM) was used to collect data on 643 face-to-face interactions during two weeks in the Flux building at Eindhoven University of Technology, the Netherlands. In general, students more often interacted in meeting rooms than teaching staff, and support staff interacted less in eat/drink areas and the hallways than other users. Unexpectedly, some of the lectures took place outside of traditional project-/lecture space. Real estate managers of university campuses could use these results to create better interactive work environments that stimulate face-to-face interactions among employees and students of different departments. Practitioner Summary: Based on longitudinal data of ftf interactions among students and employees in an academic building, results showed that ftf interaction characteristics, compared to personal characteristics, are most important for explaining the location choice of interactions. These insights could help to design academic work environments that optimise the support of interactions. Abbreviation: ABO: activity-based office; ANOVA: analyses of variance; ESM: experience sampling method; FTF: face-to-face; HR: human resources; MMNL: mixed multinomial logit model; NewWoW: new ways of working.


[Distraction osteosynthesis in occlusive diseases of the limb arteries].
The method of distraction osteosynthesis was used in 265 patients with arterial occlusions of the upper and lower limbs, which developed due to obliterative thromboangiitis and atherosclerosis. Surgical treatment of patients with distal type of occlusive disease of the arteries was followed by steady compensation of ischemic disturbances after bone chip formation and its dosage across transfer with formation of distraction regenerate transforming into mature osseous tissue. Favourable results of the treatment were observed in 236 patients (88.9%). They persisted during 3 years in 86% of patients. Analysis of outcomes after treatment demonstrated the best results in patients with grade II ischemia. Distraction osteosynthesis ensured protracted remission of the disease and elimination of its symptoms.


Mental health, violence and psychological coercion among female and male trafficking survivors in the greater Mekong sub-region: a cross-sectional study.
Human trafficking is a pervasive global crime with important public health implications that entail fundamental human rights violations in the form of severe exploitation, violence and coercion. Sex-specific associations between types of violence or coercion and mental illness in survivors of trafficking have not been established. We conducted a cross-sectional study with 1015 female and male survivors of trafficking (adults, adolescents and children) who received post-trafficking assistance services in Cambodia, Thailand or Vietnam and had been exploited in various labor sectors. We assessed anxiety and depression with the Hopkins Symptoms Checklist (HSCL-25) and post-traumatic stress disorder (PTSD) symptoms with the Harvard Trauma Questionnaire (HTQ), and used validated questions from the World Health Organization International Study on Women's Health and Domestic Violence to measure physical and sexual violence. Sex-specific modified Poisson regression models were estimated to obtain prevalence ratios (PRs) and their 95% confidence intervals (CI) for the association between violence (sexual, physical or both), coercion, and mental health conditions (anxiety, depression and PTSD). Adjusted models indicated that for females, experiencing both physical and sexual violence, compared to not being exposed to violence, was a strong predictor of symptoms of anxiety (PR = 2.08; 95% CI: 1.64-2.64), PTSD (PR = 1.55; 95% CI: 1.37-1.74), and depression (PR = 1.57; 95% CI: 1.33-1.85). Among males, experiencing physical violence with additional threats made with weapons, compared to not being exposed to violence, was associated with PTSD (PR = 1.59; 95% CI: 1.05-2.42) after adjustment. Coercion during the trafficking experience was strongly associated with anxiety, depression, and PTSD in both females and males. For females in particular, exposure to both personal and family threats was associated with a 96% elevated prevalence of PTSD (PR = 1.96; 95% CI: 1.32-2.91) and more than doubling of the prevalence of anxiety (PR = 2.11; 95% CI: 1.57-2.83). The experiences of violence and coercion in female and male trafficking survivors differed and were associated with an elevated prevalence of anxiety, depression, and PTSD in both females and males. Mental health services must be an integral part of service provision, recovery and re-integration for trafficked females and males.


