Activity-dependent tuning of inhibitory neurotransmission based on GABAAR diffusion dynamics.
An activity-dependent change in synaptic efficacy is a central tenet in learning, memory, and pathological states of neuronal excitability. The lateral diffusion dynamics of neurotransmitter receptors are one of the important parameters regulating synaptic efficacy. We report here that neuronal activity modifies diffusion properties of type-A GABA receptors (GABA(A)R) in cultured hippocampal neurons: enhanced excitatory synaptic activity decreases the cluster size of GABA(A)Rs and reduces GABAergic mIPSC. Single-particle tracking of the GABA(A)R gamma2 subunit labeled with quantum dots reveals that the diffusion coefficient and the synaptic confinement domain size of GABA(A)R increases in parallel with neuronal activity, depending on Ca(2+) influx and calcineurin activity. These results indicate that GABA(A)R diffusion dynamics are directly linked to rapid and plastic modifications of inhibitory synaptic transmission in response to changes in intracellular Ca(2+) concentration. This transient activity-dependent reduction of inhibition would favor the onset of LTP during conditioning.


[Cochleovestibular disorders after diving accidents].
Cochleo-vestibular lesions may result from decompression accidents and barotrauma. The etiology of both clinical syndromes as well as their therapy is discussed on the basis of seven personal observations.


DVV: a taxonomy for mixed reality visualization in image guided surgery.
Mixed reality visualizations are increasingly studied for use in image guided surgery (IGS) systems, yet few mixed reality systems have been introduced for daily use into the operating room (OR). This may be the result of several factors: the systems are developed from a technical perspective, are rarely evaluated in the field, and/or lack consideration of the end user and the constraints of the OR. We introduce the Data, Visualization processing, View (DVV) taxonomy which defines each of the major components required to implement a mixed reality IGS system. We propose that these components be considered and used as validation criteria for introducing a mixed reality IGS system into the OR. A taxonomy of IGS visualization systems is a step toward developing a common language that will help developers and end users discuss and understand the constituents of a mixed reality visualization system, facilitating a greater presence of future systems in the OR. We evaluate the DVV taxonomy based on its goodness of fit and completeness. We demonstrate the utility of the DVV taxonomy by classifying 17 state-of-the-art research papers in the domain of mixed reality visualization IGS systems. Our classification shows that few IGS visualization systems' components have been validated and even fewer are evaluated.


Early cellular responses to intradermal injection of Kveim suspension in normal subjects and those with sarcoidosis.
In a detailed controlled study of the cellular response to Kveim suspension in vivo we used immunohistological and histochemical methods to examine cryostat sections of immature Kveim biopsy specimens in subjects with sarcoidosis and normal controls. Changes seen at 48 hours, at which time papular reactions have sometimes been reported, are described. Eight cases of sarcoidosis previously confirmed by a positive Kveim test were studied, in five of whom the test remained positive; plus two subjects with sarcoidosis studied prospectively; and four healthy controls. There were two main features of the 48 hour response: collagen disruption with associated histiocytes, which showed increased acid phosphatase activity; and perivascular infiltrates of lymphocytes and small groups of dendritic cells. The T4:T8 ratios in the infiltrates were similar to those found in the peripheral blood of the subjects, and few lymphocytes showed evidence of activation. T lymphocytes were also seen free in the dermis and migrating to the epidermis. Small juxtacapillary clumps of dendritic cells, identified by NA1/34 (= OKT6; Langerhans' cells) and RFD1 (interdigitating cell) monoclonal antibodies, were found. The Langerhans' cells in the epidermis were, however, normal in number and distribution. These features, which were found in all groups, are not consistent with pre-existing hypersensitivity to Kveim suspension in sarcoidosis. Subsequent differences between sarcoid and normal subjects in the development of granulomas in the Kveim response may therefore relate to the different handling of the foreign material by the cells affected, rather than to differences in the early non-specific recruitment of the cells to the test site.


In vitro generation of human high-density-lipoprotein-resistant Trypanosoma brucei brucei.
The host range of African trypanosomes is influenced by innate protective molecules in the blood of primates. A subfraction of human high-density lipoprotein (HDL) containing apolipoprotein A-I, apolipoprotein L-I, and haptoglobin-related protein is toxic to Trypanosoma brucei brucei but not the human sleeping sickness parasite Trypanosoma brucei rhodesiense. It is thought that T. b. rhodesiense evolved from a T. b. brucei-like ancestor and expresses a defense protein that ablates the antitrypanosomal activity of human HDL. To directly investigate this possibility, we developed an in vitro selection to generate human HDL-resistant T. b. brucei. Here we show that conversion of T. b. brucei from human HDL sensitive to resistant correlates with changes in the expression of the variant surface glycoprotein (VSG) and abolished uptake of the cytotoxic human HDLs. Complete transcriptome analysis of the HDL-susceptible and -resistant trypanosomes confirmed that VSG switching had occurred but failed to reveal the expression of other genes specifically associated with human HDL resistance, including the serum resistance-associated gene (SRA) of T. b. rhodesiense. In addition, we found that while the original active expression site was still utilized, expression of three expression site-associated genes (ESAG) was altered in the HDL-resistant trypanosomes. These findings demonstrate that resistance to human HDLs can be acquired by T. b. brucei.


Natural antibodies to interferon-alpha and interferon-beta are a common feature of inbred mouse strains.
Neutralizing antibodies to murine interferon-alpha (MuIFN-alpha) and MuIFN-beta of the immunoglobulin (Ig)G and IgM class were naturally present in sera of BALB/c and C57BL/6 mice of four different age groups, ranging from 1 to 15 mo. These antibodies not only neutralized IFN made by C-243 cells (Swiss genotype), but also isogenic IFN-alpha and IFN-beta, and can therefore be considered to be real autoantibodies. Natural anti-MuIFN antibodies were also found in the sera of mice from 12 other inbred strains, and seem to be a common feature of inbred mouse strains.


Demonstration of a mid-infrared cavity enhanced absorption spectrometer for breath acetone detection.
A high-resolution absorption spectrum of gaseous acetone near 8.2 μm has been taken using both Fourier transform and quantum cascade laser (QCL)-based infrared spectrometers. Absolute absorption cross sections within the 1215-1222 cm(-1) range have been determined, and the spectral window around 1216.5 cm(-1) (σ = 3.4 × 10(-19) cm(2) molecule(-1)) has been chosen for monitoring trace acetone in exhaled breath. Acetone at sub parts-per-million (ppm) levels has been measured in a breath sample with a precision of 0.17 ppm (1σ) by utilizing a cavity enhanced absorption spectrometer constructed from the QCL source and a linear, low-volume, optical cavity. The use of a water vapor trap ensured the accuracy of the results, which have been corroborated by mass spectrometric measurements.


Genotype and interleukin-10 responses after cardiopulmonary bypass.
The pro- and anti-inflammatory cytokine balance has been implicated in outcome from inflammatory conditions, and cardiopulmonary bypass is associated with a marked inflammatory response. Interleukin-10 (IL-10) is an anti-inflammatory cytokine and levels have been shown to be highest in those patients who develop sepsis after trauma or surgery. IL-10 levels vary between individuals and genotype may dictate the IL-10 response. We therefore investigated IL-10 genotype, circulating IL-10 concentrations and outcome in terms of organ dysfunction 24 h after cardiopulmonary bypass. Blood samples were obtained from 150 patients before, and 3, and 24 h after cardiopulmonary bypass. IL-10 was measured by enzyme immunoassay. The single nucleotide polymorphism at -1082 base pairs was detected by restriction fragment length polymorphism analysis. Post-bypass organ system dysfunction was defined prospectively. IL-10 concentrations were increased 3 h after bypass (P<0.0001) and were still increased at 24 h (P<0.0001). Homozygosity for the G allele was associated with lower median (range) maximal IL-10 levels at 3 h (44 (13-136) pg ml(-1)) compared with the A allele (118 (39-472) pg ml(-1); P=0.042). Those patients who developed at least one organ dysfunction (n=33) had higher IL-10 levels 3 h after surgery (242 (18-694) pg ml(-1)) compared with those without organ dysfunction (77 (7-586) pg ml(-1); P=0.001, n=117). The G allele of the -1082 base pair single nucleotide polymorphism in the IL-10 gene is associated with lower IL-10 release after cardiopulmonary bypass. High levels of IL-10 secretion are associated with organ dysfunction 24 h after surgery.


Sodium cromoglycate in asthma therapy. A retrospective survey.
The data obtained in a retrospective survey of the use and effectiveness of sodium cromoglycate (Lomudal; Fisons) therapy in 635 children and young adults support the findings of other long-term studies that this drug is effective and safe in the treatment of children and young adults with clear evidence of allergy contributing towards their asthma. Its administration by Spinhaler can be commenced successfully at 4 years of age or even younger and continued for as long as is required. The drug is worth a therapeutic trial in any patient with asthma requiring regular medication to control symptoms. The absence of demonstrable allergy does not entirely preclude a favourable response. Once a patient's asthma has been stabilized, sodium cromoglycate enables concomitant therapy (especially with beta 2-adrenergic stimulants and corticosteroids) to be reduced.


Spontaneous Behavioural Recovery Following Stroke Relates to the Integrity of Parietal and Temporal Regions.
Stroke is a devastating disease that results in neurological deficits and represents a leading cause of death and disability worldwide. Following a stroke, there is a degree of spontaneous recovery of function, the neural basis of which is of great interest among clinicians in their efforts to reduce disability following stroke and enhance rehabilitation. Conventionally, work on spontaneous recovery has tended to focus on the neural reorganization of motor cortical regions, with comparably little attention being paid to changes in non-motor regions and how these relate to recovery. Here we show, using structural neuroimaging in a macaque stroke model (N = 31) and by exploiting individual differences in spontaneous behavioural recovery, that the preservation of regions in the parietal and temporal cortices predict animal recovery. To characterize recovery, we performed a clustering analysis using Non-Human Primate Stroke Scale (NHPSS) scores and identified a good versus poor recovery group. By comparing the preservation of brain volumes in the two groups, we found that brain areas in integrity of brain areas in parietal, temporal and somatosensory cortex were associated with better recovery. In addition, a decoding approach performed across all subjects revealed that the preservation of specific brain regions in the parietal, somatosensory and medial frontal cortex predicted recovery. Together, these findings highlight the importance of parietal and temporal regions in spontaneous behavioural recovery.


