Quantitative immunocytochemical characterization of mononuclear phagocytes. I. Monoblasts, promonocytes, monocytes, and peritoneal and alveolar macrophages.
The purpose of the present study was to compare the phenotype of tissue macrophages with that of their precursors in the bone marrow and blood. The phenotype was determined on the basis of the quantitative binding of monoclonal antibodies to cell-surface antigens (antigen F4/80, complement receptor III, Fc receptor II, Ia antigen, common leukocyte antigen, and Mac-2 and Mac-3 antigens) on individual mononuclear phagocytes. Monoclonal antibody binding to cells, detected by the biotin-avidin immunoperoxidase procedure, was quantitated by cytophotometric determination of the amount of enzyme reaction product on cells. The results of this quantitation are expressed as the median of the specific absorbance per unit of cell-surface area (0.25 micron2) and per cell. Shortly after collection of the mononuclear phagocytes, binding of all monoclonal antibodies except those directed against the common leukocyte and Mac-2 antigens to peritoneal macrophages was enhanced compared with binding to blood monocytes; for alveolar macrophages we found reduced binding of monoclonal antibodies F4/80 and M1/70 (complement receptor III) and enhanced binding of monoclonal antibodies with specificity for the common leukocyte antigen and Mac-2 and Mac-3 antigens. The results obtained with cultured mononuclear phagocytes show that during the development from monoblast to tissue macrophages, monoclonal antibody binding to the various types of mononuclear phagocyte, expressed per unit of cell-surface area, was not significantly altered except that of M3/38 (Mac-2 antigen) to peritoneal macrophages and that of F4/80 and M1/70 (complement receptor III) to alveolar macrophages. Expressed on a per cell basis, the results show an increase in the binding of all monoclonal antibodies except those directed against the Fc receptor II and Mac-3 antigen during the development from promonocytes to peritoneal macrophages; binding of most monoclonal antibodies to alveolar macrophages was considerably lower than that to blood monocytes. It is concluded that the expression of the various cell-surface antigens alters during mononuclear phagocyte differentiation. The expression changed also during culture, although distinct patterns of alteration could not be distinguished.


Analysis of the p21 gene in gliomas.
The p21 gene encodes a cyclin dependent kinase inhibitor protein (p21) which has a tumor suppressive activity in a variety of tumor cell lines. Since, the p21 gene is up-regulated by the p53 tumor suppressor gene, which is frequently mutated in gliomas, acting therefore in the same control pathway, it constitutes a good candidate gene to be also inactivated in these tumors. To test this hypothesis, DNAs from 81 gliomas (48 glioblastomas, 11 anaplastic astrocytomas, 10 low-grade astrocytomas, 12 oligodendrogliomas and mixed gliomas), were investigated for mutations in the p21 coding sequence by denaturant gradient gel electrophoresis followed by sequencing. All these tumors have been previously screened for p53 mutations. Three different DNA variants were identified on codon 31 (17 cases), 27 (1 case) and 117 (1 case) and shown to be also present in matching constitutional DNA, suggesting they were polymorphisms. None of the tumors demonstrated a somatic mutation. No significant correlation between the presence of a p21 variant and the p53 mutation tumor status was observed. In conclusion, mutation in the p21 gene unlikely contributes to the development of gliomas.


Opposite behavior of plasma levels surfactant protein type B and receptor for advanced glycation end products in pulmonary sarcoidosis.
No biological marker is currently available for evaluating pulmonary involvement and/or for monitoring the clinical course of sarcoidosis. The present pilot study focused on possible relationships between circulating plasma levels of surfactant protein type B (SP-B) and plasma receptor for advanced glycation end products (RAGE) and lung function abnormalities in patients with pulmonary sarcoidosis, since both SP-B and RAGE have been previously suggested as lung injury markers. The plasmatic levels of these two proteins were also investigated with respect to functional capacity, as assessed by a cardiopulmonary exercise test (CPET). Thirty pulmonary sarcoidosis outpatients and fifteen volunteers (Control Group) underwent lung function tests and CPET. Resting SP-B and RAGE plasma levels were also determined. Patients were then categorized according to the severity of their pulmonary involvement, as assessed in terms of lung diffusion for carbon monoxide (DLCO) values. Group B showed SP-B levels higher and RAGE levels lower than Group A and Control Group (p < 0.01). Group A showed lower RAGE levels than Control Group (p < 0.01), whereas SP-B levels did not differ between these two groups. A significant univariate relationship was found between both SP-B and RAGE and several lung function data, particularly with DLCO (SP-B Vs DLCO: r: -0.437, p = 0.016; RAGE Vs DLCO: r: -0.451, p = 0.012). Circulating plasma levels of SP-B and RAGE showed an opposite behavior in patients with pulmonary sarcoidosis. SP-B values are directly related to alveolar unit damage, supporting a possible role of SP-B as a marker of disease severity in these patients. Differently, RAGE decreases in severe sarcoidosis, suggesting more complex underlying mechanisms.


On new collections of Scaphidiinae (Coleoptera: Staphylinidae) from China, with description of two new species.
Two new species of Scaphidiinae, Scaphisoma konvicka sp. nov. and Toxidium hartmanni sp. nov. are described from China. New records are given for eight other Chinese scaphidiine species.


Cyclopiazonic acid and aflatoxins production byAspergillus flavus isolated from Argentinian corn.
Thirty-four isolates ofAspergillus flavus obtained from the main Argentinian corn production area were tested for their ability to produce both cyclopiazonic acid (CPA) on corn and on liquid media and aflatoxins on corn.Aflatoxins and CPA were quantified by comparison with standards. The last one was confirmed by mass spectrometry.All but one of the isolates produced CPA on liquid medium in a range between 3120 to 62500 μg/kg, 27/34 isolates produced CPA on corn at levels ranging from 833 to 10000 μg/kg and 5/34 isolates produced aflatoxin B1 in a range between 29 to 115 μg/kg. According to these findings, the percentage ofAspergillus flavus isolates with CPA production ability and their levels of CPA production were higher than the observed elsewhere.It was observed significant differences (p<0,01) between CPA production on corn (median: 1761 μg/Kg) and in liquid medium (median: 27950 μg/Kg). These data represent the first report of the co-production of CPA and aflatoxin B1 by isolates ofAspergillus flavus obtained from corn in Argentina.


Regulation of type 2 nitric oxide synthase by type 1 interferons in macrophages infected with Leishmania major.
We recently reported that the infection of macrophages with Leishmania major led to the release of type 1 interferons (IFN-alpha /beta ). Moreover, at day 1 of infection of mice with L. major, IFN-alpha /beta was required for the expression of type 2 (inducible) NO synthase (NOS2 or iNOS) which, however, was restricted to a few macrophages in the dermis. Here, we further characterized the regulation of NOS2 by IFN-alpha /beta. Macrophages that were either simultaneously or sequentially exposed to L. major promastigotes and IFN-alpha /beta expressed NOS2 and anti-leishmanial activity. In contrast, when high amounts of IFN-alpha /beta were used or when IFN-alpha /beta was added to the macrophages 2 h prior to the parasites, almost no induction of NOS2 was observed. After pretreatment with IFN-alpha /beta, tyrosine phosphorylation and nuclear DNA binding of Stat1alpha, the degradation of the NF-kappaB inhibitor (IkappaBalpha and beta), and the nuclear translocation of NF-kappaB were strongly impaired compared with macrophages exposed to IFN-alpha /beta and L. major simultaneously. Thus, IFN-alpha /beta exerts agonistic or antagonistic effects on the expression of NOS2 in macrophages infected with a microbial pathogen, depending on the sequence of the stimuli and the amount of IFN-alpha /beta added. The limited number of NOS2-positive macrophages at day 1 of infection in vivo might result from a blockage of non-infected macrophages by IFN-alpha /beta that is released by neighboring infected cells.


A new experimental method for the dynamic study of the antibody transfer mechanism from mother to fetus in the rat.
The yolk-sac is known to be a route for the transport of passive immunity from mother to fetus in the rat. The main purpose of the present paper is to describe an experimental system for ultrastructurally studying the kinetics of the uptake and transport of immunoglobulin by rat yolk-sac. This system has the advantage of enabling the membrane to be externalized and then exposed to protein under controlled environmental conditions whilst at the same time maintaining the conceptus in connection with the in situ placenta. Preliminary investigations have utilized homologous anti-horseradish peroxidase (HRP) IgG (detected as antibody by application of HRP) or HRP alone. Comparison has been made with the localization of endogenous IgG transmitted in vivo after immunization of the female rat with HRP. The results show the rapid binding of IgG to membrane since only 30 sec were sufficient for this attachment to occur. Moreover, the endocytic process also appears to be very fast as localization of IgG in clusters or patches, caveolae or pits and even rare microvesicles is observed within 8 min. On the other hand, no binding of HRP to microvilli was observed and, unlike IgG, HRP became located in the apico-tubulocanalicular system.


