The ClC-3 chloride channel promotes acidification of lysosomes in CHO-K1 and Huh-7 cells.
ClC-3 is a voltage-gated Cl- channel that is highly conserved and widely expressed, although its function, localization, and properties remain a matter of considerable debate. In this study, we have shown that heterologous expression of ClC-3 in either Chinese hamster ovary (CHO-K1) or human hepatoma (Huh-7) cells results in the formation of large, acidic vesicular structures within cells. Vesicle formation is prevented by bafilomycin, an inhibitor of the vacuolar ATPase, and is not induced by an E224A mutant of ClC-3 with altered channel activity. This demonstrates that vesicle formation requires both proton pumping and Cl- channel activity. Manipulation of the intracellular Cl- concentration demonstrated that the ClC-3-associated vesicles shrink and swell consistent with a highly Cl--permeable membrane. The ClC-3 vesicles were identified as lysosomes based on their colocalization with the lysosome-associated proteins lamp-1, lamp-2, and cathepsin D and on their failure to colocalize with fluorescently labeled endosomes. We conclude that ClC-3 is an intracellular channel that conducts Cl- when it is present in intracellular vesicles. Its overexpression results in its appearance in enlarged lysosome-like structures where it contributes to acidification by charge neutralization.


[Problems of male contraception. 2. Male hormonal contraception, post-testicular points of attack, immunization and enzyme inhibition].
In principle control of male fertility is possible by mechanical, surgical and pharmacological methods. However, up to now only the following mechanical and surgical procedures are practicable: coitus interruptus, coitus condomatus and vasectomy. Since the safety of the latter is comparable to the pill, vasectomy is a major alternative to the contraceptive methods used by the female partner. Practical aspects of vasectomy and possible complications are reviewed. Finally, pharmacological approaches and problems involved in the control of male fertility are discussed with special reference to antispermatogenic substances, steroid hormones, immunization and enzyme inhibitors. The concept of antienzymatic contraception is based on the inhibition of penetration of enzymes (acrosin) localized within the acrosomal cap. Unfortunately, penetration of most of the acrosin inhibitors into the acrosome to inactivate acrosin is prevented by the high selectivity of the acrosomal membranes. Thus, pharmacological control of male fertility is still far from being practicable.


[Continuous EEG monitoring for aneurysmal subarachnoid hemorrhage].
Subarachnoid hemorrhage (SAH) still carries a high morbidity and mortality, despite improvement in surgical and medical management. Seizures and delayed cerebral ischemia (DCI) secondary to vasospasm or cortical spreading depression are frequent after SAH. Continuous EEG allows early detection of non-convulsive seizures or delayed cerebral ischemia and may become a promissing tool in the monitoring of SAH patients. However, its use in clinical practice is still limited because many resources are required for recording and analyzing continuous EEG. Moreover, we require more data to confirm the relationship between aggressive treatment of non-convulsive seizure or delayed cerebral ischemia triggered by continuous EEG and outcome.


Surgeons' Interactions With and Attitudes Toward E-Patients: Questionnaire Study in Germany and Oman.
Doctors' interactions with and attitudes toward e-patients have an overall impact on health care delivery. This study aimed to gauge surgeons' interactions with e-patients, their attitudes toward those e-patient activities, the possible impact on the delivery of health care, and the reasons behind those activities and attitudes. We created a paper-based and electronic survey form based on pertinent variables identified in the literature, and from March 2018 to July 2018 we surveyed 49 surgeons in Germany and 59 surgeons in Oman, asking them about their interactions with and attitudes toward e-patients. Data were stored in Microsoft Excel and SPSS, and descriptive statistics, Pearson correlations, and chi-square tests were performed on the data. Of our sample, 71% (35/49) of the German surgeons and 56% (33/59) of the Omani surgeons communicated electronically with their patients. Although the German surgeons spent a greater percentage of Internet usage time on work-related activities (χ218=32.5; P=.02) than the Omani surgeons, there were many similarities in their activities. An outstanding difference was that the German surgeons used email with their patients more than the Omani surgeons (χ21=9.0; P=.003), and the Omani surgeons used social media, specifically WhatsApp, more than the German surgeons (χ21=18.6; P<.001). Overall, the surgeons were equally positive about the most common e-patient activities such as bringing material from the internet to the consultation (mean 4.11, SD 1.6), although the German surgeons (mean 3.43, SD 1.9) were more concerned (P=.001) than the Omani surgeons (mean 2.32, SD 1.3) about the potential loss of control and time consumption (German: mean 5.10, SD 1.4 and Omani: mean 3.92, SD 1.6; P<.001). The interactions show a high degree of engagement with e-patients. The differences between the German and the Omani surgeons in the preferred methods of communication are possibly closely linked to cultural differences and recent historical events. These differences may, moreover, indicate e-patients' desired method of electronic communication to include social media. The low impact of surgeons' attitudes on the activities may also result from a normalization of many e-patient activities, irrespective of the doctors' attitudes and influences.


[Applicability of daytime downward longwave radiation parameterized models in Changbai Mountains, Northeast China].
A total of eight clear-sky and eight cloudy-sky parameterized models for estimating daytime downward longwave radiation were evaluated by using the meteorological data measured in the Changbai Mountains region, Northeast China. The results indicated that the Satterlund model performed better in estimating clear-sky downward longwave radiation, and the bias (BIAS) and root mean square error (RMSE) were -23.34 and 28.55 W· m(-2), respectively. Although the coefficients were not significantly changed, the performance of Satterlund model was significantly improved after the locally calibrated, and the BIAS and RMSE decreased to -6.33 and 18.08 W · m(-2), respectively. Jacobs model was found to be best for modeling cloudy-sky downward long-wave radiation and the BIAS and RMSE were 0.38 and 29.29 W · m(-2), respectively. Sensitivity analysis showed that the vapor pressure was the most sensitive variable to the atmospheric emissivity and the temperature was not sensitive to it. The predicted results of the optimal model (clear- and cloudy-sky) were consistent with the measured data, while the performance of these models was affected by a sudden change of cloudy cover.


The Greatest Right of Them All: The Debate on the Right to Petition in the Netherlands from the Dutch Republic to the Kingdom (c. 1750-1830).
Between 1750 and 1830 the Dutch state developed from an oligarchic republic into an enlightened autocratic monarchy via a brief experiment with representative democracy. During this period, there was an ongoing debate about the right to petition. Political actors and opinion-makers addressed the questions to what and to whom this right extended, and what it meant to have such a right. While theorists of the different types of government had sharply contrasting views on the place of the people in the political process, ideas about petitioning, which throughout the period under discussion remained the principal instrument for popular involvement in politics, stayed remarkably stable. Through an investigation of the debate on the right to petition in the crucial transitional phase from the Dutch Republic to the Kingdom of the Netherlands, this article contributes to bridging the divide between petition research of the early modern period and that of the modern era.


Mouse glycine N-methyltransferase is sexually dimorphic and regulated by growth hormone.
The secretion pattern of putuitary growth hormone (GH) in rodents is sexually dimorphic and regulates the expression of some hepatic genes. We report cloning of mouse glycine N-methyltransferase (GNMT) cDNA. The mouse GNMT mRNA is expressed at much lower levels in male than in female livers. Hypophysectomy in male mice resulted in as a high level of the expression as in female mice. Treatment with GH of hypophysectomized male mice twice a day for 5 days decreased the expression. These results indicate that the sex difference of the mouse GNMT expression is regulated by GH secretion patterns.


Protocol for a systematic review on the experience of informal caregivers for people with a moderate to advanced dementia within a domestic home setting.
The knowledge about the experience of informal caregivers who provide care to people with moderate to advanced dementia in a domestic home setting is limited. A consequence of long hours of caregiving in addition to dealing with normal challenges of daily living is their experience of a poor quality of life. Some of their experiences may be described in terms of a feeling of powerlessness to make changes during care provision. This feeling may also suggest an experience of moral distress. The aim of this systematic review is to synthesise qualitative evidence relating to these experiences. This review adopts a narrative synthesis approach. A search will be conducted for studies written in the English language in the bibliographic databases MEDLINE Complete, CINAHL, EMBASE, PsycINFO, Web of Science and Academic Search Complete covering periods from 1984 to present. Included studies will be qualitative or mixed-methods designs. The search terms will be related to dementia and caregivers, and the process will be focused on dementia at the moderate to the advanced stages within the domestic home setting. Reference lists of included papers will also be searched for additional relevant citations. Search terms and strategies will be checked by two independent reviewers. The identification of abstracts and full texts of studies will be done by the author, while the quality and the risk of bias will also be checked by the two independent reviewers. Psychological distress is cited as an experience reported within informal caregiving. For the caregiver, it is associated with a negative impact on general health. To date, no synthesis exists on the specific experience of informal caregiving for people with moderate to advanced dementia within the domestic home setting. This review considers that variation of accounts contributes to how the informal caregivers' general experience is explored in future research. This may enable gaps in current knowledge to be highlighted within the wider context of caregiving in the domestic home setting. This review is registered with PROSPERO ( CRD42020183649 ).


Reference Equations for Spirometry in the Canadian Population.
Spirometry plays a major role in the diagnosis and assessment of severity of lung disease. Determining which lung function values are normal and which are below the lower limit of normal depends on reference equations derived from an appropriate population. The purpose of this study was to derive spirometric reference equations for the Canadian population. The Canadian Health Measures Survey consisted of a respiratory questionnaire, urinary cotinine measurements, and spirometry performed in the sitting position with rigorous quality control standards. Of the 16,606 respondents between 6 and 79 years of age, 11,145 were eliminated for positive responses to the respiratory questionnaire, tobacco exposure, or inability to provide high-quality spirograms. Of the remaining 5,461, roughly half were less than 18 years of age. Quantile regression was used to derive predictive (median) and lower limit of normal equations for males and females for FEV1, FVC, and FEV1/FVC ratio for those with ages greater and less than 18 years. The resulting equations were compared with those from the Global Lung Initiative (GLI) and National Health and Nutrition Examination Survey (NHANES) III by using an ideal subject on the 50th percentile for height and between the ages of 6 and 79 years; the comparison showed minor and inconsistent discrepancies among the predictive equations. A plot of residuals (predicted minus measured value for each subject) suggested a marginally better fit compared with the GLI and NHANES III equations, although differences among the equations were small and unlikely to have any clinical significance. This study provides spirometric reference equations for the Canadian population that were measured under the recommended clinical conditions and with rigorous quality control.