[Psychiatrist's burnout. A survey].
This work seeks to know the state of the Burnout syndrome among Spanish psychiatrists' with the purpose of having some national reference values. It also sought to elucidate the sociodemographic and work-related variables which can influence the burnout syndrome. A survey (which includes sociodemographic, work-related variables and the Maslach Burnout Inventory) is applied to a sample of psychiatrists assisting at the National Congress of Psychiatry held in Valencia. The statistical analysis comprises parametric, non parametric and multivariate methods. The results of the three subscales included in the Maslach Burnout Inventory is offered along their relationship with the sociodemographic and work-related variables studied. The multivariate analysis shows that only the variable sex can influence the probability of obtaining high scores in the depersonalization subscale. The scores obtained in the three subscales of the of Maslach Burnout Inventory are higher than those published to date. These prior studies were carried out with autonomous samples. The possible relationship of these high scores with the current spanish situation is considered.


Outcomes of Infants and Young Children With Relapsed Medulloblastoma After Initial Craniospinal Irradiation-Sparing Approaches: An International Cohort Study.
Infant and young childhood medulloblastoma (iMB) is usually treated without craniospinal irradiation (CSI) to avoid neurocognitive late effects. Unfortunately, many children relapse. The purpose of this study was to assess salvage strategies and prognostic features of patients with iMB who relapse after CSI-sparing therapy. We assembled a large international cohort of 380 patients with relapsed iMB, age younger than 6 years, and initially treated without CSI. Univariable and multivariable Cox models of postrelapse survival (PRS) were conducted for those treated with curative intent using propensity score analyses to account for confounding factors. The 3-year PRS, for 294 patients treated with curative intent, was 52.4% (95% CI, 46.4 to 58.3) with a median time to relapse from diagnosis of 11 months. Molecular subgrouping was available for 150 patients treated with curative intent, and 3-year PRS for sonic hedgehog (SHH), group 4, and group 3 were 60%, 84%, and 18% (P = .0187), respectively. In multivariable analysis, localized relapse (P = .0073), SHH molecular subgroup (P = .0103), CSI use after relapse (P = .0161), and age ≥ 36 months at initial diagnosis (P = .0494) were associated with improved survival. Most patients (73%) received salvage CSI, and although salvage chemotherapy was not significant in multivariable analysis, its use might be beneficial for a subset of children receiving salvage CSI < 35 Gy (P = .007). A substantial proportion of patients with relapsed iMB are salvaged after initial CSI-sparing approaches. Patients with SHH subgroup, localized relapse, older age at initial diagnosis, and those receiving salvage CSI show improved PRS. Future prospective studies should investigate optimal CSI doses and the role of salvage chemotherapy in this population.


Adolescents' Perspectives on Personalized E-Feedback in the Context of Health Risk Behavior Screening for Primary Care: Qualitative Study.
Electronic health screening tools for primary care present an opportunity to go beyond data collection to provide education and feedback to adolescents in order to motivate behavior change. However, there is limited research to guide feedback message development. The aim of this study was to explore youth perceptions of and preferences for receiving personalized feedback for multiple health risk behaviors and reinforcement for health promoting behaviors from an electronic health screening tool for primary care settings, using qualitative methodology. In total, 31 adolescents aged 13-18 years completed the screening tool, received the electronic feedback, and subsequently participated in individual, semistructured, qualitative interviews lasting approximately 60 min. Participants were queried about their overall impressions of the tool, perceptions regarding various types of feedback messages, and additional features that would help motivate health behavior change. Using thematic analysis, interview transcripts were coded to identify common themes expressed across participants. Overall, the tool was well-received by participants who perceived it as a way to enhance-but not replace-their interactions with providers. They appreciated receiving nonjudgmental feedback from the tool and responded positively to information regarding the consequences of behaviors, comparisons with peer norms and health guidelines, tips for behavior change, and reinforcement of healthy choices. A small but noteworthy minority of participants dismissed the peer norms as not real or relevant and national guidelines as not valid or reasonable. When prompted for possible adaptations to the tool, adolescents expressed interest in receiving follow-up information, setting health-related goals, tracking their behaviors over time, and communicating with providers electronically between appointments. Adolescents in this qualitative study desired feedback that validates their healthy behavior choices and supports them as independent decision makers by neutrally presenting health information, facilitating goal setting, and offering ongoing technological supports.


Upregulated expression of PDGF receptor beta in endocrine pancreatic tumors and metastases compared to normal endocrine pancreas.
Platelet-derived growth factor receptor (PDGFR) beta signaling is involved in autocrine growth stimulation of tumor cells, tumor angiogenesis and regulation of tumor interstitial fluid pressure. Development of PDGFR antagonists has further increased the interest for PDGFR as targets for anticancer treatments. Malignant endocrine pancreatic tumors (EPTs) express PDGFR beta both in stroma and on tumor cells. To investigate the role of PDGFR beta signaling in EPTs we compared PDGFR beta expression in normal endocrine pancreas to malignant EPTs and metastases. PDGFR beta expression was examined by immunohistochemistry using specific polyclonal antibodies in ten tissue samples from normal endocrine pancreas, 21 from primary EPTs and 19 from metastases. In eight patients we compared the expression in normal endocrine pancreas to the corresponding primary tumor and metastases, in two patients normal tissue to the primary tumor and in 11 patients primary tumors to the corresponding metastases. Six of ten tissues containing normal pancreas stained negative for PDGFR beta on endocrine cells, while seven of ten stained positive in the stroma. Eighteen of 21 (86%) primary tumors stained positive for PDGFR beta on tumor cells and all had positive stroma stainings. All 19 metastases stained positive for PDGFR beta on tumor cells and in evaluable stroma (n=16). We have found that PDGFR beta is more frequently expressed in primary EPTs and metastases as compared to normal endocrine pancreatic tissue. This is also true for PDGFR beta expression in the corresponding stroma. We suggest that new therapeutic options to inhibit the growth and spread of EPTs could include targeting of PDGFR beta.


Cloning and characterization of human phosphomannomutase, a mammalian homologue of yeast SEC53.
Phosphomannomutase (PMM) catalyzes the conversion of mannose-6-phosphate to mannose-1-phosphate, which is a substrate for the synthesis of GDP-mannose. This nucleotide sugar is then used in the synthesis of dolichol-phosphate-mannose, which is essential for N-linked glycosylation and thus the secretion of several glycoproteins as well as for the synthesis of glycosyl-phosphatidyl-inositol (GPI) anchored proteins. In the yeast Saccharomyces cerevisiae, SEC53, a gene required for viability, encodes PMM. Given the importance of PMM in glycoprotein synthesis, it is surprising that very little is known about the enzyme in higher eukaryotes. Recently, an autosomal recessive human disease, Carbohydrate-deficient glycoprotein syndrome type I (CDGS-I) has been correlated with severely reduced PMM activity. Here we report the isolation of a cDNA encoding human PMM, a protein of 29 kDa that is 55% identical and 66% similar to yeast Sec53p. Northern blot analysis shows a single 1.4 kb transcript that is ubiquitously expressed, although levels vary markedly among tissues. Expression of the human cDNA in a temperature-sensitive mutant sec53 yeast strain confers growth at the restrictive temperature, strongly suggesting that this gene encodes a functional PMM. Finally, when expressed in BHK cells, PMM is localized exclusively to the cytosol corresponding to its localization in yeast.


The functional outcome of operative treatment of ununited fractures of the humeral diaphysis in older patients.
Twenty-two elderly patients (average age, seventy-two years) who had an atrophic, unstable, ununited fracture of the humeral diaphysis were managed with plate-and-screw fixation and application of an autogenous bone graft from the iliac crest. Fifteen of the patients had had at least one previous operation in an attempt to obtain union of the fracture. One patient had an active infection and two had a quiescent infection, all with Staphylococcus epidermidis. The average duration of nonunion before the patients were first seen by us was two years and four months (range, five months to sixteen years). Fifteen of the nonunions were synovial. In each patient, at least one modification of the standard technique of plate-and-screw fixation was needed as a result of osteopenia. In order to enhance fixation, the standard protocol incorporated the use of a long plate (with an average of eleven holes and an average length that was 76 percent of that of the bone), a plate with a blade (used in thirteen patients), and replacement of loose, 4.5-millimeter cortical-bone screws with 6.5-millimeter cancellous-bone screws (twelve patients). Spiked nuts (Schuhli nut; Synthes, Paoli, Pennsylvania) that lock the screws to the plate, creating a solid point of fixation analogous to a blade, were incorporated into the protocol when they became available (used in six patients). In five limbs, the nonunion was associated with an osseous defect that could not be addressed by shortening of the bone alone. Three of these limbs were stabilized with a bridge plate that had been contoured to stand away from the bone at the site of nonunion (so-called wave-plate osteosynthesis), and the remaining two limbs were stabilized with a combination of intramedullary and extramedullary plates. In one of these two limbs, the extramedullary plate was contoured (that is, a wave plate). The fracture united in twenty (91 percent) of the patients. There was no progressive loosening or breakage of a fixation device, even in two patients who had radiographs that were suggestive of an incomplete union. Five of the patients were followed for a limited duration (average, one year and six months) as a result of death or illness. They had two excellent results, two good results, and one poor result according to a modification of the rating system of Constant and Murley. The remaining seventeen patients, including the two who had a persistent nonunion, were followed for an average of three years and one month (range, two years to five years and ten months). They had significant improvements in all of the functional scores at the most recent follow-up evaluation: the average score according to the modified system of Constant and Murley increased from 9 to 72 points (p < 0.001), the average score according to the Enforced Social Dependency Scale decreased from 39 to 9 points (p < 0.001), and the average score based on the Disabilities of the Arm, Shoulder, and Hand Questionnaire decreased from 77 to 24 points (p < 0.001). According to the scores based on the Disabilities of the Arm, Shoulder, and Hand Questionnaire, nine of the seventeen patients who had been followed for more than two years had an excellent result, four had a good result, two had a fair result, and the two who had a persistent nonunion had a poor result. Complications included postoperative delirium, a stitch abscess, transient radial nerve palsy, a fracture distal to the plate, and the need for a blood transfusion, in one patient each. Two patients had a fibrous union. There were no major medical complications. An unstable, united fracture of the humeral diaphysis can be extremely disabling and may threaten the ability of an elderly patient to function independently. Operative treatment can be very successful when the techniques of plate-and-screw fixation are modified to address osteopenia and relative or absolute loss of bone. Healing of the fracture substantially improves function and the degree of independence