ACTH receptor: ectopic expression, activity and signaling.
Failure in obtaining expression of functional adrenocorticotropic hormone receptor (ACTHR, or melanocortin 2 receptor, MC2R) in non-adrenal cells has hindered molecular analysis of ACTH signaling pathways. Here, we ectopically expressed the mouse ACTHR in Balb/c mouse 3T3 fibroblasts to analyze ACTH signaling pathways involved in induction of fos and jun genes. Natural constitutive expression of the MC2R accessory protein (MRAP) in Balb3T3 and other mouse 3T3 fibroblasts (NIH, Swiss and 3T3-L1) renders these fibroblastic lines suitable for ectopic expression of ACTHR in its active form properly inserted into the plasma membrane at levels similar to those found in mouse Y1 adrenocortical tumor cells. The Y1 cell line is a cultured cell system well known for stably displaying normal adrenal specific metabolic pathways, ACTHR expression and ACTH functional responses. Thirty-nine sub-lines expressing ACTHR (3T3-AR transfectants) were selected for geneticin-resistance and clonally isolated after transfection of ACTHR-cDNA (in the pSVK3 mammalian plasmidial vector) into Balb3T3 fibroblasts. In addition, sixteen clonal sub-lines of Balb3T3 (3T3-0 transfectants) carrying the pSVK3 empty vector were likewise isolated. Fourteen 3T3-AR and four 3T3-0 clones were screened for response to ACTH(39) in comparison with Y1 adrenocortical cells. Eight 3T3-AR clones responded to ACTH(39) with activation of adenylate cyclase and induction of c-Fos protein, but the levels of, respectively, activation and induction were not strictly correlated. Other fos and jun genes were also induced by ACTH(39) in 3T3-AR transfectants, which express levels of ACTHR protein similar to parental Y1 cells. Signaling pathways relevant to c-Fos induction was extensively investigated in 3 clones: 3T3-AR01 and -07 and 3T3-04. In Y1 cells, specific inhibitors (H89/PKA; PD98059/MEK; Go6983/PKC and SP600125/JNK) show that signals initiated in the ACTH/ACTHR-system activate 4 pathways to induce the c-fos gene, namely: (a) cAMP/PKA/CREB; (b) MEK/ERK1/2; (c) PKC and d) JNK1/2. In 3T3-AR transfectants, both inhibitors PD98059 and Go6983 proved completely ineffective to inhibit c-Fos induction by ACTH(39), implying that MEK/ERK and PKC pathways are not involved in this process. On the other hand, SP600125 caused 85% inhibition of c-Fos induction by ACTH(39) and, in addition, ACTH(39) promotes JNK1/2 phosphorylation, suggesting that JNK is a major signaling pathway mediating c-Fos induction by ACTH(39) in these cells. In addiction, PKA inhibitor H89 also inhibits c-Fos induction in 3T3-AR7 cells by ACTH(39), implicating activation of the cAMP/PKA/CREB pathway in c-Fos induction by ACTH(39). However, the cAMP derivatives db-cAMP and 8Br-cAMP, do not promote CREB phosphorylation and c-Fos induction in parental Balb3T3 and 3T3-AR transfectants, confirming previous report by others. In conclusion, expression of active ACTHR in Balb3T3 fibroblasts renders these cells responsive to ACTH with activation of cAMP/PKA/CREB and JNK pathways and, also, induction of genes from the fos and jun families. These results show that Balb 3T3-AR sublines are useful cellular systems for genetic analysis of ACTH-signaling pathways. However, activation of cAMP/PKA/CREB and JNK pathways and induction of fos and jun genes are not yet sufficient to enable ACTH for interference in morphology, migration and proliferation of Balb3T3 fibroblasts as it does in Y1 adrenocortical cells.


Prolonged exposure to salt stress affects specialized metabolites-artemisinin and essential oil accumulation in Artemisia annua L.: metabolic acclimation in preferential favour of enhanced terpenoid accumulation accompanying vegetative to reproductive phase transition.
Artemisia annua accumulates substantial quantities of unique and highly useful antimalarial sesquiternoid artemisinin and related phytomolecules as well as its characteristic essential oil in its glandular trichomes. The phytomolecules are mainly produced in its leaves and inflorescences. Artemisia annua plants were grown under NaCl salinity (50, 100 and 200 mM) stress conditions imposed throughout the entire life cycle of the plant. Results revealed that specialized metabolites like artemisinin, arteannuin-B, artemisinic acid + dihydroartemisinic acid and essential oil accumulation were positively modulated by NaCl salinity stress. Interestingly, total content of monoterpenoids and sesquiterpenoids of essential oil was induced by NaCl salinity treatment, contrary to previous observations. Production of camphor, the major essential oil constituent was induced under the influence of treatment. The metabolic acclimation and manifestations specific to terpenoid pathway are analysed vis-a-vis vegetative to reproductive periods and control of the modulation. WRKY and CYP71AV1 play a key role in mediating the responses through metabolism in glandular trichomes. The distinctness of the salinity induced responses is discussed in light of differential mechanism of adaptation to abiotic stresses and their impact on terpenoid-specific metabolic adjustments in A. annua. Results provide potential indications of possible adaptation of A. annua under saline conditions for agrarian techno-economic benefaction.


Living with cystic fibrosis: impact on global quality of life.
Because of better health care, most people with cystic fibrosis (CF) now survive into adulthood, raising issues related to quality of life. Few studies have focused on satisfaction with broader life domains in people with CF. The aim of this article was to examine the impact of living with CF from a global quality of life perspective. The sample consisted of 86 adults with CF recruited from the Norwegian Competence Centre for Cystic Fibrosis and a control group of 1021 individuals from the general population (GP). The Norwegian version of the Quality of Life Scale was used to measure global quality of life (satisfaction with broader life domains) in both groups. The mean global quality of life score was 84.88 for the CF group and 83.33 for the GP group. This difference was significant after controlling for age, sex, educational level, and marital status, indicating that people with CF have better global quality of life. The groups also differed significantly on other specific measures of life satisfaction. Persons with forced expiratory volume in 1 second values below 30% of predicted values reported the lowest mean global quality of life value. These results suggest the existence of a type of response shift in the CF group through changes in life standards and goals. However, people with CF who have low forced expiratory volume in 1 second values may need special attention from health care professionals on issues related to global quality of life.


Decision-Support Tools Used in the Baltic Sea Area: Performance and End-User Preferences.
Decision-support tools (DSTs) synthesize complex information to assist environmental managers in the decision-making process. Here, we review DSTs applied in the Baltic Sea area, to investigate how well the ecosystem approach is reflected in them, how different environmental problems are covered, and how well the tools meet the needs of the end users. The DSTs were evaluated based on (i) a set of performance criteria, (ii) information on end user preferences, (iii) how end users had been involved in tool development, and (iv) what experiences developers/hosts had on the use of the tools. We found that DSTs frequently addressed management needs related to eutrophication, biodiversity loss, or contaminant pollution. The majority of the DSTs addressed human activities, their pressures, or environmental status changes, but they seldom provided solutions for a complete ecosystem approach. In general, the DSTs were scientifically documented and transparent, but confidence in the outputs was poorly communicated. End user preferences were, apart from the shortcomings in communicating uncertainty, well accounted for in the DSTs. Although end users were commonly consulted during the DST development phase, they were not usually part of the development team. Answers from developers/hosts indicate that DSTs are not applied to their full potential. Deeper involvement of end users in the development phase could potentially increase the value and impact of DSTs. As a way forward, we propose streamlining the outputs of specific DSTs, so that they can be combined to a holistic insight of the consequences of management actions and serve the ecosystem approach in a better manner.


Non-invasive Assessment by B-Mode Ultrasound of Arterial Pulse Wave Intensity and Its Reduction During Ventricular Dysfunction.
Arterial pulse waves contain clinically useful information about cardiac performance, arterial stiffness and vessel tone. Here we describe a novel method for non-invasively assessing wave properties, based on measuring changes in blood flow velocity and arterial wall diameter during the cardiac cycle. Velocity and diameter were determined by tracking speckles in successive B-mode images acquired with an ultrafast scanner and plane-wave transmission. Blood speckle was separated from tissue by singular value decomposition and processed to correct biases in ultrasound imaging velocimetry. Results obtained in the rabbit aorta were compared with a conventional analysis based on blood velocity and pressure, employing measurements obtained with a clinical intra-arterial catheter system. This system had a poorer frequency response and greater lags but the pattern of net forward-traveling and backward-traveling waves was consistent between the two methods. Errors in wave speed were also similar in magnitude, and comparable reductions in wave intensity and delays in wave arrival were detected during ventricular dysfunction. The non-invasive method was applied to the carotid artery of a healthy human participant and gave a wave speed and patterns of wave intensity consistent with earlier measurements. The new system may have clinical utility in screening for heart failure.


Effect of the Wii Sports Resort on the improvement in attention, processing speed and working memory in moderate stroke.
Stroke is the most common neurological disease in the world. After the stroke, some people suffer a cognitive disability. Commercial videogames have been used after stroke for physical rehabilitation; however, their use in cognitive rehabilitation has hardly been studied. The objectives of this study were to analyze attention, processing speed, and working memory in patients with moderate stroke after an intervention with Wii Sports Resort and compared these results with a control group. A pre-post design study was conducted with 30 moderate stroke patients aged 65 ± 15. The study lasted eight weeks. 15 participated in the intervention group and 15 belong to the control group. They were assessed in attention and processing speed (TMT-A and B) and working memory (Digit Span of WAIS-III). Parametric and effect size tests were used to analyze the improvement of those outcomes and compared both groups. At the baseline, there was no difference between TMT-A and B. A difference was found in the scalar score of TMT-B, as well as in Digit Backward Span and Total Digit Task. In TMT-A and B, the intervention group had better scores than the control group. The intervention group in the Digit Forward Span and the Total Digit obtained a moderate effect size and the control group also obtained a moderate effect size in Total Digit. In the Digit scalar scores, the control group achieved better results than the intervention group. The results on attention, processing speed and working memory improved in both groups. However, according to the effect sizes, the intervention group achieved better results than the control group. In addition, the attention and processing speed improved more than the working memory after the intervention. Although more studies are needed in this area, the results are encouraging for cognitive rehabilitation after stroke.


Design and assessment of an energy self-supply process producing tetraethyl orthosilicate using rice husk.
Combusting rice husk (RH) generates energy and rice husk ash (RHA) containing high amount of silica. Recent studies showed RHA can directly react with ethanol for producing tetraethyl orthosilicate (TEOS), an important substance for different industries. Nevertheless, this process requires an intensive energy supply. This study aims to design and evaluate an energy self-supply process producing TEOS using RH for feasibility. A process simulator was used to design the target process. The simulation results revealed that RH combustion can completely meet the RHA and high energy demands of TEOS production. The economic and environmental benefits were thoroughly evaluated and compared with processes using conventional raw materials (i.e., Simg and silica). The evaluation results showed that using RH for TEOS production could reduce CO2 emissions substantially. Large economic benefit was gained when renewable electricity was co-generated and sold to the power grid as a surplus.


Measurement of glycated albumin in diabetic patients by biospecific affinity chromatography.
The percentage of glycated plasma albumin was measured by a procedure involving ammonium sulphate precipitation and Affi-Gel-Blue and phenylboronate chromatographies. The value correlates well with the amount of ketoamine-bound sugars determined by colorimetric assay (r = 0.98, n = 39). The normal mean value is 3.9 +/- 0.3% (mean +/- S.D., coefficient of variation = 7.7%, n = 32) and varies from 3.9 to 21% in diabetics (n = 54). A good correlation is found with the mean blood glucose value of the preceding twenty days (r = 0.92, n = 57). Because of its relative ease of determination, glycated albumin constitutes a good short-term glycemic index and an alternative to glycated haemoglobin in some specific cases.


