Genetic counselling for affective disease.
Classic genetic studies utilizing twin methods, adoption and cross-fostering data and pedigree analysis, have confirmed the hypothesis that the familial component in affective disease is biological, rather than social or environmental. Only recently, however, has it been possible to clarify the mode of inheritance of affective disorder or to identify specific DNA segments which are physically linked to genes conferring susceptibility to affective illness. At present, two susceptibility genes have been identified in individual families, and it is certain that additional vulnerability genes remain to be identified. Thus, affective illness is said to be genetically heterogeneous; i.e., different genes confer susceptibility in different families. Empiric risk figures suggest that the first degree relatives of a person with affective illness have about a 25-30% liability of major depression or manic-depressive illness, as compared to a population risk of 3-6%. Second degree relatives have a 12-15% risk, and third degree relatives a 6-9% risk. Pedigree studies strongly suggest that these figures represent the arithmetic mean of a high risk in families "loaded" for affective disorder, and a low risk in families with sporadic occurrence of the same illness. These data would argue that it is incumbent upon any physician to determine whether individual patients have a familial or non-familial form of affective illness, and to inform the patient accordingly.


The revolution of PDMS microfluidics in cellular biology.
Microfluidics is revolutionizing the way research on cellular biology has been traditionally conducted. The ability to control the cell physicochemical environment by adjusting flow conditions, while performing cellular analysis at single-cell resolution and high-throughput, has made microfluidics the ideal choice to replace traditional in vitro models. However, such a revolution only truly started with the advent of polydimethylsiloxane (PDMS) as a microfluidic structural material and soft-lithography as a rapid manufacturing technology. Indeed, before the "PDMS age," microfluidic technologies were: costly, time-consuming and, more importantly, accessible only to specialized laboratories and users. The simplicity of molding PDMS in various shapes along with its inherent properties (transparency, biocompatibility, and gas permeability) has spread the applications of innovative microfluidic devices to diverse and important biological fields and clinical studies. This review highlights how PDMS-based microfluidic systems are innovating pre-clinical biological research on cells and organs. These devices were able to cultivate different cell lines, enhance the sensitivity and diagnostic effectiveness of numerous cell-based assays by maintaining consistent chemical gradients, utilizing and detecting the smallest number of analytes while being high-throughput. This review will also assist in identifying the pitfalls in current PDMS-based microfluidic systems to facilitate breakthroughs and advancements in healthcare research.


Theta-phase closed-loop stimulation induces motor paradoxical responses in the rat model of Parkinson disease.
High-frequency deep brain stimulation (DBS) has become a widespread therapy used in the treatment of Parkinson's Disease (PD) and other diseases. Although it has proved beneficial, much recent attention has been centered around the potential of new closed-loop DBS implementations. Here we present a new closed-loop DBS scheme based on the phase of the theta activity recorded from the motor cortex. By testing the implementation on freely moving 6-OHDA lesioned and control rats, we assessed the behavioral and neurophysiologic effects of this implementation and compared it against the classical high-frequency DBS. Results show that both stimulation modalities produce significant and opposite changes on the movement and neurophysiological activity. Close-loop stimulation, far from improving the animals' behavior, exert contrary effects to those of high-frequency DBS which reverts the parkinsonian symptoms. Motor improvement during open-loop, high-frequency DBS was accompanied by a reduction in the amount of cortical beta oscillations while akinetic and disturbed behavior during close-loop stimulation coincided with an increase in the amplitude of beta activity. Cortical-phase-dependent close-loop stimulation of the STN exerts significant behavioral and oscillatory changes in the rat model of PD. Open-loop and close-loop stimulation outcomes differed dramatically, thus suggesting that the scheme of stimulation determines the output of the modulation even if the target structure is maintained. The current framework could be extended in future studies to identify the correct parameters that would provide a suitable control signal to the system. It may well be that with other stimulation parameters, this sort of DBS could be beneficial.


[Epilepsy as a symptom].
Epilepsy and seizures are often misinterpreted as identical phenomenon and treatment of seizures confused with treatment of epilepsy. In this paper the concept of both epilepsy and seizures are revised and also the more new etiopathogenics hypothesis, stressing the importance of etiological diagnosis for a suitable therapy and prognosis, which depends more of the etiology than the morphology of seizures.


Evaluation of column carryover of phosphorylated peptides and fumonisins by duplicated solvent gradient method in liquid chromatography/tandem mass spectrometry.
Columns made of three different materials were evaluated with regard to the carryover of phosphorylated peptides and fumonisins in liquid chromatography/tandem mass spectrometry (LC/MS/MS). In order to eliminate carryover caused by the injection operation in the autosampler, the column carryover was calculated using the duplicated solvent gradient method. A column made of a glass-lined stainless-steel tube and polyethylene frits (GL-PE column) yielded the most significant improvements in the peak shape and the carryover as compared to the other columns. The carryover of fumonisin B1 (FB1) and HLADLSpK (T19p) in the GL-PE column could be reduced; the lower limit of quantitation of T19p, and the range of the calibration curve were also improved. Since carryover peaks with the GL-PE column were symmetrical peaks of the samples, carryover in the column did not occur. The carryover calculated by the duplicated solvent gradient method corresponded to those in the flow path from the injection port to the inlet frit of the column. The carryover value of FB1 in the column with a stainless-steel tube and stainless-steel frits (S-S column) was 1.70%, and that of the flow path was 0.23%. We found that the majority of the carryover in our system occurred in the S-S column.


Active infective endocarditis: Clinical characteristics and factors related to hospital mortality.
Little information exists on the clinical characteristics and factors related to hospital mortality in patients with active infective endocarditis referred for surgery. Between January 1, 2003 and December 31, 2006, 86 patients (56 males, 30 females, mean age 59.2 years) with active infective endocarditis were referred to our Department (2.8% of overall hospitalizations). The relation of several clinical, laboratory and echocardiographic findings at admission with hospital mortality was evaluated. A native valve (NVE) was involved in 50/86; the other 30 had a prosthetic valve endocarditis (PVE). Six had pacemaker endocarditis. The aortic valve was involved more frequently than the mitral valve, both in NVE and PVE. The tricuspid valve was involved in four drug addicts; 51% of patients were in NYHA class III-IV. Staphylococci and streptococci were isolated in 69% of patients (39% vs 30%). Blood cultures were negative in 24%. Overall hospital mortality has been 11.6%. Two patients died before surgery, eight in the perioperative period. Hospital mortality was closely related to age, clinical and laboratory evidence of advanced septic condition (temperature > 38°C, leukocytosis and creatinine > 2.0 mg/dL) and hemodynamic impairment. Active infective endocarditis is a significant cause of referral to heart surgery departments and hospital mortality is still > 10%. Clinical and laboratory parameters easily available at admission suggest that severe sepsis and/or hemodynamic impairment may be helpful in predicting the clinical outcome in this group of high risk patients.


Secrets and social networks.
Secrets are information kept from others; they are relational. They shape the intimacy of our relationships, what we know of others and what we infer about the world. Recent research has promoted two models of voluntary secret disclosure. The first highlights deliberate and strategic disclosure to garner support and to avoid judgment. The second maintains strategic action but foregrounds that disclosures are made in contexts which shape who is in one's social network and who may be the recipient of a disclosure. Work outside of this main vein examines the mechanisms and motivations to share others' secrets as well as the potential consequences of doing so. The final avenue of inquiry in this review considers how keeping secrets can change (or avoid changing) the size and composition of the secret-keeper's social network and what information is shared within it. Understanding how secrets spread within and form social networks informs work from public health to criminology to organizational management.


Intracranial intraparenchymal schwannomas: a series of eight cases.
Intraparenchymal schwannomas of the CNS are extremely rare. Between 1979 and 1993 400 cases of intracranial schwannomas were operated on and among them were eight patients with intraparenchymal schwannomas comprising 2% of intracranial benign nerve sheath tumours. Four of them were located in the cerebral hemispheres and two each in the brain stem and in the cerebellum. In two cases, there was associated neurofibromatosis (NF-1 and NF-2, one case each). The age ranged from 6 months to 45 years with a male/female ratio of 3:1 and, surprisingly, six of them were in the left cerebral or cerebellar hemisphere.


[Ten years of alcohol intoxication among young people in The Netherlands].
To provide insight into the number of adolescents admitted to hospital with alcohol intoxication in the period 2007-2016. Observational cross-sectional study. We used data from the hospital admissions for alcohol intoxication among minors that had been reported to the Dutch Paediatric Surveillance System (NSCK). We also used data on minors who had been admitted to hospital for other alcohol-related reasons. In the period 2007-2016, 4,674 minors were admitted to a hospital in the Netherlands with alcohol intoxication. The number of minors admitted with alcohol intoxication increased annually from 2007 to 2011, after which it became reasonably stable, with a peak in 2015. The average age at which a minor was admitted increased (2007: 14.9 years; 2016: 15.5 years), and this also applied to the duration of loss of consciousness (2007: 2.24 hours; 2016: 3.12 hours). Parents of young people admitted with alcohol intoxication have become markedly stricter; 69% had given permission for consumption of alcohol in 2011; in 2016 that had decreased to 19%. Raising the minimum age for obtaining low-alcohol drinks, the creation of better awareness of the consequences of alcohol use and the opening of the Outpatient Adolescents and Alcohol Clinics seem to have had an effect. The average age of young people with alcohol intoxication has increased and parents have become stricter, but binge-drinking is still a serious problem. Further research is needed into characteristics and treatment of the group of young people who end up in hospital.


[Acute post-cholecystitis bilioma. Based on two cases].
A bilioma is an encapsulated collection of bile outside the biliary tree, which, in general, appears after a laceration in the biliary tract secondary to traumatism or to an iatrogenic lesion. Rarely, it occurs without a traumatic factor. We present two cases of bilioma as a complication of acute alithiasic cholecystitis. A review of the literature and the role of the ultrasonography in the diagnosis and management of biliomas is performed.