Pre-registration nursing student's quality of practice learning: Clinical learning environment inventory (actual) questionnaire.
Clinical learning is a vital component of nurse education and assessing student's experiences can provide useful insights for development. Whilst most research in this area has focused on the acute setting little attention has been given to all pre-registration nurses' experience across the clinical placements arenas. To examine of pre-registration nursing students (first, second and third year) assessment of their actual experiences of their most recent clinical learning clinical learning experience. A cross sectional survey involving a descriptive online anonymous questionnaire based on the clinical learning environment inventory tool. One higher education institution in the United Kingdom. Nursing students (n=147) enrolled in an undergraduate nursing degree. This questionnaire included demographic questions and the Clinical Learning Environment Inventory (CLEI) a 42 item tool measuring student's satisfaction with clinical placement. SPPS version 22 was employed to analyse data with descriptive and inferential statistics. Overall students were satisfied with their clinical learning experience across all placement areas. This was linked to the 6 constructs of the clinical learning environment inventory; personalization, innovation, individualization, task orientation, involvement, satisfaction. Significant differences in student experience were noted between age groups and student year but there was no difference noted between placement type, age and gender. Nursing students had a positive perception of their clinical learning experience, although there remains room for improvement. Enabling a greater understanding of students' perspective on the quality of clinical education is important for nursing education and future research.


Resveratrol downregulates type-1 glutamate transporter expression and microglia activation in the hippocampus following cerebral ischemia reperfusion in rats.
The naturally occurring polyphenol phytoalexin resveratrol (RSV) regulates neuronal inflammation in various disease models and protects the brain against ischemic injury. Cell surface glutamate transporters on perisynaptic astrocytes are important regulators of extracellular glutamate levels and synaptic activation. Following cerebral ischemia, reduced astroglial type-1 glutamate transporter (GLT-1) expression in the CA1 pyramidal layers of the hippocampus contribute to neurotoxic glutamate levels. The current study examined the effects of 21-day RSV pretreatment (1 or 10mg/kg dose; i.p.) on microglia and astrocyte activation and characterized GLT-1 expression in the dentate gyrus (DG), CA1 and CA3 layers of the hippocampus 7 days following 10min global ischemia. Male Wistar rats were divided into five groups; sham/saline, ischemia/saline, ischemia/1mg/kg RSV, ischemia/10mg/kg RSV and sham/10mg/kg RSV. Immunohistochemical detection of GLT-1, CD11b/c, glial fibrillary acidic protein (GFAP) assessed type 1 glutamate transporter expression and microglial/glial cell activation following sham surgery or global ischemia. Our findings demonstrate prevention by RSV of ischemia-induced reduction of GLT-1 expression in the vulnerable CA1 layer 7 days following global ischemia, which was accompanied by the polyphenol's inhibition of post ischemic increase in CD11b/c and GFAP expression. RSV also conferred significant CA1 neuronal protection positively correlated with attenuation of glutamate transporter activation. These findings support beneficial effects of RSV in modulation of the excitotoxic cascade postischemia, which are congruent with anti-inflammatory effects observed in various pathological models.


Parenteral amino acid intakes in critically ill children: a matter of convenience.
Parenteral and enteral amino acid requirements for nutrition balance and function have not been defined in critically ill children or adults. In addition to playing a role in protein synthesis, amino acids trigger signaling cascades that regulate various aspects of fuel and energy metabolism and serve as precursors for important substrates. Amino acids can also be toxic. In this study, parenteral intakes of essential and nonessential amino acids (EAAs and NEAAs) supplied to critically ill children were assessed as an initial step for further studies aimed at establishing parenteral amino acid requirements. A retrospective review was conducted to assess intakes of parenteral amino acid for 116 critically ill children, and these intakes were compared with EAA intakes recommended by the Institute of Medicine. Because there are no recommended intakes for NEAA, NEAA intakes were compared with mixed muscle protein content in the older children and breast milk amino acid content in the infants. Parenteral EAAs were provided in amounts that exceeded recommended intakes for healthy children, except for phenylalanine and methionine, which although excessive, were given in less generous amounts. NEAAs were supplied in lower or higher amounts than the content of mixed muscle proteins or breast milk. Parenteral amino acid formulas are limited in taurine, glutamine, and asparagine despite the fact that inflammatory/immune proteins are rich in these amino acids. Amino acid composition of parenteral formulas is variable and lacks scientific support. Parenteral amino acid intakes should be based on measured requirements to maintain nutrition and functional balance and on knowledge of toxicity.


Impact of gold nanoparticles (AuNPs) on eosinophils isolated from male and female individuals.
It is well established that some differences exist between the male and female immune systems. Despites this, a sex-based analysis is not frequently performed in most scientific published reports. Knowing that inflammation is a common undesired effect observed resulting from nanoparticle (NP) exposure, we investigate here how in vitro treatment of gold NPs with a primary size of 20 and 70 nm (AuNP20 and AuNP70, respectively) will alter the biology of human eosinophils isolated from men and women blood. We found that treatment of AuNP70, but not AuNP20, significantly delay apoptosis only in eosinophils isolated from women. AuNPs were found to decrease eosinophil phagocytosis, however, significance was only observed in AuNP20-induced eosinophils isolated from women. The production of IL-8 was significantly increased in response to both AuNPs but only in eosinophils isolated from men and the production of IL-1β was increased in AuNPs-induced eosinophils, although significance was observed only in AuNP70-induced eosinophils isolated from women. We conclude that future studies investigating the toxicity of AuNPs (or other NPs) should include a sex-based analysis, especially if the tested NPs have potential medical applications knowing the increased interest in the development of personalized precision medicine.


The role of diffusion- and perfusion-weighted magnetic resonance imaging in drug development for ischemic stroke: from laboratory to clinics.
Ischemic stroke is a major cause of mortality and morbidity in industrialized countries and is almost always caused by occlusion of a cerebral artery by a clot. As the reversibly injured brain tissue evolves into irreversible infarction within a short period of time after onset of ischemia, it is extremely important and urgent to reverse the serious consequences of brain ischemia in the hyperacute phase when the ischemic brain tissue is still salvageable. Numerous thrombolytic and potentially neuroprotective agents have been studied in stroke patients with little success as the only approved therapy is thrombolysis with recombinant tissue plasminogen activator (r-tPA) within 3 h of stroke onset in highly selected patients (approximately 5 to 10 % of all acute stroke patients). One major obstacle in the development of effective therapies for ischemic stroke has been the lack of versatile imaging techniques. New magnetic resonance imaging (MRI) modalities, specially diffusion- and perfusion-weighted MRI (DWI and PWI, respectively) have been used in experimental studies with great success for over a decade and now are gradually entering clinical use. DWI and PWI can detect brain ischemia in the early phase in its full extent thus ensuring a definite diagnosis, allowing for follow-up of the ischemic lesion size over time with good spatial and temporal resolution, demonstrating perfusion deficit and reperfusion and the existence and the extent of penumbra while only requiring a few minutes of imaging time. DWI and PWI do not just give us the correct diagnosis of ischemic stroke, but allow us to acquire in vivo lesion size before therapeutic regimen is started and monitor the therapeutic efficacy thereafter, thus overcoming the potential pretreatment bias. We used DWI and PWI to evaluate novel therapeutic approaches for ischemic stroke in numerous experimental studies and lately in humans. With DWI and PWI, we are able to determine the in vivo efficacy (or lack of efficacy) of new therapeutic regiments (both neuroprotective and thrombolytic agents, or combination therapies) in a rapid, safe, and reliable way and in a relatively small number of well-selected, well-defined, and homogeneous patients. This approach may, therefore, significantly accelerate the development of new remedies for stroke patients.


Examining the Etiology of Reading Disability as Conceptualized by the Hybrid Model.
A fairly recent definition of reading disability (RD) is that in the form of a hybrid model. The model views RD as a latent construct that is manifested through various observable unexpected impairments in reading related skills and through inadequate response to intervention. The current report evaluated this new conceptualization of RD from an etiological perspective. The sample consisted of 2737 twin pairs in first through fourth grade (Mage = 8.52) from the Florida Twin Project on Reading. Using twin analyses, results showed that a substantial proportion of genetic variance, a small proportion of shared environmental, and a small proportion of non-shared environmental variance was attributed to the RD factor. Findings suggest that the etiological architecture of RD closely mirrors the structure of the hybrid model of RD.


An immunoinformatic approach driven by experimental proteomics: in silico design of a subunit candidate vaccine targeting secretory proteins of Leishmania donovani amastigotes.
Visceral leishmaniasis (VL) caused by dimorphic Leishmania species is a parasitic disease with high socioeconomic burden in endemic areas worldwide. Sustaining control of VL in terms of proper and prevailing immunity development is a global necessity amid unavailability of a prophylactic vaccine. Screening of experimental proteome of the human disease propagating form of Leishmania donovani (amastigote) can be more pragmatic for in silico mining of novel vaccine candidates. By using an immunoinformatic approach, CD4+ and CD8+ T cell-specific epitopes from experimentally reported L. donovani proteins having secretory potential and increased abundance in amastigotes were screened. A chimera linked with a Toll-like receptor 4 (TLR4) peptide adjuvant was constructed and evaluated for physicochemical characteristics, binding interaction with TLR4 in simulated physiological condition and the trend of immune response following hypothetical immunization. Selected epitopes from physiologically important L. donovani proteins were found mostly conserved in L. infantum, covering theoretically more than 98% of the global population. The multi-epitope chimeric vaccine was predicted as stable, antigenic and non-allergenic. Structural analysis of vaccine-TLR4 receptor docked complex and its molecular dynamics simulation suggest sufficiently stable binding interface along with prospect of non-canonical receptor activation. Simulation dynamics of immune response following hypothetical immunization indicate active and memory B as well as CD4+ T cell generation potential, and likely chance of a more Th1 polarized response. The methodological approach and results from this study could facilitate more informed screening and selection of candidate antigenic proteins for entry into vaccine production pipeline in future to control human VL.