Studies of lipoproteins and fatty acids in maternal and cord blood of two racial groups in Trinidad.
The high mortality rate from coronary heart disease (CHD) among Indians compared to Negroes in Trinidad led us to test plasma lipid profiles to see whether dietary or genetic factors might be involved. There were no interracial differences in the composition of plasma cholesterol ester fatty acids of the tested women and neonates. This finding suggests that dietary fat does not account for the interracial difference in CHD, nor does the cause appear to be due to genetic differences in lipid profiles, as there was no significant difference between values for plasma triglycerides, total cholesterol, high density lipoprotein (HDL) cholesterol, apo-I, apo-II, apo B or cholesterol ester fatty acids in the cord blood of each racial group. Blood samples were collected from 69 nonpregnant and 71 postpartum, fasted Negro and Indian women. Also taken were 71 umbilical cord blood samples. The mean triglyceride level was significantly lower in the Negro nonpregnant and postpartum women than in the Indians. HDL cholesterol and apo-I values were lower in the Indian women. There were no significant differences in the total cholesterol and apo B measurements. The triglyceride values for postpartum women were higher than those of the nonpregnant Negroes and Indians (75% and 47%, respectively), whereas the total cholesterol and HDL cholesterol, apo A-I and apo A-II ranged from 9% to 29% higher in the postpartum women. Apo B was about 40% higher postpartum in both ethnic groups. The high CHD rate of Indians in Trinidad cannot be explained by dietary factors, plasma total cholesterol or fatty acid composition.(ABSTRACT TRUNCATED AT 250 WORDS)


Consultation as a political process.
Consultation may be viewed as a political-bargaining process in which actors seek to maximize at minimal cost expertise, organizational position, and organizational reputation. A view of consultation as political process allows for a shift in language in discussions of consultation. Such a language shift suggests shifts in the social meaning of the consulation process. Emphasis was placed on four suggested functions of consultation: definition and legitimation of a situation or of facts as "problematic"; raising the priority of an i5sue on the agenda of action in a consultee's agency; legitimation of deviant administrative behavior, and creation and sustenance of interagency linkages. A perspective is proposed that looks at the consequences as well as the intents of the process. This view of consultation can be studied emprically.


Health care professionals and use of digital technologies in patients with dementia and related disorders.
Digital technologies (digiTech) enhance benefits in healthcare and are present in elderly people lives. Healthcare professionals (HCP) who deal with dementia and related disorders face enormous challenges in delivering healthcare using digital devices, and Covid-19 pandemic showed these pitfalls should be addressed. Technology solutions in elder people are being developed, not every HCP is heard, and we lack information about what these professionals think about this. Using structured questionnaires and asynchronous electrotonic data capture, we asked experienced (84.3% postgraduate degrees) HCP with different backgrounds (8 practice areas) about what they think of technology use among their patients and carers. The average person in our sample (N = 51) was a highly skilled (22.2 study-years) female (70.6%) medical doctor (62.8%) aged 39.1 years old (27 to 77 years), highly exposed to digiTech (68.6% mobile devices usage ≥4 hours/day). Neurologists (37.3%), neuropsychologists (15.7%), psychiatrists (13.7%), geriatricians (11.8%) were the most represented groups. This observational cross-sectional study is part of a Brazilian pilot study to explore technology usage in higher risk dementia adults (DigiTAU Project). Although HCP consider asking patients and carers about technology use to be theoretically important (90.2%) and useful in practice (94.2%), they often do not do it (45.1%). There is incongruity when they feel safe to offer interventions by digital means (58.8%) but commonly agree, on average, that HCP are not prepared to use digiTech (51.0%). Despite having positive views towards benefits in adopting digiTech for patients (98.1%), they think they bring more benefits to families and caregivers than to patients themselves (56.9%). HCP consider digiTech will change their practice in the short term within 1 year (58.8%) and within 5 years (88.3%). None of the 8 options suggested as the most "promising" technology in 10 years reaches more than 25% of the choices. HCP have conflicting views about the usage of technologies by their patients and caregivers but expect some positive changes in their work practice. It suggests new digiTech must be submitted to scrutiny so that its applicability overcomes the challenges. Further studies and analysis are needed to understand better this data.


Peritoneal lavage in penetrating injuries of the small bowel and colon: value of enzyme determinations.
The value of lavage alkaline phosphatase determinations following hollow visceral injury was studied. Twenty-nine mongrel dogs were subjected to partial transection of the small intestine or colon, or underwent sham laparotomy. Sequential diagnostic peritoneal lavage was performed. At one hour post injury the lavage red cell count, white cell count, and amylase levels were within normal limits in each group. The lavage alkaline phosphatase, however, was markedly elevated in both bowel-injured groups compared with the sham group. In the five-hour lavage, red cell counts and amylase levels remained normal. White cell counts were now elevated in all three groups, but the results did not distinguish the intestinal injury from the sham model. Lavage alkaline phosphatase was again significantly increased in the injured groups versus the sham model. Serum alkaline phosphatase and amylase levels were normal in all study groups at baseline and one and five hours post injury. This canine study demonstrates the utility of peritoneal lavage alkaline phosphatase levels in identifying isolated small intestine and colon injuries that were otherwise undetected by red cell, white cell, and amylase determinations.


Effect of chlorhexidine varnish application on mutans streptococci counts in orthodontic patients.
Enamel demineralization occurs frequently around orthodontic brackets. High levels of mutans streptococci (MS) increase the risk of caries. The hypotheses tested in this study were that high-frequency application of 40% chlorhexidine varnish (40%CHX) results in lower levels of MS in plaque of orthodontic patients compared with low-frequency application, and that bimonthly application of 40%CHX results in lower levels of MS in plaque compared with 1% chlorhexidine/1% thymol varnish (1%CHX). Eighty adolescent patients were randomly allocated to 4 treatment groups: monthly, bimonthly, and quarterly 40%CHX application, and bimonthly application of 1%CHX. Plaque samples were collected monthly and processed for MS counts. Twenty patients dropped out before the evaluation time at 1 month. Data from the remaining 60 adolescents were analyzed by using linear regression models with the plaque bacterial ratio (PBRx) as the dependent variable. The trial was ended after 5 months because of a high number of dropouts. High-frequency application of 40%CHX did not provide lower mean PBRx than low-frequency application. Mean PBRx after 40%CHX application was lower than 1%CHX after 1 month (P = .002). This reduction was undiscernible 2 months after the application. 40%CHX application results in greater reduction of MS in plaque than 1%CHX a month after application. A follow-up study with a larger study population is warranted to test the first hypothesis.


Involvement of microglial RhoA/Rho-kinase pathway activation in the dopaminergic neuron death. Role of angiotensin via angiotensin type 1 receptors.
It has recently been shown that the dopaminergic cell loss induced by neurotoxins is enhanced by brain angiotensin II (AII) via type 1 receptors (AT1). However, the mechanisms involved in the dopaminergic degeneration and the brain inflammatory effects of AII have not been clarified. The RhoA-Rho-Kinase (ROCK) pathway may play a critical role in the inflammatory and oxidative effects of AII. In the substantia nigra of mice, administration of the dopaminergic neurotoxin MPTP induced an increase in the expression of RhoA and ROCK II mRNA levels and ROCK activity, which were inhibited by AT1 receptor deletion (i.e., in AT1a null mice treated with MPTP). Administration of the ROCK inhibitor Y-27632 or AT1 deletion induced a significant decrease in MPTP-induced microglial activation and dopaminergic cell death. In rat primary mesencephalic cultures treated with MPP(+), the increase in dopaminergic cell loss induced by AII administration was also inhibited by treatment with Y27632. Intense expression of ROCK II was observed in the microglial cells in the substantia nigra of mice treated with MPTP, and the major role of the microglial ROCK was confirmed by comparing mesencephalic cultures with and without microglia. Activation of the RhoA/ROCK pathway is involved in the MPTP-induced dopaminergic degeneration, and in the enhancing effect of AII/AT1 activation on the microglial response and dopaminergic degeneration. ROCK inhibitors and AT1 receptor antagonists may provide new neuroprotective strategies against the progression of Parkinson's disease.


Oncogene Convergence in Extrachromosomal DNA Hubs.
Extrachromosomal DNA circles (ecDNA) are a common mechanism for oncogene amplification and are associated with worse clinical outcomes compared with other types of oncogene amplification. Several recent discoveries of ecDNA hubs-local congregations of ecDNAs in the nucleus-highlight unique features of ecDNA biology that may contribute to higher oncogene expression and rapid tumor evolution.


Cyclobenzaprine and ethanol interaction.
The effects of cyclobenzaprine, a tricyclic compound, on the central depressant action of ethanol and on hepatic ethanol metabolizing enzymes were studied in rodents. Administration of cyclobenzaprine, 5 mg/kg, IP, 30 min prior to a narcotic dose of ethanol solution, 5 g/kg, IP, enhanced ethanol-produced narcosis in mice. This effect was greater in male than in female mice. Cyclobenzaprine inhibited endogenous rat liver alcohol dehydrogenase in vitro in the concentration range between 10(-5) M and 10(-6)M. Cyclobenzaprine exerted little effect on hepatic aldehyde dehydrogenase in vitro. The results suggest that cyclobenzaprine possesses depressant properties and inhibition of liver alcohol dehydrogenase may underlie the observed behavioral response studied. It is concluded that alteration of endogenous liver alcohol dehydrogenase by certain tricyclic antidepressant drugs may be involved in the mechanism(s) of their toxic interaction with ethanol.


Pavlovian Learning Processes in Pediatric Anxiety Disorders: A Critical Review.
Deficits in associative and Pavlovian learning are thought to lie at the center of anxiety-related disorders. However, the majority of studies have been carried out in adult populations. The aim of this review was to critically examine the behavioral and neuroimaging literature on Pavlovian learning in pediatric anxiety disorders. We conclude that although there is evidence for deficits in Pavlovian processes (e.g., heightened reactivity to safety cues in anxious samples), the extant literature suffers from key methodological and theoretical issues. We conclude with theoretical and methodological recommendations for future research in order to further elucidate the role of Pavlovian learning in the etiology, maintenance, and treatment of pediatric anxiety disorders.


Pollination and pre-dispersal seed predation: net effects on reproduction and inflorescence characteristics in Ipomopsis aggregata.
We examined net seed production for the self-incompatible, monocarp, Ipomopsis aggregata, by monitoring pre-pollination seed parasite (Hylemya sp.) oviposition and hummingbird mediated fruit set on 21 plants of variable height. Both pollination and seed predation increased with inflorescence height. Net seed production (incorporating seed predator mortality) also was positively related to height, and this would have been the case if pollination or seed predation were doubled. Although results suggest pomopsis aggregata should be under selective pressure to maximize inflorescence height, generation time and resource limits could result in advantages for inflorescences of intermediate height.