Identification of cell membrane proteins that bind visna virus.
Visna virus infects cells of ovine origin by attaching to a cell surface receptor via its envelope glycoprotein. The identity of the visna virus receptor is not known. To identify the molecule responsible for binding the virus to target cells, virus overlay protein blot assays were used to examine the molecular weights of cell surface molecules which bind purified virus. Molecules on the surface of goat synovial membrane (GSM) cells and sheep choroid plexus (SCP) cells of approximately 15, 30, and 50 kDa bound to visna virus. The binding of visna virus to these proteins was reduced by preincubating virus with neutralizing antibodies. 125I-labeled cell membrane preparations of GSM and SCP cells were used to affinity purify these virus-binding proteins. These proteins were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and had molecular masses of 15, 30, and 50 kDa. Antibodies to the 50-kDa protein bound to the surface of both live SCP and GSM cells in immunofluorescence assays. In addition, antibodies to the 50-kDa protein blocked the binding of [35S]methionine-labeled visna virus to SCP cells in culture. Antibodies raised against the 15- and 30-kDa proteins did not block virus binding to cells. The blocking activity of antibody of the 50-kDa protein provided data that this protein is the molecule which visna virus recognizes and binds to on the surface of target cells.


Detection of an earlier tubulopathy in diabetic nephropathy among children with normoalbuminuria.
Diabetic nephropathy is a major cause of morbidity and mortality among young adults with type 1 diabetes mellitus (DM). Albuminuria, the gold standard for early diagnosis, cannot always detect early diabetic nephropathy. We aimed at evaluating the level of urine neutrophil gelatinase-associated lipocalin (NGAL) as a marker of tubulointerstitial damage in children and adolescents with type 1 DM in relation to the level of albuminuria and other parameters. Fifty children with type 1 DM for more than 5 years were included in this study (mean age, 13.8 ± 4.0 years), and 18 healthy children served as controls. Patients with overt albuminuria (> 300 mg/g creatinine) or inflammatory states were excluded. Urine NGAL, microalbuminuria, and urine albumin-creatinine ratio were measured in patients and controls as well as other parameters. Urine NGAL was significantly higher in microalbuminuric in comparison with normoalbuminuric patients and controls, and correlated positively with urine albumin-creatinine ratio. A positive urine NGAL was observed in 12 of 38 normoalbuminuric patients (31.6%) compared to 9 of 12 microalbuminuric patients (75%). A positive correlation was reported between urine NGAL and both Hemoglobin A1c and duration of DM, but not with estimated glomerular filtration rate or hypertension. Diabetic children, even some normoalbuminurics, showed increased urine NGAL. This finding may support the hypothesis of a "tubular phase" of diabetic disease preceding overt diabetic nephropathy, and hence, the use of urine NGAL measurement for early evaluation of renal involvement.


Central neuropeptide Y infusion and melanocortin 4 receptor antagonism inhibit thyrotropic function by divergent pathways.
Weight loss inhibits thyrotropic function and reduces metabolic rate, thereby contributing to weight regain. Under negative energy balance there is an increase in the hypothalamic expression of both neuropeptide Y (NPY) and agouti related peptide (AgRP), the endogenous antagonist of melanocortin 4 (MC4) receptors. Both NPY and MC4 receptor antagonism reduce thyrotropic function centrally, but it is not known whether these pathways operate by similar or distinct mechanisms. We compared the time-course of effects of acute or chronic intracerebroventricular (ICV) administration of NPY (1.2 nmol acute bolus, or 3.5 nmol/day for 6 days) or the MC4 receptor antagonist HS014 (1.5 nmol bolus, or 4.8 nmol/day) on plasma concentrations of thyroid stimulating hormone (TSH) or free thyroxine (T4) in male rats pair-fed with vehicle-infused controls. These doses equipotently induced hyperphagia in acute studies, reduced latency to feed, and increased white adipose tissue mass after 6 days of infusion. Acute central NPY but not HS014 administration significantly reduced plasma TSH concentrations within 30-60 min and plasma free T4 levels within 90-120 min. These inhibitory effects were sustained for up to 5-6 days of continuous NPY infusion. HS014 induced a transient decrease in plasma free T4 levels that was observed only after 1-2 days of continuous ICV infusion. While both NPY and HS014 significantly increased corticosteronemia within an hour after ICV injection, the effect of NPY was significantly more pronounced and was sustained for up to 4 days of administration. Both NPY and HS014 significantly decreased the brown adipose tissue protein levels of uncoupling protein-3. We conclude that central NPY and MC4 antagonism decrease thyrotropic function via partially distinct mechanisms with different time courses, possibly involving glucocorticoid effects of NPY. MC4 receptor antagonism increases adiposity via pathways independent of increased food intake or changes in circulating concentrations of TSH, free T4 or corticosterone.


Acute L-DOPA administration reverses changes in firing pattern and low frequency oscillatory activity in the entopeduncular nucleus from long term L-DOPA treated 6-OHDA-lesioned rats.
The pathophysiology of Parkinson's disease (PD) and L-DOPA-induced dyskinesia (LID) is associated with aberrant neuronal activity and abnormal high levels of oscillatory activity and synchronization in several basal ganglia nuclei and the cortex. Previously, we have shown that the firing activity of neurons in the substantia nigra pars reticulata (SNr) is relevant in dyskinesia and may be driven by subthalamic nucleus (STN) hyperactivity. Conversely, low frequency oscillatory activity and synchronization in these structures seem to be more important in PD because they are not influenced by prolonged L-DOPA administration. The aim of the present study was to assess (through single-unit extracellular recording techniques under urethane anaesthesia) the neuronal activity of the entopeduncular nucleus (EPN) and its relationship with LID and STN hyperactivity, together with the oscillatory activity and synchronization between these nuclei and the cerebral cortex in 6-OHDA-lesioned rats that received long term L-DOPA treatment (or not). Twenty-four hours after the last L-DOPA injection the firing activity of EPN neurons in long term L-DOPA treated 6-OHDA-lesioned rats was more irregular and bursting compared to sham rats, being those alterations partially reversed by the acute challenge of L-DOPA. No correlation between EPN neurons firing activity and abnormal involuntary movements score was found. However, there was a significant correlation between the firing activity parameters of EPN and STN neurons recorded from long term L-DOPA treated 6-OHDA-lesioned rats. Low frequency oscillatory activity and synchronization both within the EPN and with the cerebral cortex were enhanced in 6-OHDA-lesioned animals. These changes were reversed by the acute L-DOPA challenge only in long term L-DOPA treated 6-OHDA-lesioned rats. Altogether, these results obtained from long term L-DOPA treated 6-OHDA-lesioned rats suggest (1) a likely relationship between STN and EPN firing patterns and spiking phases induced by changes after prolonged L-DOPA administration and (2) that the effect of L-DOPA on the firing pattern, low frequency oscillatory activity and synchronization in the EPN may have a relevant role in LID.


Longitudinal changes in cognitive and physical function and health-related quality of life in older adults with acute myeloid leukemia.
Many older adults with acute myeloid leukemia (AML) do not receive chemotherapy because of physicians' and patients' concern for toxicities and functional decline. This highlights the critical and urgent need to generate knowledge of functional changes following new treatments. As a part of a pragmatic single-center trial, 59 older adults ≥60 years with AML completed geriatric assessment and health-related quality of life measures before treatment and at one month and three months after chemotherapy initiation. Changes in scores of various geriatric assessment measures were computed by subtracting the baseline score from the one-month and three-month scores for each patient. Established cut-offs were used to determine a clinically meaningful change (improvement or worsening). This study provides results of descriptive exploratory analyses. Patients experienced significant comorbidity burden and a high prevalence of functional impairments before treatment, with 56% of patients having ≥2 comorbid conditions, 69% having abnormal cognitive function (using Montreal Cognitive Assessment), 69% having impaired objective physical function (using Short Physical Performance Battery), and 64% having a positive depression screen (Patient Health Questionnaire-9). Patients (n = 53) received treatment with predominantly low-intensity chemotherapy; six patients received intensive chemotherapy. Among those who completed some or all of the three-month evaluation (N = 43), from baseline before treatment to three months later, cognitive function improved (38.7%) or remained stable (38.7%), objective physical function improved (51.6%) or remained stable (22.6%), and depression scores improved (9.4%) or remained stable (53.1%). Global health status score and role functioning moderately improved by a score of >16. An exploratory analysis of our phase 2 trial demonstrated improvement or stabilization of cognitive and physical function and depression score at three months in a high proportion of older survivors of AML, despite a high prevalence of frailty and significant comorbidity burden at baseline. These results demonstrate success of treatment in improving cognitive and physical function and depression score, and, if confirmed in larger studies, should encourage oncologists to offer chemotherapy to older adults with AML. The study is registered in the ClinicalTrials.gov ID: NCT03226418.


A new living cell-based assay system for monitoring genome-length hepatitis C virus RNA replication.
We previously developed a cell-based luciferase reporter assay system for monitoring genome-length hepatitis C virus (HCV) RNA replication (OR6 assay system). Here, we aimed to develop a new living cell-based reporter assay system using enhanced green fluorescent protein (EGFP). Genome-length HCV RNAs encoding EGFP were introduced into a subline of HuH-7 cells and G418 selection was performed. One cloned cell line, OGF7, was successfully selected from among the several G418-resistant cell lines obtained, and the robust expression of HCV RNA and proteins in OGF7 cells was confirmed. The fluorescent intensity of OGF7 cells was decreased by interferon-alpha treatment in a dose-dependent manner, and it correlated well with the HCV RNA concentration. We demonstrated that the interferon-alpha sensitivity in the OGF7 assay system measuring the fluorescent intensity was equivalent to that of the OR6 assay system, and that the OGF7 assay system was useful for quantitative evaluation of anti-HCV reagents. The OGF7 assay system is expected to be the most time-saving and inexpensive assay system for high-throughput screening of anti-HCV reagents.