Redox regulation of matrix metalloproteinase gene family in small cell lung cancer cells.
It has been implicated that reactive oxygen species (ROS) play important roles in modulating tumor progression. However, the mechanisms by which redox-regulated tumor progression are largely unknown. We previously demonstrated that reduced intracellular redox conditions could be achieved in stably transfected small cell lung cancer cells with gamma-glutamylcysteine synthetase (gamma-GCSh) cDNA which encodes a rate-limiting enzyme in the biosynthesis of glutathione (GSH), a major physiological redox regulator. In the present study, using DNA microarray analyses, we compared the expression profiles between the gamma-GCSh-transfected cells and their nontransfected counterpart. We observed downregulation of several matrix metalloproteinases (MMPs), i.e., MMPI and MMP3, and MMP10 in the transfected cells. Dot blot and Northern blot hybridizations confirmed that, among the 18 MMP gene family members and four tissue inhibitors of matrix metalloprotein family (TIMP) analyzed, the expression levels of these three MMPs were consistently reduced. Transiently increased gamma-GCSh expression using tetracycline-inducible gamma-GCSh adenoviral expression system also showed down-regulation of MMP3 and MMP10, but not MMP1. Our results demonstrated that redox regulation of MMP1, MMP3 and MMP10 expression depend upon different modes of redox manipulation. These results bear implication that antioxidant modulation of antitumor progression may be contributed at least in part by the downregulation of a subset of metrix metalloproteins.


Flow-Rate-Insensitive Plasma Extraction by the Stabilization and Acceleration of Secondary Flow in the Ultralow Aspect Ratio Spiral Channel.
Although microfluidic devices have made remarkable strides in blood cell separation, there is still a need for further development and improvement in this area. Herein, we present a novel ultralow aspect ratio (H/W = 1:36) spiral channel microfluidic device with ordered micro-obstacles for sheathless and flow-rate-insensitive blood cell separation. By introducing ordered micro-obstacles into the spiral microchannels, reduced magnitude fluctuations in secondary flow across different loops can be obtained through geometric confinement. As a result, the unique Dean-like secondary flow can effectively enhance the separation efficiency of particles in different sizes ranging from 3 to 15 μm. Compared to most existing microfluidic devices, our system offers several advantages of easy manufacturing, convenient operation, long-term stability, highly efficient performance (up to 99.70% rejection efficiency, including platelets), and most importantly, insensitivity to cell sizes as well as flow rates (allowing for efficient separation of different-sized blood cells in a wide flow rate from 1.00 to 2.50 mL/min). The unique characteristics, such as ultralow aspect ratio, sequential micro-obstacles, and controlled secondary flow, make our device a promising solution for practical plasma extraction in biomedical research and clinical applications.


Fetal health shocks and early inequalities in health capital accumulation.
Several studies report socioeconomic inequalities in child health and consequences of early disease. However, not much is known about inequalities in health capital accumulation in the womb in response to fetal health shocks, which is essential for finding the earliest sensitive periods for interventions to reduce inequalities. We identify inequalities in birth weight accumulation as a result of fetal health shocks from the occurrence of one of the most common birth defects, oral clefts, within the first 9 weeks of pregnancy, using quantile regression and two datasets from South America and the USA. Infants born at lower birth weight quantiles are significantly more adversely affected by the health shock compared with those born at higher birth weight quantiles, with overall comparable results between the South American and US samples. These results suggest that fetal health shocks increase child health disparities by widening the spread of the birth weight distribution and that health inequalities begin in the womb, requiring interventions before pregnancy.


[The clinical features and natural history of post-transfusion hepatitis C].
To study the clinical features and natural history of post-transfusion hepatitis C (PTHC). Ninety-nine post-transfusion hepatitis C patients were analyzed using retrospective and prospective study and follow-up. (1) Ninety-nine post-transfusion HCV patients were infected during 1989-1994, mostly between 1990-1992. (2) Ninety patients were diagnosed as chronic hepatitis C, and 9 as hepatic cirrhosis (period of compensation). (3) The intervals between their transfusions and their initial diagnoses of PTHC were 7.4+/-6.6 years in all 99 patients, and the intervals in 9 cirrhosis patients were 12.7+/-5.8 years. (4) Among 63 male patients, 59 cases were chronic hepatitis C and 4 were cirrhosis while among 36 female patients, 31 were chronic hepatitis C and 5 were cirrhosis. There was no significant difference of the ratio for hepatitis C and cirrhosis between the male and female patients (P>0.05). (5) Repeat abnormal liver function occurred accompanied with a fluctuation of ALT elevation in those patients with cirrhosis. (6) No patient developed hepatic carcinoma during the study period. (1) The possibility of HCV infection by transfusion has declined greatly since 1995 in Guangzhou. (2) Nine of the 99 (9.1%) chronic HCV-infected patients developed a compensated cirrhosis after 12.7+/-5.8 years. (3) For those PTHC patients with repeat abnormal liver functions, interferon combined with ribavirin is recommended to prevent the development of cirrhosis.


Left Atrium Wall-mapping Application for Wall Thickness Visualisation.
The measurement method for the LA wall thickness (WT) using cardiac computed tomography (CT) is observer dependent and cannot provide a rapid and comprehensive visualisation of the global LA WT. We aim to develop a LA wall-mapping application to display the global LA WT on a coplanar plane. The accuracy, intra-observer, and inter-observer reproducibility of the application were validated using digital/physical phantoms, and CT images of eight patients. This application on CT-based LA WT measures were further validated by testing six pig cardiac specimens. To evaluate its accuracy, the expanded maps of the physical phantom and pig LA were generated from the CT images and compared with the expanded map of the digital phantom and LA wall of pig heart. No significant differences (p > 0.05) were found between physical phantom and digital phantom as well as pig heart specimen and CT images using our application. Moreover, the analysis was based on the LA physical phantom or images of clinical patients; the results consistently demonstrated high intra-observer reproducibility (ICC > 0.9) and inter-observer reproducibility (ICC > 0.8) and showed good correlation between measures of pig heart specimen and CT data (r = 0.96, p < 0.001). The application can process and analyse the LA architecture for further visualisation and quantification.


The impact of COVID-19 misinformation and trust in institutions on preventive behaviors.
Misinformation related to coronavirus disease 2019 (COVID-19) has the potential to suppress preventive behaviors that mitigate the spread of COVID-19. Early research on the behavioral consequences of COVID-19 misinformation is mixed, and most rely on cross-sectional data. We examined whether believing in COVID-19 misinformation at one time point influences engaging in preventive behaviors later. In addition, we investigated the role of trust in institutions. We conducted a two-wave survey in South Korea and examined the association between belief in COVID-19 misinformation at Wave 1 and preventive behaviors at Wave 2 controlling for preventive behaviors at Wave 1. We also analyzed whether there is an interaction between belief in COVID-19 misinformation and trust in institutions. Belief in COVID-19 misinformation at Wave 1 significantly increased avoidance of preventive behaviors at Wave 2, but after accounting for trust in institutions, this effect disappeared. Rather, trust in institutions significantly decreased avoidance of preventive behaviors. In addition, misinformation increased avoidance of preventive behaviors among those who trusted institutions the most. Results suggest that building trust in institutions is essential in promoting COVID-19 preventive behaviors. Belief in COVID-19 misinformation may have harmful effects, but these effects were pronounced for those who highly trust institutions.


Development, dissemination, implementation and evaluation of a clinical pathway for oxygen therapy.
Oxygen is commonly administered to patients in hospital, but prescribing and monitoring of such therapy may be suboptimal. The objective of this study was to develop, disseminate, implement and evaluate a multidisciplinary clinical pathway for the administration of oxygen. The authors developed a clinical pathway for the ordering, titration and discontinuation of oxygen, which was disseminated through teaching sessions, in-service training sessions and information posters in a medical clinical teaching unit (CTU). Implementation of the pathway was ensured by means of reminders and patient-centred audit and feedback to CTU nurses and house staff. During a 3-month intervention phase, consecutive patients requiring supplemental oxygen were treated according to the pathway. During a 1-month "wash-out" phase followed by a 3-month non-intervention phase, patients were treated at the discretion of the CTU team. Clinical and economic data were collected in both phases. In the 2 phases, patient characteristics, the concentration and duration of oxygen prescribed, the frequency of oxygen saturation monitoring, the frequency of arterial blood gas testing and the clinical outcomes were similar. However, there were more discontinuation orders in the intervention phase (p < 0.001). In the intervention phase, costs were higher for monitoring of oxygen saturation ($44.95/patient v. $36.17/patient, p = 0.048) and for order transcription ($2.71/patient v. $1.28/patient, p < 0.001); total costs, including those for personnel, were also higher in the intervention phase ($76.93/patient v. $56.67/patient, p = 0.02). The cost of education about the oxygen pathway was $45.71/patient. When the education cost was included, the total cost of oxygen therapy during the intervention phase was $122.64/patient; this was significantly higher than the total cost of oxygen therapy during the non-intervention phase ($56.67/patient) (p < 0.001). This multidisciplinary, multimethod oxygen pathway led to changes in oxygen-prescribing behaviour, consumed more resources than standard management and was not associated with changes in patient outcome. Appropriate management of oxygen prescribing and monitoring by physicians and nurses take time and costs money.


Cancer incidence in the Zuni Indians of New Mexico.
The total age-adjusted incidence of cancer in the Zuni Indians of New Mexico was significantly lower than that of the New Mexico Anglo population during the period 1969-1982. Specific sites at which the Zunis had a significantly lower number of cases than expected, based on the rates for Anglos, are: colon, rectum and anus, lung, breast, endometrium, melanoma of the skin, pancreas, and the leukemias. Sites at which the Zunis had a higher number of cases than expected are stomach and gallbladder. The Zunis have a pattern of occurrence of cancer that is similar to other American Indians of New Mexico (Navajo, Apache, and Pueblo); however, rates of lung, colonic, and pancreatic cancer among the Zunis are significantly lower. The occurrence and anatomic distribution of cancer among the Zunis may be the result of cultural and environmental conditions or genetic influences. Further studies may clarify the risk factors which contribute to this pattern of disease.