Acetaminophen-induced fulminant hepatic failure in dogs.
Results concerning morphological and biochemical changes following intravenous administration of different doses of acetaminophen in dogs are reported. Acetaminophen infusion, as a parenteral solution (500 mg per kg per 90 min), produced fulminant hepatitis characterized by a good correlation between Portmann's grade of lesion and percentage of necrosis. All animals died within 76 hr after intoxication. Analysis of biochemical parameters revealed positive correlation between serum bilirubin levels and severity of the hepatic lesion. The experimental model of acetaminophen-induced hepatotoxicity is proposed as a model for evaluation of the therapeutic efficacy of new medical and surgical procedures.


Structure and mechanics of starfish body wall.
The structure of the dorsal body wall of the starfish Echinaster spinulosus was studied using polarized light microscopy of frozen tissues, scanning electron microscopy and histology. The collagen fibres of the body wall form a three-dimensional orthogonal web. Voids in the web contain ossicles and papulae. The orthogonal web delivers dimensional stability but allows shear necessary for ray torsion. The ossicles and fibres interact to load the fibres in tension and the ossicles in compression. Strain rates of the dorsal body wall were measured on live animals during typical movements. Uniaxial tension tests of the body wall yielded Young's moduli of 267 MPa (longitudinal), 249 MPa (transverse) and 353 MPa (bias); curves were essentially linear. The body wall was approximately linearly viscoelastic and showed hysteresis at 0.01 Hz. Stress relaxation over five decades of time (in seconds) yielded relaxation spectra with peaks in relaxation time at 2.96-3.35, depending on test direction. Stress relaxation caused the connective tissue to soften. The surface of fractured stress-relaxed tissue revealed wispy, dissociated fibril tufts, whereas unrelaxed fractures produced blunt-ended fibre bundles. Neural control was necessary for body wall integrity.


Reconstruction of the Second Metatarsal with Non-vascularised Fibular Graft following En-bloc Resection for Giant Cell Tumour: A Case Report.
Giant cell tumour in the metatarsal of a skeletally immature person is uncommon. Adequate surgical resection in this region can be difficult to achieve as there is little space between the rays of the foot. Furthermore, there is the challenge of restoring the metatarsophalangeal articulation after resection of the tumor. We describe a technique using non-vascularised fibular graft for reconstructing the 2nd metatarsal after en-bloc resection for giant cell tumour in a 14 year old adolescent female. Metatarsal, Giant cell tumour, Non-vascularised fibular graft, En-bloc resection.


[Virus-like particles as a source for obtaining antigens for diagnosing rubella].
Rubella diagnostic agents for hemagglutination inhibition (HI) and enzyme immunoassay (EIA) based on rubella virus-like particles (RVLP) have been developed. Noninfectious RVLPs containing three structural E1, E2, and C proteins were expressed in transfected CHO24S cell culture. HI titer in culture medium was 1:256. Tween-80 treatment and ether increased HI titer 4-6-fold and rendered the antigen higher stability. Immunogenic properties of RVLPs were similar to the native rubella virus in HI test with international reference human rubella serum and sera from convalescents after rubella. Serum of mice immunized with RVLP reacted similarly with RVLP antigens and native virus. Antigen for EIA from RVLP was prepared by concentrating RVLP from culture fluid and partial purification by ultracentrifugation. The results of human sera testing by HI and EIA with RVLP and native virus antigens coincided. RVLP are identical to antigenic structure of the virus, are stable and easily purified, and can therefore be used for commercial production of HI and EIA antigens.


Strain-Induced Electronic Structure Modulation on MnO2 Nanosheet by Ir Incorporation for Efficient Water Oxidation in Acid.
Oxygen electrochemistry plays a key role in renewable energy technologies, such as fuel cells and electrolyzers, but its slow kinetics limits the performance and the commercialization of such devices. Here, a strained MnO2 nanosheet induced by Ir incorporation is developed with optimized electronic structure by a simple hydrothermal method. With the incorporation of Ir, the strain induces elongated Mn─O bond length, and thereby tuning the electronic structure to favor the oxygen evolution reaction (OER) performance. The obtained catalyst exhibits an excellent mass activity of 5681 A g-1 at an overpotential of 300 mV in 0.5 m H2 SO4 , and reaches 50 and 100 mA cm-2 at overpotentials of only 240 and 277 mV, respectively. The catalyst is also stable even at 300 mA cm-2 in 0.5 m H2 SO4 . Using the nanosheet as the OER catalyst and the Pt/C as the hydrogen evolution reaction catalyst, a two-electrode electrolyzer achieves 10 mA cm-2 with only a cell voltage of 1.453 V for overall water splitting in 0.5 m H2 SO4 . This strategy enables the material with high feasibility for practical applications on hydrogen production.