Phylogeography and demographic history of Amblyomma variegatum (Fabricius) (Acari: Ixodidae), the tropical bont tick.
The genetic diversity of Amblyomma variegatum (Fabricius) from four Caribbean islands and five African countries was compared by analyzing the sequences of three gene fragments, two mitochondrial (12SrDNA and D-Loop-DL), and one nuclear (intergenic transcribed spacer 2 [ITS2]). Genetic variability of the ITS2 DNA fragment consisted of only uninformative single nucleotide mutations, and therefore this gene was excluded from further analyses. Mitochondrial gene divergences among African populations and between Caribbean and African populations were very low. Nevertheless, the data suggest that A. variegatum is divided into distinct East and West African groups, the western group including all Caribbean samples. Phylogenetic analyses of the 12SrDNA and DL gene sequences showed that the West African A. variegatum clustered in a well-supported monophyletic clade, distinct from eastern paraphyletic lineages. Sequences of A. variegatum from the Caribbean were embedded in the West African clade, which supports the known West African historical origin for these ticks.


Creating De Novo Overlapped Genes.
Future applications of synthetic biology will rely on deploying engineered cells outside of lab environments for long periods of time. Currently, a significant roadblock to this application is the potential for deactivating mutations in engineered genes. A recently developed method to protect engineered coding sequences from mutation is called Constraining Adaptive Mutations using Engineered Overlapping Sequences (CAMEOS). In this chapter we provide a workflow for utilizing CAMEOS to create synthetic overlaps between two genes, one essential (infA) and one non-essential (aroB), to protect the non-essential gene from mutation and loss of protein function. In this workflow we detail the methods to collect large numbers of related protein sequences, produce multiple sequence alignments (MSAs), use the MSAs to generate hidden Markov models and Markov random field models, and finally generate a library of overlapping coding sequences through CAMEOS scripts. To assist practitioners with basic coding skills to try out the CAMEOS method, we have created a virtual machine containing all the required packages already installed that can be downloaded and run locally.


Selecting the best index for following the temporal evolution of apparent diffusion coefficient and diffusion anisotropy after hypoxic-ischemic white matter injury in neonates.
Diffusion-weighted (DW) MR imaging is a useful technique for detecting ischemia. In adults and neonates, however, temporal changes on DW images after ischemia complicate interpretation. Our purpose was to investigate the temporal evolution of the apparent diffusion coefficient (ADC), diffusion-tensor (DT) imaging components, and anisotropy in neonatal brain after hypoxic-ischemic white matter injury and to determine which anisotropy index is preferable. DT images were obtained with single-shot echo-planar imaging, by using pulsed field gradients in six directions. Sixteen volunteers and 10 term neonates with normal MR images were evaluated to obtain reference values. Among the anisotropy indexes of fractional anisotropy (FA), relative anisotropy (RA), volume ratio (VR), linear (CI) and planar (Cp) diffusion, and axial anisotropy (Am), simulations were performed to select the most appropriate indexes for clinical practice. The ADC, DT imaging components, and anisotropy were evaluated as a function of time after onset of symptoms in 11 neonates with hypoxic ischemia. In neonates, changes in lesions were characterized by a large decrease (40%) in all eigenvalues, with a stronger decrease in the direction perpendicular to the fibers, resulting in increased anisotropy indexes. The temporal evolution of the relative change in ADC did not show a significant trend (P >.05). The relative change in anisotropy decreased linearly with time (P <.05), with the strongest trend in anisotropy index Am. In clinical practice, anisotropy indexes FA, RA and Am appear to be useful. Am is the best index to monitor anisotropy changes. DT imaging provides information about diffusion parallel and perpendicular to white matter fibers, which helps the interpretation of physiologic changes after hypoxic-ischemic injury.


Nucleotide polymorphism at the alcohol dehydrogenase locus of pocket gophers, genus Geomys.
Using the strictly neutral model as a null hypothesis, we tested for deviations from expected levels of nucleotide polymorphism at the alcohol dehydrogenase locus (Adh-1) within and among four species of pocket gophers (Geomys bursarius major, G. knoxjonesi, G. texensis llanensis, and G. attwateri). The complete protein-encoding region was examined, and 10 unique alleles, representing both electromorphic and cryptic alleles, were used to test hypotheses (e.g., the neutral model) concerning the maintenance of genetic variation. Nineteen variable sites were identified among the 10 alleles examined, including 9 segregating sites occurring in synonymous positions and 10 that were nonsynonymous. Several statistical methods, including those that test for within-species variation as well as those that examine variation within and among species, failed to reject the null hypothesis that variation (both within and between species of Geomys) at the Adh locus is consistent with the neutral theory. However, there was significant heterogeneity in the ratio of polymorphism to divergence across the gene, with polymorphisms clustered in the first half of the coding region and fixed differences clustered in the second half of the gene. Two alternative hypotheses are discussed as possible explanations for this heterogeneity: an old balanced polymorphism in the first half of the gene or a recent selective sweep in the second half of the gene.


Photoacoustic lifetime imaging for direct in vivo tissue oxygen monitoring.
Measuring the partial pressure of oxygen (pO₂) in tissue may provide physicians with essential information about the physiological state of tissue. However, currently available methods for measuring or imaging tissue pO₂ have significant limitations, preventing them from being widely used in clinics. Recently, we have reported a direct and noninvasive in vivo imaging modality based on the photoacoustic lifetime which overcomes certain drawbacks of the existing methods. The technique maps the excited triplet state of oxygen-sensitive dye, thus reflecting the spatial and temporal distributions of tissue oxygen. Here, we present two studies which apply photoacoustic lifetime imaging (PALI) to monitor changes of tissue oxygen induced by external modulations. The first study modulates tissue oxygen by controlling the percentage of oxygen a normal mouse inhales. We demonstrate that PALI is able to reflect the change in oxygen level with respect to normal, oxygen-rich, and oxygen-poor breathing conditions. The second study involves an acute ischemia model using a thin thread tied around the hindlimb of a normal mouse to reduce the blood flow. PALI images were acquired before, during, and after the restriction. The drop of tissue pO₂ and recovery from hypoxia due to reperfusion were tracked and observed by PALI.


Strain differences in the expression of the Epa-1-restricting element.
Epa-1-specific cytotoxic T lymphocytes (CTL) lyse epidermal cells (EC) of different Epa-1+ H-2k strains, such a AKR, CBA, C58, and RF, at different levels. We used an H-2Kk-specific monoclonal antibody (mAb) to test the hypothesis that this phenomenon is due to differences in the H-2-restricting element. Initially, we established the specificity of this mAb for the Epa-1-restricting element by demonstrating its capacity to inhibit the lysis of CBA EC by Epa-1-specific CTL. We then used it as the probe in a cellular radioimmunoassay to quantify the expression of the restricting element by EC of different H-2k strains. We found that C58 and RF EC bound significantly less of the mAb than did CBA EC. Although AKR also bound less of the mAb than did CBA EC, the difference was not statistically significant. To examine the generality of this phenomenon, we quantified the expression of Kk antigens on spleen cells (SC) of the same four strains. We found that RF SC, but not AKR or C58 SC, bound significantly less of the Kk mAb than did CBA SC. Thus, the differential CTL lysis of Epa-1+ EC of different strains probably reflects differences in expression of the H-2-restricting element rather than of the nominal antigen.


Furin cleavage of the SARS-CoV-2 spike is modulated by O-glycosylation.
The SARS-CoV-2 coronavirus responsible for the global pandemic contains a unique furin cleavage site in the spike protein (S) that increases viral infectivity and syncytia formation. Here, we show that O-glycosylation near the furin cleavage site is mediated by specific members of the GALNT enzyme family and is dependent on the novel proline at position 681 (P681). We further demonstrate that O-glycosylation of S decreases furin cleavage. Finally, we show that GALNT family members capable of glycosylating S are expressed in human respiratory cells that are targets for SARS-CoV-2 infection. Our results suggest that O-glycosylation may influence viral infectivity/tropism by modulating furin cleavage of S and provide mechanistic insight into the potential role of P681 mutations in the recently identified, highly transmissible B.1.1.7 variant.


Rapid and simultaneous detection of Mycobacterium tuberculosis complex and Beijing/W genotype in sputum by an optimized DNA extraction protocol and a novel multiplex real-time PCR.
Rapid diagnosis and genotyping of Mycobacterium tuberculosis by molecular methods are often limited by the amount and purity of DNA extracted from body fluids. In this study, we evaluated 12 DNA extraction methods and developed a highly sensitive protocol for mycobacterial DNA extraction directly from sputa using surface-coated magnetic particles. We have also developed a novel multiplex real-time PCR for simultaneous identification of M. tuberculosis complex and the Beijing/W genotype (a hypervirulent sublineage of M. tuberculosis) by using multiple fluorogenic probes targeting both the M. tuberculosis IS6110 and the Rv0927c-pstS3 intergenic region. With reference strains and clinical isolates, our real-time PCR accurately identified 20 non-Beijing/W and 20 Beijing/W M. tuberculosis strains from 17 different species of nontuberculosis Mycobacterium (NTM). Further assessment of our DNA extraction protocol and real-time PCR with 335 nonduplicate sputum specimens correctly identified all 74 M. tuberculosis culture-positive specimens. In addition, 15 culture-negative specimens from patients with confirmed tuberculosis were also identified. No cross-reactivity was detected with NTM specimens (n = 31). The detection limit of the assay is 10 M. tuberculosis bacilli, as determined by endpoint dilution analysis. In conclusion, an optimized DNA extraction protocol coupled with a novel multiprobe multiplex real-time PCR for the direct detection of M. tuberculosis, including Beijing/W M. tuberculosis, was found to confer high sensitivity and specificity. The combined procedure has the potential to compensate for the drawbacks of conventional mycobacterial culture in routine clinical laboratory setting, such as the lengthy incubation period and the limitation to viable organisms.


Kv3.3b expression defines the shape of the complex spike in the Purkinje cell.
The complex spike (CS) in cerebellar Purkinje Cells (PC) is not an all-or-nothing phenomena as originally proposed, but shows variability depending on the spiking behavior of the Inferior Olive and intrinsic variability in the number and shape of spikelets. The potassium channel Kv3.3b, which has been proposed to undergo developmental changes during the postnatal PC maturation, has been shown to be crucial for the repolarization of the spikelets in the CS. We address here the regulation of the intrinsic CS variability by the expression of inactivating Kv3.3 channels in PCs by combining patch-clamp recordings and single-cell PCR methods on the same neurons, using a technique that we recently optimized to correlate single cell transcription levels with membrane ion channel electrophysiology. We show that while the inactivating TEA sensitive Kv3.3 current peak intensity increases with postnatal age, the channel density does not, arguing against postnatal developmental changes of Kv3.3b expression. Real time PCR of Kv3.3b showed a high variability from cell to cell, correlated with the Kv3.3 current density, and suggesting that there are no mechanisms regulating these currents beyond the mRNA pool. We show a significant correlation between normalized quantity of Kv3.3b mRNA and both the number of CS spikelets and their rate of voltage fluctuation, linking the intrinsic CS shape directly to the Kv3.3b mRNA pool. Comparing the observed cell-to-cell variance with studies on transcriptional noise suggests that fluctuations of the Kv3.3b mRNA pool are possibly not regulated but represent merely transcriptional noise, resulting in intrinsic variability of the CS.