Influenza Vaccine Effectiveness in the Inpatient Setting: Evaluation of Potential Bias in the Test-Negative Design by Use of Alternate Control Groups.
The test-negative design is validated in outpatient, but not inpatient, studies of influenza vaccine effectiveness. The prevalence of chronic pulmonary disease among inpatients can lead to nonrepresentative controls. Test-negative design estimates are biased if vaccine administration is associated with incidence of noninfluenza viruses. We evaluated whether control group selection and effects of vaccination on noninfluenza viruses biased vaccine effectiveness in our study. Subjects were enrolled at the University of Michigan and Henry Ford hospitals during the 2014-2015 and 2015-2016 influenza seasons. Patients presenting with acute respiratory infection were enrolled and tested for respiratory viruses. Vaccine effectiveness was estimated using 3 control groups: negative for influenza, positive for other respiratory virus, and pan-negative individuals; it was also estimated for other common respiratory viruses. In 2014-2015, vaccine effectiveness was 41.1% (95% CI: 1.7, 64.7) using influenza-negative controls, 24.5% (95% CI: -42.6, 60.1) using controls positive for other virus, and 45.8% (95% CI: 5.7, 68.9) using pan-negative controls. In 2015-2016, vaccine effectiveness was 68.7% (95% CI: 44.6, 82.5) using influenza-negative controls, 63.1% (95% CI: 25.0, 82.2) using controls positive for other virus, and 71.1% (95% CI: 46.2, 84.8) using pan-negative controls. Vaccination did not alter odds of other respiratory viruses. Results support use of the test-negative design among inpatients.


Protein S: A multifunctional anticoagulant vitamin K-dependent protein at the crossroads of coagulation, inflammation, angiogenesis, and cancer.
Since its discovery in 1970, protein S (PS) has emerged as a key vitamin K-dependent natural anticoagulant protein at the crossroads of multiple biological processes, including coagulation, apoptosis, atherosclerosis, angiogenesis/vasculogenesis, and cancer progression. Following the binding to a unique family of protein tyrosine kinase receptors referred to as Tyro-3, Axl and Mer (TAM) receptors, PS can lead to regulation of coagulation, phagocytosis of apoptotic cells, cell survival, activation of innate immunity, vessel integrity and angiogenesis, and local invasion and metastasis. Because of these dynamics and multiple functions of PS, which are largely lost following invalidation of the mouse PROS1 gene, this molecule is currently intensively studied in biomedical research. The purpose of this review is to provide a brief chronicle of the discovery and current understanding of the mechanisms of PS signaling, and how PS and their signaling partners regulate various cellular functions, with a particular focus on TAM receptors.


Screening for cytomegalovirus shedding in vagina and saliva: Significant differences between biological fluids, swab types and storage durations in DNA recovery.
Cytomegalovirus (CMV) shedding in genital and oral secretions during pregnancy is associated with adverse outcomes. Sample collection methods between studies are not uniform and currently there are limited data on the impact of biological fluids, swab types and storage durations on the detection of CMV DNA. To evaluate the absorption efficiency and CMV DNA recovery of various commercially available swabs in vaginal and saliva fluids. The absorption volume of different swab types was evaluated. The recovery of CMV DNA over time from vaginal and saliva fluids, and vaginal and saliva swabs, was evaluated by PCR measurements of samples spiked with CMV standard. Absorption efficiency of swabs varied significantly. The duration of storage did not affect CMV DNA recovery from vaginal fluid or swabs, but did significantly affect CMV DNA recovery from neat saliva fluid and saliva swabs stored dry or in viral transport media (VTM). Flocked swab/eNAT media recovered the highest amount of CMV DNA from both fluids. In saliva, flocked swab/eNAT media and polyester swab/cobas media demonstrated a higher CMV DNA recovery than foam swab/VTM. 25% of dry saliva foam swabs were falsely negative for CMV DNA. Recovery of CMV DNA is dependent on sample type and swab type used. Flocked swab/eNAT media and polyester swab/cobas media appear acceptable, though flocked swab/eNAT media was superior demonstrating the best recovery of CMV DNA. For saliva, foam swabs stored dry or in VTM was shown to be inferior to flocked or polyester swabs.


Peripheral ischemia due to retroperitoneal fibrosis.
Two patients with retroperitoneal fibrosis demonstrated symptoms of peripheral vascular ischemia. Arteriolysis to free the distal aorta and iliac vessels was successful in both patients. Aortography in the posteroanterior view was equivocal, but the accompanying urologic abnormalities and a history of methysergide ingestion helped establish the etiology of the ischemia. The most direct approach to the treatment of the vascular obstruction caused by retroperitoneal fibrosis is complete arteriolysis. Most previous reports indicate that it is relatively easy to establish a dissection plane between the fibrotic plaque and the vessel wall. We found that the fibrotic process invaded the vessel wall. Accordingly, the surgeon must anticipate a difficult, tedious dissection when performing arteriolysis for the treatment of vascular compression secondary to retroperitoneal fibrosis.


Assessing soil erosion risk in a peri-urban catchment of the Lake Victoria basin.
Soil erosion and sedimentation contribute to deteriorating water quality, adverse alterations in basin hydrology and overall ecosystem biogeochemistry. Thus, understanding soil erosion patterns in catchments is critical for conservation planning. This study was conducted in a peri-urban Inner Murchison Bay (IMB) catchment on the northern shores of Lake Victoria since most soil erosion studies in Sub-Saharan Africa have been focused on rural landscapes. The study sought to identify sediment sources by mapping erosion hotspots using the revised universal soil loss equation (RUSLE) model in appendage with field walks. RUSLE model was built in ArcGIS 10.5 software with factors including: rainfall erosivity, soil erodibility, slope length and steepness, land cover and support practices. The model was run, producing an erosion risk map and field assessments conducted to ground-truth findings and identify other hotspots. The percentage areas for RUSLE modelled erosion rates were: 66.8% for 0-2 t ha-1 year-1; 10.8% for 2-5 t ha-1 year-1; 10.1% for 5-10 t ha-1 year-1; 9% for 10-50 t ha-1 year-1 and 3.3% for 50-100 t ha-1 year-1. Average erosion risk was 7 t ha-1 year-1 and the total watershed erosion risk was 197,400 t year-1, with croplands and steep areas (slope factor > 20) as the major hotspots (> 5 t ha-1 year-1). Field walks revealed exposed soils, marrum (gravel) roads and unlined drainage channels as other sediment sources. This study provided the first assessment of erosion risk in this peri-urban catchment, to serve as a basis for identifying mitigation priorities. It is recommended that tailored soil and water conservation measures be integrated into physical planning, focusing on identified non-conventional hotspots to ameliorate sediment pollution in Lake Victoria.


Enhancement of lead uptake by alfalfa (Medicago sativa) using EDTA and a plant growth promoter.
Phytoremediation is a novel cleanup technology for the removal of contaminants from polluted waters and soils. In phytoremediation, the plant uptake capability and the availability of the pollutant in the media are important. Here we show the results of a study on the combined effects of ethylenediaminetetraacetic acid (EDTA) and the phytohormone indole-3-acetic acid (IAA) on Pb uptake by Medicago sativa (alfalfa). Plants were grown in hydroponics media containing a nutrient solution amended with Pb at 0.2mM and different combinations of EDTA, and IAA. After 10d of treatment, the Pb content in plant tissues was quantified using an Inductively Coupled Plasma Optical Emission Spectrometer (ICP/OES). The results showed that the combination of 100 microM IAA/0.2 mM EDTA increased the Pb accumulation in leaves by about 2800% and by about 600%, as compared to Pb content in leaves of plants exposed to Pb alone and those cultivated with Pb/EDTA, respectively. These results indicate that non-metal hyperaccumulator plants could increase their hyperaccumulating potential without genetic manipulation.


The Pekin duck IL-10R2 common chain: cDNA cloning, genomic structure, molecular characterization and mRNA expression analysis.
The interleukin-10 receptor 2 (IL-10R2, IL-10Rβ) is required for the signalling of the class 2 cytokines IL-10, IL-22, IL-26 and IFN-λ1-3 . Here, we describe the identification of the Pekin duck IL-10R2 (duIL-10R2) common chain and its gene structure. The duIL-10R2 cDNA encodes a 343 amino acid protein that has an amino acid identity of 76% and 42% with chicken and human IL-10R2, respectively. Binding residues of human IL-10R2 for IL-10 and IL-22 were mostly conserved in the avian IL-10R2 proteins within loops L3 and L5, but not within loops L2 and L6. Homology modelling of the duIL-10R2 extracellular domain structure using soluble human IL-10R2 (shIL-10R2, PDB ID: 3LQM) as a template revealed a protruding loop L5 and two distinct clefts between loops L2/L3 and L3/L5, similar to shIL-10R2. However, in contrast to the three amino acid β-hairpin loop L2 of shIL-10R2, loop L2 of duIL-10R2 is five residues longer. Residues within a putative Tyk2 binding site were highly conserved across all vertebrate IL-10R2 proteins examined. The duIL-10R2 gene shares a seven exon-six intron structure with chicken and human IL-10R2 genes, but avian genes are more compact. DuIL-10R2 mRNA was constitutively expressed in all tissues. Mitogen stimulation of duck peripheral blood mononuclear cells (PBMC) did not alter transcript levels. Our observations suggest that genomic organization and structural features implicated in multiple cytokine-binding properties of human IL-10R2 are conserved in duck IL-10R2, but the evolutionary changes that appear to have lead to low-affinity cytokine interaction within loop L2 are distinct to mammalian species.


Understanding the synergistic mechanism of PAM-FeCl3 for improved sludge dewaterability.
Hybrid flocculant polyacrylamide-ferric chloride (PAM-FeCl3) was developed to improve the dewaterability of sewage sludge and the dewatering performance, properties of treated sludge, composition and morphology distribution of extracellular polymeric substance (EPS) were investigated. The physicochemical properties of the PAM-FeCl3 were characterized, and its effectiveness as a sludge conditioner was evaluated. The results indicated that PAM-FeCl3 conditioning was able to promote sludge dewaterability. Simultaneously, PAM-FeCl3 neutralized the negative charges on the surface of sludge particles and increased the sludge floc size. Besides, PAM-FeCl3 also formed a rough and porous floc structure that reduced sludge compressibility. Meanwhile, the exciting emission matrix analysis suggested that PAM-FeCl3 can effectively disintegrate of EPS fraction in sludge and decompose the aromatic protein-like substances as well as the humic acid-like substances in EPS. Additionally, the larger sludge floc formation, electrostatic interaction and adsorption bridging effect resulted in compression of sludge structure and the decomposition of EPS fractions and improved sludge dewatering performance.


Visual assessment with preferential looking techniques in mentally retarded children.
An operant preferential looking technique was successfully used for testing of visual acuity in mentally retarded children with visual dysfunction.