Acute Flaccid Myelitis: A Clinical Review.
Acute flaccid myelitis (AFM) is an emerging disorder primarily affecting children that is characterized by acute flaccid paralysis accompanied by abnormalities of the spinal cord gray matter on magnetic resonance imaging. In most cases, prodromal fever or respiratory symptoms occur, followed by acute-onset flaccid limb weakness. Respiratory, axial, bulbar, facial, and extraocular muscles may also be affected. The clinical manifestations have been described as "polio-like," due to striking similarities to cases of poliomyelitis. The primary site of injury in AFM is the anterior horn cells of the spinal cord, resulting in a motor neuronopathy. Seasonal peaks of cases have occurred in the United States every 2 years since 2012. However, AFM remains a rare disease, which can make it challenging for physicians to recognize and differentiate from other causes of acute flaccid paralysis such as Guillain-Barre syndrome, spinal cord stroke, and transverse myelitis. Epidemiological evidence suggests that AFM is linked to a viral etiology, with nonpolio enteroviruses (in particular enterovirus D68) demonstrating a plausible association. The epidemiology, possible etiological factors, clinical features, differential diagnosis, treatment, and outcomes of AFM are discussed in this review.


Assessing Endogenous and Exogenous Hormone Exposures and Breast Development in a Migrant Study of Bangladeshi and British Girls.
Timing of breast development (or thelarche) and its endogenous and exogenous determinants may underlie global variation in breast cancer incidence. The study objectives were to characterize endogenous estrogen levels and bisphenol A (BPA) exposure using a migrant study of adolescent girls and test whether concentrations explained differences in thelarche by birthplace and growth environment. Estrogen metabolites (EM) and BPA-glucuronide (BPA-G) were quantified in urine spot samples using liquid chromatography tandem mass spectrometry (LC-MS/MS) from a cross-sectional study of Bangladeshi, first- and second-generation Bangladeshi migrants to the UK, and white British girls aged 5-16 years (n = 348). Thelarche status at the time of interview was self-reported and defined equivalent to Tanner Stage ≥2. We compared geometric means (and 95% confidence interval (CIs)) of EM and BPA-G using linear regression and assessed whether EM and BPA-G explained any of the association between exposure to the UK and the age at thelarche using hazard ratios and 95% confidence intervals. Average EM decreased with exposure to the UK, whereas BPA-G increased and was significantly higher among white British (0.007 ng/mL, 95% CI: 0.0024-0.0217) and second-generation British-Bangladeshi girls (0.009 ng/mL, 95% CI: 0.0040-0.0187) compared to Bangladeshi girls (0.002 ng/mL, 95% CI: 0.0018-0.0034). Two of four EM ratios (16-pathway/parent and parent/all pathways) were significantly associated with thelarche. The relationship between exposure to the UK and thelarche did not change appreciably after adding EM and BPA-G to the models. While BPA-G is often considered a ubiquitous exposure, our findings suggest it can vary based on birthplace and growth environment, with increasing levels for girls who were born in or moved to the UK. Our study did not provide statistically significant evidence that BPA-G or EM concentrations explained earlier thelarche among girls who were born or raised in the UK.


Endoscopic ultrasound guided fine-needle aspiration diagnosis of duodenal high grade neuroendocrine carcinoma underlying a villous adenoma: report of a case.
Endoscopic ultrasound guided fine-needle aspiration biopsy is a reliable and accurate method for the diagnosis of submucosal lesions of the gastrointestinal tract. We report the cytopathologic findings of a case of duodenal high-grade neuroendocrine carcinoma in a 68-year-old woman who presented with melena and marked anemia, 45 years after kidney transplantation. Imaging studies performed in the work-up of melena showed a duodenal mass, which on endoscopy proved to be an exophytic, villous duodenal lesion, 3 cm from the ampulla. Forceps biopsy of the exophytic lesion showed a villous adenoma. Endoscopic ultrasound additionally revealed an underlying submucosal lesion and EUS-guided fine needle aspiration of this submucosal mass and of the enlarged mesenteric lymph nodes was diagnostic of a high-grade neuroendocrine carcinoma. The aspirates showed abundant cellularity with tumor cells arranged in sheets and occasional loose clusters. The neoplastic cells had a moderate amount of pale cytoplasm and large round to oval hyperchromatic nuclei with focally prominent nucleoli. Mitoses, apoptotic bodies and necrotic debris were also present. The tumor cells were strongly and diffusely positive for cytokeratin AE1/AE3, synaptophysin and chromogranin and showed a very high proliferative fraction on Ki67 staining, supporting the diagnosis of a high-grade neuroendocrine carcinoma. This is to our knowledge the first case of high-grade neuroendocrine carcinoma of the duodenum diagnosed by EUS-FNA. This case also emphasizes the diagnostic value of EUS-FNA sampling of the submucosal and intramural component of villous tumors of the gastrointestinal tract when mucosal forceps biopsies show only benign findings.


Transmission Dynamics and Rare Clustered Transmission Within an Urban University Population Before Widespread Vaccination.
Universities returned to in-person learning in 2021 while SARS-CoV-2 spread remained high. At the time, it was not clear whether in-person learning would be a source of disease spread. We combined surveillance testing, universal contact tracing, and viral genome sequencing to quantify introductions and identify likely on-campus spread. Ninety-one percent of viral genotypes occurred once, indicating no follow-on transmission. Less than 5% of introductions resulted in >3 cases, with 2 notable exceptions of 40 and 47 cases. Both partially overlapped with outbreaks defined by contact tracing. In both cases, viral genomics eliminated over half the epidemiologically linked cases but added an equivalent or greater number of individuals to the transmission cluster. Public health interventions prevented within-university transmission for most SARS-CoV-2 introductions, with only 2 major outbreaks being identified January to May 2021. The genetically linked cases overlap with outbreaks identified by contact tracing; however, they persisted in the university population for fewer days and rounds of transmission than estimated via contact tracing. This underscores the effectiveness of test-trace-isolate strategies in controlling undetected spread of emerging respiratory infectious diseases. These approaches limit follow-on transmission in both outside-in and internal transmission conditions.


Construction of improved tools for protein localization studies in Streptococcus pneumoniae.
We have constructed a set of plasmids that allow efficient expression of both N- and C-terminal fusions of proteins of interest to fluorescent proteins mCherry, Citrine, CFP and GFP in the Gram-positive pathogen Streptococcus pneumoniae. In order to improve expression of the fluorescent fusions to levels that allow their detection by fluorescence microscopy, we have introduced a 10 amino acid tag, named i-tag, at the N-terminal end of the fluorescent proteins. This caused increased expression due to improved translation efficiency and did not interfere with the protein localization in pneumococcal bacteria. Localizing fluorescent derivatives of FtsZ, Wzd and Wze in dividing bacteria validated the developed tools. The availability of the new plasmids described in this work should greatly facilitate studies of protein localization in an important clinical pathogen.


Cryptocandin, a potent antimycotic from the endophytic fungus Cryptosporiopsis cf. quercina.
A unique lipopeptide antimycotic, termed cryptocandin, is described from Cryptosporiopsis cf. quercina, an endophytic fungus. Cryptocandin, with a molecular mass of 1079 Da, contains equimolar amounts of 3,4-dihydroxyhomotyrosine, 4-hydroxyproline, threonine, glutamine, 3-hydroxy-4-hydroxymethylproline, 4,5-dihydroxyornithine and palmitic acid. Cryptocandin is chemically related to well-known antimycotics, the echinocandins and pneumocandins, which are produced by such fungi as Zalerion arboricola, Pezicula spp. and Aspergillus spp. Cryptocandin has minimal inhibitory concentration values of 0.03-0.07 microgram ml-1 against isolates of Candida albicans, Trichophyton mentagrophytes and Trichophyton rubrum. Cryptocandin is also active against a number of plant-pathogenic fungi including Sclerotinia sclerotiorum and Botrytis cinerea.


Medical Sensors for the Diagnosis and Management of Disease: The Physician Perspective.
The objective of this paper is to assist developers of medical sensors to better formulate the clinically relevant design criteria and required performance characteristics of their novel sensor based on an understanding of how these devices will be used by physicians. Sensor technologies play a central role in medicine, and the most critical aspect of the sensor's clinical utility relates to these design decisions. Clinically, sensors are used by health care providers to make both diagnostic and management decisions, and the sensors that aid in these decisions are evaluated by certain clinical, as well as analytical, criteria. Failure to adequately address these end-user requirements can lead to the development of sensors without clinical utility.


[Regeneration of sanitaro-hygienic water during long space flights by reverse osmosis].
Experiments on the purification of wash water by means of reverse osmosis membranes MGA-100 were performed. The membrane selectivity with respect to the major components (catamine AB, amine oxide and sodium chloride) was close to 100% whereas membrane permeability was not lower than 70% that of distilled water. The results of a field study in which 330 1 of repeatedly used wash water were reclaimed showed that reverse osmosis can be effectively employed for that purpose.


Temperature and injection water source influence microbial community structure in four Alaskan North Slope hydrocarbon reservoirs.
A fundamental knowledge of microbial community structure in petroleum reservoirs can improve predictive modeling of these environments. We used hydrocarbon profiles, stable isotopes, and high-density DNA microarray analysis to characterize microbial communities in produced water from four Alaskan North Slope hydrocarbon reservoirs. Produced fluids from Schrader Bluff (24-27°C), Kuparuk (47-70°C), Sag River (80°C), and Ivishak (80-83°C) reservoirs were collected, with paired soured/non-soured wells sampled from Kuparuk and Ivishak. Chemical and stable isotope data suggested Schrader Bluff had substantial biogenic methane, whereas methane was mostly thermogenic in deeper reservoirs. Acetoclastic methanogens (Methanosaeta) were most prominent in Schrader Bluff samples, and the combined δD and δ(13)C values of methane also indicated acetoclastic methanogenesis could be a primary route for biogenic methane. Conversely, hydrogenotrophic methanogens (e.g., Methanobacteriaceae) and sulfide-producing Archaeoglobus and Thermococcus were more prominent in Kuparuk samples. Sulfide-producing microbes were detected in all reservoirs, uncoupled from souring status (e.g., the non-soured Kuparuk samples had higher relative abundances of many sulfate-reducers compared to the soured sample, suggesting sulfate-reducers may be living fermentatively/syntrophically when sulfate is limited). Sulfate abundance via long-term seawater injection resulted in greater relative abundances of Desulfonauticus, Desulfomicrobium, and Desulfuromonas in the soured Ivishak well compared to the non-soured well. In the non-soured Ivishak sample, several taxa affiliated with Thermoanaerobacter and Halomonas predominated. Archaea were not detected in the deepest reservoirs. Functional group taxa differed in relative abundance among reservoirs, likely reflecting differing thermal and/or geochemical influences.