Multiple spinal cord melanoma: case report with emphasis on the difficult preoperative diagnosis.
Primary spinal melanoma (PSM) is rare and usually occurs in the cervical and thoracic regions. Here, we present a case of multiple spinal cord melanoma located at the level of cervical and upper thoracic spine in a 24-year-old male. The patient underwent a T6-T7 thoracic laminectomy and gross-total resection of the duty lesion. The histological findings of the tumor were consistent with an intermediate-grade melanocytoma. To the best of our knowledge, only two cases of multifocal primary melanocytic tumor have been reported in the literature. In this case study we review the pertinent literature and emphasize the importance and difficulties concerning the preoperative diagnosis of multiple PSM.


Progressive systemic sclerosis (scleroderma), carcinoma breast and valvular heart disease: an unusual combination.
Systemic sclerosis is a multi system disorder characterised by fibrosis of skin and internal organs. There are reports of relation between cancer and polymyositis/dermatomyositis, but no overall association with systemic sclerosis. Reports of the coexistence of cancer and systemic sclerosis, however, emphasise a close temporal relation in their occurrence. Cardiac involvement in the form of myocardial fibrosis and pericarditis occurs frequently in systemic sclerosis, while valvular involvement has been reported only sporadically. We report a patient, admitted for adenocarcinoma of left breast who was found to have features of systemic sclerosis, pulmonary hypertension, gangrene of toes, and stenotic mitral valve disease. The possible mechanisms of the coincidence of the three disorders are discussed.


Rat skeletal muscle phosphorylase kinase: turnover and control of isozyme levels in culture.
The expression of phosphorylase kinase was investigated in rat skeletal muscle cells developing in vitro. The enzyme was immunoprecipitated from cells cultured in the presence of [35S]methionine, and the 35S-labeled alpha-, alpha'-, and beta-subunits of the kinase were resolved by polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate. Fusion of myoblasts into myotubes was associated with marked increases in the amounts of kinase activity and the three 35S-labeled subunits. In 2-wk-old myotubes, the net amount of alpha'-subunit represented less than 20% of the total alpha-subunits (alpha + alpha'); however, alpha'-subunits appeared to be synthesized at least as rapidly as alpha-subunits. That alpha'-subunits were degraded more rapidly was confirmed by pulse-chase experiments, which also indicated that alpha'-subunits were not formed by proteolytic processing of the larger alpha-subunit. Inhibition of the spontaneous contractile activity of the myotubes with lidocaine markedly increased both phosphorylase kinase activity and the amounts of the 35S-labeled subunits. The divalent cation ionophore, A23187, decreased the alpha-subunits by 60%, but did not change levels of the alpha'-subunits. Taken together, the present results indicate that rat myotubes synthesize the two isozymes of phosphorylase kinase, and that levels of both are controlled by differentiation and muscle activity.


Axial Spondyloarthritis: Mimics and Pitfalls of Imaging Assessment.
Axial spondyloarthritis (axSpA) is a chronic inflammatory disorder that predominantly involves the axial skeleton. Imaging findings of axSpA can be divided into active changes, which include bone marrow edema, synovitis, enthesitis, capsulitis, and intra-articular effusion, and structural changes, which include erosions, sclerosis, bone fatty infiltration, fat deposition in an erosion cavity, and bone bridging or ankylosis. The ability to distinguish between imaging lesions suggestive of axSpA and artifacts or lesions suggestive of other disorders is critical for the accurate diagnosis of axSpA. Diagnosis may be challenging, particularly in early-stage disease and magnetic resonance imaging (MRI) plays a key role in the detection of subtle or inflammatory changes. MRI also allows the detection of structural changes in the subchondral bone marrow that are not visible on conventional radiography and is of prognostic and monitoring value. However, bone structural changes are more accurately depicted using computed tomography. Conventional radiography, on the other hand, has limitations, but it is easily accessible and may provide insight on gross changes as well as rule out other pathological features of the axial skeleton. This review outlines the imaging evaluation of axSpA with a focus on imaging mimics and potential pitfalls when assessing the axial skeleton.


The Potential of Trichoderma-Mediated Nanotechnology Application in Sustainable Development Scopes.
The environmental impact of industrial development has been well-documented. The use of physical and chemical methods in industrial development has negative consequences for the environment, raising concerns about the sustainability of this approach. There is a growing need for advanced technologies that are compatible with preserving the environment. The use of fungi products for nanoparticle (NP) synthesis is a promising approach that has the potential to meet this need. The genus Trichoderma is a non-pathogenic filamentous fungus with a high degree of genetic diversity. Different strains of this genus have a variety of important environmental, agricultural, and industrial applications. Species of Trichoderma can be used to synthesize metallic NPs using a biological method that is environmentally friendly, low cost, energy saving, and non-toxic. In this review, we provide an overview of the role of Trichoderma metabolism in the synthesis of metallic NPs. We discuss the different metabolic pathways involved in NP synthesis, as well as the role of metabolic metabolites in stabilizing NPs and promoting their synergistic effects. In addition, the future perspective of NPs synthesized by extracts of Trichoderma is discussed, as well as their potential applications in biomedicine, agriculture, and environmental health.


Successful port-a-cath salvage using linezolid in children with acute leukemia.
Central venous catheter (CVC) removal is indicated when persistent catheter-related bloodstream infection (CRBSI) occurs. This is a retrospective study to analyze the use of linezolid as a salvage therapy for CRBSIs due to coagulase-negative Staphylococci in children diagnosed with acute leukemia. Seven treatment courses of linezolid were administrated to six patients with port-type-CRBSI after non-effective intravenous vancomycin or teicoplanin treatment. Simultaneous lock and systemic therapy with linezolid avoided the removal of port-type-CVC in all cases. Treatment with linezolid was an alternative to catheter removal in these patients. Prospective studies are needed to confirm linezolid effectiveness as a salvage treatment in CRBSI.


Acrylamide-induced peripheral neuropathy: manifestations, mechanisms, and potential treatment modalities.
Acrylamide is a chemical monomer; its polymer compounds are used in the manufacture of plastic, papers, adhesive tapes, dyes, and food packaging. Lately, scientists found that cooking (mainly roasting, baking, and frying) yields acrylamide. In addition to fried/baked potatoes, coffee and bakery products still contain substantial amounts of acrylamide. Acrylamide has toxic effects on different body systems include genitourinary, reproductive, nervous system, along with being a carcinogenic substance. The neurotoxicity of acrylamide includes central and peripheral neuropathy. In humans, the clinical manifestations include sensory or motor peripheral neuropathy, drowsiness, or cerebellar ataxia. Likewise, it presents with skeletal muscle weakness, hindlimb dysfunction, ataxia, and weight loss in animals. The suggested mechanisms for acrylamide neurotoxicity include direct inhibition of neurotransmission, cellular changes, inhibition of key cellular enzymes, and bonding of kinesin-based fast axonal transport. Moreover, it is suggested that acrylamide's molecular effect on SNARE core kinetics is carried out through the adduction of NSF and/or SNARE proteins. Lately, scientists showed disruption of focal adhesion kinase (FAK) and proline-rich tyrosine kinase 2 (Pyk2) cell signaling pathways in human differentiating neuroblastoma SH-SY5Y cells, exposed to acrylamide. Different treatment modalities have been revealed to shield against or hasten recovery from acrylamide-induced neuropathy in preclinical studies, including phytochemical, biological, and vitamin-based compounds. Still, additional studies are needed to elucidate the pathogenesis and to identify the best treatment modality.


Global Transcriptomic Effects of Environmentally Relevant Concentrations of the Neonicotinoids Clothianidin, Imidacloprid, and Thiamethoxam in the Brain of Honey Bees ( Apis mellifera).
Neonicotinoids are implicated in the decline of honey bees, but the molecular basis underlying adverse effects is poorly known. Here we describe global transcriptomic profiles in the brain of honey bee workers exposed for 48 h at one environmentally realistic and one sublethal concentration of 0.3 and 3.0 ng/bee clothianidin and imidacloprid, respectively, and 0.1 and 1.0 ng/bee thiamethoxam (1-30 ng/mL sucrose solution) by high-throughput RNA-sequencing (RNA-seq). All neonicotinoids led to significant alteration (mainly down-regulation) of gene expression, generally with a concentration-dependent effect. Among many others, genes related to metabolism and detoxification were differently expressed. Gene ontology (GO) enrichment analysis of biological processes revealed catabolic carbohydrate metabolism (regulation of enzyme activities such as amylase), lipid metabolism, and transport mechanisms as shared terms between all neonicotinoids at high concentrations. KEGG pathway analysis indicated that at least two neonicotinoids induced changes in expression of various metabolic pathways: pentose phosphate pathways, starch and sucrose metabolism, and sulfur metabolism, in which glucose 1-dehydrogenase and alpha-amylase were down-regulated and 3'(2'), 5'-bisphosphate nucleotidase was up-regulated. RT-qPCR analysis confirmed the down-regulation of major royal jelly proteins, hbg3, and cyp9e2 found by RNA-seq. Our study highlights the comparative molecular effects of neonicotinoid exposure to bees. Further studies should link these effects with physiological outcomes for a better understanding of effects of neonicotinoids.


Estimating the conditional false-positive rate for semi-latent data.
When comparing tests for a disease, it is necessary to know whether individuals are diseased or nondiseased. In practice, the confirmatory (gold standard) procedure is often limited to individuals with positive test results, because the confirmatory procedure is not applied to individuals with negative test results. We present a model for estimating the sensitivity and specificity when two tests are compared and the gold standard classification is unavailable (semi-latent) for those individuals with negative results on both tests. The model does not assume independent error rates, and estimates of specificity conditional on a false-positive result for another test are derived. We use a Bayes approach for estimating the distributions of the performance parameters.