Isolation of Actinobacillus pleuropneumoniae serotype 2 by immunomagnetic separation.
In Denmark porcine pleuropneumonia is most frequently caused by Actinobacillus pleuropneumoniae serotype 2 (60%). Isolation of A. pleuropneumoniae from nasal cavities or tonsils from carrier animals is complicated due to the mixed bacterial flora present. An immunomagnetic separation technique (IMS) using immunomagnetic beads (Dynabeads((R))) was developed for isolation of A. pleuropneumoniae serotype 2 from pure cultures and from heterogeneous suspensions. Different coating and washing procedures were evaluated in pure and mixed cultures using polyclonal (PAb) and monoclonal antibodies. The highest reisolation yield was achieved when the beads were coated with 1.5 microg PAb IgG/10(7) beads. After washing the beads for four times 9-24% of the bacteria could be reisolated depending on the amount of IgG attached to the beads and the number of beads used. The recovery was increased to 19-61% when only two washing steps were performed. The IMS was further evaluated using dilutions of A. pleuropneumoniae with added Pasteurella multocida (10(9) CFU/ml). After two washing steps 15% of the A. pleuropneumoniae cells and no P. multocida was reisolated. A detection limit of 10 CFU/ml was found in this heterogeneous suspension. No significant difference was observed when comparing the recovery of A. pleuropneumoniae from pure culture, from mixed cultures and from artificially inoculated tonsils. From 12 pigs inoculated with an aerosol of A. pleuropneumoniae serotype 2 the bacterium could not be detected from the nasal cavity or tonsils by cultivation or PCR 6 weeks later. By using IMS A. pleuropneumoniae serotype 2 could be reisolated from the tonsils of three pigs. The IMS method represents a valuable tool for isolation of A. pleuropneumoniae from tissue samples.


Dual angiopoietin-2 and VEGFA inhibition elicits antitumor immunity that is enhanced by PD-1 checkpoint blockade.
Pathological angiogenesis is a hallmark of cancer and a therapeutic target. Vascular endothelial growth factor A (VEGFA) and angiopoietin-2 (ANGPT2; also known as ANG2) are proangiogenic cytokines that sustain tumor angiogenesis and limit antitumor immunity. We show that combined ANGPT2 and VEGFA blockade by a bispecific antibody (A2V) provided superior therapeutic benefits, as compared to the single agents, in both genetically engineered and transplant tumor models, including metastatic breast cancer (MMTV-PyMT), pancreatic neuroendocrine tumor (RIP1-Tag2), and melanoma. Mechanistically, A2V promoted vascular regression, tumor necrosis, and antigen presentation by intratumoral phagocytes. A2V also normalized the remaining blood vessels and facilitated the extravasation and perivascular accumulation of activated, interferon-γ (IFNγ)-expressing CD8+ cytotoxic T lymphocytes (CTLs). Whereas the antitumoral activity of A2V was, at least partly, CTL-dependent, perivascular T cells concurrently up-regulated the expression of the immune checkpoint ligand programmed cell death ligand 1 (PD-L1) in tumor endothelial cells. IFNγ neutralization blunted this adaptive response, and PD-1 blockade improved tumor control by A2V in different cancer models. These findings position immune cells as key effectors of antiangiogenic therapy and support the rationale for cotargeting angiogenesis and immune checkpoints in cancer therapy.


Computing the stochastic dynamics of phosphorylation networks.
Cells of all organisms share the ability to respond to various extracellular signals. Depending on the cell type and the organism, these signals may include hormones secreted by other cells or changes in nutrient concentrations. The signals are processed by an intricate network of protein-protein interactions, including phosphorylation and de-phosphorylation events. As some signaling proteins are only present in low concentrations, random fluctuations may affect the dynamics of the network. The mathematical modeling of networks with significant random fluctuations requires the use of stochastic methods. The stochastic dynamics of a chemical reaction system are described by the Chemical Master Equation. Often the numerical evaluation of this equation is problematic. The first problem is that many systems have an infinite number of possible states; leaving simulations of individual trajectories as the only alternative. To circumvent this problem, we focus on a class of systems that have a finite state space. More specifically, we focus on networks of phosphorylation cycles without taking into account the synthesis and degradation of proteins. The second problem is that memory requirements cause a practical limit to the size of systems that can be evaluated. In this paper, we discuss how these limitations can be overcome using parallel computation and methods dealing efficiently with the available memory. These methods were implemented in a parallel C++ program. We discuss two networks for which the stochastic dynamics were evaluated using this program: a single phosphorylation cycle and an oscillating MAP-kinase cascade.


[Adolescents with chronic pain: practical assessment and management].
More than 20% of adolescents in the general population suffer of chronic pain mainly headaches, abdominal or musculoskeletal pain. Often, these complaints are self limited without impact on adolescent development but the cause of emergency consultations, unnecessary costs or inappropriate prescriptions. For a small number of adolescents, chronic pain can express psychological suffering or impact on growth and physical, cognitive or social development. Continuity of care and collaboration among professionals is central. Primary care phyisicians play a crucial role (private practice, adolescent clinic...). Efforts should be made to increase access to modern approaches of chronic pain taking into account family, development as well as aspects related to pain in itself.


The effect of military motion-assisted memory desensitization and reprocessing treatment on the symptoms of combat-related post traumatic stress disorder: first preliminary results.
Although the symptoms of Post-Traumatic Stress Disorder (PTSD) in the general and military population seem very similar, combat-related PTSD (cr-PTSD) is typically thought to be more severe due to the repeated and prolonged exposure of traumatic events. Therapeutic adherence is reported a problem in military populations compromising treatment efficacy. Therefore, a new potential supplementary treatment is specially designed for patients with cr-PTSD. This intervention is called Military Motion Memory Desensitization and Reprocessing (3MDR). The treatment incorporates key elements of successful treatments as Virtual Reality Exposure (VRE) and Eye Movement Desensitization Reprocessing (EMDR) and adds motion to the condition. We aimed at designing a treatment procedure that preserved dual task processing principle, yet introduced new engagement by performing the desensitization during motion by to walking on a treadmill. Moreover, we aimed at exposure to real high-affect pictures of deployment setting. Subjects walk a repetitive cycle while walking and viewing high affect pictures of deployment scenes. Dual task processing was maintained by an oscillating ball. Aspects of presence are adhered to, to maximize possible positive outcome. Two veterans with chronic PTSD, received four weekly sessions of 3MDR therapy. The indicator of effectiveness was difference in CAPS (Clinical Administrated PTSD Scale)-score. The treatment was designed on the Computer Assisted Rehabilitation Environment (CAREN) facility. The 3MDR treatment did further decrease PTSD symptoms. Patients were highly satisfied about the treatment and had no attention to drop out. The results of the two cases suggest that the 3MDR treatment is a successful, more additional treatment that goes further into the patients affect where other treatment may stagnate. The presence was highly appreciated. Further research with more patients needs to be performed to obtain more reliable results.


[Familial myasthenia gravis: report of 2 brothers].
The familial form of myasthenia gravis is a relatively rare condition, occurring in about 3.4 per cent of myasthenic patients. Two familial cases with ocular myasthenia gravis are reported. They had a third brother who died probably with the same disease. Their parents are cousins. The authors made a brief approach of genetic, clinical, statistical and therapeutic aspects of the disease.


Measurement of magnetic excitations in the two-dimensional antiferromagnetic Sr₂CuO₂Cl₂ insulator using resonant x-ray scattering: evidence for extended interactions.
We measured the momentum dependence of magnetic excitations in the model spin-1/2 2D antiferromagnetic insulator Sr2CuO2Cl2 (SCOC). We identify a single-spin-wave feature and a multimagnon continuum, with different polarization dependences. The spin waves display a large (70 meV) dispersion between the zone-boundary points (π, 0) and (π/2, π/2). Employing an extended t-t'-t''-U one-band Hubbard model, we find significant electronic hopping beyond nearest-neighbor Cu ions, indicative of extended magnetic interactions. The spectral line shape at (π, 0) indicates sizable quantum effects in SCOC and probably more generally in the cuprates.


DNA methylation modulated genetic variant effect on gene transcriptional regulation.
Expression quantitative trait locus (eQTL) analysis has emerged as an important tool in elucidating the link between genetic variants and gene expression, thereby bridging the gap between risk SNPs and associated diseases. We recently identified and validated a specific case where the methylation of a CpG site influences the relationship between the genetic variant and gene expression. Here, to systematically evaluate this regulatory mechanism, we develop an extended eQTL mapping method, termed DNA methylation modulated eQTL (memo-eQTL). Applying this memo-eQTL mapping method to 128 normal prostate samples enables identification of 1063 memo-eQTLs, the majority of which are not recognized as conventional eQTLs in the same cohort. We observe that the methylation of the memo-eQTL CpG sites can either enhance or insulate the interaction between SNP and gene expression by altering CTCF-based chromatin 3D structure. This study demonstrates the prevalence of memo-eQTLs paving the way to identify novel causal genes for traits or diseases associated with genetic variations.


Molecular Profiling of Shiga Toxin-Producing Escherichia coli and Enteropathogenic E. coli Strains Isolated from French Coastal Environments.
Shiga toxin-producing Escherichia coli (STEC) and enteropathogenic E. coli (EPEC) strains may be responsible for food-borne infections in humans. Twenty-eight STEC and 75 EPEC strains previously isolated from French shellfish-harvesting areas and their watersheds and belonging to 68 distinguishable serotypes were characterized in this study. High-throughput real-time PCR was used to search for the presence of 75 E. coli virulence-associated gene targets, and genes encoding Shiga toxin (stx) and intimin (eae) were subtyped using PCR tests and DNA sequencing, respectively. The results showed a high level of diversity between strains, with 17 unique virulence gene profiles for STEC and 56 for EPEC. Seven STEC and 15 EPEC strains were found to display a large number or a particular combination of genetic markers of virulence and the presence of stx and/or eae variants, suggesting their potential pathogenicity for humans. Among these, an O26:H11 stx1a eae-β1 strain was associated with a large number of virulence-associated genes (n = 47), including genes carried on the locus of enterocyte effacement (LEE) or other pathogenicity islands, such as OI-122, OI-71, OI-43/48, OI-50, OI-57, and the high-pathogenicity island (HPI). One O91:H21 STEC strain containing 4 stx variants (stx1a, stx2a, stx2c, and stx2d) was found to possess genes associated with pathogenicity islands OI-122, OI-43/48, and OI-15. Among EPEC strains harboring a large number of virulence genes (n, 34 to 50), eight belonged to serotype O26:H11, O103:H2, O103:H25, O145:H28, O157:H7, or O153:H2. The species E. coli includes a wide variety of strains, some of which may be responsible for severe infections. This study, a molecular risk assessment study of E. coli strains isolated from the coastal environment, was conducted to evaluate the potential risk for shellfish consumers. This report describes the characterization of virulence gene profiles and stx/eae polymorphisms of E. coli isolates and clearly highlights the finding that the majority of strains isolated from coastal environment are potentially weakly pathogenic, while some are likely to be more pathogenic.