Nitrogen sources for neotropical nasute termites: Fixation and selective foraging.
Studies with Nasutitermes and Rhynchotermes (Isoptera, Nasutitermitinae) in a Costa Rican rainforest suggest that nitrogen fixation by gut symbionts may play a significant role in termite nutrition. Leaf-litter feeders ingest more and fix less nitrogen than do wood-litter feeders; both species feed preferentially on N-rich foods in their foraging repertoire. Unusually high N2 fixation (acetylene reduction) rates were found in Nasutitermes soldiers, thereby reducing their nutritional dependence on workers. Finally, N2 fixation rates of termites diminished rapidly within 24 h after removal of a colony from the field, underlining the importance of conducting future N2 fixation studies under field conditions.


Who gets treated where. A study of patients transferred and not transferred from a consultation-liaison service to a general hospital psychiatry unit.
The decision to transfer a patient from a general hospital unit to a psychiatric unit is multifactorial. The most salient patient characteristics include the degree of dysfunctional behavior, the absence of social supports, and the presence of Axis II pathology. Severity of medical illness does not discriminate between the two groups. Awareness of these characteristics could aid physicians in formulating transfer decisions.


An association study of asthma and total serum immunoglobin E levels for Toll-like receptor polymorphisms in a Japanese population.
The prevalence of atopic diseases has been increasing in developed countries. This could be explained by the hygiene hypothesis, which states that exposure to specific infections or endotoxins during infancy drives the maturing immune system towards a Th1 phenotype and away from the Th2 phenotype, which is associated with allergic diseases. Toll-like receptors (TLRs) play important roles in the signalling of many pathogen-related molecules and endogenous proteins associated with immune activation. The aim of the present study was to investigate whether polymorphisms in genes encoding TLRs are associated with asthma or total serum IgE levels. We screened the 5' flanking and coding regions of the TLR2,TLR3, TLR4, and TLR9 genes for polymorphisms by direct sequencing of DNA from 32 asthmatics, and analysed the effect of the polymorphisms on the development of atopic asthma and on total serum IgE levels. We identified 16 variants in TLRs. The transmission disequilibrium test of the families revealed that none of the alleles or haplotypes were associated with asthma or total IgE levels (P>0.05). However, we found an insertion/deletion polymorphism in the 5' untranslated region of TLR2, and an expression construct containing the deletion allele showed lower luciferase activity than the wild-type alleles, suggesting that the deletion allele has reduced transcriptional activity. Our results indicate that polymorphisms in TLRs are not likely to be associated with the development of atopy-related phenotypes in a Japanese population.


Different brain functional network abnormalities between attention-deficit/hyperactivity disorder youth with and without familial risk for bipolar disorder.
Attention-deficit/hyperactivity disorder (ADHD) commonly precedes the initial onset of mania in youth with familial risk for bipolar disorder (BD). Although ADHD youth with and without BD familial risk exhibit different clinical features, associated neuropathophysiological mechanisms remain poorly understood. This study aimed to identify brain functional network abnormalities associated with ADHD in youth with and without familial risk for BD. Resting-state functional magnetic resonance imaging scans were acquired from 37 ADHD youth with a family history of BD (high-risk), 45 ADHD youth without a family history of BD (low-risk), and 32 healthy controls (HC). Individual whole-brain functional networks were constructed, and graph theory analysis was applied to estimate network topological metrics. Topological metrics, including network efficiency, small-worldness and nodal centrality, were compared across groups, and associations between topological metrics and clinical ratings were evaluated. Compared to HC, low-risk ADHD youth exhibited weaker global integration (i.e., decreased global efficiency and increased characteristic path length), while high-risk ADHD youth showed a disruption of localized network components with decreased frontoparietal and frontolimbic connectivity. Common topological deficits were observed in the medial superior frontal gyrus between low- and high-risk ADHD. Distinct network deficits were found in the inferior parietal lobule and corticostriatal circuitry. Associations between global topological metrics and externalizing symptoms differed significantly between the two ADHD groups. Different patterns of functional network topological abnormalities were found in high- as compared to low-risk ADHD, suggesting that ADHD in youth with BD familial risk may represent a phenotype that is different from ADHD alone.


Imaging strategies for diagnosing calcaneal and cuboid stress fractures.
Stress fractures of the calcaneus and cuboid bones are not among the most common pathologies seen by the foot and ankle specialist on a day-to-day basis. Therefore, the clinician must be suspicious of these particular entities when conservative therapy has failed to alleviate the pain of a more common condition. The purpose of this article is to discuss the signs, symptoms, MRI findings, nuclear imaging characteristics, and radiographic features associated with calcaneal and cuboid stress fractures. Case studies of both conditions are also discussed within this article.


High stability of immobilized β-D-galactosidase for lactose hydrolysis and galactooligosaccharides synthesis.
β-D-Galactosidase from Kluyveromyces lactis was immobilized on glutaraldehyde-activated chitosan and used in a packed-bed reactor for the continuous hydrolysis of lactose and the synthesis of galactooligosaccharides (GOS). The biocatalyst was tested for its optima pH and temperature, thermal stability in the presence of substrate and products, and operational stability. Immobilization increased the range of operational pH and temperature, and the enzyme thermal stability was sharply increased in the presence of lactose. Almost complete lactose hydrolysis was achieved for both milk whey and lactose solution at 37 °C at flow rates up to 2.6 mL min(-1). Maximal GOS concentration of 26 g L(-1) was obtained at a flow rate of 3.1 mL min(-1), with a productivity of 186 g L(-1) h(-1). Steady-state operation for 15 days showed the reactor stability concerning lactose hydrolysis.


Influence of substrate carbon on the metabolism of Clostridium thermohydrosulfuricum.
The concentration of carbon sources has a significant influence on the growth, carbohydrate uptake and metabolite distribution in Clostridium thermohydrosulfuricum. The growing concentrations of glucose or starch increase the production of ethanol and lactate, the intracellular fructose-1,6-diphosphate (FDP) and the specific activity of lactate dehydrogenase (LDH), but decrease the ethanol/lactate ratio.


FLT-PET/CT diagnosis of primary and metastatic nodal lesions of gastric cancer: comparison with FDG-PET/CT.
To examine the diagnostic performance of (18)F-fluorothymidine (FLT)-PET/CT of primary and metastatic nodal lesions of gastric cancer by comparing with (18)F-fluorodeoxyglucose (FDG)-PET/CT. The enrolled study population comprised 17 patients with 17 newly diagnosed gastric cancers who underwent surgery of the primary lesion and regional nodes after both FDG- and FLT-PET/CT scans. Visual detectability of the primary gastric lesions was correlated with pathological factors using the Fisher exact or Mann-Whitney U test. The sensitivity, specificity, and accuracy in detecting nodal lesions were compared between both PET/CT scans using the McNemar exact or χ (2) test. Fourteen of 17 (82.4%) primary cancers were visualized by both FDG- and FLT-PET/CT scans. Although FDG or FLT visibility was not significantly associated with tumor size (p = 0.16) or histological type (p = 1.00), the 3 nonvisible lesions were pathologically early (T1) cancers. The sensitivity, specificity, and accuracy for detecting nodal metastasis were 44.8% (13/29), 98.7% (164/166), and 90.8% (177/195) for FDG-PET/CT, and 31.0% (9/29), 100% (166/166), and 89.7% (175/195) for FLT-PET/CT, respectively. No significant difference was found between the two scans in sensitivity (p = 0.13), specificity (p = 0.48), or accuracy (p = 1.00). FLT-PET/CT may have the same diagnostic value as FDG-PET/CT for detection of primary and nodal lesions of gastric cancer.


Outcomes for surgical correction for stages 2 and 3 tibialis posterior dysfunction.
Between the years 1991 and 1996, 13 patients with stage 2 or 3 tibialis posterior dysfunction were evaluated following surgical reconstruction. Those patients with posterior tibial tendon tendinitis with a progressive flatfoot were categorized as having a stage 2 deformity according to Mueller's developmental stages of tibialis posterior dysfunction. Those patients with increasing severity of symptoms including a forefoot abductus component were classified as stage 3. There were five patients in stage 2, ranging in age from 53 to 80 years old; and there were eight patients in stage 3, ranging in age from 41 to 73 years old. Standard conservative care was utilized prior to surgical intervention in all cases. Follow-up was 12 months to 63 months. Patients in stage 2 underwent a Cobb reconstruction utilizing a split tibialis anterior tenodesis, and patients in stage 3 underwent an Evans lateral column-lengthening procedure combined with a Cobb procedure. Utilizing retrospective radiographic evaluation and patient interviews, results indicated that patients in stage 2 had a better patient satisfaction than those patients in stage 3. Although both patient groups had a 6-point average decrease in pain according to the 0- to 10-point visual analog pain scale, 50% of the patients undergoing a Cobb-Evans procedure felt that the procedure did not meet their expectations. Only one out of the five Cobb procedure patients felt that the procedure did not meet his expectations. The results of this limited study of patients with stage 3 tibialis posterior dysfunction suggest that although the lateral column lengthening with tendon augmentation renders good radiographic correction, many patients develop protracted lateral column pain and felt that surgery did not meet their expectations. Additional calcaneal osteotomies and arthrodesing procedures of the hindfoot may render a more satisfactory outcome.


Synthesis, spectral characterization and bio-analysis of some organotin(IV) complexes.
Five novel organotin(IV) derivatives have been synthesized by refluxing trimethyl, triethyl, tributyl, and triphenyl and tribenzyltin chloride with Schiff base derived from salicylaldehyde and adenine. These compounds were characterized by spectroscopic (IR, (1)H, (13)C, (119)Sn-NMR, (119m)Sn Mössbauer) techniques and elemental analysis. Based on these results, trigonal bipyramidal geometry is suggested. The synthesized compounds were also treated with various microorganisms and found to be active.


Peripheral glutamate receptors participate in interleukin-1beta-induced mechanical allodynia in the orofacial area of rats.
The present study was performed to examine peripheral cytokine-induced mechanical allodynia in the orofacial area and to investigate whether peripheral excitatory amino acids participate in the cytokine-induced mechanical allodynia. Experiments were carried out on male Sprague-Dawley rats. After interleukin-1beta (IL-1beta) was applied subcutaneously to the orofacial area, we examined withdrawal responses produced by air puffs applied to the IL-1beta injection site. The threshold of air puffs that produced withdrawal behavioral responses decreased significantly in a dose-dependent manner after injection of IL-1beta. Pretreatment with an IL-1 receptor antagonist abolished the decrease in the threshold of air puffs. Pretreatment with dl-2-amino-5-phosphonvaleric acid, an N-methyl-d-aspartic acid (NMDA) receptor antagonist, did not affect IL-1beta-induced mechanical allodynia. However, pretreatment with 6,7-dinitroquinoxaline-2,3-dione, a non-NMDA receptor antagonist, abolished the decrease in the threshold of air puffs. These results suggest that peripheral cytokine can produce mechanical allodynia in the orofacial area and that excitatory amino acids can modulate IL-1beta-induced mechanical allodynia via non-NMDA receptors.