Geometric phases for mixed states in interferometry.
We provide a physical prescription based on interferometry for introducing the total phase of a mixed state undergoing unitary evolution, which has been an elusive concept in the past. We define the parallel transport condition that provides a connection form for obtaining the geometric phase for mixed states. The expression for the geometric phase for mixed state reduces to well known formulas in the pure state case when a system undergoes noncyclic and unitary quantum evolution.


Constitutive activation of STAT5 by the BCR-ABL oncogene in chronic myelogenous leukemia.
Using chronic myelogenous leukemia (CML) as a model, we tested the hypothesis that cytokine-independent growth of leukemia cells results from aberrant activation of cytokine signaling pathways. The STAT5 (signal transducer and activator of transcription) protein, which is activated transiently in normal myeloid cells by cytokines such as GM-CSF (granulocyte-macrophage colony stimulating factor), was constitutively activated in cell lines derived from CML patients, even in the absence of GM-CSF. STAT5 was also activated in primary mouse bone marrow cells acutely transformed by the CML-specific BCR-ABL oncogene, but not by the serine kinase oncogene v-MOS. Reconstitution experiments in non-hematopoietic cells show that STAT5 activation by BCR-ABL occurs independent of cytokines. Results using BCR-ABL mutants which specifically uncouple connections to known signal transduction pathways show that STAT5 activation is kinase dependent and correlates directly with ability to confer cytokine independent growth in hematopoietic cells. BCR-ABL also activates JAK kinases, which may provide a mechanism for STAT activation. These findings are consistent with a role for STAT5 in hematopoietic transformation by BCR-ABL.


Comparative cytogenetics of tapirs, genus tapirus (Perissodactyla, tapiridae).
Chromosomes of the four species of Tapirus were 2n = 52 in T. indicus, 2n = 76 in T. pinchaque, 2n = 80 in T. bairdii, and 2n = 80 in T. terrestris. The number of autosomal arms was 80-94. G-banded karyotypes indicated that a heterochromatic addition/deletion distinguished chromosomes 2 and 3 of T. bairdii and T. pinchaque, respectively. There were at least 13 conserved autosomes between the karyotypes of T. bairdii and T. terrestris, and at least 15 were conserved between T. bairdii and T. pinchaque. In G- and C-banded preparations, the X chromosomes of T. bairdii, T. indicus, and T. terrestris were identical, whereas the X chromosome of T. pinchaque differed from the X of the other species by a heterochromatic addition/deletion. The Y chromosome was a medium-sized to small acrocentric in T. bairdii, T. indicus, and T. pinchaque, but it was not positively identified in T. terrestris. There appeared to be fewer homologies between T. indicus and the three species occurring in Central and South America. Future cytogenetic studies of tapirs from the entire range of each of the four species might provide additional insight into their evolutionary biology and aid wildlife conservation efforts aimed at these threatened mammals.


Cognitive skills and reading in adults with Usher syndrome type 2.
To investigate working memory (WM), phonological skills, lexical skills, and reading comprehension in adults with Usher syndrome type 2 (USH2). The participants performed tests of phonological processing, lexical access, WM, and reading comprehension. The design of the test situation and tests was specifically considered for use with persons with low vision in combination with hearing impairment. The performance of the group with USH2 on the different cognitive measures was compared to that of a matched control group with normal hearing and vision (NVH). Thirteen participants with USH2 aged 21-60 years and a control group of 10 individuals with NVH, matched on age and level of education. The group with USH2 displayed significantly lower performance on tests of phonological processing, and on measures requiring both fast visual judgment and phonological processing. There was a larger variation in performance among the individuals with USH2 than in the matched control group. The performance of the group with USH2 indicated similar problems with phonological processing skills and phonological WM as in individuals with long-term hearing loss. The group with USH2 also had significantly longer reaction times, indicating that processing of visual stimuli is difficult due to the visual impairment. These findings point toward the difficulties in accessing information that persons with USH2 experience, and could be part of the explanation of why individuals with USH2 report high levels of fatigue and feelings of stress (Wahlqvist et al., 2013).


Symptom manifestations and expressions among Korean immigrant women suffering with depression.
This paper is a report of a study to investigate the culture-specific symptom manifestation and expression of depression in Korean immigrant women in the United States of America (USA). The literature indicates that Korean immigrants to the USA report higher levels of depressive symptoms than immigrants from other Asian ethnic groups, and women immigrants tend to encounter more gender-related emotional difficulties. However, because of traditional cultural beliefs, immigrants may not seek treatment for depression and, when they do, tend to leave treatment prematurely. We used an ethnographic method based on Spradley's Developmental Research Sequence with a sample of 17 Korean immigrant women who scored mild-to-severe depression on a Korean version of the Hamilton Depression Inventory Scale. Data were collected from 2005 to 2006, and consisted of tape-recorded interviews, field notes and diaries. Depressive symptoms were complex, intertwined and felt in all domains of the person's existence. They were expressed mainly through emotional and physical suffering, and the expression of suffering as a way of life. Themes elicited were emotional entrapment, shame and failure as women, disappointment at not being able to live a normal life and emotional restraint, as emotions were not generally expressed verbally but instead were expressed somatically, bodily and metaphorically. Being aware of culture-specific symptoms of depression among Korean immigrant women can assist nurses in caring for them and thus improving their therapeutic alliances and preventing them from prematurely terminating treatment.


Inhibition of Carbonic Anhydrase IX Suppresses Breast Cancer Cell Motility at the Single-Cell Level.
Protein Carbonic Anhydrase IX (CA IX), which is expressed in various hypoxic solid tumors in order to maintain proper pH, is also related to cancer cell adhesion, invasion, and metastasis processes. Here, we investigated whether CA IX inhibition by a highly CA IX selective agent benzenesulfonamide VD11-4-2 triggers changes in individual cell motility. We seeded breast cancer cells on an extracellular matrix-coated glass-bottomed dish and in a microfluidic device with a gradient flow of epidermal growth factor (EGF), tracked individual cell movement, calculated their migration speeds, and/or followed movement direction. Our results showed that the inhibitor VD11-4-2 decreased the speed of CA IX positive breast cancer cells by 20-26% while not affecting non-cancerous cell migration. The inhibitor suppressed the cell migration velocity increment and hindered cells from reaching their maximum speed. VD11-4-2 also reduced CA IX, expressing cell movement towards the growth factor as a chemoattractant. Such a single cell-based migration assay enabled the comprehensive investigation of the cell motility and revealed that VD11-4-2 shows the ability to suppress breast cancer cell migration at a lower concentration than previously tested CA IX inhibitors.


Stress-related modulation of hippocampal long-term potentiation in rats: Involvement of adrenal steroid receptors.
Stress is usually correlated with an increased release of glucocorticoids from the adrenal glands. Within the hippocampus, a structure long known to be involved in spatial learning, two corticosterone-binding receptors are identified: the glucocorticoid receptor (GR) and the mineralocorticoid receptor (MR). Activation of these receptors impairs or facilitates hippocampal long-term potentiation (LTP), respectively. Stress elicited by behavioral manipulations may interfere with cognitive modulations of LTP during learning experiments. Here, we explore the influence of two stress-inducing procedures, handling and swimming, on the maintenance of dentate gyrus LTP in the rat induced by a weak tetanization of the perforant path. Manipulations started 15 min after tetanization. Handling alone resulted in a complete reversal of LTP. Handling followed by a 2 min swim in a water tank elicited prolonged protein synthesis but not beta-adrenergic-dependent LTP compared with either control or handled animals. Blockade of the GRs but not of the MRs prevented the reversal of LTP by handling. Inactivation of the MRs but not of the GRs hindered LTP prolongation by swimming. Because the activated receptor complexes act as transcription factors, MR- and GR-related proteins may play a role in the maintenance of LTP. The data suggest a complex interplay of corticosterone-binding receptors on modulations of hippocampal LTP and thus, of stress on learning and functional plasticity in general.


Unconventional oil and gas chemicals and wastewater-impacted water samples promote adipogenesis via PPARγ-dependent and independent mechanisms in 3T3-L1 cells.
Unconventional oil and natural gas (UOG) operations have contributed to a surge in domestic oil and natural gas production in the United States, combining horizontal drilling with hydraulic fracturing to unlock previously inaccessible fossil fuel deposits. >1000 organic chemicals are used in the production process, and wastewater is produced following injection and for the life of the producing well. This wastewater is typically disposed of via injecting into disposal wells for long-term storage, treatment and discharge from wastewater treatment plants, and/or storage in open evaporation pits; however, wastewater spill rates are reported at 2-20% of active well sites across regions, increasing concerns about the environmental impacts of these wastewaters. This study assessed adipogenic activity (both triglyceride accumulation and pre-adipocyte proliferation) for a mixture of 23 commonly used UOG chemicals and a small subset of UOG wastewater-impacted surface water extracts from Colorado and West Virginia, using 3T3-L1 cells and a peroxisome proliferator activated receptor gamma (PPARγ) reporter assay. We report potent and efficacious adipogenic activity induced by both a laboratory-created UOG chemical mixture and UOG-impacted water samples at concentrations below environmental levels. We further report activation of PPARγ at similar concentrations for some samples, suggesting a causative molecular pathway for the observed effects, but not for other adipogenic samples, implicating PPARγ-dependent and independent effects from UOG associated chemicals. Taken together, these results suggest that UOG wastewater has the potential to impact metabolic health at environmentally relevant concentrations.


Unusual muscle pathology in McLeod syndrome.
Muscle pathology in McLeod syndrome is usually mild; patchy necrotic or regenerating fibres, occasional internal nuclei, and the absence of an inflammatory cell infiltrate are the usual findings. We report on a 29 year old man presenting with chronic fatiguability and excessive sweating in whom an open quadriceps muscle biopsy demonstrated grouped necrotic fibres accompanied by striking patchy mononuclear cell infiltrates. The diagnosis of McLeod syndrome was made on the basis of red blood cell acanthocytosis, raised serum creatine kinase, and weak expression of Kell blood group antigens. The quadriceps muscle infiltrate consisted principally of histologically typical macrophages. These cells had prominent nucleoli, displayed numerous mitoses, and were strongly CD68+. A small population of typical CD3+, CD43+ lymphocytes was also present. In addition, a small population of large atypical CD3+ cells was noted. Immunoperoxidase stains for CD20, CD30, CD79a, and CD56 were negative. Immunocytochemical studies for the common muscular dystrophies were normal. The muscle biopsy findings highlight a potential for confusion of this condition with idiopathic polymyositis. The expanding range of muscle pathology reported in McLeod syndrome, to which this case adds, may reflect variable involvement of the XK gene on chromosome Xp21, or of the adjacent loci of Duchenne muscular dystrophy and chronic granulomatous disease.