In vitro studies on guar gum based formulation for the colon targeted delivery of Sennosides.
The objective of the present study is to develop colon targeted drug delivery systems for sennosides using guar gum as a carrier. Matrix tablets containing various proportions of guar gum were prepared by wet granulation technique using starch paste as a binder. The tablets were evaluated for content uniformity and in vitro drug release study as per BP method. T(50) % value from the dissolution studies was taken for selecting the best formulation. Guar gum matrix tablets released 4-18% sennosides in the physiological environment of gastrointestinal tract depending on the proportion of the guar gum used in the formulation. The matrix tablets containing 50% of guar gum were found to be suitable for targeting of sennosides for local action in the colon. Compared to tablets having 30% and 40% of guar gum, those with 50% guar gum gave better T(50)% (11.7 h) le and fewer amounts (5-8%) of drug release in upper GIT. These tablets with 50% guar gum released 43% and 96% sennosides with and without rat caecal fluids. This suggests the susceptibility of matrix to the colonic micro flora. The similarity factor (f2 value) for drug release with and without rat caecal fluids was found to be less than 30. When hydroxy propyl methylcellulose phthalate (10%) was used as a coat material on the matrix tablets, the initial loss of 5-8% sennosides in stomach could be completely averted. These tablets showed no change in physical appearance, content and dissolution profile upon storage at 45 degrees C / 75% relative humidity for 3 months. The results of our study indicates that matrix tablets containing 50% guar gum and coated with 10% hydroxy propyl methylcellulose phthalate are most suitable for drugs like sennosides which are mainly active in the lower GIT.


Premature termination of hair follicle morphogenesis and accelerated hair follicle cycling in Iasi congenital atrichia (fzica) mice points to fuzzy as a key element of hair cycle control.
Inbred laboratory mice have proven to be useful model systems for studying hair biology and pathomechanisms of hair loss. Fuzzy (fz) is an autosomal recessive mutation that results in hair coat abnormalities. Though this mutant has long been known, its cutaneous abnormalities still await systematic analysis. Here, we provide a systematic skin phenotype analysis of mice that are homozygous for Iasi congenital atrichia (fzica/fzica), which is allelic to fz. Homozygous mice exhibit a sparse hair coat after birth and completely loose their hair at around postnatal day 120. Although early and mid stages of hair follicle morphogenesis are normal, late hair follicle morphogenesis reveals multifocal cell degeneration within the Huxley layer of the inner root sheath (IRS) and a complete lack of the hair shaft medulla. In addition, hair follicle development is prematurely terminated by induction of the first postnatal hair cycle with premature entry into catagen. Subsequently, a dramatically shortened telogen is immediately followed by premature anagen development, resulting in a marked, generalized acceleration of hair follicle cycling. This suggests that fuzzy is not only involved in structural hair shaft integrity and differentiation of the IRS and medulla, but also plays an important role in the control of hair follicle cycling. Our data show that fuzzy is involved in controlling both catagen and anagen initiation, designating fuzzy an exciting target for characterizing the intracutaneous oscillator system that drives hair follicle cycling.


The utilization of molecular microscope in management of heart transplant recipients in the era of noninvasive monitoring.
Monitoring for graft rejection is a fundamental tenet of post-transplant follow-up. In heart transplantation (HT) in particular, rejection has been traditionally assessed with endomyocardial biopsy (EMB). EMB has potential complications and noted limitations, including interobserver variability in interpretation. Additional tests, such as basic cardiac biomarkers, cardiac imaging, gene expression profiling (GEP) scores, donor-derived cell-free DNA (dd-cfDNA) and the novel molecular microscope diagnostic system (MMDx) have become critical tools in rejection surveillance beyond standard EMB. This paper describes an illustrative case followed by a review of MMDx within the context of other noninvasive screening modalities for rejection. We suggest MMDx be used to assist with early detection of rejection in cases of discordance between EMB and other noninvasive studies.


[New findings on the geographic distribution of the verrucarum group (Diptera: Psychodidae) in Colombia].
The incrimination of sand flies belonging to verrucarum species group in the leishmaniasis transmission underscores the need for a detailed information on the geographical distribution of these species. The current listing adds 34 new records that extend significantly the knowledge of the geographical distribution of the verrucarum group in Colombia. The most important new records pertain to Lutzomyia spinicrassa in the tropical dry forest of the Atlantic coast, Lutzomyia ovallesi in the Orinoco and Amazon River watersheds regions of Colombia, and the sympatric occurrence of Lutzomyia spinicrassa and Lutzomyia quasitownsendi in the eastern chain of the Andes mountains. Additionally, the municipal and altitudinal distributions are compiled for 19 new species recorded for Colombia. The series townsendi of the verrucarum group generally restricted to premontane and lower montane zones based on distribution data on Lutzomyia longiflocosa, L. quasitownsendi, Lutzomyia sauroida, L. spinicrassa, Lutzomyia torvida, Lutzomyia townsendi and Lutzomyia youngi. The series verrucarum is distributed from the tropical lowland to the montane zones, and includes species with wide geographical range (Lutzomyia evansi, Lutzomyia columbiana, L. ovallesi, Lutzomyia nuneztovari, Lutzomyia nevesi), and species of highly endemic distribution (Lutzomyia andina, Lutzomyia disiuncta, Lutzomyia moralesi, Lutzomyia antioquiensis). Members of the series pia (Lutzomyia pia, Lutzomyia limafalcaoae) and the series serrana (Lutzomyia serrana) occur from the tropical lowlands to the lower montane zones. The altitudinal divergences may be intrinsically tied to speciation process, especially as it relates to the climatic and geologic events that have affected the flora and fauna of the Andean region.


JAK/STAT signaling and human in vitro myogenesis.
A population of satellite cells exists in skeletal muscle. These cells are thought to be primarily responsible for postnatal muscle growth and injury-induced muscle regeneration. The Janus kinase/signal transducers and activators of transcription (JAK/STAT) signaling cascade has a crucial role in regulating myogenesis. In rodent skeletal muscle, STAT3 is essential for satellite cell migration and myogenic differentiation, regulating the expression of myogenic factors. The aim of the present study was to investigate and compare the expression profile of JAK/STAT family members, using cultured primary human skeletal muscle cells. Near confluent proliferating myoblasts were induced to differentiate for 1, 5 or 10 days. During these developmental stages, members of the JAK/STAT family were examined, along with factors known to regulate myogenesis. We demonstrate the phosphorylation of JAK1 and STAT1 only during myoblast proliferation, while JAK2 and STAT3 phosphorylation increases during differentiation. These increases were correlated with the upregulation of genes associated with muscle maturation and hypertrophy. Taken together, these results provide insight into JAK/STAT signaling in human skeletal muscle development, and confirm recent observations in rodents.


Research Review: Outcomes of 24- to 36-month-old children with autism spectrum disorder vary by ascertainment strategy: a systematic review and meta-analysis.
Despite widespread recommendations for early surveillance of risk for autism spectrum disorder (ASD), no research to date has shown that early surveillance leads to better clinical outcomes. Preliminary research has suggested that children with ASD ascertained via prospective follow-up have better outcomes than those ascertained via community referral. Because prospective studies include early surveillance, by comparing outcomes of children with ASD across ascertainment strategies, we may gain insight into the effects of early surveillance relative to its absence. A systematic review was conducted to identify studies reporting outcomes of 24- to 36-month-olds with ASD ascertained via prospective follow-up, community referral, or universal screening. A meta-analysis using a random effects model was used to calculate overall effect size estimates for developmental level and symptom severity across ascertainment cohorts. Eleven prospective, ten community referral, and eight universal screening studies were identified, reporting on 1,658 toddlers with ASD. We found no differences in outcomes between community referral and universal screening studies. Relative to both, prospective studies reported significantly higher developmental levels and lower symptom severities. Outcomes of young children with ASD ascertained via prospective follow-up are better than those of children with ASD recruited via community referral or universal screening. Although we discuss why sampling bias is not likely the driving force behind these findings, we cannot rule out the possibility that sampling bias contributes to the observed differences; future studies should probe the effects of sociodemographic variables on clinical outcomes as a function of ascertainment strategy. This limitation notwithstanding, our results raise the possibility that prospective follow-up may confer a 'surveillance effect' that contributes to improved developmental and diagnostic outcomes in children with ASD. Future research should test this hypothesis and determine the specific mechanism by which surveillance may improve outcomes.


Neural regions essential for distinct cognitive processes underlying picture naming.
We hypothesized that distinct cognitive processes underlying oral and written picture naming depend on intact function of different, but overlapping, regions of the left hemisphere cortex, such that the distribution of tissue dysfunction in various areas can predict the component of the naming process that is disrupted. To test this hypothesis, we evaluated 116 individuals within 24 h of acute ischaemic stroke using a battery of oral and written naming and other lexical tests, and with magnetic resonance diffusion and perfusion imaging to identify the areas of tissue dysfunction. Discriminant function analysis, using the degree of hypoperfusion in various Brodmann's areas--BA 22 (including Wernicke's area), BA 44 (part of Broca's area), BA 45 (part of Broca's area), BA 21 (inferior temporal cortex), BA 37 (posterior, inferior temporal/fusiform gyrus), BA 38 (anterior temporal cortex) and BA 39 (angular gyrus)--as discriminant variables, classified patients on the basis of the primary component of the naming process that was impaired (defined as visual, semantics, modality-independent lexical access, phonological word form, orthographic word form and motor speech by the pattern of performance and types of errors across lexical tasks). Additionally, linear regression analysis demonstrated that the areas contributing the most information to the identification of patients with particular levels of impairment in the naming process were largely consistent with evidence for the roles of these regions from functional imaging. This study provides evidence that the level of impairment in the naming process reflects the distribution of tissue dysfunction in particular regions of the left anterior, inferior and posterior middle/superior temporal cortex, posterior inferior frontal and inferior parietal cortex. While occipital cortex is also critical for picture naming, it is likely that bilateral occipital damage is necessary to disrupt visual recognition. These findings provide new evidence that a network of brain regions supports naming, but separate components of this network are differentially required for distinct cognitive processes or representations underlying the complex task of naming pictures.