Some current issues in the pharmacokinetics/pharmacodynamics of antimicrobials in intensive care.
Infections, particularly those caused by resistant pathogens, are a common cause of morbidity and mortality in critically ill patients. However, the availability of effective antimicrobial agents is limited. Critical illness itself can influence the pharmacokinetic/pharmacodynamic (PK/PD) parameters of antimicrobials by altering their volume of distribution and the rate of their excretion and elimination and by impairing their penetration into tissues. Therefore, when designing a treatment regimen, the intensivist should consider and take advantage of antibiotic PK/PD properties. There is significant but inconclusive evidence that critically ill patients may benefit more when antibiotics with time-dependent action are administered in a continuous/prolonged infusion regimen. On the other hand, aminoglycosides exhibit a concentration-dependent pattern of killing and should be administered at high doses once daily or at extended intervals, and their levels in the plasma should by strictly monitored to avoid both underexposure and toxicity. The problem of antimicrobial resistance now involves agents traditionally considered reliable in that aspect, such as vancomycin. Strict monitoring of vancomycin MIC for methicillin-resistant Staphylococcus aureus and the prudent use of the available alternative agents as well as de-escalation strategies might be reasonable strategies for dealing with this problem.


Bayesian model of Hamilton Depression Rating Score (HDRS) with memantine augmentation in bipolar depression.
Presynaptic and post-synaptic glutamatergic modulation is associated with antidepressant activity that takes several weeks to reach a maximal full effect. Limiting mood elevating effects after single drug administration may be the result of compensatory synaptic processes. Therefore, using augmentation treatment with agents having presynaptic and post-synaptic effects on the glutamatergic system, this study aims to evaluate the effect of augmentation therapy on the rate of change in mood elevation in patients with bipolar depression. In a pilot study, 29 outpatients with bipolar depression on a stable lamotrigine dose regimen received placebo or memantine pills daily (titrated up by 5 mg week⁻¹ to 20 mg) in a randomized, double-blind, parallel group, 8 week study. Patients were evaluated weekly using the 17-item Hamilton Depression Rating Score (HDRS) and all data were analyzed simultaneously. Linear, exponential, maximal effect, Gompertz and inverse Bateman functions were evaluated using a Bayesian approach population pharmacodynamic model framework. In these models, differences in parameters were examined across the memantine and placebo augmentation groups. A Gompertz function with a treatment switch on the parameter describing the speed of HDRS decline (γ, 95% confidence interval [CI]) best described the data (γ(memantine) = 1.8, 95% CI 0.9, 3.6), γ(placebo) = 1.2, 95% CI 0.5, 3.5)). Between subject variability was identified on baseline HDRS (2.9, 95% CI 1.5, 4.4) and amplitude of score improvement (4.3, 95% CI 2.7, 6.5). This pharmacodynamic approach identified an increased speed of response after memantine augmentation, compared with placebo augmentation in bipolar depression patients.


Peri-operative outcomes for pancreatoduodenectomy in India: a multi-centric study.
There have been an increasing number of reports world-wide relating improved outcomes after pancreatic resections to high volumes thereby supporting the idea of centralization of pancreatic resectional surgery. To date there has been no collective attempt from India at addressing this issue. This cohort study analysed peri-operative outcomes after pancreatoduodenectomy (PD) at seven major Indian centres. Between January 2005 and December 2007, retrospective data on PDs, including intra-operative and post-operative factors, were obtained from seven major centres for pancreatic surgery in India. Between January 2005 and December 2007, a total of 718 PDs were performed in India at the seven centres. The median number of PDs performed per year was 34 (range 9-54). The median number of PDs per surgeon per year was 16 (range 7-38). Ninety-four per cent of surgeries were performed for suspected malignancy in the pancreatic head and periampullary region. The median mortality rate per centre was four (range 2-5%). Wound infections were the commonest complication with a median incidence per centre of 18% (range 9.3-32.2%), and the median post-operative duration of hospital stay was 16 days (range 4-100 days). This is the first multi-centric report of peri-operative outcomes of PD from India. The results from these specialist centers are very acceptable, and appear to support the thrust towards centralization.


Methylprednisolone induces reversible clinical and pathological remission and loss of lymphocyte reactivity to myelin oligodendrocyte glycoprotein in experimental autoimmune encephalomyelitis.
Experimental autoimmune encephalomyelitis (EAE) is an animal model of human multiple sclerosis (MS). EAE, induced by immunisation with myelin-associated autoantigens, is characterised by an inflammatory infiltrate in the central nervous system (CNS) associated with axonal degeneration, demyelination and damage. We have recently shown in an experimental mouse model of autoimmune gastritis that methylprednisolone treatment induces a reversible remission of gastritis with regeneration of the gastric mucosa. Here, we examined the effect of oral methylprednisolone on the mouse EAE model of human MS induced by immunisation with myelin oligodendrocyte glycoprotein peptide (MOG(35-55)). We examined the clinical scores, CNS pathology and lymphocyte reactivity to MOG(35-55) following treatment and withdrawal of the steroid. Methylprednisolone remitted the clinical signs of EAE and the inflammatory infiltrate in the CNS, accompanied by loss of lymphocyte reactivity to MOG(35-55) peptide. Methylprednisolone withdrawal initiated relapse of the clinical features, a return of the CNS inflammatory infiltrate and lymphocyte reactivity to MOG(35-55) peptide. This is the first study to show that methylprednisolone induced a reversible remission in the clinical and pathological features of EAE in mice accompanied by loss of lymphocyte reactivity to the encephalitogen. This model will be useful for studies directed at a better understanding of mechanisms associated with steroid-induced disease remission, relapse and remyelination and also as an essential adjunct to an overall curative strategy.


Dissemination beyond publication.
The mantra 'publish or perish' has meant disseminating work in peer-reviewed literature. Digital has changed the way readers access content and has created a need to disseminate work widely for maximum impact. Research metrics used internationally to evaluate research are now based on the number of citations of a paper.


β-Amyloid and tau biomarkers and clinical phenotype in dementia with Lewy bodies.
In a multicenter cohort of probable dementia with Lewy bodies (DLB), we tested the hypothesis that β-amyloid and tau biomarker positivity increases with age, which is modified by APOE genotype and sex, and that there are isolated and synergistic associations with the clinical phenotype. We included 417 patients with DLB (age 45-93 years, 31% women). Positivity on β-amyloid (A+) and tau (T+) biomarkers was determined by CSF β-amyloid1-42 and phosphorylated tau in the European cohort and by Pittsburgh compound B and AV-1451 PET in the Mayo Clinic cohort. Patients were stratified into 4 groups: A-T-, A+T-, A-T+, and A+T+. A-T- was the largest group (39%), followed by A+T- (32%), A+T+ (15%), and A-T+ (13%). The percentage of A-T- decreased with age, and A+ and T+ increased with age in both women and men. A+ increased more in APOE ε4 carriers with age than in noncarriers. A+ was the main predictor of lower cognitive performance when considered together with T+. T+ was associated with a lower frequency of parkinsonism and probable REM sleep behavior disorder. There were no significant interactions between A+ and T+ in relation to the clinical phenotype. Alzheimer disease pathologic changes are common in DLB and are associated with the clinical phenotype. β-Amyloid is associated with cognitive impairment, and tau pathology is associated with lower frequency of clinical features of DLB. These findings have important implications for diagnosis, prognosis, and disease monitoring, as well as for clinical trials targeting disease-specific proteins in DLB. This study provides Class II evidence that in patients with probable DLB, β-amyloid is associated with lower cognitive performance and tau pathology is associated with lower frequency of clinical features of DLB.


Helicobacter hepaticus cholesterol-α-glucosyltransferase is essential for establishing colonization in male A/JCr mice.
Helicobacter pylori cholesterol-α-glucosyltransferase (cgt) is essential for survival of H. pylori in mice. Enterohepatic H. hepaticus, the cause of colonic and hepatocellular carcinoma in susceptible mouse strains, contains an ortholog of the H. pylori cgt. However, the role of cgt in the pathogenesis of H. hepaticus has not been investigated. Two cgt-deficient isogenic mutants of wild-type H. hepaticus (WT) 3B1 were generated and used to inoculate male A/JCr mice. Cecal and hepatic colonization levels of the mutants and WT 3B1 as well as select inflammation-associated cytokines were measured by qPCR at 4 months postinoculation. Both mutants were undetectable in the cecum of any inoculated mice (10 per mutant) but were detected in two livers (one for each mutant); by contrast, 9 and 7 of 10 mice inoculated with WT 3B1 were qPCR positive in the ceca and livers, respectively. The mice inoculated with the mutants developed significantly less severe hepatic inflammation (p < .05) and also produced significantly lower hepatic mRNA levels of proinflammatory cytokines Ifn-γ (p < .01) and Tnf-α (p ≤ .02) as well as anti-inflammatory factors Il10 and Foxp3 compared with the WT 3B1-inoculated mice. Additionally, the WT 3B1-inoculated mice developed significantly higher Th1-associated IgG2a (p < .0001) and Th2-associated IgG1 responses (p < .0001) to H. hepaticus infection than mice dosed with isogenic cgt mutants. Our data indicate that the cholesterol-α-glucosyltransferase is required for establishing colonization of the intestine and liver and therefore plays a critical role in the pathogenesis of H. hepaticus.


Impact of a depression care management program for hospitalized cardiac patients.
Depression is independently associated with poor outcomes among patients with acute cardiac disease. Collaborative care depression management programs have been used in outpatients to improve depression outcomes, but such a program had never been initiated in the hospital or used for patients with a wide range of cardiac illnesses. This was a prospective, randomized trial of a low-intensity, 12-week collaborative care program versus usual care for 175 depressed patients hospitalized for acute coronary syndrome, arrhythmia, or heart failure. Study outcomes, assessed using mixed regression models to compare groups at 6 weeks, 12 weeks, and 6 months, included mental health (depression, cognitive symptoms of depression, anxiety, and mental health-related quality of life) and medical (physical health-related quality of life, adherence to medical recommendations, and cardiac symptoms) outcomes. Collaborative care subjects (n=90) had significantly greater improvements on all mental health outcomes at 6 and 12 weeks, including rates of depression response (collaborative care, 59.7% versus usual care 33.7%; odds ratio, 2.91; P=0.003 at 6 weeks; 51.5% versus 34.4%; odds ratio, 2.02; P=0.04 at 12 weeks), though these effects decreased after intervention. At 6 months, intervention subjects had significantly greater self-reported adherence and significantly reduced number and intensity of cardiac symptoms. Among patients with a broad range of cardiac diagnoses, a collaborative care depression management program initiated during hospitalization led to significant improvements in multiple clinically important mental health outcomes and had promising effects on relevant medical outcomes after intervention. Clinical Trial Registration- URL: http://www.clinicaltrials.gov. Unique identifier: NCT00847132.