Photophysical characterization of anticancer drug valrubicin in rHDL nanoparticles and its use as an imaging agent.
Nanoparticles are target-specific drug delivery agents that are increasingly used in cancer therapy to enhance bioavailability and to reduce off target toxicity of anti-cancer agents. Valrubicin is an anti-cancer drug, currently approved only for vesicular bladder cancer treatment because of its poor water solubility. On the other hand, valrubicin carrying reconstituted high density lipoprotein (rHDL) nanoparticles appear ideally suited for extended applications, including systemic cancer chemotherapy. We determined selected fluorescence properties of the free (unencapsulated) drug vs. valrubicin incorporated into rHDL nanoparticles. We have found that upon encapsulation into rHDL nanoparticles the quantum yield of valrubicin fluorescence increased six fold while its fluorescence lifetime increased about 2 fold. Accordingly, these and potassium iodide (KI) quenching data suggest that upon incorporation, valrubicin is localized deep in the interior of the nanoparticle, inside the lipid matrix. Fluorescence anisotropy of the rHDL valrubicin nanoparticles was also found to be high along with extended rotational correlation time. The fluorescence of valrubicin could also be utilized to assess its distribution upon delivery to prostate cancer (PC3) cells. Overall the fluorescence properties of the rHDL: valrubicin complex reveal valuable novel characteristics of this drug delivery vehicle that may be particularly applicable when used in systemic (intravenous) therapy.


Injection of Peptide nanogels preserves postinfarct diastolic function and prolongs efficacy of cell therapy in pigs.
Accumulating evidence suggests that the benefits of cell therapy for cardiac repair are modest and transient due to progressive harmful cardiac remodeling as well as loss of transplanted cells. We previously demonstrated that injection of peptide nanofibers (NFs) reduces ventricular remodeling and facilitates cell retention at 1 month after acute myocardial infarction (MI) in pigs. However, it remains unclear whether these benefits still persist as the material is being degraded. In this study, 2 mL of placebo or NFs, with or without 1×10(8) mononuclear cells (MNCs), was injected into the pig myocardium after MI (n≥5 in each group), and cardiac function was assessed by echocardiography, including myocardial deformation analyses and catheterization at 3 months post-MI. Our results reveal that MNC-only injection slightly improved cardiac systolic function at 1 month post-MI, but this benefit was lost at later time points (ejection fraction: 42.0±2.3 in MI+normal saline [NS] and 43.5±1.1 in MI+MNCs). In contrast, NF-only injection resulted in improved cardiac diastolic function and reduced pathological remodeling at 3 months post-MI. Furthermore, combined injection of MNCs/NFs provided a greater and longer term cardiac performance (52.1±1.2 in MI+MNCs/NFs, p<0.001 versus MI+NS and MI+MNCs) and 11.3-fold transplanted cell retention. We also found that about 30% NFs remained at 3 months after injection; however, endogenous myofibroblasts were recruited to the NF-injected microenvironment to replace the degraded NFs and preserved cardiac dimensions and mechanics. In conclusion, we demonstrated that injection of NFs contributes to preservation of ventricular mechanical integrity and sustains MNC efficacy at 3 months postinjection.


Comparison of four methods for DNA extraction from Echinococcus granulosus protoscoleces.
Cystic echinococcosis caused by the dog tapeworm Echinococcus granulosus, is a global zoonotic infection which is economically important and constitutes a major threat to public health in many countries, especially in Middle Eastern countries. Strain characterization is essential for the establishment of a preventive and control strategy in every endemic area. Before all molecular studies, it is necessary to achieve DNA of the parasite. The aim of this study was to compare four simple methods of DNA extraction from protoscolecses of E. granulosus in samples from the West Azerbaijan province of Iran. After collecting sheep and cow hydatid cysts from several slaughterhouses of the province, DNA samples were extracted using four different methods involving the use of glass beads, commercial DNA extraction kit, boiling and crushing. For all DNA samples extracted, electrophoresis on 1.3% agarose gel was performed. The methods were compared regarding DNA, time and cost effectiveness and facility. All methods showed some advantages and disadvantages. The commercial kit method, which was the most time and cost effectiveness method, achieved no bands and glass beads method had the best results on gel electrophoresis. Glass beads and boiling methods were the most suitable methods regarding their ease, quickness and cost effectiveness.


Colonic smooth muscle cells and colonic motility patterns as a target for irritable bowel syndrome therapy: mechanisms of action of otilonium bromide.
Otilonium bromide (OB) is a spasmolytic compound of the family of quaternary ammonium derivatives and has been successfully used in the treatment of patients with irritable bowel syndrome (IBS) due to its specific pharmacodynamic effects on motility patterns in the human colon and the contractility of colonic smooth muscle cells. This article examines how. OB inhibits the main patterns of human sigmoid motility in vitro, which are spontaneous rhythmic phasic contractions, smooth muscle tone, contractions induced by stimulation of excitatory motor neurons and contractions induced by direct effect of excitatory neurotransmitters. It does this mainly by blocking calcium influx through L-type calcium channels and interfering with mobilization of cellular calcium required for smooth muscle contraction, thereby limiting excessive intestinal contractility and abdominal cramping. OB also inhibits T-type calcium channels and muscarinic responses. Finally, OB inhibits tachykinin receptors on smooth muscle and primary afferent neurons which may have the joint effect of reducing motility and abdominal pain. All these mechanisms mediate the therapeutic effects of OB in patients with IBS and might be useful in patients with other spastic colonic motility disorders such as diverticular disease.


Direct analysis of biological samples using extractive electrospray ionization mass spectrometry (EESI-MS).
Mass spectrometry (MS) is one of the most widely used techniques for the analysis of biological samples. In the past decade, a novel improvement in MS was the invention of ambient ionization which stands out owing to its unique capability of direct analysis of complex samples with no or minimal pretreatment. In this review, extractive electrospray ionization (EESI), a representative ambient ionization technique, is introduced focusing on its mechanism, instrumentation, and applications in biological analysis. EESI uses a traditional ESI channel to produce primary ions which subsequently ionize neutral chemicals from the sample introduction channel through an online extraction process. When analyzing biological samples, EESI has advantages of rapid analysis, high matrix tolerance, and the ability to perform in vivo analysis. According to previous studies, EESI is able to directly analyze various chemicals in complex biological specimens in liquid, gas, and solid states. EESI can provide a sensitive and selective measurement of biological samples for both qualitative and quantitative purposes. Therefore, it is anticipated that EESI will have promising applications, especially in fields which require the fast and/or in vivo analysis of biological samples with complicated matrixes.


Serotonin transporter (5-HTTLPR) and monoamine oxidase (MAOA) promoter polymorphisms in women with severe alcoholism.
The serotonin system is known to play a pivotal role for mood, behaviour and psychic illness as e.g. alcoholism. Alcoholism in both males and females has been associated with polymorphisms in genes encoding for proteins of importance for central serotonergic function. Genotyping of two functional polymorphisms in the promoter region of the serotonin transporter and monoamine oxidase-A, respectively, (5-HTT-LPR and MAOA-VNTR), was performed in a group of women with severe alcohol addiction. A large sample of adolescent females from a normal population was used as controls. A significantly higher frequency of the LL 5-HTT genotype (high activity) was found in female addicts without a known co-morbid psychiatric disorder than in the controls. Genotype of the MAOA-VNTR polymorphism did not differ significantly between addicts and controls. However, within the group of alcoholics, when the patients with known co-morbid psychiatric disorders were excluded, aggressive anti-social behaviour was significantly linked to the presence of the high activity MAOA allele. The pattern of associations between genotypes of 5-HTT-LPR and MAOA-VNTR in women with severe alcoholism differs from most corresponding studies on males.


Unexpected long-term protection of adult offspring born to high-fat fed dams against obesity induced by a sucrose-rich diet.
Metabolic and endocrine environment during early life is crucial for metabolic imprinting. When dams were fed a high fat diet (HF diet), rat offspring developed hypothalamic leptin resistance with lean phenotype when weaned on a normal diet. Interestingly, when grown on the HF diet, they appeared to be protected against the effects of HF diet as compared to offspring of normally fed dams. The mechanisms involved in the protective effect of maternal HF diet are unclear. We thus investigated the impact of maternal high fat diet on offspring subjected to normal or high palatable diet (P diet) on metabolic and endocrine parameters. We compared offspring born to dams fed P or HF diet. Offspring born to dams fed control or P diet, when fed P diet exhibited a higher body weight, altered hypothalamic leptin sensitivity and metabolic parameters suggesting that maternal P diet has no protective effect on offspring. Whereas, maternal HF diet reduces body weight gain and circulating triglycerides, and ameliorates corpulence index of offspring, even when subjected to P diet. Interestingly, this protective effect is differently expressed in male and female offspring. Male offspring exhibited higher energy expenditure as mirrored by increased hypothalamic UCP-2 and liver AdipoR1/R2 expression, and a profound change in the arcuate nucleus astrocytic organization. In female offspring, the most striking impact of maternal HF diet is the reduced hypothalamic expression of NPY and POMC. HF diet given during gestation and lactation protects, at least partially, offspring from excessive weight gain through several mechanisms depending upon gender including changes in arcuate nucleus astrocytic organization and increased hypothalamic UCP-2 and liver AdipoR1/2 expression in males and reduced hypothalamic expression of NPY and POMC in females. Taken together our results reveal new mechanisms involved in the protective effect of maternal HF diet.