Resource document on mandatory outpatient treatment.
Mandatory outpatient treatment, or outpatient commitment, refers to court-ordered treatment for patients who suffer from severe mental illness and who are unlikely to be compliant with such treatment without a court order. Many states already have commitment statutes that permit mandatory outpatient treatment, and others are considering enacting new legislation or amending existing statutes. This Resource Document was prepared under the auspices of the American Psychiatric Association's Council on Psychiatry and Law to provide information to those who are drafting mandatory outpatient treatment legislation. It begins with a review of the history of mandatory outpatient treatment and recent empirical findings, followed by a detailed discussion of the salient issues in mandatory outpatient treatment. The document concludes with a statement of recommendations concerning key provisions in statutory schemes of mandatory outpatient treatment programs. This Resource Document endorses the view that mandatory outpatient treatment can be a useful intervention for a small subset of noncompliant patients with severe and chronic mental illness who go in and out of psychiatric hospitals through the so-called "revolving door."


Retrospective study of turkey viral hepatitis in California turkey flocks, 2000-2012.
Turkey viral hepatitis (TVH) is a disease characterized by an inflammation of the liver, and occasionally of the pancreas, of turkeys. Little is known about the occurrence of TVH in turkey flocks; thus, the aim of the present article is to summarize retrospectively 76 cases of TVH diagnosed at the California Animal Health and Food Safety System (CAHFS), University of California, Davis, in the years 2000 through 2012. Flocks diagnosed with TVH were between 7 and 61 days old, with an average age of 29.4 days and a median age of 28 days. In the majority of cases, increased mortality was reported. In 55 cases, gross lesions were present in the liver; the most common lesions were a few to numerous pale white foci in 35 cases. In livers of 74 cases, histologic lesions were observed. Multifocal necrosis with inflammation was observed in livers of 42 cases, inflammation but no necrosis in 22 cases, and necrosis without inflammation in 8 cases. In 17 cases, pale white foci were found in the pancreas; in 4 cases, larger areas of the organ were pale. In 33 cases, histologic changes were observed in the pancreas. Necrosis with inflammation was observed in the pancreas of 17 cases, necrosis without inflammation in 7 cases, and inflammation without necrosis in 9 cases. No Salmonella was isolated from any of the livers cultured. Transmission electron microscopy of liver and pancreas demonstrated viral particles between 23 and 25 nm in two cases. Concurrent diseases were mostly poult enteritis (65 cases), but also respiratory diseases and colibacillosis. TVH occurred more often in turkeys during winter months. In conclusion, TVH was observedon a regular basis in California turkey flocks during the last 12 yr.


Enzymatic Degradation of p-Nitrophenyl Esters, Polyethylene Terephthalate, Cutin, and Suberin by Sub1, a Suberinase Encoded by the Plant Pathogen Streptomyces scabies.
The genome of Streptomyces scabies, the predominant causal agent of potato common scab, encodes a potential cutinase, the protein Sub1, which was previously shown to be specifically induced in the presence of suberin. The sub1 gene was expressed in Escherichia coli and the recombinant protein Sub1 was purified and characterized. The enzyme was shown to be versatile because it hydrolyzes a number of natural and synthetic substrates. Sub1 hydrolyzed p-nitrophenyl esters, with the hydrolysis of those harboring short carbon chains being the most effective. The Vmax and Km values of Sub1 for p-nitrophenyl butyrate were 2.36 mol g-1 min-1 and 5.7 10-4 M, respectively. Sub1 hydrolyzed the recalcitrant polymers cutin and suberin because the release of fatty acids from these substrates was observed following the incubation of the enzyme with these polymers. Furthermore, the hydrolyzing activity of the esterase Sub1 on the synthetic polymer polyethylene terephthalate (PET) was demonstrated by the release of terephthalic acid (TA). Sub1 activity on PET was markedly enhanced by the addition of Triton and was shown to be stable at 37°C for at least 20 d.