Novel biallelic ASTL variants are associated with polyspermy and female infertility: A successful live birth following ICSI treatment.
Fertilization of the egg by the sperm is the first vital stage of embryogenesis. In mammals, only one sperm is incorporated into the oocyte. Polyspermy is a key anomaly of fertilization that is generally lethal to the embryo. To date, only a few causative genes for polyspermy have been reported. In a recent study, a homozygous variant in astacin-like metalloendopeptidase (ASTL), which encodes the ovastacin enzyme that cleaves ZP2 to prevent polyspermy, was found to be associated with female infertility characterized by polyspermy in vitro. Herein, we identified two ASTL variants in a Chinese woman likely responsible for her primary infertility and polyspermy in in vitro fertilization. Both variants were located within the key catalytic domain and predicted to alter hydrogen bonds, potentially impairing protein stability. Moreover, expression and immunoblot analyses in CHO-K1 cells indicated abnormal ovastacin zymogen activation or decreased enzyme stability. Intracytoplasmic sperm injection treatment successfully bypassed the defect in polyspermy blocking and resulted in a live birth. Our study associates ASTL variants with human infertility and further supports the contribution of this gene to blocking polyspermy in humans. Our findings expand the spectrum of ASTL mutations and should facilitate the diagnosis of oocyte-borne polyspermy.


Picture archiving and communication system activities at the Mayo Clinic Rochester.
This report summarizes Mayo Clinic Rochester's experience with a picture archiving and communication system (PACS) to date and discusses the principles that have guided implementation.


Audit of physicians' adherence to the antibiotic policy guidelines in Kuwait.
To audit physicians' adherence to the local antibiotic policy guidelines in government hospitals in Kuwait. The study was a retrospective review of patient records in nine hospitals between July 1 and December 31, 2008. Clinical notes and medication charts of the latest hospital admissions were checked for antibiotic prescribing. On the audit form, aspects of the prescribed antibiotic were benchmarked to the hospital antibiotic policy guidelines to evaluate adherence. Of 2,232 reviewed records, 1,112 (49.8%) patients had 1,528 antibiotic prescriptions. Patients who received antibiotics were significantly younger than those who did not (median age: 26.3 vs. 29.8 years, p < 0.001) and their hospital stay was significantly longer (median: 4 vs. 2 days, p < 0.001). The choice of an antibiotic was appropriate and matched the policy in 806 (52.7%) prescriptions. Of such appropriate antibiotics, adherence to route of administration was observed in 768/806 (95.3%), to dose in 758 (94%), to frequency in 746 (92.6%) and to duration in 608 (75.4%). Full adherence to all aspects of antibiotic choice, dose, route, frequency and duration was achieved in 464 (30.4%) prescriptions. In 382 (25%), the antibiotics administered were not indicated. There was low adherence to the local antibiotic policy guidelines. Physicians' antibiotic prescribing practices should be optimized. Adherence to, and update of, the policy is recommended.


Cell cycle modifications of breast cancers during neoadjuvant chemotherapy: a flow cytometry study on fine needle aspirates.
Breast cancer cells from 92 patients were obtained by repeated fine needle sampling and analysed by flow cytometry for cell cycle modifications during neoadjuvant chemotherapy. Modifications of the histograms were observed for 47 of the 71 informative cases (66%), the most frequent concerning S-phase (increase or decrease) and G2M accumulation. These modifications correlated well with the efficacy of cytotoxic chemotherapy (P < 0.0001). A significant relationship between clinical regression and pretreatment proliferative activity was also observed, with 31/35 (89%) responders in the high proliferation group (S-phase fraction > 5% or BrdU labelling index > 3.3%) compared to 20/36 (56%) in the low proliferation group (P < 0.002). For patients undergoing chemotherapy including doxorubicin, a high incidence of G2M accumulation was observed (33%), a modification which was rare (4.5%) for a regimen with no anthracycline, for which S-phase was the most frequently modified cell cycle compartment (64%). The measurement of the pretreatment tumour proliferative activity as well as the early kinetic modifications, as indicators of response, may prove interesting parameters for the future management of neoadjuvant chemotherapy.


Triple-Branched Stent Graft Implantation for Acute Non-A-non-B Aortic Dissection.
The optimal treatment for acute non-A-non-B aortic dissection remains controversial. Triple-branched stent graft (TBSG) implantation has been used to treat acute type A aortic dissection. This study aimed to evaluate the safety and efficacy of TBSG as a treatment for acute non-A-non-B aortic dissection. Fifty patients with non-A-non-B dissection received TBSG implantation in our center between January 2014 and December 2019. Early mortality, morbidity, and dissected aorta remodeling during follow-up were calculated. There were no deaths in-hospital or within 30 days. Postoperative complications included pneumonia (n = 12), acute kidney injury (n = 6; preoperative renal malperfusion, n = 4), transient cerebral injury (n = 6; preoperative cerebral malperfusion, n = 4), pleural effusion (n = 4), and pericardial effusion (n = 2). During follow-up, 1 patient experienced a stroke, and 2 patients required secondary interventional therapy for residual dissection below the level of the TBSG. All implanted TBSGs had good positioning, and all sidearm stent graft grafts were fully patent. No retrograde aortic dissection or type I endoleak was detected. TBSG implantation for acute non-A-non-B aortic dissection had a low incidence of mortality and morbidity, featuring good remodeling of the dissected aortic wall during follow-up. The early outcomes of this technique were satisfactory.


Different binding mechanisms of myeloid leukemic cells to adhesion molecules on bone marrow stromal fibroblasts induced by TNF-alpha and IL-4.
To study the mechanisms of adhesion of myeloid leukemic cells to bone marrow stroma, we analyzed the interaction of bone marrow stromal fibroblasts with myeloid leukemic cell lines and the modulation of adhesion molecule expression on stromal fibroblasts by TNF-alpha and IL-4. Like others, we found up-regulation of VCAM-1 and ICAM-1 on fibroblasts with TNF-alpha treatment, whereby IL-4 acted synergistically with TNF-alpha. VCAM-1 expression on the cell surface was maximal after 10 h, while ICAM-1 expression increased up to 48 h. All myeloid leukemic cell lines tested (HL-60, K562, TMM, U937, ML-2, PLB-985, THP-1, KG1a) revealed weak adhesion to untreated bone marrow fibroblasts (< or =10% bound cells). TNF-alpha and IL-4 significantly enhanced adhesiveness of fibroblasts to the cell lines PLB-985, THP-1, and ML-2, with a peak between 6 and 10 h of treatment. Adhesiveness to the cell line TMM was increased up to eightfold in a time-dependent manner for up to 48 h. The enhanced binding of ML-2-, THP-1-, and PLB-985 cells to stimulated fibroblasts was due at least partially to the interaction of VLA-4 with VCAM-1. Increased adhesion of TMM cells was impaired neither by antibodies to VLA-4, LFA-1, or Mac-1 nor by antibodies to their counter-receptors VCAM-1 or ICAM-1, suggesting that adhesion molecules distinct from VCAM-1 or ICAM-1 are involved in enhanced adhesiveness of the fibroblasts to myeloid leukemic cells.


Effects of feeding and digestion on myocardial contractility and expression of calcium-handling proteins in Burmese pythons (Python molurus).
Pythons are important models of studies on postprandial metabolism because their physiological responses are exacerbated when digesting large prey. Prior studies of these animals have shown hypertrophy of the cardiac tissue 2 to 3 days after feeding, coinciding with the peak of the specific dynamic action (SDA), but the consequences of this remodeling in myocardial contractility have not been studied, which is the purpose of this work. Specimens of Python molurus were divided into two groups: a Digesting group (2 days after feeding, at the peak of SDA), and a Fasting group (28 days after feeding). When compared to the Fasting group, the Digesting group showed higher relative ventricular mass and calcium-handling protein expression such as sarcoplasmic reticulum Ca2+-ATPase (SERCA), phospholamban (PLB), and the Na+/Ca2+ exchanger (NCX). Digesting pythons also exhibited significant increases in the cardiac contraction force (Fc), rates of force development and relaxation, and cardiac pumping capacity. Therefore, the higher SERCA, PLB and NCX expression levels increased cytosolic Ca2+ transient amplitude, improving myofilament force. These changes are crucial to maintain cardiac output and a relatively high and continuous blood flow required by metabolic expenditure that occurs in postprandial animals.


Genetic identity of eutetrarhynchids from the Persian Gulf, with intraindividual and intraspecific variability of Prochristianella butlerae Beveridge, 1990.
The examination of the two batoid species, Pastinachus sephen (Forsskål) and Himantura gerrardi (Gray), resulted in a morphologic identification of the five eutetrarhynchid species, namely Parachristianella indonesiensis Palm, 2004 reported for the first time from Himantura gerrardi, Pa. monomegacantha Kruse, 1959, Prochristianella clarkeae Beveridge, 1990, Pr. butlerae Beveridge, 1990 and Dollfusiella spinulifera (Beveridge et Jones, 2000). By sequencing the partial 28S rDNA and calculating the genetic distances between the morphologically identified specimens from the Persian Gulf and their allopatric morphological conspecifics as well as using a phylogeny, the specimens of the Persian Gulf were considered as the allopatric conspecifics of Parachristianella indonesiensis, Pa. monomegacantha and Prochristianella clarkeae. Since the specimens of Dollfusiella from the Persian Gulf and D. spinulifera from Australia had a high genetic distance and placed in two separate clades, the Iranian specimens of Dollfusiella were treated as the cryptic species of D. spinulifera. Following the treatment of Prochristianella macracantha Palm, 2004 as the synonym of Pr. butlerae Beveridge, 1990, it was also expressed that although Pr. butlerae appeared to be a variable species in terms of its metric data, there was an identical oncotaxy in all the conspecifics. A detail examination of the specimens from the Persian Gulf as well as Australian voucher specimens revealed a great intraindividual and intraspecific variability in this species. Since, the sequence information is only available for the Iranian specimens and there is no possibility to evaluate the genetic distance and their monophyly, we consider these two allopatric populations as one species.


Nurses' attitudes to sexuality in caring for cancer patients.
The attitudes of hospice nurses towards psychosexual issues in palliative care were surveyed. Sexuality was acknowledged to be both important and relevant in hospice nursing. A high level of awareness of sexual and relationship issues was apparent, reflecting broadly liberal attitudes.