Technological developments and approaches to improving service quality.
In summary, major paradigm shifts in the health care industry are altering the way technology is maintained and supported. Service organizations are now responsible for maintaining a broader base of technology within the health care delivery network and must to this on an extremely rapid, efficient, and productive basis. A number of new technologies are coming on-line, which can allow a health care technology service organization to experience significant improvements in profitability, efficiency, and productivity. To realize maximum benefit from these technologies, service organizations may find themselves re-engineering their service processes. The author believes that this is a requirement for many service organizations, regardless of whether new technology is implemented. The traditional approaches to service delivery are ineffective in managing the new structural realities and service requirements of today's health care environment. New strategies and tactics are required for ensuring that these requirements are met. These approaches will no doubt improve the overall quality, productivity, and efficiency of service and are based on best practices utilized by leading OEMs and ISOs in the medical electronics and other high technology service industry such as information technology and telecommunications, where the service organization is responsible for supporting a broad array of the technology over a large geography with a densely populated installed base, not unlike the typical health care delivery service environment. Once operational improvements are made, a service organization can take advantage of the productivity and efficiency gains brought on by new technology. Organizations interested in doing so are urged to thoroughly research the current state-of-the-art and best practices, because there are numerous systems currently available off-the-shelf. The author believes that new technology will be a basic requirement for competing in the health care technology service marketplace, because it can significantly affect the profitability of service organizations. This technology will help level the playing field between ISOs, OEMs, and biomedical personnel. As our research suggests, efficiently operating biomedical personnel can achieve a significantly higher utilization and profitability than efficiently operating OEMs, due to the advantages of lower overhead and operating cost structure. In general, the process to improve service productivity and efficiency involves a review of current service operations and understanding of the customer environment perceptions as well as understanding of key service factors parameters. From there, service organizations should identify the current state-of-the-art service and infrastructure systems and technology. Based on this assessment, a service organization can evaluate best practices and identify new strategies and tactics for improving service delivery. Through better service management control and education of users on the improvement in service, which the new processes and technologies provide, the service organization can realize real, quantifiable improvements in service quality, productivity, and profitability.


miR-9a prevents apoptosis during wing development by repressing Drosophila LIM-only.
Loss of Drosophila mir-9a induces a subtle increase in sensory bristles, but a substantial loss of wing tissue. Here, we establish that the latter phenotype is largely due to ectopic apoptosis in the dorsal wing primordium, and we could rescue wing development in the absence of this microRNA by dorsal-specific inhibition of apoptosis. Such apoptosis was a consequence of de-repressing Drosophila LIM-only (dLMO), which encodes a transcriptional regulator of wing and neural development. We observed cell-autonomous elevation of endogenous dLMO and a GFP-dLMO 3'UTR sensor in mir-9a mutant wing clones, and heterozygosity for dLMO rescued the apoptosis and wing defects of mir-9a mutants. We also provide evidence that dLMO, in addition to senseless, contributes to the bristle defects of the mir-9a mutant. Unexpectedly, the upregulation of dLMO, loss of Cut, and adult wing margin defects seen with mir-9a mutant clones were not recapitulated by clonal loss of the miRNA biogenesis factors Dicer-1 or Pasha, even though these mutant conditions similarly de-repressed miR-9a and dLMO sensor transgenes. Therefore, the failure to observe a phenotype upon conditional knockout of a miRNA processing factor does not reliably indicate the lack of critical roles of miRNAs in a given setting.


Weighted least squares training of support vector classifiers leading to compact and adaptive schemes.
An iterative block training method for support vector classifiers (SVCs) based on weighted least squares (WLS) optimization is presented. The algorithm, which minimizes structural risk in the primal space, is applicable to both linear and nonlinear machines. In some nonlinear cases, it is necessary to previously find a projection of data onto an intermediate-dimensional space by means of either principal component analysis or clustering techniques. The proposed approach yields very compact machines, the complexity reduction with respect to the SVC solution is especially notable in problems with highly overlapped classes. Furthermore, the formulation in terms of WLS minimization makes the development of adaptive SVCs straightforward, opening up new fields of application for this type of model, mainly online processing of large amounts of (static/stationary) data, as well as online update in nonstationary scenarios (adaptive solutions). The performance of this new type of algorithm is analyzed by means of several simulations.


[Deficits in management of patients with colorectal carcinoma in Germany. Results of multicenter documentation of therapy algorithms].
Adjuvant chemotherapy for patients with UICC III (Dukes C) colorectal cancer (consensus statements NIH 1990, German Cancer Society 1994) and palliative chemotherapy for metastatic disease have long been recognized to provide a survival benefit in colorectal cancer. However, it remains unclear if these concepts have made inroads into clinical practice. Therefore, we asked 74 institutions treating colorectal cancer patients in Germany to document the treatment algorithms of all patients with colorectal cancer seen in the third quarter of 1998. Clinical careers of 1,001 patients (m/f 465/536; median age 62.9 years [28-93]; colon 596, rectum 405; UICC I 117, II 206, III 407, IV 218) were documented. Only 63.4% of patients with UICC III colorectal cancer received adjuvant therapy with a significant difference between hospitals with (67.1%) and without (42.6%) oncological departments (p < 0.01). Higher age appeared to be the most important factor for withholding treatment since 196 of 286 (68.5%) patients < 70 years, but only 57 of 121 (47.1%) > 70 years underwent adjuvant therapy. 78.4% of patients with UICC IV colorectal cancer (91.8% university hospitals, 76.8% hospital with, 50% without oncological departments, 66.7% rehabilitation clinics, 82.4% private practices) received palliative chemotherapy (first line: 5-FU/FA bolus 57%, 5-FU/FA infusion 20%, 5-FU mono 15%). Considering an annual incidence of colorectal cancer in Germany of 52,000 with 30% UICC III, discounting patients > 80 years or ECOG status > 2, and estimating a survival benefit of 10% after adjuvant chemotherapy, approximately 530 lifes are lost annually in Germany due to insufficient treatment of UICC III colorectal cancer based on our survey. In addition, substantial financial demand is generated by the subsequent palliative treatment of potentially curable patients.--In conclusion, survey-based analysis of treatment algorithms can provide valuable insights into clinical practice in oncology and can disclose deficits in patient care as demonstrated here in colorectal cancer.


An insertion/deletion polymorphism of the dihydrofolate reductase (DHFR) gene is associated with serum and red blood cell folate concentrations in women.
A low serum folate and high homocysteine phenotype is associated with an increased risk of neural tube defects (NTDs), cardiovascular diseases and other pathologies. Thus defining both genetic and non-genetic factors that may impact folate/homocysteine metabolism will enhance our understanding of the etiologic mechanisms underlying these conditions and facilitate risk assessment. Dihydrofolate reductase catalyzes the reduction of folic acid to dihydrofolate and thereafter to tetrahydrofolate. The impact of the dihydrofolate reductase (DHFR) c.86 + 60_78 insertion/deletion (ins/del) polymorphism on folate and homocysteine concentrations was analyzed using data from healthy young adults from Northern Ireland, collected as part of visit three of the Young Hearts Project. Among men the DHFR c.86 + 60_78 polymorphism was not significantly associated with serum or red blood cell folate concentrations, or with homocysteine concentrations. Among women the DHFR c.86 + 60_78 polymorphism explained 2% of the variation in RBC folate levels and 5% of the variation in serum folate levels, but did not appear to have an independent effect on homocysteine. Relative to women with the DHFR c.86 + 60_78 ins/ins and ins/del genotypes, del/del homozygotes had increased serum and red blood cell folate concentrations and may therefore be at decreased risk of having offspring affected by NTDs and of other adverse reproductive and health outcomes attributable to low folate.


Prophylactic antibiotic use: hardwiring of physician behavior, not education, leads to compliance.
Surgical wound infections can be significantly reduced by appropriate administration of prophylactic antibiotics (ProAbx). The purpose of this study was to determine the effect of interventions, specifically, forced functions, designed to improve administration of ProAbx in a university hospital setting. We retrospectively reviewed charts from 1,622 consecutive patients, seen between March 2005 and June 2007, for evaluation of ProAbx administration to determine correct choice, appropriate timing of administration, and appropriate postoperative cessation of antibiotics. Processes designed to improve compliance were devising orders to limit ProAbx choice; requiring those orders in preadmission testing; administering ProAbx in the preparation and hold area, subsequently, at the time of the universal timeout; and standardizing postoperative orders in the computerized physician order entry. Specialty-specific standardized orders improved compliance from 76% to 91% (p < 0.001) concerning ProAbx choice. Regarding timing of preoperative administration, a baseline compliance of 55% improved to: 78% (p < 0.001) on requiring orders in the preadmission testing area; 90% (p < 0.008) on administering ProAbx in the preparation and hold area; and 95% (p < 0.07) when ProAbx were administered at the universal timeout. Standardization of postoperative orders in the computerized physician order entry improved compliance with cessation from 60% to 86%, p < 0.001. Despite hospital-wide education, improving compliance with evidence-based recommendations for ProAbx required processes that "forced" physician behavior, specifically: specialty-specific, preprinted physician orders limiting ProAbx choice; linking administration of preoperative ProAbx administration to the universal timeout; and standardization of the postoperative ProAbx order in the computerized physician order entry.


Cerebral palsy in a term population: risk factors and neuroimaging findings.
The purpose of this work was to study risk factors and neuroimaging characteristics of cerebral palsy in term and near-term infants. Among a cohort of 334,339 infants > or = 36 weeks' gestation born at Kaiser Permanente Medical Care Program in northern California in 1991-2003, we identified infants with cerebral palsy and obtained clinical data from electronic and medical charts. Risk factors for cerebral palsy among infants with different brain abnormalities were compared using polytomous logistic regression. Of 377 infants with cerebral palsy (prevalence: 1.1 per 1000), 273 (72%) received a head computed tomography or MRI. Abnormalities included focal arterial infarction (22%), brain malformation (14%), and periventricular white matter abnormalities (12%). Independent risk factors for cerebral palsy were maternal age > 35, black race, and intrauterine growth restriction. Intrauterine growth restriction was more strongly associated with periventricular white matter injury than with other neuroimaging findings. Nighttime delivery was associated with cerebral palsy accompanied by generalized brain atrophy but not with cerebral palsy accompanied by other brain lesions. Cerebral palsy is a heterogeneous syndrome with focal arterial infarction and brain malformation representing the most common neuroimaging abnormalities in term and near-term infants. Risk factors for cerebral palsy differ depending on the type of underlying brain abnormality.