Discriminant quantitation of posttransplant hepatic reticuloendothelial function. The impact of ischemic preservation.
This study focuses upon two discrete components of posttransplant hepatic reticuloendothelial system (RES) function-phagocytosis and killing of bacteria-under various conditions of ischemic preservation. We had previously reported that, following intravenous injection of rats with 51Cr and 125I double-labeled Escherichia coli, hepatic 51Cr levels can be used to reliably quantify hepatic phagocytic clearance of the bacteria from the blood (HPC), while the subsequent release of 125I from the liver accurately parallels hepatic bacterial killing. Here, Wistar rats were transplanted with syngeneic livers perfused with either normal saline (NS) or University of Wisconsin solution (UW) and stored at 4 degrees C for 1, 2, or 3 hr prior to implantation. Control rats underwent laparotomy and hepatic artery ligation. Using the double-labeled E coli assay, HPC was decreased in all transplanted animals when compared with controls, reaching a nadir on the third postoperative day (P < 0.05). In rats transplanted with livers preserved in NS, the fraction of phagocytosed organisms that were subsequently killed (hepatic killing efficiency=HKE) was increased to 142%, 129%, or 112% of normal following 1, 2, or 3 hr of cold ischemia, respectively; P < 0.05). Conversely, preservation of donor allografts in UW was associated with marked depression of HKE. Moreover, rats receiving NS- or UW-preserved livers tolerated an intravenous challenge with Streptococcus pneumoniae poorly (50% mortality) compared with hepatic artery ligated controls (12% mortality) at 7 days. Ischemic preservation of rat livers in NS resulted in a dose (of ischemia)-dependent reduction of hepatic phagocytosis coupled with a potentiation of HKE. Preservation in UW, however, produced a striking suppression of both components of hepatic RES function. Following a septic challenge survival was reduced in both groups of transplanted rats.


Target of Rapamycin Regulates Photosynthesis and Cell Growth in Auxenochlorella pyrenoidosa.
Auxenochlorella pyrenoidosa is an efficient photosynthetic microalga with autotrophic growth and reproduction, which has the advantages of rich nutrition and high protein content. Target of rapamycin (TOR) is a conserved protein kinase in eukaryotes both structurally and functionally, but little is known about the TOR signalling in Auxenochlorella pyrenoidosa. Here, we found a conserved ApTOR protein in Auxenochlorella pyrenoidosa, and the key components of TOR complex 1 (TORC1) were present, while the components RICTOR and SIN1 of the TORC2 were absent in Auxenochlorella pyrenoidosa. Drug sensitivity experiments showed that AZD8055 could effectively inhibit the growth of Auxenochlorella pyrenoidosa, whereas rapamycin, Torin1 and KU0063794 had no obvious effect on the growth of Auxenochlorella pyrenoidosaa. Transcriptome data results indicated that Auxenochlorella pyrenoidosa TOR (ApTOR) regulates various intracellular metabolism and signaling pathways in Auxenochlorella pyrenoidosa. Most genes related to chloroplast development and photosynthesis were significantly down-regulated under ApTOR inhibition by AZD8055. In addition, ApTOR was involved in regulating protein synthesis and catabolism by multiple metabolic pathways in Auxenochlorella pyrenoidosa. Importantly, the inhibition of ApTOR by AZD8055 disrupted the normal carbon and nitrogen metabolism, protein and fatty acid metabolism, and TCA cycle of Auxenochlorella pyrenoidosa cells, thus inhibiting the growth of Auxenochlorella pyrenoidosa. These RNA-seq results indicated that ApTOR plays important roles in photosynthesis, intracellular metabolism and cell growth, and provided some insights into the function of ApTOR in Auxenochlorella pyrenoidosa.