[Rehabilitation following anterior acromioplasty].
The shoulder region is one of the most challenging part of the human body to rehabilitate. Postoperative days following anterior acromioplasty are painful, with a high potential course of developing soft tissue fibrosis and joint contracture. In the past, following anterior acromioplasty and/or rotator cuff repair, the rehabilitation process was usually delayed for up to four to six weeks to allow healing of the repaired site. This conservative rehabilitation approach resulted in significant restrictions in glenohumeral and scapular joint mobility, weakness of the shoulder girdle musculature, prolonged pain, and functional limitations. Currently the trend of rehabilitation following anterior acromioplasty shifted toward a more earlier and aggressive approach thanks to improved surgical and soft tissue fixation techniques and advances in arthroscopic procedures. This has allowed active-assisted shoulder motion immediately after subacromial decompression and rotator cuff surgery. The rehabilitation team following anterior acromioplasty of the shoulder should consist of the physiatrist, orthopedic surgeon, physical therapist, and the patient.


Hypertension May Reduce the Infection Risk but Increase the Severity of COVID-19: Based on the Current Data in China.
Increasing evidence has shown an unusual relationship between hypertension and COVID-19, which may not be as simple as previously thought. The purpose of our study was to determine the association of hypertension with the onset and development of COVID-19. A meta-analysis was performed to summarize the prevalence of hypertension in COVID-19 patients, as well as the usage of ACEIs/ARBs. Metaregression analyses were used to evaluate the association of hypertension with disease severity and mortality. PubMed and Google Scholar were searched for relevant studies. A total of 42 studies including 14138 patients were enrolled in the study. The proportion of hypertension in COVID-19 patients in China was 17.7% according to the enrolled studies, while it was 6.0% in a study containing 72314 confirmed cases, which are both much lower than in the general population. All of the data from the 11 provinces in China showed the same tendency. The proportions of hypertension were higher in severe/ICU patients and nonsurvivors than in nonsevere/ICU patients and survivors. The metaregression analyses suggested that both disease severity and risk of death were associated with the incidence of hypertension. A total of 27.6% of COVID-19 patients with hypertension received ACEI/ARB therapy. The proportion of deaths in COVID-19 patients with hypertension treated with ACEIs/ARBs was significantly lower than that in nonuse patients treated with ACEIs/ARBs. In conclusion, hypertension may reduce the infection risk of COVID-19 but increase the risk of developing worse clinical outcomes. The use of ACEIs/ARBs may benefit COVID-19 patients with hypertension.


Driving time and compliance with post-vasectomy semen analysis drop-off.
Current literature demonstrates low rates of compliance with post vasectomy semen analysis (PVSA). This study sought to determine factors that correlate with non-compliance with PVSA. A retrospective chart review was conducted for patients who underwent vasectomy within our institution. ArcGIS was used to securely calculate the shortest driving time from each patient's home to the single PVSA drop-off site. Kruskal-Wallis and chi-squared tests analyzed characteristics of patients who did and did not submit PVSA samples, and odds ratios were calculated via multivariable logistic regression. Overall, 515 of 850 patients met inclusion criteria and 219 (42.5%) of these had no recorded PVSA. Of those with a PVSA, 59% were completed within 16 weeks. Compliance with PVSA was associated with a shorter median driving time (30.6min vs 34.2min), more vasectomy in the operating room (19% vs 10%), and attending a follow-up appointment (40% vs 17%) (p<0.005 for all). Age at vasectomy, race, ethnicity, BMI, paternity, and location of pre-operative consultation did not significantly differ between the groups. Each 30 minutes of driving time was associated with a 48% reduction in the odds of a patient submitting PVSA at any time (OR 0.52 [0.37, 0.73]). As driving time to a drop-off center appears to be a significant barrier to PVSA compliance, providers should consider alternative collection methods such as at-home or in-office semen analysis.


Double-stranded RNA induces similar pulmonary dysfunction to respiratory syncytial virus in BALB/c mice.
Both respiratory syncytial virus (RSV) and influenza A virus induce nucleotide/P2Y purinergic receptor-mediated impairment of alveolar fluid clearance (AFC), which contributes to formation of lung edema. Although genetically dissimilar, both viruses generate double-stranded RNA replication intermediates, which act as Toll-like receptor (TLR)-3 ligands. We hypothesized that double-stranded RNA/TLR-3 signaling underlies nucleotide-mediated inhibition of amiloride-sensitive AFC in both infections. We found that addition of the synthetic double-stranded RNA analog poly-inosinic-cytidylic acid [poly(I:C)] (500 ng/ml) to the AFC instillate resulted in nucleotide/P2Y purinergic receptor-mediated inhibition of amiloride-sensitive AFC in BALB/c mice but had no effect on cystic fibrosis transmembrane regulator (CFTR)-mediated Cl(-) transport. Poly(I:C) also induced acute keratinocyte cytokine-mediated AFC insensitivity to stimulation by the β-adrenergic agonist terbutaline. Inhibitory effects of poly(I:C) on AFC were absent in TLR-3(-/-) mice and were not replicated by addition to the AFC instillate of ligands for other TLRs except TLR-2. Intranasal poly(I:C) administration (250 μg/mouse) similarly induced nucleotide-dependent AFC inhibition 2-3 days later, together with increased lung water content and neutrophilic inflammation. Intranasal treatment of BALB/c mice with poly(I:C) did not induce airway hyperresponsiveness at day 2 but did result in insensitivity to airway bronchodilation by β-adrenergic agonists. These findings suggest that viral double-stranded RNA replication intermediates induce nucleotide-mediated impairment of amiloride-sensitive AFC in both infections, together with β-adrenergic agonist insensitivity. Both of these effects also occur in RSV infection. However, double-stranded RNA replication intermediates do not appear to be sufficient to induce either adenosine-mediated, CFTR-dependent Cl(-) secretion in the lung or severe, lethal hypoxemia, both of which are features of influenza infection.


[Characteristics of oxygen balance and acid-base equilibrium of the blood in patients with hereditary erythrocytosis].
Blood oxygen balance and acid-base equilibrium were studied in 22 Chuvash patients with hereditary erythrocytosis. Despite the lack of the anomalous types of hemoglobin in the given type of erythrocytosis a number of the patients showed secondary tissue hypoxia associated with hyperoxia of the peripheral venous blood. It is shown that tissues are capable of extracting oxygen from the blood. It is assumed that the development of tissue hypoxia in the patients is determined by a fall in the rate of oxygen diffussion from the capillaries to the tissues or by blood shunting via the arteriolo-venular anastomoses because of microcirculatory disorders. The lowering of hematocrit to the level close to normal with the aid of bloodlettings makes oxygen balance return to normal and reduces tissue hypoxia.


'As above, so below' examining the interplay between emotion and the immune system.
While the concept of a palpable relationship between our mental and physical well-being is certainly not new, it is only in the light of modern scientific research that we have begun to realize how deeply connected our emotional and immune states may be. We begin this review with a series of studies demonstrating how four fundamental emotional responses: anger, anxiety, mirth and relaxation are able modulate cytokine production and cellular responses to a variety of immune stimuli. These modulations are shown to be either detrimental or beneficial to a patient's health dependent on the context and duration of the emotion. We also discuss the reverse, highlighting research demonstrating how the loss of key immune cells such as T lymphocytes in clinical and animal studies can negatively impact both emotional well-being and cognition. Additionally, to give a more complete picture of the manifold pathways that link emotion and the immune system, we give a brief overview of the influence the digestive system has upon mental and immunological health. Finally, throughout this review we attempt to highlight the therapeutic potential of this burgeoning field of research in both the diagnosis and treatment of immune and disorders. As well as identifying some of the key obstacles the field must address in order to put this potential into practice.


Planning to change diet: a controlled trial of an implementation intentions training intervention to reduce saturated fat intake among patients after myocardial infarction.
This article investigates the effects of a brief psychological intervention-implementation intentions training-on the reduction of saturated fat intake among patients after myocardial infarction (MI). One hundred fourteen patients who had experienced a first uncomplicated MI took part in the study. Data were collected at approximately 1 week after MI, 2 weeks after short-term Phase 2 cardiac rehabilitation (approximately 2 months after MI), and 6 months after rehabilitation (8 months after MI). After data collection at 2 weeks after rehabilitation, patients were randomly assigned to the control group or the intervention group (an individually delivered implementation intentions training). Daily saturated fat intake was used as the primary outcome; total fat intake and percentage of calories from fat were secondary outcomes. Repeated-measures analysis of variance showed a significant TimexGroup interaction: Compared to time before MI, patients in both groups reported a decrease in saturated fat intake at 2 weeks after rehabilitation. Those who participated in the implementation intentions intervention were able to further decrease saturated fat intake from 22.88 g at 2 months after MI to 19.71 g at 8 months after MI. Patients from the control group maintained the same level of saturated fat intake at 2 months after MI (mean=22.30) and 6 months later (mean=22.47). An individually delivered implementation intentions intervention may reduce saturated fat intake among patients after MI.