[Historical dynamics of age-related mortality: 1. Evolution of the specific lifespan of the Finnish population].
Historical dynamics of parameters of age-related mortality of Finnish men and women for the period 1952 to 2000 is studied. The Gompertz equation is found to describe age-related mortality of men quite exactly during the historical interval 1952 to 1989. From 1990 to 2000 for men and during the entire interval for women the Gompertz-Makeham equation is more exact. The constant component of mortality is found to be increasing since 1990 for men and since 1980 for women. The presence of the Strehler-Mildvan correlation is shown. There are 3 evidently separated periods: 1) from 1952 to 1971, 2) from 1972 to 1980, 3) from 1981 to 2000. The correlation has the highest value in the 1st and 3rd periods. During the 2nd, transitional, period the correlation value is minimal. An opinion is expressed that it's not correct to treat the incline coefficient of correlation dependence as a specific lifespan since it takes on different values even for a single country, though the values are stable during certain historical periods.


Hitting a Moving Target: Simulation and Crystallography Study of ATAD2 Bromodomain Blockers.
Small molecule ligand binding to the ATAD2 bromodomain is investigated here through the synergistic combination of molecular dynamics and protein crystallography. A previously unexplored conformation of the binding pocket upon rearrangement of the gatekeeper residue Ile1074 has been found. Further, our investigations reveal how minor structural differences in the ligands result in binding with different plasticity of the ZA loop for this difficult-to-drug bromodomain.


Epilepsy as a manifestation of neurocysticercosis.
Epilepsy is the commonest manifestation of neurocysticercosis. Epilepsy was observed in 127 of 150 cases (84.7%) of neurocysticercosis seen over a period of 17 years. The basis of diagnosis was clinical presentation and concomitant evidence of extraneural cysticercosis in the pre-computed tomography (CT) scan era, and typical CT findings in later years. Eighty one cases (54%) who primarily presented as epilepsy without any neurological deficit have been analysed in detail. In the pre-CT period the occurrence of epilepsy as a presenting feature in neurocysticercosis was 43.5% whereas in later years it was 61.4%. Primary generalised seizure (49 cases) was more common than partial seizure (29) and partial complex seizure (3). Status epilepticus was seen in 6 cases. Magnetic resonance imaging, done in 8 cases, proved to be more sensitive in demonstrating various stages in the development of noncalcified cysticercosis. The new larvicidal drugs offer a potential cure and cysticercosis should be entertained as a cause of epilepsy especially in endemic areas.


Switch from antagonist to agonist after addition of a DOTA chelator to a somatostatin analog.
Peptide receptor targeting has become an increasingly attractive method to target tumors diagnostically and radiotherapeutically. Peptides linked to a variety of chelators have been developed for this purpose. They have, however, rarely been tested for their agonistic or antagonistic properties. We report here on a somatostatin antagonist that switched to an agonist upon coupling to a DOTA chelator. Two novel somatostatin analogs, 406-040-15 and its DOTA-coupled counterpart 406-051-20, with and without cold Indium labeling, were tested for their somatostatin receptor subtypes 1-5 (sst(1)-sst(5)) binding affinity using receptor autoradiography. Moreover, they were tested functionally for their ability to affect sst(2) and sst(3) internalization in vitro in HEK293 cells stably expressing the human sst(2) or sst(3) receptor, using an immunofluorescence microscopy-based internalization assay. All three compounds were characterized as pan-somatostatin analogs having a high affinity for all five sst. In the sst(2) internalization assay, all three compounds showed an identical behavior, namely, a weak agonistic effect complemented by a weak antagonistic effect, compatible with the behavior of a partial agonist. Conversely, in the sst(3) internalization assay, 406-040-15 was a full antagonist whereas its DOTA-coupled counterpart, 406-051-20, with and without Indium labeling, switched to a full agonist. Adding the DOTA chelator to the somatostatin analog 406-040-15 triggers a switch at sst(3) receptor from an antagonist to an agonist. This indicates that potential radioligands for tumor targeting should always be tested functionally before further development, in particular if a chelator is added.


Describing the pattern of the COVID-19 epidemic in Vietnam.
Given the rapid spread of the COVID-19 pandemic and the huge negative impacts it is causing, researching on COVID-19-related issues is very important for designing proactive and comprehensive public health interventions to fight against the pandemic. We describe the characteristics of COVID-19 patients detected in the two phases of the epidemic in Vietnam. Data used in this paper were mainly obtained from the official database of the Ministry of Health of Vietnam. Descriptive statistics were carried out using Stata 16 software. As of 18 May 2020, the cumulative number of COVID-19 cases detected in Vietnam was 324, 16 cases from 4 cities and provinces in the first phase (during 20 days, 0.8 cases detected per day) and 308 cases from 35 cities, provinces in the second phase (during 76 days, 4.1 cases detected per day). Vietnam has mobilized its entire political system to fight the COVID-19 and achieved some initial successes. We found both similarities and differences between the two phases of the COVID-19 epidemic in Vietnam. We demonstrated that the situation of the COVID-19 epidemic in Vietnam is getting more complicated and unpredictable.


Preliminary field evaluation of the combined adulticidal, larvicidal, and wall residual activity of ULV-applied bifenthrin against mosquitoes.
The combined adulticidal, larvicidal, and wall residual activity of ULV-applied bifenthrin, a synthetic pyrethroid, was evaluated in houses in Kuala Lumpur, Malaysia, against larvae and adults of lab-bred Aedes aegypti and Culex quinquefasciatus. A portable ULV sprayer was used to disperse a ULV formulation of bifenthrin at a discharge rate of 45 ml/min. The results indicated that bifenthrin sprayed at this rate exhibited all the three activities against the test mosquitoes. Complete adult mortalities were achieved, while very high larvicidal activity was also effected, which persisted for seven days. Wall bioassay with adults of Ae aegypti also resulted in very high mortality, which also persisted for one week. The combined mosquitocidal activities of bifenthrin is considered more effective especially in the control of dengue vectors.


Impaired Cerebrovascular Reactivity Predicts Recurrent Symptoms in Patients with Carotid Artery Occlusion: A Hypercapnia BOLD fMRI Study.
A key factor in predicting recurrent ischemic episodes in patients with carotid artery occlusion is the presence of hemodynamic impairment. There is, however, no consensus on how to best assess this risk in terms of imaging modalities or thresholds used. Here we investigated whether a predefined threshold of hemispheric asymmetry in hypercapnia fMRI predicts recurrent symptoms in patients with carotid artery occlusion. We studied 23 patients (2 women) with a mean age of 67.5 ± 9 years. Patients were assessed for recurrent ischemic events until lost to follow-up, study end, death, or recurrent ischemic event. Hypercapnia fMRI was used to assess the cerebrovascular reserve and quantify the percentage signal change in GM in the MCA territory and the hemispheric asymmetry index. Kaplan-Meier survival analysis and log-rank tests were performed to assess differences between patients with normal or abnormal hemispheric indices. The median follow-up was 20 months. During this period, 8 patients experienced recurrent events, and 15 did not. The percentage signal change in GM in the MCA territory was significantly decreased in those patients with recurrent events compared with those without (2.39 ± 0.22 versus 2.70 ± 0.42, P = .032). The normal hemispheric index predicted event-free survival during follow-up (median, 20 months) for both the combined outcome (recurrent events and/or death, log-rank, P = .034) and recurrent retinal or ipsilateral ischemic events only (log-rank, P = .012). The hemispheric asymmetry index derived from hypercapnia fMRI showed hemodynamic impairment in more than half of the studied patients with carotid occlusion, and those patients showed a higher risk of recurrent ischemic symptoms.


Epidemiologic indicators of health status to guide health care management decision making.
Health objectives, developed by the United States Department of Health and Human Services, were published recently in the document entitled Healthy People 2000: National Health Promotion and Disease Prevention Objectives. They were developed to guide national and local health policy toward actions to increase the health of the nation. To effectively apply these objectives locally, epidemiologists and health planners must work together. Through collaboration, the Healthy People 2000 objectives can be prioritized to guide health policy and planning on a regional basis. The purpose of this study was to assess certain health status indicators in southwestern Pennsylvania to determine whether it was likely that the year 2000 targets would be met if trends from the past 20 years were to continue. The following mortality rates were analyzed: heart disease, homicide, breast cancer, colon cancer, lung cancer, suicide, motor vehicle accidents, work-related injury, and infant mortality. In addition, incidence of the following diseases was evaluated against the year 2000 targets: gonorrhea, primary and secondary syphilis, measles, tuberculosis, and AIDS. By employing epidemiological principles and considering strategic planning needs, it is possible to prioritize some of the health care needs in local areas for the next decade.


Disruption of the elastin gene in adult Williams syndrome is accompanied by a paradoxical reduction in arterial stiffness.
Although the aetiology of Williams syndrome (WS) is related to elastin gene disruption, its pathogenesis remains unknown, particularly that of vascular lesions. The aim of the present study was to compare the elastic properties of three WS patients with age- and gender-matched normotensive and hypertensive controls. Common carotid arteries of WS patients had a higher distensibility, a thicker intima-media and a lower elastic modulus. Electron microscopy studies of one WS renal artery showed major abnormalities of the elastic fibres, which displayed a reticular structure and a thickening of the internal elastic lamina, whereas the ultrastructure of elastic fibres was normal in a control subadventitial muscular fibrodysplasia. In this WS arterial stenosis, we studied the expression patterns of several major smooth muscle (SM) phenotypic markers using immunofluorescence and used a normal renal artery as a control. In WS, SM-alpha-actin- and myosin-heavy-chain-positive cells contained low amounts of heavy caldesmon, and laminin-beta1 chain was expressed into the basement membranes, indicating a less differentiated phenotype. In conclusion, in WS patients, the carotid artery wall was abnormally distensible and thick, and major ultrastructural abnormalities of elastic fibres were observed in association with smooth muscle cell de-differentiation. These results indicate that the haplo-insufficiency of the elastin gene in WS patients leads to abnormal elastic fibre assembly within the media. Arterial wall hypertrophy found with a primary defect in elastin may represent a major factor responsible for increased distensibility. We suggest that, in WS, the increased proliferative response and the associated de-differentiation process represent two important mechanisms underlying the matrix accumulation and the development of arterial stenosis.