New species of the Spiny Mouse genus Neacomys (Cricetidae, Sigmodontinae) from northwestern Ecuador.
Neacomys is a genus of small spiny or bristly sigmodontine rodents that are common components of mammalian faunas in multiple biomes on Central and South America. Recent studies on this group have demonstrated that there is cryptic diversity yet to be discovered within currently recognized species that have not received comprehensive revisions, as well as in areas that have been overlooked. Here we ratify this assertion by describing a new species previously misidentified as the Narrow-footed Spiny Mouse (Neacomystenuipes) from the Chocó biogeographic region in northwestern Ecuador, Neacomysmarci Brito & Tinoco, sp. nov. Distinctiveness of this entity is supported by the combination of the following morphological characters: small size (head-body length 65-85 mm); long tail (69-126% longer than head-body length); pale buff-colored but gray-based belly fur; white throat; hypothenar pad usually absent; long nasals; and a condylar process higher than the coronoid process. Likewise genetic distance analyses and phylogenetic reconstructions based on cytochrome-b (Cytb) sequence data indicate a clear divergence from typical populations of N.tenuipes, and a sister relationship between them. The results presented here increase the diversity of Neacomys to 24 species, placing it among the most diverse genera within the sigmodontine rodents.


Immunotherapy in breast cancer.
The idea of using the immune system to fight cancer is over 100 years old. A new molecular approach led to a better understanding of the immune system. Checkpoint regulation, understanding the roles of Tregs, Th1, and Th2, development of Chimeric antigen receptor (CAR)-T cells, as well as regulation of dendritic cells and macrophages, are just a few examples of our understating that has also led to the discovery of immune checkpoint inhibitors (ICIs) and modulators. This led the Nobel Prize committee in 2018, to award Dr. James P. Allison the Nobel Prize in medicine for the discovery of Cytotoxic T-lymphocyte-associated antigen-4, and Dr. Tasuku Honjo for the discovery of programmed cell death-1 (PD-1)/PD-1-ligand (PDL-1). Several ICIs are already approved by the regulatory authorities, and many more are currently used in studies of several solid tumors and hematologic malignancies. Positive studies have led to the US Food and Drug Administration (FDA) and European Medicines Agency approval of a number of these compounds, but none to date are approved in breast cancer (BC). Moreover, PD-1/PDL-1, MSI high (and dMMR), and tumor mutational burden are the currently "best" predictive markers for benefit from immunotherapy. BCs have some of these markers positive only in subsets but less frequently expressed than most other solid tumors, for example, malignant melanoma or non-small cell lung cancer. To improve the potential efficacy of ICI in BC, the addition of chemotherapy was one of the strategies. Many early and large clinical trials in all phases are underway in BC. We will discuss the role of immune system in BC editing, and the potential impact of immunotherapy in BC outcomes.


Clinical burden, risk factor impact and outcomes following myocardial infarction and stroke: A 25-year individual patient level linkage study.
Understanding trends in the incidence and outcomes of myocardial infarction and stroke, and how these are influenced by changes in cardiovascular risk factors can inform health policy and healthcare provision. We identified all patients 30 years or older with myocardial infarction or stroke in Scotland. Risk factor levels were determined from national health surveys. Incidence, potential impact fractions and burden attributable to risk factor changes were calculated. Risk of subsequent fatal and non-fatal events (myocardial infarction, stroke, bleeding and heart failure hospitalization) were calculated with multi-state models. From 1990 to 2014, there were 372,873 (71±13 years) myocardial infarctions and 290,927 (74±13 years) ischemic or hemorrhagic strokes. Age-standardized incidence per 100,000 fell from 1,069 (95% confidence interval, 1,024-1,116) to 276 (263-290) for myocardial infarction and from 608 (581-636) to 188 (178-197) for ischemic stroke. Systolic blood pressure, smoking and cholesterol decreased, but body-mass index increased, and diabetes prevalence doubled. Changes in risk factors accounted for a 74% (57-91%) reduction in myocardial infarction and 68% (55-83%) reduction in ischemic stroke. Following myocardial infarction, the risk of death decreased (30% to 20%), but non-fatal events increased (20% to 24%) whereas the risk of both death (47% to 34%) and non-fatal events (22% to 17%) decreased following stroke. Over the last 25 years, substantial reductions in myocardial infarction and ischemic stroke incidence are attributable to major shifts in risk factor levels. Deaths following the index event decreased for both myocardial infarction and stroke, but rates remained substantially higher for stroke. British heart foundation.