Promising preclinical patient-derived organoid (PDO) and xenograft (PDX) models in upper gastrointestinal cancers: progress and challenges.
Gastrointestinal (GI) cancers (gastric cancer, oesophageal cancer, liver cancer, colorectal cancer, etc.) are the most common cancers with the highest morbidity and mortality in the world. The therapy for most GI cancers is difficult and is associated with a poor prognosis. In China, upper GI cancers, mainly gastric cancer (GC) and oesophageal cancer (EC), are very common due to Chinese people's characteristics, and more than half of patients are diagnosed with distant metastatic or locally advanced disease. Compared to other solid cancers, such as lung cancer and breast cancer, personalized therapies, especially targeted therapy and immunotherapy, in GC and EC are relatively lacking, leading to poor prognosis. For a long time, most studies were carried out by using in vitro cancer cell lines or in vivo cell line-derived xenograft models, which are unable to reproduce the characteristics of tumours derived from patients, leading to the possible misguidance of subsequent clinical validation. The patient-derived models represented by patient-derived organoid (PDO) and xenograft (PDX) models, known for their high preservation of patient tumour features, have emerged as a very popular platform that has been widely used in numerous studies, especially in the research and development of antitumour drugs and personalized medicine. Herein, based on some of the available published literature, we review the research and application status of PDO and PDX models in GC and EC, as well as detail their future challenges and prospects, to promote their use in basic and translational studies or personalized therapy.


Insulin-like growth factor II signaling through the insulin-like growth factor II/mannose-6-phosphate receptor promotes exocytosis in insulin-secreting cells.
The insulin-like growth factor II (IGF-II)/mannose-6-phosphate (M-6-P) receptor is known to participate in endocytosis as well as sorting of lysosomal enzymes and is involved in membrane trafficking through rapid cycling between cytosolic membrane compartments and the plasma membrane. Here we demonstrate that IGF-II, acting through the IGF-II/M-6-P receptor, promotes exocytosis of insulin in the pancreatic beta cell. The effect of IGF-II was evoked at nonstimulatory concentrations of glucose, was mediated by a pertussis toxin sensitive GTP-binding protein, was dependent on protein kinase C-induced phosphorylation, and was independent of changes in cytoplasmic free Ca2+ concentration. Since the applied concentration of IGF-II is within the range normally found free in circulation in humans, this novel signaling pathway for the IGF-II/M-6-P receptor is likely to be involved in modulation of insulin exocytosis under physiological conditions.


Mitochondria-targeted antioxidants protect pancreatic β-cells against oxidative stress and improve insulin secretion in glucotoxicity and glucolipotoxicity.
Mitochondrial oxidative damage is thought to play a key role in pancreatic β-cell failure in the pathogenesis of type 2 diabetes. Despite this, the potential of mitochondria-targeted antioxidants to protect pancreatic β-cells against oxidative stress has not yet been studied. Therefore, we investigated if mitochondria-targeted antioxidants protect pancreatic β-cells such as RINm5F and HIT-T15 cells against oxidative stress under glucotoxic and glucolipotoxic conditions. When β-cells were incubated under these conditions, the expression levels of mitochondrial electron transport chain complex subunits, mitochondrial antioxidant enzymes (such as MnSOD and Prx3), β-cell apoptosis, lipogenic enzymes (such as ACC, FAS and ABCA1), intracellular lipid accumulation, oxidative stress, ER stress, mitochondrial membrane depolarization, nuclear NF- κB and sterol regulatory element binding protein 1c (SREBP1c) were all increased, in parallel with decreases in intracellular ATP content, citrate synthase enzymatic activity and glucose-stimulated insulin secretion. These changes were consistent with elevated mitochondrial oxidative stress, and incubation with the mitochondria-targeted antioxidants, MitoTempol or Mitoquinone (MitoQ), prevented these effects. In conclusion, mitochondria-targeted antioxidants protect pancreatic β-cells against oxidative stress, promote their survival, and increase insulin secretion in cell models of the glucotoxicity and glucolipotoxicity associated with Type 2 diabetes.


Losartan Rescues Inflammation-related Mucociliary Dysfunction in Relevant Models of Cystic Fibrosis.
Rationale: Despite therapeutic progress in treating cystic fibrosis (CF) airway disease, airway inflammation with associated mucociliary dysfunction remains largely unaddressed. Inflammation reduces the activity of apically expressed large-conductance Ca2+-activated and voltage-dependent K+ (BK) channels, critical for mucociliary function in the absence of CFTR (CF transmembrane conductance regulator).Objectives: To test losartan as an antiinflammatory therapy in CF using CF human bronchial epithelial cells and an ovine model of CF-like airway disease.Methods: Losartan's antiinflammatory effectiveness to rescue BK activity and thus mucociliary function was tested in vitro using primary, fully redifferentiated human airway epithelial cells homozygous for F508del and in vivo using a previously validated, now expanded pharmacologic sheep model of CF-like, inflammation-associated mucociliary dysfunction.Measurements and Main Results: Nasal scrapings from patients with CF showed that neutrophilic inflammation correlated with reduced expression of LRRC26 (leucine rich repeat containing 26), the γ subunit mandatory for BK function in the airways. TGF-β1 (transforming growth factor β1), downstream of neutrophil elastase, decreased mucociliary parameters in vitro. These were rescued by losartan at concentrations achieved by nebulization in the airway and oral application in the bloodstream: BK dysfunction recovered acutely and over time (the latter via an increase in LRRC26 expression), ciliary beat frequency and airway surface liquid volume improved, and mucus hyperconcentration and cellular inflammation decreased. These effects did not depend on angiotensin receptor blockade. Expanding on a validated and published nongenetic, CF-like sheep model, ewes inhaled CFTRinh172 and neutrophil elastase for 3 days, which resulted in prolonged tracheal mucus velocity reduction, mucus hyperconcentration, and increased TGF-β1. Nebulized losartan rescued both mucus transport and mucus hyperconcentration and reduced TGF-β1.Conclusions: Losartan effectively reversed CF- and inflammation-associated mucociliary dysfunction, independent of its angiotensin receptor blockade.


HPLC-mass spectrometry analysis of isoflavones.
The current interest in the role of dietary isoflavonoids, particularly the soy isoflavone genistein, in lowering the risk of several chronic diseases, has led to the need for rapid, sensitive and precise assays for isoflavones and their metabolites in food matrices and in various physiological fluids and tissues. HPLC has the advantage over GC-based methods in that all the conjugated and unconjugated isoflavonoids and their metabolites can be separated and analyzed without the need for derivatization. An important advance in mass spectrometry has been the introduction of effective interfaces between the HPLC and the mass spectrometer, namely the electrospray ionization (ESI) and the heated nebulizer-atmospheric pressure chemical ionization (HN-APCI) interfaces. Because of the isoflavonoid concentrations in fluids such as bile or urine, preliminary extraction, so essential for GC-MS and many other analytical methods, is not necessary. This immediately overcomes the thorny issue of finding an effective solid-phase extraction procedure. Using reversed-phase HPLC-ESI-MS, it is possible to obtain a mass/intensity map of all isoflavonoid metabolites in a single 20 min analysis. Analysis of isoflavonoid conjugates in serum/plasma samples requires initial extraction, but the conjugates can be measured intact either by capillary reversed-phase HPLC-ESI-MS or on regular reversed-phase columns by HPLC-HN-APCI-MS. In both cases, specificity is obtained by causing the parent isoflavonoid molecular ions to undergo collision-induced dissociation to form specific daughter ions in a triple quadrupole MS instrument. When it is only necessary to measure the total isoflavonoids and their metabolites in blood, hydrolysis can be performed directly in serum/plasma samples and isoflavonoids recovered by ether or ethyl acetate solvent extraction. The isoflavone aglucones can be analyzed by HPLC-MS under isocratic solvent conditions, thereby drastically shortening analysis time and opening up prospects for automation. Therefore, HPLC-MS is a technique that is broadly applicable to the major issues in phytoestrogen research.


Milk exosomes: Nature's abundant nanoplatform for theranostic applications.
Exosomes are a unique subpopulation of naturally occurring extracellular vesicles which are smaller intracellular membrane nanoparticle vesicles. Exosomes have proven to be excellent nanocarriers for carrying lipids, proteins, mRNAs, non-coding RNAs, and DNAs, and disseminating long-distance intercellular communications in various biological processes. Among various cell-line or biological fluid derived exosomes, milk exosomes are abundant in nature and exhibit many nanocarrier characteristics favorable for theranostic applications. To be an effective delivery carrier for their clinical translation, exosomes must inbuilt loading, release, targeting, and imaging/tracking characteristics. Considering the unmet gaps of milk exosomes in theranostic technology it is essential to focus the current review on drug delivery and imaging applications. This review delineates the efficiency of exosomes to load therapeutic or imaging agents and their successful delivery approaches. It is emphasized on possible modifications of exosomes towards increasing the stability and delivery of agents. This article also summarizes the specific applications and the process of developing milk exosomes as a future pharmaceutical drug delivery vehicle.


Synthesis of interleukin-5 by activated eosinophils in patients with eosinophilic heart diseases.
Eosinophilic endomyocardial disease represents a major evolutive risk in chronic eosinophilia-associated disorders. Eosinophil granule proteins appear to be involved in cardiac injury, but the mechanisms leading to eosinophil infiltration and degranulation are not clear. Interleukin-5 (IL-5) has been recently shown to be produced by eosinophils and might play a role in both chemoattraction and degranulation of eosinophils. In four cases of eosinophilic diseases with severe cardiac failure, we evaluated the proportion of eosinophil phenotypes and the serum levels of eosinophil cationic protein (ECP) and soluble IL-2 receptor (sIL-2R), markers of disease activity in the hypereosinophilic syndromes. All four patients showed a markedly increased proportion of hypodense eosinophils with elevated serum ECP and sIL-2R levels. In all four patients, extracellular deposition of eosinophil granule proteins and features of eosinophil activation were observed in cardiac tissues. The synthesis of IL-5 by eosinophils was detected in myocardial sections and blood cells by in situ hybridization and by immunostaining with a monoclonal antibody against human IL-5. Sixty percent to 90% of tissue eosinophils expressed IL-5 mRNA and IL-5 protein. These data suggest that IL-5 can be produced by eosinophils at the sites of myocardial tissue damage and might participate in local eosinophil activation.