Recommendations for medical training: a Native Hawaiian patient perspective.
Culturally competent health care providers are needed to eliminate healthcare disparities. In the State of Hawai'i, Native Hawaiians suffer some of the worst health disparities. Prior to implementing a cultural competency curriculum to address these disparities, the John A. Burns School of Medicine's Department of Native Hawaiian Health Cultural Competency Curriculum Development team asked Native Hawaiian patients about their experiences and recommendations. We conducted four focus groups of Native Hawaiians to obtain recommendations on physician training, to be incorporated into the curriculum. Participants came from both rural and urban areas. Classical qualitative analysis of data identified recurrent themes. Five primary themes, arising in all four groups, were: (1) customer service; (2) respect for the patient; (3) inter-personal skills; (4) thoroughness of care; and (5) costs of medical care. Secondary themes, occurring in three of the four groups, were: (1) cultural competency training; (2) the training of medical office staff; (3) continuity of care; and (4) the role of the patient. Participants specifically requested that medical students receive cultural competency training about the host culture, its history, values, and traditional and alternative healing practices. The emphasis participants placed on the need for cultural competency training of physicians supports the need to address the role of culture in medical education. Although most of the issues raised are not unique to Hawai'i, participants' recommendations to teach students about the host culture and traditional healing practices identify important themes not usually found in medical school curricula.


[Laparoscopic distal pancreatectomy for pancreatic cystadenomas].
Cystic neoplasms are an uncommon group among pancreatic tumors. These lesions are seen more frequently in recent surgical practice, probably because of advances in diagnostic and surgical techniques. Total tumor resection provides the best chance of cure and may remove the risk of malignant transformation of the cystadenomas, particularly of the mucinous type. Minimally invasive techniques have been revolutionary and provide clinical evidence of decreased morbidity and comparable efficacy to traditional, open surgery. However, laparoscopic pancreatic resection is not an established treatment for tumors of the pancreas. The authors present their initial experience with laparoscopic distal pancreatectomy for pancreatic cystadenomas. Three female patients (mean age, 55 years) underwent laparoscopic pancreatic resection between September 2001 and December 2003. Laparoscopic pancreatic resection was successfully performed in all patients. Operative time varied between 4 and 6 hours. Intraoperative bleeding was minimal. Due to a thick pancreas, the application of vascular endoscopic stapler was difficult in one patient. Two patients presented postoperative pancreatic leakage with spontaneous resolution. Resection of the pancreas can be safely performed via the laparoscopic approach with all the potential benefits to the patients of minimally invasive surgery.


The effects of child maltreatment on language development.
The separate effects of child neglect, abuse, and their interaction upon language development as measured by the Preschool Language Scale were examined in four groups of children (n = 79) in a quasi-experimental design. There were three groups of maltreated subjects, all drawn from a treatment center: an abused only group (n = 13), a neglected only group (n = 7), and an abused and neglected group (n = 31); a non-maltreated group (n = 28) was drawn from a day care center. Abuse, neglect, and their interaction were used to predict both auditory comprehension and verbal ability as separate criteria in two stepwise multiple regression equations, where the variance attributable to gender and mother's status on Aid to Dependent Children had been removed. Child neglect was found to predict significantly both auditory comprehension and verbal ability. Neither child abuse nor the interaction between abuse and neglect significantly predicted either dependent variable. These findings suggest that child neglect may be the critical type of maltreatment associated with language delay.


Diagnostic Accuracy of Dysphonia Classification of DSI and AVQI.
The Dysphonia Severity Index (DSI) and the Acoustic Voice Quality Index (AVQI) have been successfully investigated to quantify voice quality. The aim of the present study was to evaluate the diagnostic accuracy of both measurements in comparison with the dysphonia classification. In total, 264 subjects with vocally healthy voices (n = 105) and with various voice disorders (n = 159) were included in the study. To determine the dysphonia classification, all subjects underwent a videolaryngostroboscopy and, if necessary, a direct microlaryngoscopy plus a clinical examination to diagnose a voice disorder. Patients with a vocally healthy voice had no actual voice complaints, no history of chronic laryngeal diseases or voice disorders, no hearing problems and were determined as healthy voices by clinical voice specialists. To evaluate the diagnostic accuracy, receiver operating characteristic statistics and correct classification rate (CCR) were used. The diagnostic accuracy of DSI and AVQI showed strong sensitivity and specificity in the determination of dysphonia classification. A DSI threshold of 3.05 obtained a high sensitivity of 94.3% and specificity of 84.3%. An CCR of 88% was determined for DSI. Also, an AVQI threshold of 3.31 showed reasonable sensitivity of 71.7% and specificity of 88%. The CCR for AVQI was 79%. Although DSI and AVQI were developed to quantify voice quality, the present results showed that both measurements can evaluate the dysphonia classification as well. Particularly, the DSI might have higher potential in the evaluation of dysphonia classification. 2C Laryngoscope, 129:692-698, 2019.


Polarized super-resolution structural imaging inside amyloid fibrils using Thioflavine T.
Thioflavin T (ThT) is standardly used as a fluorescent marker to detect aggregation of amyloid fibrils by conventional fluorescence microscopy, including polarization resolved imaging that brings information on the orientational order of the fibrils. These techniques are however diffraction limited and cannot provide fine structural details at the fibrils scales of 10-100 nm, which lie beyond the diffraction limit. In this work, we evaluate the capacity of ThT to photoswitch when bound to insulin amyloids by adjusting the redox properties of its environment. We demonstrate that on-off duty cycles, intensity and photostability of the ThT fluorescence emission under adequate buffer conditions permit stochastic super-resolution imaging with a localization precision close to 20 nm. We show moreover that signal to noise conditions allow polarized orientational imaging of single ThT molecules, which reveals ultra-structure signatures related to protofilaments twisting within amyloid fibrils.


A Case of Ocular Kaposi's Sarcoma Successfully Treated with Highly Active Antiretroviral Therapy (HAART) Combined with Docetaxel.
BACKGROUND Ocular Kaposi's sarcoma (KS) involving the conjunctiva and ocular adnexa is uncommon and is usually treated with cryotherapy or surgical excision. We report a case of ocular KS successfully treated with HAART combined with 8 cycles of weekly docetaxel. CASE REPORT Our patient was a 24-year-old, treatment-naïve, HIV-positive (CD4 cell count 198 cells/mm3), homosexual man treated as having atypical hordeolum and subconjunctival hemorrhage, and later confirmed with pathology to have ocular KS with immunohistochemistry study showing KS with positive HHV8, CD34, CD31, and focal positive staining with Factor VIIIRA. HAART therapy was initiated combined with weekly docetaxel. With 2-month treatment of HAART and 8 cycles of weekly docetaxel, the KS of the bulbar conjunctiva and the eyelid partially resolved. CONCLUSIONS HAART combined with weekly docetaxel is an effective and well-tolerated option for ocular KS, which could be considered before cryotherapy or surgical excision.


Combination of Mineral Trioxide Aggregate and Platelet-rich Fibrin Promotes the Odontoblastic Differentiation and Mineralization of Human Dental Pulp Cells via BMP/Smad Signaling Pathway.
Recent reports have shown that the combined use of platelet-rich fibrin (PRF), an autologous fibrin matrix, and mineral trioxide aggregate (MTA) as root filling material is beneficial for the endodontic management of an open apex. However, the potential of the combination of MTA and PRF as an odontogenic inducer in human dental pulp cells (HDPCs) in vitro has not yet been studied. The purpose of this study was to evaluate the effect of the combination of MTA and PRF on odontoblastic maturation in HDPCs. HDPCs extracted from third molars were directly cultured with MTA and PRF extract (PRFe). Odontoblastic differentiation of HDPCs was evaluated by measuring the alkaline phosphatase (ALP) activity, and the expression of odontogenesis-related genes was detected using reverse-transcription polymerase chain reaction or Western blot. Mineralization formation was assessed by alizarin red staining. HDPCs treated with MTA and PRFe significantly up-regulated the expression of dentin sialoprotein and dentin matrix protein-1 and enhanced ALP activity and mineralization compared with those with MTA or PRFe treatment alone. In addition, the combination of MTA and PRFe induced the activation of bone morphogenic proteins (BMP)/Smad, whereas LDN193189, the bone morphogenic protein inhibitor, attenuated dentin sialophosphoprotein and dentin matrix protein-1 expression, ALP activity, and mineralization enhanced by MTA and PRFe treatment. This study shows that the combination of MTA and PRF has a synergistic effect on the stimulation of odontoblastic differentiation of HDPCs via the modulation of the BMP/Smad signaling pathway.


Synthesis, in vitro anti-human immunodeficiency virus structure-activity relationships and biological stability of 5'-O-myristoyl analogue derivatives of 3'-azido-2',3'-dideoxythymidine (AZT) as potential prodrugs.
5'-O-Myristoyl analogue derivatives of 3'-azido-2',3'-dideoxythymidine (AZT), designed as potential double-barrelled prodrugs to AZT and the myristic acid analogues, were synthesized. Their ability to protect CEM cells against human immunodeficiency virus (HIV)-induced cytopathogenicity was determined and structure-activity paradigms were developed. 3'-Azido-2',3'-dideoxy-5'-O-(4-oxatetradecanoyl)thymidine (EC50 = 1.4 nM) and 3'-azido-2',3'-deoxy-5'-O-(12-bromododecanoyl)thymidine (EC50 = 3.2 nM) were the most effective anti-HIV-1 agents, relative to AZT (EC50 = 10 nM). These myristoyl analogue derivatives were more lipophilic (calculated log P = 4.5-8.1 range) than the parent compound AZT (log P = 0.06), and a linear correlation between their log P and HPLC log retention times was observed. The ester cleavage half-lives (t1/2) for esters upon in vitro incubation with porcine liver esterase, rat plasma or rat brain homogenate was dependent on the steric bulk, and electronegative inductive effect of the alpha-substituent (H, Br, F), of the 5'-O-myristoyl analogue moiety. 3'-Azido-2',3'-dideoxy-5'-O-(11-(4-iodophenoxy) undecanoyl)-thymidine exhibited t1/2 values of 80.4, 3.7 and 150.0 min upon incubation with porcine liver esterase, rat plasma and rat brain homogenate, respectively.