Comparing appropriateness and equivalence of email interviews to phone interviews in qualitative research on reproductive decisions.
Despite an increasing use of qualitative email interviews by nurse researchers, there is little understanding about the appropriateness and equivalence of email interviews to other qualitative data collection methods, especially on sensitive topics research. The purpose is to describe our procedures for completing asynchronous, email interviews and to evaluate the appropriateness and equivalency of email interviews to phone interviews in two qualitative research studies that examined reproductive decisions. Content analysis guided the methodological appraisal of appropriateness and equivalency of in-depth, asynchronous email interviews to single phone interviews. Appropriateness was determined by: (a) participants' willingness to engage in email or phone interviews, (b) completing data collection in a timely period, and (c) participants' satisfaction with the interview. Equivalency was evaluated by: (a) completeness of the interview data, and (b) insight obtained from the data. Of the combined sample in the two studies (N=71), 31% of participants chose to participate via an email interview over a phone interview. The time needed to complete the email interviews averaged 27 to 28days and the number of investigator probe-participant response interchanges was 4 to 5cycles on average. In contrast, the phone interviews averaged 59 to 61min in duration. Most participants in both the email and phone interviews reported they were satisfied or very satisfied with their ability to express their true feelings throughout the interview. Regarding equivalence, 100% of the email and phone interviews provided insight into decision processes. Although insightful, two of the email and one phone interview had short answers or, at times, underdeveloped responses. Participants' quotes and behaviors cited within four published articles, a novel evaluation of equivalency, revealed that 20% to 37.5% of the citations about decision processes were from email participants, which is consistent with the percent of email participants. In-depth, asynchronous email interviews were appropriate and garnered rich, insightful data that augmented the phone interviews. Awareness of the procedures, appropriateness, and nuances when carrying out email interviews on sensitive topics may provide nurse researchers with the ability to obtain thick, rich data that can best advance clinical practice and direct future research.


Congruence of spatial organization of tactile projections to granule cell and Purkinje cell layers of cerebellar hemispheres of the albino rat: vertical organization of cerebellar cortex.
1. We compared the spatial pattern of shortest latency somatosensory (tactile) projections to the Purkinje cell (PC) layer and to the underlying granule cell (GC) layer in tactile areas of rat cerebellar cortex. Micro-mapping methods were used to sample single units in the PC layer and multiple units in the GC layer of both anesthetized and unanesthetized rats. Mechanical and electrical stimulation of the body surface were employed. Responsiveness of PCs to cutaneous stimulation was assessed by constructing histograms of simple spike activity and statistically comparing poststimulus activity to nonstimulated base-line PC activity. 2. We found that PCs respond to tactile stimulation with increases (7-10 ms) followed by decreases (8-15 ms) in simple spike activity. Increases in simple spike activity followed activation of the underlying GC layer by 1-4 ms, while decreases in simple spike activity were found 2-5 ms after GC layer activation. 3. PCs were found to have both excitatory and inhibitory receptive fields (RFs). Excitatory RFs were restricted to small areas of a single body part and for each PC were very similar or identical to the RFs of neurons in the immediately subjacent GC layer. Inhibitory PC RFs were larger, often containing more than one body part and for each PC, were only partially similar to the RFs of subjacent GCs. PC inhibitory RFs also often included body surfaces projecting to the nearby but not to the underlying GC layer. 4. Stimulation of a single peripheral locus resulted in small, distinct regions of PC layer excitation and inhibition. Areas of PC excitation overlie activated regions of the GC layer, while inhibited PCs overlie both activated and nonactivated GC regions. 5. We found PCs to be organized in groups or patches with respect to the specific body region that was capable of activating them (upper lip, lower lip, etc.). Adjacent patches of PCs often represented widely separated body parts. This pattern of PC layer activating RF projections was congruent with the pattern of excitatory RF projections to the underlying GC layer. 6. These results indicate that there is a vertical organization in GC-PC excitatory relations, while GC-induced PC inhibition is slightly more widely distributed. 7. Our finding that the patchlike activation of PCs is congruent with that of the underlying GC layer contrasts with the classical concept that PCs are activated by parallel fibers in a "beamlike" fashion from a patch of GCs. Thus, a reevaluation of the role of parallel fibers seems to us to be in order. 8. In conclusion, our results support the view that short-latency afferent tactile projections to both the GC and PC layers of cerebellar cortex are highly organized spatially. This specificity of body surface projections must be incorporated into modern views of the functional organization of cerebellar cortex.