Multivariate segregation analysis using the mixed model.
Most major genes involved in the etiology of complex diseases are likely to have pleiotropic effects on a number of intervening quantitative traits. Methods of segregation analysis that incorporate the additional information from such multiple traits will exhibit greater power for detecting the effects of major genes and allow explicit tests of major locus pleiotropy hypotheses. In this study, we present a new method for multivariate segregation analysis that utilizes a multivariate generalization of Hasstedt's [1982] technique for calculating approximate mixed model likelihoods on pedigrees. The method is based on a simplification of the multivariate conditional likelihood via a transformation that simultaneously orthogonalizes the residual additive genetic and environmental covariance matrices. This transformation allows the multivariate conditional likelihood to be factored into the product of independent univariate conditional likelihoods. Resulting computations are relatively fast, making it feasible to analyze multiple traits in extended pedigrees. We demonstrate our method with a bivariate analysis of high-density lipoprotein cholesterol (HDL-C) and apolipoprotein AI (apo AI) serum levels in 585 pedigreed baboons.


Alcopops in Sweden--a supply side initiative.
This paper describes the introduction of alcopops and sweet ciders in Sweden in 1996, as a result of the membership of the European Union. Special attention is given to its effect on youth drinking. Surveys of young people show that alcopops and sweet ciders are perceived to be easy to drink and to have a sweet and pleasant taste, which tends to conceal the taste of alcohol. These results suggest that alcopops and sweet ciders should be regarded as one category of drinks. Alcopops and sweet ciders have penetrated the youth market rapidly. The introduction of these beverages accounted for approximately half of the increase in alcohol consumption by 15-16-year-old boys from 1996 to 1999, and two-thirds of the increase among girls, according to the surveys. A large proportion of the youth answered that alcopops and ciders did not replace any other alcoholic beverage. The paper also describes the public debate on alcopops and cider. The introduction of alcopops was criticized by the media and the responsible minister. Prominent spokesmen of the alcohol industry put themselves at a distance from these, primarily imported, products. The sales of alcopops have been declining since 1997. Sweet ciders, on the other hand, which in reality are synonymous with alcopops, have continued to increase during the whole period since 1996, and have not been subject to the same public criticism.


Gene expression model (in)validation by Fourier analysis.
The determination of the right model structure describing a gene regulation network and the identification of its parameters are major goals in systems biology. The task is often hampered by the lack of relevant experimental data with sufficiently low noise level, but the subset of genes whose concentration levels exhibit an oscillatory behavior in time can readily be analyzed on the basis of their Fourier spectrum, known to turn complex signals into few relatively noise-free parameters. Such genes therefore offer opportunities of understanding gene regulation quantitatively. Fourier analysis is applied to data on gene expression levels in mouse liver cells that oscillate according to the circadian rhythm. Several model structures in the form of linear and nonlinear differential equations are matched to the data and it is shown that although the considered models can reproduce many features of the oscillatory patterns, some can be excluded on the basis of Fourier analysis without appeal to prior knowledge of regulatory pathways. A systematic method for testing models is also proposed based on measuring the effects of variations in gene copy-number on the expression levels of coupled genes. Fourier analysis is a technique that is well-adapted to the study of biological oscillators and can be used instead or in addition to conventional modeling techniques. Its usefulness will increase as more high-resolution data become available.


Regulation of ovarian ornithine decarboxylase by human chorionic gonadotrophin.
Treatment of 29-day-old female Sprague-Dawley rats with human chorionic gonadotrophin (hCG) produced a large and rapid increase in the activity of ornithine decarboxylase. Measurements that use a specific radioimmunoassay showed that the increased activity could be accounted for by a parallel change in the amount of ornithine decarboxylase protein. The increased protein content was caused by an increased rate of synthesis, since the half-life of ornithine decarboxylase was not changed by the hormone treatment. The content of mRNA for ornithine decarboxylase was determined by hybridization with a cDNA probe, and it was found that the increased amount of protein was correlated with a change in the amount of mRNA. These results indicate that treatment with hCG induces ornithine decarboxylase in the rat ovary by increasing the production or the stability of the mRNA for this enzyme. The increased amount of ornithine decarboxylase led to an increase in putrescine in the ovary but did not increase the content of the polyamines spermidine and spermine. These findings show that, despite its rapid and large scale induction, ornithine decarboxylase is not the rate-limiting step that determines the content of these polyamines in this tissue. They also suggest that putrescine itself may play an important role in the ovary.


Towards a Wireless Smart Polysomnograph Using Symbolic Fusion.
Polysomnography is the gold standard test for sleep disorders among which the Sleep Apnea Syndrome (SAS) is considered a public health issue because of the increase of the cardio-and cerebro-vascular risk it is associated with. However, the reliability of this test is questioned since sleep scoring is a time-consuming task performed by medical experts with a high inter- and intra-scorers variability, and because data are collected from 15 sensors distributed over a patient's body surface area, using a wired connection which may be a source of artefacts for the patient's sleep. We have used symbolic fusion to support the automated diagnosis of SAS on the basis of the international guidelines of the AASM for the scoring of sleep events. On a sample of 70 patients, and for the Apnea-Hypopnea Index, symbolic fusion performed at the level of sleep experts (97.1% of agreement). The next step is to confirm these preliminary results and move forward to a smart wireless polysomnograph.


RISK OF RHEGMATOGENOUS RETINAL DETACHMENT WITH CENTRAL SEROUS CHORIORETINOPATHY.
To investigate the risk of rhegmatogenous retinal detachment (RRD) after central serous chorioretinopathy (CSCR). The study included 2,830 patients with CSCR and 16,980 control patients matched using a propensity score for age, sex, and comorbidities including status after cataract operation, blunt trauma, myopia, diabetes mellitus, hypertension, and hyperlipidemia from January 2001 through December 2010 from the Taiwan Longitudinal Health Insurance Database 2000. Information of each patient was collected until December 2011. Cox proportional hazard regression analysis was used to obtain the adjusted hazard ratio for RRD. The RRD-free survival rate was calculated using Kaplan-Meier analysis. Thirty-five patients with CSCR (1.24%) and 27 controls (0.16%) had RRD (P < 0.0001) during follow-up, resulting in a significantly higher risk of RRD in the patients with CSCR (incidence rate ratio = 7.83, 95% confidence interval = 4.74-12.93). After adjustment for potential confounders, the adjusted hazard ratio for developing RRD was increased 7.85 times in the cohort of total sample (adjusted hazard ratio = 7.85, 95% confidence interval = 4.75-12.97). It was found that CSCR increased the risk of RRD even after adjustment for age, sex, and comorbidities including status after cataract operation, blunt trauma, myopia, diabetes mellitus, hypertension, and hyperlipidemia.


Strategies to model the near-solute solvent molecular density/polarization.
The solvent molecular distribution significantly affects the behavior of the solute molecules and is thus important in studying many biological phenomena. It can be described by the solvent molecular density distribution, g, and the solvent electric dipole distribution, p. The g and p can be computed directly by counting the number of solvent molecules/dipoles in a microscopic volume centered at r during a simulation or indirectly from the mean force F and electrostatic field E acting on the solvent molecule at r, respectively. However, it is not clear how the g and p derived from simulations depend on the solvent molecular center or the solute charge and if the g(F) and p(E) computed from the mean force and electric field acting on the solvent molecule, respectively, could reproduce the corresponding g and p obtained by direct counting. Hence, we have computed g, p, g(F), and p(E) using different water centers from simulations of a solute atom of varying charge solvated in TIP3P water. The results show that g(F) and p(E) can reproduce the g and p obtained using a given count center. This implies that rather than solving the coordinates of each water molecule by MD simulations, the distribution of water molecules could be indirectly obtained from analytical formulas for the mean force F and electrostatic field E acting on the solvent molecule at r. Furthermore, the dependence of the g and p distributions on the solute charge revealed provides an estimate of the change in g and p surrounding a biomolecule upon a change in its conformation.


Toward hyperuniform disordered plasmonic nanostructures for reproducible surface-enhanced Raman spectroscopy.
We report on the self-assembling of clusters of gold-nanoparticles (Au-NPs) directed by the phase separation of poly(styrene)-b-poly(methylmethacrylate) (PS-b-PMMA) block-copolymer (BCP) on indium tin oxide coated glass, which induces the onset of vertical lamellar domains. After thermal evaporation of gold on BCP, Au-NPs of 4 nm are selectively included into PS-nanodomains by thermal annealing, and then clustered with large density of hot spots (> 10(4) μm(2)) in a random two-dimensional pattern. The resulting nanostructure exhibits near-hyperuniform long-range correlations. The consequent large degree of homogeneity of this isotropic plasmonic pattern gives rise to a highly reproducible Surface-Enhanced Raman Scattering (SERS) enhancement factor over the centimeter scale (std. dev. ∼ 10% over 0.25 cm(2)). We also discuss the application of a static electric field for modulating the BCP host morphology. The electric field induces an alignment of Au-NP clusters into ordered linear chains, exhibiting a stronger SERS activity, but reduced SERS spatial reproducibility.


[Spectral indices of heart rate variability in patients with vasovagal syncopes according to evidence of 5-min ECG].
To compare autonomic nervous system activity estimated by power spectral analysis of heart rate variability in patients with vasovagal syncopes and healthy volunteers. Seventeen health volunteers and 25 patients with vasovagal syncope were included in the study. In 16 cases faints were induced by head-up tilt table test (HTT), in 9 cases--by bicycle exercise test (BET). Power spectral analysis of heart rate variability in 5-th min ECG records was performed in supine and upright position while breath was fixed. In supine position the data of low frequency (LF) and high frequency (HF) of spectral power in both groups of patients were significantly higher than in volunteers. During upright the HF of spectral power significantly decreased in patients and volunteers. In upright position LF of spectral power significantly decreased in patients with syncope induced by HTT, did not change in patients with syncope induced by BET, but significantly increased in volunteers. Patients with vasovagal syncope had abnormal autonomic nervous system activity: a high level of parasympathetic influence in supine position and poor reaction of sympathetic nervous system during upright.


Mandibuloacral dysplasia and LMNA A529V mutation in Turkish patients with severe skeletal changes and absent breast development.
Mandibuloacral dysplasia (MAD) is an autosomal recessive disorder characterized by acroosteolysis (resorption of terminal phalanges), skin changes (hyperpigmentation), clavicular hypoplasia, craniofascial anomalies, a hook nose and prominent eyes, delayed closures of the cranial sutures, lipodystrophy, alopecia, and skeletal anomalies. MAD patients are classified according to lipodystrophy patterns: type A and type B. The vast majority of MAD cases are caused by LMNA gene mutations. MAD patients with type A lipodystrophy (MADA) have been reported to have LMNA R527H, A529V, or A529T mutations. In this report, we describe two MADA patients with progressive skeletal changes, absent breast development, and cataract in addition to the classical MAD phenotype. Both patients were found to be homozygous for the Ala529Val mutation of the LMNA gene. Our female patient is the oldest MADA patient (59 years old) who has ever been reported with the LMNA mutation and also the LMNA Ala529Val mutation. This study is the second report on MADA patients with a homozygous Ala529Val mutation.