Biochemical characterization of the Nocardia lactamdurans ACV synthetase.
The L-δ-(α-aminoadipoyl)-L-cysteinyl-D-valine synthetase (ACVS) is a nonribosomal peptide synthetase (NRPS) that fulfills a crucial role in the synthesis of β-lactams. Although some of the enzymological aspects of this enzyme have been elucidated, its large size, at over 400 kDa, has hampered heterologous expression and stable purification attempts. Here we have successfully overexpressed the Nocardia lactamdurans ACVS in E. coli HM0079. The protein was purified to homogeneity and characterized for tripeptide formation with a focus on the substrate specificity of the three modules. The first L-α-aminoadipic acid-activating module is highly specific, whereas the modules for L-cysteine and L-valine are more promiscuous. Engineering of the first module of ACVS confirmed the strict specificity observed towards its substrate, which can be understood in terms of the non-canonical peptide bond position.


Early-life establishment of the swine gut microbiome and impact on host phenotypes.
Early bacterial colonization and succession within the gastrointestinal tract has been suggested to be crucial in the establishment of specific microbiota composition and the shaping of host phenotype. Here, the composition and dynamics of faecal microbiomes were studied for 31 healthy piglets across five age strata (days 14, 36, 48, 60 and 70 after birth) together with their mothers. Faecal microbiome composition was assessed by 16S rRNA gene 454-pyrosequencing. Bacteroidetes and Firmicutes were the predominant phyla present at each age. For all piglets, luminal secretory IgA concentration was measured at day 70, and body weight was recorded until day 70. The microbiota of suckling piglets was mainly represented by Bacteroides, Oscillibacter, Escherichia/Shigella, Lactobacillus and unclassified Ruminococcaceae genera. This pattern contrasted with that of Acetivibrio, Dialister, Oribacterium, Succinivibrio and Prevotella genera, which appeared increased after weaning. Lactobacillus fermentum might be vertically transferred via breast milk or faeces. The microbiota composition coevolved with their hosts towards two different clusters after weaning, primarily distinguished by unclassified Ruminococcaceae and Prevotella abundances. Prevotella was positively correlated with luminal secretory IgA concentrations, and body weight. Our study opens up new possibilities for health and feed efficiency manipulation via genetic selection and nutrition in the agricultural domain.


Prognostic factors of alveolar rhabdomyosarcoma in childhood. An International Society of Pediatric Oncology study.
Pretreatment characteristics of 57 patients with alveolar rhabdomyosarcoma (RMS) registered by the International Society of Pediatric Oncology (SIOP) between January 1975 and December 1983 were identified to perform a prognostic factor study. Living status and disease-free survival (DFS) time were assessed for all patients on January 1, 1988. By multi-variate analyses, lymph node involvement (P = 0.0003) and tumor invasiveness (P = 0.007) were identified as the most significant covariates correlated with survival. A model including N stage (P = 0.001) and age (P = 0.03) was selected for predicting DFS rates. Children between 5 and 10 years of age had better DFS rates than those younger than 5 years of age or those older than 10 years of age. The authors suggest a prognostic classification that could allow treatment to be adjusted according to clinical staging.


A proposal for a new arteriovenous malformation grading scale for neuroendovascular procedures and literature review.
Traditionally, decisions regarding treatment and outcomes for patients with cerebral arteriovenous malformations (AVM) have made use of the Spetzler-Martin grading scale. The latter has withstood the test of time in clinical practice for AVM patients managed surgically and even when comparing studies involving other modalities of treatment. Recent awareness on the applicability of the grading system for risk assessment and outcome determination in cases of treatment by neuroendovascular means has emerged. We propose a preliminary grading system for neuroendovascular procedures based on a revision of the available literature. A literature search using the keywords 'arteriovenous malformation', 'embolization' and 'outcome' was done. Articles studying the factors involved in complications and outcome determination for endovascular cerebral AVM patients were reviewed. These were tabulated and those dealing with anatomical, radiological and hemodynamic descriptions that were noted as significant determinants of risk or clinical outcome were used for development of a preliminary grading system to be used in a follow-up validation study. A grading system similar to the Spetzler-Martin grading scale was developed using factors deemed in the literature as significant determinants of outcome. The classification includes the number of feeding vessels into the AVM, the eloquence of adjacent areas, and the presence of fistulous components. Follow-up study is underway at our institution to validate our proposal. Yet, significant evidence exists in the literature validating those factors as stand alone determinants of outcome and risk, suggesting that this grading scale may well be applicable to endovascular embolization procedures. A grading scale similar to the Spetzler-Martin grading system for use in risk assessment and outcome determination in brain AVM patients treated by endovascular techniques seems adequate and clinically feasible. Studies on applicability and validation are underway.


Listeria monocytogenes serotype identification by PCR.
Serotyping is a universally accepted subtyping method for Listeria monocytogenes. Identification of the strain serotype permits differentiation between important food-borne strains (1/2a, 1/2b, and 4b) and provides a "gold standard" for comparing isolates analyzed in different labs and with different techniques. Although an efficient enzyme-linked immunosorbent assay serotyping protocol was described recently, identification of PCR serotyping primers would further increase the ease and accessibility of this classification system. Serotyping PCR primers were designed from variable regions of the L. monocytogenes genome. Three primer sets were used in conjunction with a previously described Division III primer set in order to classify 122 L. monocytogenes strains into five serotype groups [1/2a(3a), 1/2b, 1/2c(3c), 4b(d,e), and 4a/c]. Results of the PCR method agreed with those of the conventional slide agglutination method for 97, 100, 94, and 91% of strains belonging to serotypes 1/2a, 1/2b, 1/2c, and 4b, respectively.


Oncogenic JAK1 and JAK2-activating mutations resistant to ATP-competitive inhibitors.
Activating mutations in JAK1 and JAK2 have been described in patients with various hematologic malignancies including acute lymphoblastic leukemia and myeloproliferative neoplasms, leading to clinical trials with JAK inhibitors. While there has been a tremendous effort towards the development of specific JAK inhibitors, mutations conferring resistance to such drugs have not yet been observed. Taking advantage of a model of spontaneous cellular transformation, we sequenced JAK1 in selected tumorigenic BaF3 clones and identified 25 de novo JAK1 activating mutations, including 5 mutations already described in human leukemias. We further used this library of JAK1 mutation-positive cell lines to assess their sensitivity to ATP-competitive inhibitors. While most JAK1 mutants were sensitive to ATP-competitive JAK inhibitors, mutations targeting Phe958 and Pro960 in the hinge region of the kinase domain rendered JAK1 constitutively active but also resistant to all tested JAK inhibitors. Furthermore, mutation of the homologous Tyr931 in JAK2 wild-type or JAK2 V617F mutant found in patients with myeloproliferative neoplasms also conferred resistance to JAK inhibitors, such as INCB018424, which is currently in clinical use. Our data indicate that some activating mutations not only promote autonomous cell proliferation but also confer resistance to ATP-competitive inhibitors. In vivo, such a mutation can potentially occur as primary JAK-activating mutations but also as secondary mutations combining oncogenicity with drug resistance.


Irritant-induced occupational asthma.
A retrospective review was performed on the files of 154 consecutive workers assessed for occupational asthma to clarify the relative frequency of asthma induced by irritants in the workplace and to determine whether such asthma was clearly distinguishable from other forms of occupational asthma. Fifty-nine workers were considered to have occupational asthma. A subset of ten had a history consistent with asthma initiated by exposure to high concentrations of an irritant, had persistent symptoms for an average of five years when seen, demonstrated increased reactivity to methacholine, and gave no prior history of pulmonary complaints. These ten had a lower incidence of atopy (20 percent vs 58 percent) and a more frequent history of smoking (80 percent vs 38 percent) than the other subjects with occupational asthma but did not differ in average latency (5.9 years vs 5.7 years). Our findings suggest that irritant-induced asthma is not uncommon, and those affected may have different baseline characteristics from others with occupational asthma.


Alterations in serum lipid profile patterns in oral cancer.
Alterations in serum lipids have long been associated with cancer as lipids play an important role in maintenance of cell integrity. To evaluate alterations in plasma lipid profile in oral cancer patients, to compare and correlate the serum lipid profile in different grades of carcinoma and to evaluate the correlation of serum lipid profile between the tobacco habituates and non-habituates. Among 75 study subjects, 50 individuals were oral carcinoma patients and 25 individuals were healthy controls. The parameters assessed included total cholesterol (TC), high-density lipoprotein-cholesterol (HDLC), low-density lipoprotein-cholesterol, very low-density lipoprotein-cholesterol and triglycerides (TGL). These groups were subdivided into subjects with no habit of tobacco (NHT) and subjects with habit of tobacco (WHT). Evaluation of results and statistical analysis was carried out using Student's t-test and one-way Analysis of Variance. There was a significant decrease in TC, HDLC and TGL in the oral cancer group as compared with the control group. The lipid profile levels between histological grading of the oral cancer and between WHT and NHT had no statistical significance. There was an inverse relationship between serum lipid profile and oral cancer. The lower serum lipid status may be considered a useful indicator for initial changes occurring in the neoplastic cells.


Premutations in the FMR1 gene in Serbian patients with undetermined tremor, ataxia and parkinsonism.
Introduction: Although one of the most common monogenic late-onset neurodegenerative disorders, fragile-X-associated tremor/ataxia syndrome (FXTAS) is still underdiagnosed. The aim of the present study was to estimate the frequency of premutation carriers in patients with unexplained degenerative ataxias, action tremor or parkinsonism, and action tremor with or without associated cognitive impairment.Methods: The study comprised 100 consecutive patients with the disease onset >49 years who had any form of unexplained action tremor, cerebellar ataxia, followed by parkinsonism with or without incipient dementia, and in whom the FMR1 repeats size was determined.Results: Premutation in the FMR1 was identified in two patients (2%): the first, male patient had 83 CGG repeats and the second, female patient had 32 and 58 CGG repeats.Discussion/Conclusion: FXTAS was relatively rare among older patients with unexplained ataxia and action tremor, with or without parkinsonism and/or cognitive impairment. Tremor and ataxia were major clinical features in our two patients, although parkinsonism, autonomic dysfunction and psychiatric problems might be an important part of the spectrum. Probable FXTAS should be considered in the differential diagnosis of patients with unexplained action tremor and ataxia, and undetermined parkinsonism, especially when there was a positive family history for involuntary movement disorders in other family members and/or autism spectrum disorders in younger cousins.