Use of transendoscopic contact neodymium:yttrium aluminum garnet laser to drain dorsal epiglottic abscesses in two horses.
A submucosal abscess, located on the dorsal surface of the epiglottis, was diagnosed in 2 Thoroughbred racehorses by use of endoscopy. Both horses had exercise intolerance. One horse had intermittent dorsal displacement of the soft palate, coughed while eating and galloping, and made an abnormal respiratory noise. Both abscesses were drained transendoscopically by use of a contact neodymium:yttrium aluminum garnet laser. Eleven days after surgery, the surgical sites appeared to have healed. Clinical signs resolved permanently, and both horses returned to successful racing careers.


Safe and cost-effective cleaning of pressure-monitoring transducers.
The CDC category I recommendation for high-level disinfection of pressure-monitoring transducers between uses imposes substantial costs. This practice has not been shown to be useful when disposable transducer domes are used. A prospective, randomized, double-blind study assessed the consequences of wiping transducers with alcohol between uses rather than sterilizing with ethylene oxide. We studied 5,197 transducer courses with disposable domes involving 2,202 patients in intensive care units during 15 months. There was no significant difference between the two treatment regimens in the risk of positive cultures of fluids from monitored lines, of cultures of cannula tips or of a positive blood culture. We conclude that under normal conditions, ie, in the absence of a cluster of transducer-related bacteremias, wiping transducers with alcohol between uses is sufficient when disposable domes are used.


Phosphofructokinase from foot muscle of the whelk, Busycotypus canaliculatum: evidence for covalent modification of the enzyme during anaerobiosis.
Phosphofructokinase (PFK) was purified from foot muscle of aerobic and anaerobic (24 h of anoxia) whelks, Busycotypus canaliculatum. Fructose-6-P kinetics were sigmoidal at pH 7.0 with affinity constants, S0.5, of 2.18 +/- 0.10 (nH = 2.5 +/- 0.1) and 2.48 +/- 0.13 mM (nH = 2.7 +/- 0.1) for the enzyme from aerobic verus anaerobic muscle. Affinity for ATP, like that for fructose-6-P, did not differ for the two enzymes (0.031 +/- 0.003 for the aerobic vs 0.041 +/- 0.007 mM for the anaerobic enzyme), but S0.5 for Mg2+ was significantly different for the two enzymes (0.060 +/- 0.006 vs 0.130 +/- 0.020 mM). Whelk muscle PFK was activated by NH+4, Pi, AMP, ADP, and fructose-2,6-P2.NH+4 and fructose-2,6-P2 were less effective activators of PFK from anoxic muscle, with apparent Ka's 1.6- and 3.5-fold higher for the anaerobic vs aerobic enzyme. Activators decreased S0.5 for fructose-6-P and reduced nH. With the exception of fructose-2,6-P2, the effects of activators on S0.5 were the same for the enzyme from aerobic and anaerobic muscle; fructose-2,6-P2 at 2.5 microM reduced S0.5 by only 3.3-fold for the anaerobic enzyme compared to 5.5-fold for the aerobic enzyme. ATP was a strong substrate inhibitor of PFK; the enzyme from anaerobic muscle showed greater ATP inhibition, with I50's 1.5- to 2.0-fold lower than those for the aerobic enzyme. The kinetic differences between PFK from anaerobic versus aerobic foot muscle (stronger ATP inhibition and decreased sensitivity to activators for the anaerobic enzyme) were consistent with kinetic differences reported for the phosphorylated versus dephosphorylated forms, respectively, of PFK in other systems. Treatment of PFK from anaerobic muscle with alkaline phosphatase resulted in a decrease in the Ka for fructose-2,6-P2 to a level similar to that of the aerobic enzyme. The physiological stress of anoxia may, therefore, induce a covalent modification of PFK.