Optimization of a Benzothiazole Indolene Scaffold Targeting Bacterial Cell Wall Assembly.
The bacterial cell envelope is comprised of the cell membrane and the cell wall. The bacterial cell wall provides rigidity to the cell and protects the organism from potential harmful substances also. Cell wall biosynthesis is an important physiological process for bacterial survival and thus has been a primary target for the development of antibacterials. Antimicrobial peptides that target bacterial cell wall assembly are abundant and many bind to the essential cell wall precursor molecule Lipid II. We describe the structure-to-activity (SAR) relationship of an antimicrobial peptide-derived small molecule 7771-0701 that acts as a novel agent against cell wall biosynthesis. Derivatives of compound 7771-0701 (2-[(1E)-3-[(2E)-5,6-dimethyl-3-(prop-2-en-1-yl)-1,3-benzothiazol-2-ylidene]prop-1-en-1-yl]-1,3,3-trimethylindol-1-ium) were generated by medicinal chemistry guided by Computer-Aided Drug Design and NMR. Derivatives were tested for antibacterial activity and Lipid II binding. Our results show that the N-alkyl moiety is subject to change without affecting functionality and further show the functional importance of the sulfur in the scaffold. The greatest potency against Gram-positive bacteria and Lipid II affinity was achieved by incorporation of a bromide at the R3 position of the benzothiazole ring. We identify optimized small molecule benzothiazole indolene scaffolds that bind to Lipid II for further development as antibacterial therapeutics.


Effect of culture media on porcine embryos produced by in vitro fertilization or parthenogenetic activation after oocyte maturation with cycloheximide.
This study evaluated the effects of reversible meiotic inhibition and different culture media (PZM3 or NCSU23) on production of porcine embryos by either in vitro fertilization (IVF) or parthenogenetic activation (PA). Oocytes from abattoir-derived ovaries were allocated into two groups for maturation: CHX (5 μg/ml cycloheximide for 10 h) or Control (no CHX). The percentage of metaphase II (MII) oocytes was determined at 36, 40 or 44 h of in vitro maturation. For IVF and PA, denuded oocytes were fertilized with purified sperm for 6 h or activated by electric stimuli. Zygotes were then subdivided into two culture groups: NCSU23 or PZM3. No effect of treatment with CHX and culture media was observed on cleavage (D3) and blastocyst (D7) rates in IVF and PA groups. There are no differences of quality or development rates between IVF-derived embryos cultured in NCSU23 or PZM3. However, we observed high quality PA embryos in PZM3 compared with NCSU23. Maturation arrest with CHX decreased the average blastocyst cell number in IVF while it was increased in PA embryos. As older oocytes are more effectively activated, CHX- blocked oocytes reached the mature stage faster than the control group. In conclusion, the CHX treatment for 10 h, followed by oocyte maturation for 40 h, is an efficient protocol to produce high quality parthenote embryos, especially when they are cultured in PZM3. However, this protocol is not satisfactory for IVF embryos production. In this case, a shorter maturation period could provide better embryo quality.


Spectacle Independence for Pseudophakic Patients - Experience with a Trifocal Supplementary Add-on Intraocular Lens.
To evaluate the refractive and functional outcomes of the trifocal 1stQ AddOn® (Medicontur) supplementary intraocular lenses (IOLs) designed for implantation into the ciliary sulcus. The study included 18 eyes of 11 pseudophakic patients with uncomplicated previous implantation of monofocal capsular bag IOLs. These patients had a desire for spectacle independence. Distance, intermediate and near visual acuities were measured, and defocus curves were plotted over a period of 6 months following implantation of the add-on IOLs. Intraocular pressure (IOP), endothelial cell density measurements and biomicroscopic evaluation were also performed. In this study, 83.3% of eyes had spherical refractions within ±0.5 D from emetropia and 100% of eyes had spherical equivalent refractions that were within ±1.0 D of the target refraction. Visual acuities and defocus curves clearly confirmed trifocal optical performance (UDVA=0.03 ±0.05; UIVA=0.21 ±0.04; UNVA=0.12 ±0.04 logMAR; expressed as mean ±SD). Depth of focus showed identical results (DOF=0.486 D) compared to a trifocal capsular bag IOL, while the defocus curve was found to be superior in the intermediate and near ranges when compared to a trifocal capsular bag IOL. All patients achieved spectacle independence at all distances. All add-on IOLs were well positioned in the ciliary sulcus. No negative changes were noted in connection with endothelial cell counts, IOPs, the angle structure during surgery and during the follow-up period. The supplementary trifocal add-on IOL seems to be a safe, efficient and stable solution for achieving spectacle independence in pseudophakic patients with monofocal primary IOLs.


Body weight control by a high-carbohydrate/low-fat diet slows the progression of diabetic kidney damage in an obese, hypertensive, type 2 diabetic rat model.
Obesity is one of several factors implicated in the genesis of diabetic nephropathy (DN). Obese, hypertensive, type 2 diabetic rats SHR/NDmcr-cp were given, for 12 weeks, either a normal, middle-carbohydrate/middle-fat diet (MC/MF group) or a high-carbohydrate/low-fat diet (HC/LF group). Daily caloric intake was the same in both groups. Nevertheless, the HC/LF group gained less weight. Despite equivalent degrees of hypertension, hyperglycemia, hyperlipidemia, hyperinsulinemia, and even a poorer glycemic control, the HC/LF group had less severe renal histological abnormalities and a reduced intrarenal advanced glycation and oxidative stress. Mediators of the renoprotection, specifically linked to obesity and body weight control, include a reduced renal inflammation and TGF-beta expression, together with an enhanced level of adiponectin. Altogether, these data identify a specific role of body weight control by a high-carbohydrate/low-fat diet in the progression of DN. Body weight control thus impacts on local intrarenal advanced glycation and oxidative stress through inflammation and adiponectin levels.


Development of an in vivo model of human multiple myeloma bone disease.
Osteolytic bone destruction and its complications, bone pain, pathologic fractures, and hypercalcemia, are a major source of morbidity and mortality in patients with multiple myeloma. The bone destruction in multiple myeloma is due to increased osteoclast (OCL) activity and decreased bone formation in areas of bone adjacent to myeloma cells. The mechanisms underlying osteolysis in multiple myeloma in vivo are unclear. We used a human plasma cell leukemia cell line, ARH-77, that has disseminated growth in mice with severe combined immunodeficiency (SCID) and expresses IgG kappa, as a model for human multiple myeloma, SCID mice were irradiated with 400 rads and mice were injected either with 10(6) ARH-77 cells intravenously (ARH-77 mice) or vehicle 24 hours after irradiation. Development of bone disease was assessed by blood ionized calcium levels, x-rays, and histology. All ARH-77, but none of control mice that survived irradiation, developed hind limb paralysis 28 to 35 days after injection and developed hypercalcemia (1.35 to 1.46 mmol/L) a mean of 5 days after becoming paraplegic. Lytic bone lesions were detected using x-rays in all the hypercalcemic mice examined. No lytic lesions or hypercalcemia developed in the controls. Controls or ARH-77 mice, after developing hypercalcemia, were then killed and bone marrow plasma from the long bones were obtained, concentrated, and assayed for bone-resorbing activity. Bone marrow plasma from ARH-77 mice induced significant bone resorption in the fetal rat long bone resorption assay when compared with controls (percentage of total 45Ca released = 35% +/- 4% v 11% +/- 1%). Histologic examination of tissues from the ARH-77 mice showed infiltration of myeloma cells in the liver and spleen and marked infiltration in vertebrae and long bones, with loss of bony trabeculae and increased OCL numbers. Interestingly, cultures of ARH-77 mouse bone marrow for early OCL precursors (colony-forming unit-granulocyte-macrophage [CFU-GM]) showed a threefold increase in CFU-GM from ARH-77 marrow versus controls (185 +/- 32 v 40 +/- 3 per 2 x 10(5) cell plated). Bone-resorbing human and murine cytokines such as interleukin-6 (IL-6), IL-1 alpha or beta, TGF-alpha, lymphotoxin, and TNF alpha were not significantly increased in ARH-77 mouse sera or marrow plasma, compared with control mice, although ARH-77 cells produce IL-6 and lymphotoxin in vitro. Conditioned media from ARH-77 cells induced significant bone resorption in the fetal rat long bone resorption assay when compared with untreated media (percentage of total 45Ca released = 22% +/- 2% v 11% +/- 1%). This effect was not blocked by anti-IL-6 or antilymphotoxin (percentage of total 45Ca released = 19% +/- 1% and 22% +/- 1%, respectively). Thus, we have developed a model of human multiple myeloma bone disease that should be very useful to dissect the pathogenesis of the bone destruction in multiple myeloma.


Food insecurity and effectiveness of behavioral interventions to reduce blood pressure, New York City, 2012-2013.
Food insecurity is associated with diet-sensitive diseases and may be a barrier to successful chronic disease self-management. To evaluate the impact of food insecurity on blood pressure reduction in a pilot clinical trial, we tested the effectiveness of 2 behavioral interventions for hypertension in people with and without food security. A group of 28 men and women with type 2 diabetes and uncontrolled hypertension were randomized to either 1) home blood pressure telemonitoring alone or 2) home blood pressure telemonitoring plus telephone-based nurse case management. The primary outcome was 6-month change in systolic blood pressure. The 2 interventions resulted in modest, nonsignificant blood pressure reductions. Food-secure patients experienced clinically and statistically significant reductions in blood pressure, whereas no significant change was seen among food-insecure patients. Screening for food insecurity may help identify patients in need of tailored disease management interventions.


alpha-Ketoglutarate dehydrogenase mutant of Rhizobium meliloti.
A mutant of Rhizobium meliloti selected as unable to grow on L-arabinose also failed to grow on acetate or pyruvate. It grew, but slower than the parental strain, on many other carbon sources. Assay showed it to lack alpha-ketoglutarate dehydrogenase (kgd) activity, and revertants of normal growth phenotype contained the activity again. Other enzymes of the tricarboxylic acid cycle and of the glyoxylate cycle were present in both mutant and parent strains. Enzymes of pyruvate metabolism were also assayed. L-Arabinose degradation in R. meliloti was found to differ from the known pathway in R. japonicum, since the former strain lacked 2-keto-o-deoxy-L-arabonate aldolase but contained alpha-ketoglutarate semialdehyde dehydrogenase; thus, it is likely that R. meliloti has the L-arabinose pathway leading to alpha-ketoglutarate rather than the one to glycolaldehyde and pyruvate. This finding accounts for the L-arabinose negativity of the mutant. Resting cells of the mutant were able to metabolize the three substrates which did not allow growth.