Results from Phase I Extension Study Assessing Pexidartinib Treatment in Six Cohorts with Solid Tumors including TGCT, and Abnormal CSF1 Transcripts in TGCT.
To assess the response to pexidartinib treatment in six cohorts of adult patients with advanced, incurable solid tumors associated with colony-stimulating factor 1 receptor (CSF1R) and/or KIT proto-oncogene receptor tyrosine kinase activity. From this two-part phase I, multicenter study, pexidartinib, a small-molecule tyrosine kinase inhibitor that targets CSF1R, KIT, and FMS-like tyrosine kinase 3 (FLT3), was evaluated in six adult patient cohorts (part 2, extension) with advanced solid tumors associated with dysregulated CSF1R. Adverse events, pharmacokinetics, and tumor responses were assessed for all patients; patients with tenosynovial giant cell tumor (TGCT) were also evaluated for tumor volume score (TVS) and patient-reported outcomes (PRO). CSF1 transcripts and gene expression were explored in TGCT biopsies. Ninety-one patients were treated: TGCT patients (n = 39) had a median treatment duration of 511 days, while other solid tumor patients (n = 52) had a median treatment duration of 56 days. TGCT patients had response rates of 62% (RECIST 1.1) and 56% (TVS) for the full analysis set. PRO assessments for pain showed improvement in patient symptoms, and 76% (19/25) of TGCT tissue biopsy specimens showed evidence of abnormal CSF1 transcripts. Pexidartinib treatment of TGCT resulted in tumor regression and symptomatic benefit in most patients. Pexidartinib toxicity was manageable over the entire study. These results offer insight into outcome patterns in cancers whose biology suggests use of a CSF1R inhibitor. Pexidartinib results in tumor regression in TGCT patients, providing prolonged control with an acceptable safety profile.


Detecting, visualising, and quantifying mucins.
The extreme size, extensive glycosylation, and gel-forming nature of mucins make them a challenge to work with, and methodologies for the detection of mucins must take into consideration these features to ensure that one obtains both accurate and meaningful results. In understanding and appreciating the nature of mucins, this affords the researcher a valuable toolkit which can be used to full advantage in detecting, quantifying, and visualising mucins. The employment of a combinatorial approach to mucin detection, using antibody, chemical, and lectin detection methods, allows important information to be gleaned regarding the size, extent of glycosylation, specific mucin species, and distribution of mucins within a given sample. In this chapter, the researcher is guided through considerations into the structure of mucins and how this both affects the detection of mucins and can be used to full advantage. Techniques including ELISA, dot/slot blotting, and Western blotting, use of lectins and antibodies in mucin detection on membranes as well as immunohistochemistry and immunofluorescence on both tissues and cells grown on Transwell™ inserts are described. Notes along with each section advice the researcher on best practice and describe any associated limitations of a particular technique from which the researcher can further develop a particular protocol.


Studies on thymocyte subpopulations in guinea pigs. VI. Differentiation of precursor cells in vivo and in vitro.
The relation between six subpopulations of guinea pig thymocytes, separated on the basis of PNA binding and buoyant density, was studied. Incubation in vitro of either unseparated thymocytes or isolated large, low density cells, caused a shift towards smaller cells with high buoyant density. In order to determine whether these changes reflect a normal differentiation we pulsed labelled thymocytes with 3H-Thymidine and traced the labelling in the separate subpopulations after different intervals by autoradiography. Both in vivo and in vitro, the thymidine was initially incorporated mainly into PNA+ cells (85% of all labelled cells), particularly into PNA+ low-density cells (called PNA+Ia), which constitute 14% of all thymus cells (1). Correlation with autoradiographic sections of the thymus indicated that these cells were mainly located in the subcapsular and juxtamedullary cortex. At 24 h after labelling in vivo, more than 95% of the labelled cells were smaller, high density cells, both PNA+ and PNA-. At this time, labelled cells were present throughout the thymus with no preferential localization. Labelling and incubation in vitro were accompanied by similar changes after a 24-h time period when cells were cultured in the presence of serum. We conclude that the labelled PNA+Ia cells represent precursor cells, located in the subcapsular and juxtamedullary parts of the cortex. Within 24 h these cells differentiate into high density cells, some of which are PNA-. The corresponding results obtained in vitro indicate that these differentiation steps may also occur in the absence of a thymic microenvironment, and the study of histological sections indicate that these steps were not associated with any major relocalization within the thymus.


Differences between the effects of calcium antagonists in the pithed rat preparation.
The drugs classed as calcium antagonists display marked differences in vitro and it has been proposed that different subclasses of antagonists exist (Spedding M. Naunyn Schmiedebergs Arch Pharmacol 1982; 318:234-40). The effects of these drugs have now been compared in vivo using the pithed rat preparation. Whereas the hypotensive effects of verapamil (0.1-3 mumol/kg i.v.) and diltiazem (0.1-10 mumol/kg i.v.) were accompanied by prolongation of PR intervals and second-degree atrioventricular block with little bradycardia, fendiline (0.1-30 mumol/kg i.v.), cinnarizine (0.1-10 mumol/kg i.v.), and pimozide (0.1-10 mumol/kg i.v.) reduced heart rate at hypotensive doses but had less marked effects on AV conduction, thus implying different selectivities among calcium antagonists for the sinus or AV nodes. These differences were not secondary to effects on the autonomic nervous system. In contrast, nifedipine (0.01-1 mumol/kg i.v.) had no effect on the ECG in doses which markedly reduced blood pressure elevated by an infusion of angiotensin II and thus showed specificity for vascular smooth muscle. The marked differences between the effects of the calcium antagonists in vivo confirm the disparate nature of this group of drugs. The reasons for these differences are discussed.


Management of complications of forearm fractures.
Forearm fractures may be complicated by the disruption of the distal radioulnar, proximal radioulnar, or radiocapitellar joints. The key principles in treating this unique subset of fractures include early recognition and management of the injury and restoration and maintenance of the anatomic alignment. This articles addresses radial diaphyseal fractures with distal radioulnar joint disruption, proximal ulnar fractures with radiocapitellar disruption, and disruption of the forearm longitudinal axis and how to properly recognize and manage these forearm fracture-dislocations.


Accurate photodensitometric measurement of the lecithin: sphingomyelin ration after elimination of the acidic phospholipids from extracts of amniotic fluid specimens.
Treatment of human amniotic fluid lipids dissolved in a chloroform/methanol (9:1, v/v) mixture by batchwise addition of diethylaminoethyl cellulose in a dry state proved to be an easy and rapid procedure for the removal of the acidic phospholipids which may interfere in the photodensitometric evaluation of the lecithin: spingomyelin ratio on a thin-layer chromatogram. This method was used for the measurement of the lecithin: sphingomyelin ratio in a series of normal and abnormal pregnancies. Ratios higher than expected from gestational age were observed in stressed pregnancies. A significant elevation of the ratio was also observed under treatment by dexamethasone.


Material flow cost accounting (MFCA) to enhance environmental entrepreneurship in the meat sector: Challenges and opportunities.
The material flow cost accounting (MFCA) is one of the most broadly standardized tools accepted in environmental, social and economic research, which traces and quantifies material flows and stock in physical and economic units. Although its application has been recently developed in the field of resource and waste management, few academic articles have investigated its value towards food waste management, which represents a topical concern on a global scale. The present research applies the MFCA to investigate the material, energetic and economic costs associated with the Italian beef, pork and poultry production, exploring related challenges and opportunities towards the enhancement of the environmental entrepreneurship in the meat sector. The present countryside analysis is based on literature and empirical data collected during the Covid-19 pandemic. It highlights the need to improve knowledge on food waste issue under the economic perspective and its dual impact: when it is generated, in terms of income losses due to by-products and finished products sales failure, and when it is disposed, in terms of disposal costs sustained by farms, processing plants and distribution and sales centers. It is estimated that more than 0.45-0.50 Mt of fresh meat is wasted along the entire Italian agri-food chain, equal to more than 242-268 million euros, to which additional energy and water losses should be added (435-481 million euros). MFCA results are useful for business decisions, highlighting quantities, qualities and costs otherwise not considered in common financial reports.


A Case of Breakthrough COVID-19 during Hydroxychloroquine Maintenance.
There have been controversies on the prophylactic effect of hydroxychloroquine against severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2). We describe a patient, 60-year old Korean woman, with coronavirus disease 2019 (COVID-19) who had been taking hydroxychloroquine for 6 months. Her serum and saliva concentrations of hydroxychloroquine were 280 μg/L and 4,890 μg/L, respectively. The present case raises concerns on hydroxychloroquine's role as a prophylactic agent for COVID-19.


[Temperature measurements and course in a kidney preservation unit].
It is reported on two models of organ containers in which the course of the temperature was measured and registered on different conditions. The principle of construction used allowed the resignation of a special isolation. The results show that behaviour and course of the temperature in the organ containers meet all requirements.


An unusual case of trimethylaminuria.
A case is reported of trimethylaminuria that first developed in adulthood without any apparent cause. The patient developed a characteristic fish odour of his sweat, urine and to his breath after the consumption of choline-rich foods. Elevated levels of trimethylamine were present in the urine after dietary tests and identified by means of gas chromatography.


Validity of clinically significant change classifications yielded by Jacobson-Truax and Hageman-Arrindell methods.
Reporting of the clinical significance of observed changes is recommended when publishing mental health treatment outcome studies and is increasingly used in routine outcomes monitoring systems. Since recovery rates vary with the method chosen, we investigated the validity of classifications of clinically significant change when the Jacobson-Truax method and the Hageman-Arrindell method were used. Of 718 inpatients who completed the Depression Anxiety Stress Scales (DASS-21) and Quality of Life Enjoyment and Satisfaction Questionnaire at admission and discharge to a psychiatric clinic, 355 were invited (and 119 agreed) to complete the questionnaires and the Recovery Assessment Scale six weeks post discharge. Both the JT and HA methods showed comparably good validity when referenced against the other indices. Clinically significant change on the DASS-21 was related to a greater consumer-based sense of recovery, greater perceived quality of life, and fewer readmissions to hospital within 28 days of discharge. Since there was found to be no advantage to using one method over another when recovery is of interest, the simpler JT method is recommended for routine usage.


Prospects for controlling future pandemics of influenza.
Pandemic influenza remains one of the most serious threats to global public health and continued global vigilance to monitor emerging threats is crucial. Of the weapons available to control a pandemic, vaccination is potentially the most powerful, but there are currently serious limitations to timely availability of vaccine supply in an emergency. Many novel influenza vaccines are in development, some of which have the potential to deliver the massive quantities of vaccine that would be required in a pandemic in a short period of time. However, for the foreseeable future, it is likely that the principal vaccine that will be deployed in a pandemic will be an inactivated egg-derived vaccine of the kind that has been available for several decades. This review will focus on the practical hurdles that need to be surmounted to deliver large amounts of safe and effective pandemic vaccine to the general public. There needs to be a continued focus on improvement to the vaccine response system that will require close collaboration between influenza and vaccine experts, manufacturers, regulators and public health authorities around the world.