Relationship Between Preoperative 25-Hydroxy Vitamin D and Surgical Site Infection.
Surgical site infection (SSI) is one of the most important and costly complications of surgical operations. Vitamin D antimicrobial and wound healing effects have been recently shown in animal models and in laboratory settings. Furthermore, potential effects of vitamin D in mitigating nosocomial infections and SSIs have been examined at a limited scale. To our knowledge, no comprehensive study has been performed to show the relationship between preoperative level of vitamin D and incidence of SSI. The present study was designed and implemented to investigate this relationship. We performed a prospective cross-sectional study involving 300 adult patients who were admitted to undergo surgery in our tertiary care unit from January 2016 to January 2018. Cutoff point was considered at a level of 30 (ng/mL) in defining vitamin D deficiency. The presence of any SSI was investigated and recorded at the time of discharge and at postoperative visits up to 30 d after the surgery. Cross-tabulation and bivariate and multivariate logistic regression with unadjusted and adjusted odd ratio were used to determine the association between dependent and independent variables and to identify factors associated with SSIs. Overall, of 300 patients who were investigated, 39% had preoperative vitamin D deficiency and 11% developed SSI. In univariate logistic regressions, 20 predictors were selected to be included in the multivariate analysis. Finally preoperative level of 25-hydroxy vitamin D, history of recent infection, preoperative and postoperative hospital length of stay, and postoperative blood transfusions were confirmed as statistically significant independent predictors of SSI. Preoperative 25-hydroxy vitamin D level has a strong effect on postoperative SSI. Prospective double-blinded randomized clinical trials are required to confirm such strong relationship and to settle preoperative vitamin D measurement as a standard approach to reduce postoperative complications including SSI. Preoperative patient optimization, limiting hospital length of stay, and blood transfusion are other strategies to reduce SSI.


Cell-free transport from the trans-golgi network to late endosome requires factors involved in formation and consumption of clathrin-coated vesicles.
Transport between the trans-Golgi network (TGN) and late endosome represents a conserved, clathrin-dependent sorting event that separates lysosomal from secretory cargo molecules and is also required for localization of integral membrane proteins to the TGN. Previously, we reported a cell-free reaction that reconstitutes transport from the yeast TGN to the late endosome/prevacuolar compartment (PVC) and requires the PVC t-SNARE Pep12p. Here, we report that factors required both for formation of clathrin-coated vesicles at the TGN (the Chc1p clathrin heavy chain and the Vps1p dynamin homolog) and for vesicle fusion at the PVC (the Vps21p rab protein and Vps45p SM (Sec1/Munc18) protein) are required for cell-free transport. The marker for TGN-PVC transport, Kex2p, is initially present in a clathrin-containing membrane compartment that is competent for delivery of Kex2p to the PVC. A Kex2p chimera containing the cytosolic tail (C-tail) of the vacuolar protein sorting receptor, Vps10p, is also efficiently transported to the PVC. Antibodies against the Kex2p and Vps10p C-tails selectively block transport of Kex2p and the Kex2-Vps10p chimera. The requirements for factors involved in vesicle formation and fusion, the identification of the donor compartment as a clathrin-containing membrane, and the need for accessibility of C-tail sequences argue that the TGN-PVC transport reaction involves selective incorporation of TGN cargo molecules into clathrin-coated vesicle intermediates. Further biochemical dissection of this reaction should help elucidate the molecular requirements and hierarchy of events in TGN-to-PVC sorting and transport.