Images of edge current in InAs/GaSb quantum wells.
Quantum spin Hall devices with edges much longer than several microns do not display ballistic transport; that is, their measured conductances are much less than e(2)/h per edge. We imaged edge currents in InAs/GaSb quantum wells with long edges and determined an effective edge resistance. Surprisingly, although the effective edge resistance is much greater than h/e(2), it is independent of temperature up to 30 K within experimental resolution. Known candidate scattering mechanisms do not explain our observation of an effective edge resistance that is large yet temperature independent.


Seven perspectives on GPCR H/D-exchange proteomics methods.
Recent research shows surging interest to visualize human G protein-coupled receptor (GPCR) dynamic structures using the bottom-up H/D-exchange (HDX) proteomics technology. This opinion article clarifies critical technical nuances and logical thinking behind the GPCR HDX proteomics method, to help scientists overcome cross-discipline pitfalls, and understand and reproduce the protocol at high quality. The 2010 89% HDX structural coverage of GPCR was achieved with both structural and analytical rigor. This article emphasizes systematically considering membrane protein structure stability and compatibility with chromatography and mass spectrometry (MS) throughout the pipeline, including the effects of metal ions, zero-detergent shock, and freeze-thaws on HDX result rigor. This article proposes to view bottom-up HDX as two steps to guide choices of detergent buffers and chromatography settings: (I) protein HDX labeling in native buffers, and (II) peptide-centric analysis of HDX labels, which applies (a) bottom-up MS/MS to construct peptide matrix and (b) HDX MS to locate and quantify H/D labels. The detergent-low-TCEP digestion method demystified the challenge of HDX-grade GPCR digestion. GPCR HDX proteomics is a structural approach, thus its choice of experimental conditions should let structure lead and digestion follow, not the opposite.


Efficacy Endpoints Conference on Acute Rejection in Kidney Transplantation: summary report of the database.
An international database of rejection episodes and their characteristics with regard to definition and follow-up parameters was developed to improve the approach to protocol development for clinical trials. Nineteen North American, European, and Australian centers uniformly reported on 50 consecutive transplant rejection episodes. Data collected included patient demographic parameters, induction and maintenance immunosuppression therapies, rejection agents (drug, dose, duration), clinical signs (50% decrease in urine, fever), serum creatinine (nadir, at rejection, daily during antirejection therapy to 15 days, and days 30, 90, 180, and 365 after rejection date), histopathological findings, morbidity, recurrence of rejection, and function at 1 year. The centers contributed a total of 953 presumed rejection episodes, of which 842 were confirmed as acute rejection episodes. The majority of cases were first rejections (81%), and rejection occurred 119 +/- 345 days following transplantation. The Banff Schema of histological grading was used in 38% of biopsy-proven rejection episodes (2% Borderline, 42% Grade I, 38% Grade II, 18% Grade III). Only 30% of all rejections showed clinical signs that were most likely to occur in Grade III rejection episodes (P < 0.006). The serum creatinine response was well below the rejection creatinine level by day 15 following initiation of antirejection therapy in the majority of cases. A significant number of the cases (34%) reported in this database experienced a recurrent rejection approximately 85 days following the first rejection. Graft survival 1 year following the rejection episode was good (83%). These findings will facilitate development of clinical trial design and beneficially impact approaches to antirejection therapies for kidney allograft recipients.


Body sway induced by a low frequency alternating current.
A sine wave current stimulus, applied between electrodes placed about one ear and an indifferent electrode, produced a cyclical sway predominantly in the coronal plane. In 12 normal subjects, stimulus frequency (0.1-0.6 Hz) and intensity (+/-0.05 - +/-1.5 mA) had no significant effect on the amplitude of the postural response, but phase lag increased from 0.7 rad at 0.1 Hz to 3.8 rad at 0.6 Hz. Stimulation at 0.3 Hz, +/-1.0 mA evoked detectable responses in all but one of 50 normal subjects. Seven patients with end organ lesions all gave positive responses with the same test procedure. Likewise 13 out of 16 patients with destruction of the VIII nerve also gave positive responses. In the other 3 patients the response on the side of the lesion was absent, as it was in another 3 individuals with lesions of the vestibular nuclei. It was concluded that the sway induced by a low frequency alternating current is the manifestation of the stimulation of structures within the CNS central to the vestibular nerve or its end organ.


Commodifying Vaccines to Curtail Antibiotic Resistance Impact in Malaria Endemic Countries.
Rampant and prevalent deployment of an efficient malaria vaccine in Pakistan, together with basic control and preventive measures, could significantly decrease the economic and healthcare burden caused by drug-resistant malaria. Moreover, RTS, S/AS01 vaccine has attained a much-needed breakthrough after decades of growth, as an innovative vaccine for malaria in Phase III clinical trials, and presently undergoing implementation studies. So far Gavi, WHO, and other stakeholders are contemplating on the practical issues, risk-benefit, and cost-effectiveness in resource-limited settings of vaccine implementation capacity. Imminent advances, like using a delayed as well as enhanced protection, divided schedule for dosing, and alternate adjuvants are likely to attain the vital goal of eradication of malaria. Vaccination is a potentially critical component of efforts to arrest the development and dissemination of antimicrobial resistance; though little is known about the impact vaccination may have within low-and-middle-income countries. Key Words: Antimicrobial resistance, Malaria, Vaccine.


Toxicity of chlorine to zebrafish embryos.
Surface disinfection of fertilized fish eggs is widely used in aquaculture to reduce extraovum pathogens that may be released from brood fish during spawning, and this is routinely used in zebrafish Danio rerio research laboratories. Most laboratories use approximately 25 to 50 ppm unbuffered chlorine solution for 5 to 10 min. Treatment of embryos with chlorine has significant germicidal effects for many Gram-negative bacteria, viruses, and trophozoite stages of protozoa, but is less effective against cyst or spore stages of protozoa and certain Mycobacterium spp. Therefore, we evaluated the toxicity of unbuffered and buffered chlorine solutions to embryos exposed at 6 or 24 h post-fertilization (hpf) to determine whether higher concentrations can be used for treating zebrafish embryos. Most of our experiments entailed using an outbred line (5D), with both mortality and malformations as endpoints. We found that 6 hpf embryos consistently were more resistant than 24 hpf embryos to the toxic effects of chlorine. Chlorine is more toxic and germicidal at lower pH, and chlorine causes elevated pH. Consistent with this, we found that unbuffered chlorine solutions (pH ca. 8-9) were less toxic at corresponding concentrations than solutions buffered to pH 7. Based on our findings here, we recommend treating 6 hpf embryos for 10 min and 24 hpf embryos for 5 min with unbuffered chlorine solution at 100 ppm.


Immune response pattern across the asymptomatic, symptomatic and convalescent periods of COVID-19.
We present an integrated analysis of urine and serum proteomics and clinical measurements in asymptomatic, mild/moderate, severe and convalescent cases of COVID-19. We identify the pattern of immune response during COVID-19 infection. The immune response is activated in asymptomatic infection, but is dysregulated in mild and severe COVID-19 patients. Our data suggest that the turning point depends on the function of myeloid cells and neutrophils. In addition, immune defects persist into the recovery stage, until 12 months after diagnosis. Moreover, disorders of cholesterol metabolism span the entire progression of the disease, starting from asymptomatic infection and lasting to recovery. Our data suggest that prolonged dysregulation of the immune response and cholesterol metabolism might be the pivotal causative agent of other potential sequelae. Our study provides a comprehensive understanding of COVID-19 immunopathogenesis, which is instructive for the development of early intervention strategies to ameliorate complex disease sequelae.


Endoscopic versus surgical treatment for a rare form of gallstone ileus: Bouveret's syndrome. Two case reports and review of the literature.
Bouveret's syndrome is a gastric outlet obstruction due to a gallstone impacted in the duodenal lumen or in the stomach via a bilioenteric fistula. It's the rarest form of gallstone ileus, with few cases reported in medical literature. Typically, this condition affects elderly people and causes high morbidity and mortality rates. We present a review of the published cases of Bouveret's syndrome between 2006, year of publication of the largest case series, and 2015 with particular regard to the operative management and treatment options. Two demanding cases of Bouveret's syndrome reported in our Hospital are also described. Bouveret's syndrome is a rare clinical condition and data reported in medical literature only derive from single case reports and case series. Therefore no clear management indications have been ruled out and treatment options are still matter of debate. Bouveret's syndrome is a rare clinical entity with a challenging operative management. Surgical or endoscopic treatments should be tailored on patient medical conditions and clinical presentation. Bouveret's Syndrome, Cholelithiasis, Gallstone Ileus, Gastric Outlet Obstruction, Intestinal Obstruction.


A light and electron microscopic study with comments on their relation to malignant fibrous histiocytoma and angiosarcoma.
Analysis of 365 cases of malignant soft tissue tumors revealed 83 cases of liposarcoma. The ages of the patients were distributed between 18 and 86 years with a mean average of 54.3 years. Approximately 50% of the cases were located in the lower extremity. Histologically, they consisted of 5 types; well differentiated (17 cases), myxoid (49 cases), round cell (3 cases), pleomorphic (11 cases), and mixed (3 cases). The appearance of lipoblasts of either the signet-ring type or mulberry (multivacuolated) type with displaced nuclei was the common characteristic feature for all types. Electron microscopically, lipid droplets were found inside of endoplasmic reticulum in lipoblasts, and abundant glycogen granules were seen in the cells containing only few lipid droplets. The lipoblasts were usually located close to the vascular wall in the interstitium, and findings suggesting an intimate relation between pericytes and lipoblasts were encountered. Although the differential diagnosis of malignant fibrous histiocytoma and liposarcoma was made possible by the appearance of lipoblasts in the latter, storiform pattern and histiocyte-like cells sometimes appeared in poorly differentiated liposarcoma. This tendency was also found in angiosarcoma. A close follow-up of recurrent cases revealed that there are actually cases of liposarcoma with malignant fibrous histiocytoma-like pattern and angiosarcoma with malignant fibrous histiocytoma-like pattern. It was considered that malignant fibrous histiocytoma-like figures can appear in various poorly differentiated sarcomas.