Lupus anticoagulant-hypoprothrombinemia syndrome and immunoglobulin-A vasculitis: a report of Japanese sibling cases and review of the literature.
Lupus anticoagulant-hypoprothrombinemia syndrome (LAHPS) is a rare bleeding disorder caused by antiprothrombin antibodies. LAHPS is associated with systemic lupus erythematosus (SLE) or infections. We describe two Japanese brothers with immunoglobulin-A vasculitis (IgAV) who met the diagnostic criteria of LAHPS. They presented with palpable purpura and abdominal pain, and had a prolonged activated partial thromboplastin time (APTT) and prothrombin deficiency with the presence of lupus anticoagulant. Pediatric LAHPS was reviewed in abstracts from the Japan Medical Abstracts Society that were written in Japanese and PubMed or Web of Science-listed articles in English between 1996 and 2019. Including our cases, pediatric LAHPS has been reported in 40 Japanese and 46 non-Japanese patients. We summarized the clinical and laboratory characteristics of all 86 cases, and found only one Japanese LAHPS case with IgAV, except for our cases. Of the 86 cases, most were associated with infections followed by SLE. The presence of SLE, older age, lower prothrombin levels, severe bleeding symptoms, and positivity of immunoglobulin G anticardiolipin antibodies and anticardiolipin/β2-glycoprotein I antibodies and/or β2-glycoprotein I-dependent anticardiolipin antibodies had higher odds of requiring treatment. Measuring the APTT and prothrombin time (PT) might be required in patients with IgAV when they do not have a typical clinical course or distinctive symptoms. LAHPS should be considered with prolongation of the APTT and/or PT. Additionally, it is important to maintain a balance between the risk of thrombosis and hemorrhage when normalization of the PT and FII levels occurs in LAHPS cases under treatment.


Signals from Single-Opponent Cortical Cells in the Human cVEP.
We used the chromatic visual evoked potential (cVEP) to study responses in human visual cortex evoked by equiluminant color stimuli for 6 male and 11 female observers. Large-area, colored squares were used to stimulate Single-Opponent cells preferentially, and fine color-checkerboard stimuli were used to activate Double-Opponent responses preferentially. Stimuli were modulated along the following two directions in color space: (1) the cardinal direction, L-M or M-L of DKL (Derrington, Krauskopf, and Lennie) space; and (2) the line from the white point to the color of the Red LED in the display screen, which was approximately intermediate between the L-M and -S directions in DKL space in cone-contrast coordinates. The amplitudes of cVEPs to large squares were smaller than those to checkerboards, and the latency of the cVEP response to squares was significantly less than the checkerboard latency. The latency of cVEP responses to the squares varied little with cone-contrast unlike the steep reduction of latency with cone-contrast observed in responses to color checkerboard patterns. The dynamic differences between cVEPs to squares and checkerboards support the hypothesis that a distinct neuronal mechanism responded to squares: Single-Opponent cells. Response amplitude, latency, and transientness-and their dependence on cone-contrast-were similar in the responses in the L-M and Red color directions. The similarity supports the hypothesis that the Single-Opponent signals in the cVEP come from a distinct population of cells that receives subtractive inputs from L and M cones, either L-M or M-L.SIGNIFICANCE STATEMENT This article is about characterizing the visual behavior of a distinct population of neurons in the human visual cortex, the Single-Opponent color cells. Based on single-cell results in the visual cortex of macaque monkeys, we used large uniformly colored stimuli to isolate the responses of Single-Opponent cells in the chromatic visual evoked potential (cVEP) recorded on the scalp of human observers. VEP signals recorded under conditions believed to reveal Single-Opponent responses are small and transient. Their time course is relatively unaffected by cone-contrast, and they are relatively insensitive to stimulus modulation of short wavelength-sensitive S cones. Because Single-Opponent cells convey signals that can be used to judge the color of scene illumination, knowing their visual properties is important for understanding color vision.


Isolation and characterization of Corynebacterium diphtheriae nontandem double lysogens hyperproducing CRM197.
Phage beta 197tox-, which codes for CRM197, a nontoxic protein immunochemically indistinguishable from diphtheria toxin, was UV induced from a culture of the C7(beta 197)tox- strain. A total of 191 C7(beta 197)tox- lysogens were isolated and selected according to the halo produced on TYE agar containing antidiphtheria toxin serum and were further characterized by Southern blots of their chromosomal DNA. Most of the isolates turned out to be monolysogens, but some tandem and nontandem double lysogens were also found. The nontandem double lysogens were stable and capable of giving high yields of CRM197, up to threefold higher than monolysogens. They are, therefore, suitable for large-scale industrial production.


Complexity of Delivering Precision Medicine: Opportunities and Challenges.
Precision medicine has emerged as a tool to match patients with the appropriate treatment based on the precise molecular features of an individual patient's tumor. Although examples of targeted therapies exist resulting in dramatic improvements in patient outcomes, comprehensive genomic profiling of tumors has also demonstrated the incredible complexity of molecular alterations in tissue and blood. These sequencing methods provide opportunities to study the landscape of tumors at baseline and serially in response to treatment. These tools also serve as important biomarkers to detect resistance to treatment and determine higher likelihood of responding to particular treatments, such as immune checkpoint blockade. Federally funded and publicly available data repositories have emerged as mechanisms for data sharing. In addition, novel clinical trials are emerging to develop new ways of incorporating molecular matched therapy into clinical trials. Various challenges to delivery of precision oncology include understanding the complexity of advanced tumors based on evolving "omics" and treatment resistance. For physicians, determining when and how to incorporate genetic and molecular tools into clinic in a cost-effective manner is critical. Finally, we discuss the importance of well-designed prospective clinical trials, biomarkers such as liquid biopsies, the use of multidisciplinary tumor boards, and data sharing as evidence-based medicine tools to optimally study and deliver precision oncology to our patients.


Relationship of adiponectin with markers of systemic inflammation, atherogenic dyslipidemia, and heart failure in patients with coronary heart disease.
Adiponectin, an adipocyte-derived hormone, appears to be a modulator of lipid metabolism and systemic inflammation and is present in particularly low concentrations in patients with coronary heart disease (CHD). However, the clinical importance of adiponectin in individuals at markedly high risk for future cardiovascular morbidity and mortality has not been fully elucidated. We examined the associations between serum adiponectin and several biomarkers related to cardiovascular disease and heart failure in a large high-risk population comprising patients with prevalent CHD. We measured fasting adiponectin, interleukin-6 (IL-6), high-sensitivity C-reactive protein (hsCRP), N-terminal pro-B-type natriuretic peptide (NT-proBNP), and markers of lipoprotein metabolism in 1174 patients with CHD. After adjustment for age and sex, adiponectin was associated with HDL-cholesterol (HDL-C; r = 0.25; P <0.0001), NT-proBNP (r = 0.17; P <0.0001), and plasma triglyceride (r = -0.21; P <0.0001) concentrations. There was, however, no statistically significant association between adiponectin and markers of systemic inflammation. In partial correlation analyses further adjusted for body mass index, alcohol intake, smoking status, presence of diabetes and/or hypertension, lipid-lowering drug therapy, and fasting plasma glucose, adiponectin remained significantly associated with HDL-C (r = 0.21; P <0.0001), NT-proBNP (r = 0.15; P <0.0001), and plasma triglycerides (r = -0.16; P <0.0001). Serum adiponectin is associated with the presence of atherogenic dyslipidemia and with NT-proBNP concentration but not with markers of systemic inflammation in patients with manifest CHD. Thus, atherogenic dyslipidemia may link adiponectin with the progression of atherosclerosis. Moreover, serum adiponectin may be related to BNP in patients with CHD.


The schizophrenia susceptibility gene dysbindin controls synaptic homeostasis.
The molecular mechanisms that achieve homeostatic stabilization of neural function remain largely unknown. To better understand how neural function is stabilized during development and throughout life, we used an electrophysiology-based forward genetic screen and assessed the function of more than 250 neuronally expressed genes for a role in the homeostatic modulation of synaptic transmission in Drosophila. This screen ruled out the involvement of numerous synaptic proteins and identified a critical function for dysbindin, a gene linked to schizophrenia in humans. We found that dysbindin is required presynaptically for the retrograde, homeostatic modulation of neurotransmission, and functions in a dose-dependent manner downstream or independently of calcium influx. Thus, dysbindin is essential for adaptive neural plasticity and may link altered homeostatic signaling with a complex neurological disease.


The hormonal control of sexual development.
The formation of the testis or ovary is a critical step in development. The pioneering studies of Professor Alfred Jost showed that the hormones produced by the embryonic rabbit testis are essential for development of the male phenotype. Sexually dimorphic hormones play a key role in the transition from an undifferentiated gonad into the mature testis and ovary. Marsupials, with their altricial young, provide an accessible model for the study of sexual differentiation because most of these events occur postnatally, while the young are attached to teats within their mothers' pouches. The relatively long time-course for the marsupial sexual differentiation has provided an excellent opportunity to correlate morphological changes with the genes and hormones that control them. Using this model species we have demonstrated that not all sexual dimorphisms are controlled by hormones. Virilization of the prostate and phallus is androgen dependent but appears to rely on circulating 5alpha-androstane-3alpha, 17beta-diol which is converted to dihydrotestosterone in these target tissues. Collectively these studies have led to the development of new paradigms to explain the hormonal mechanisms mediating sexual differentiation.


Levels of plasma-soluble triggering receptor expressed on myeloid cells-1 (sTREM-1) are correlated with disease activity in rheumatoid arthritis.
To determine whether levels of plasma-soluble triggering receptor expressed on myeloid cells-1 (sTREM-1) are elevated in patients with rheumatoid arthritis (RA) and whether levels are correlated with disease activity and other variables. Our study included 71 patients with RA and 50 age- and sex-matched healthy controls. Clinical characteristics and laboratory measures, including erythrocyte sedimentation rate (ESR), C-reactive protein (CRP), and 28-joint Disease Activity Score (DAS28) were assessed. Plasma levels of sTREM-1 and tumor necrosis factor-α (TNF-α) were measured by ELISA. Patients with RA had significantly higher plasma sTREM-1 levels than healthy controls (170.10 ± 84.71 pg/ml vs 97.41 ± 40.64 pg/ml; p < 0.001). In patients with RA, plasma sTREM-1 levels were found to be correlated with DAS28, ESR, CRP, white blood cell counts, neutrophil counts, and plasma TNF-α levels (r = 0.329, p = 0.005; r = 0.241, p = 0.043; r = 0.314, p < 0.001; r = 0.261, p = 0.028; r = 0.278, p = 0.019; and r = 0.313, p = 0.009, respectively). Plasma sTREM-1 levels in patients with active disease status (DAS28 > 3.2) were significantly higher than in those with low disease status (DAS28 ≤ 3.2; 208.89 ± 100.14 pg/ml vs 150.29 ± 68.70 pg/ml; p = 0.005). Patients with RA had higher plasma sTREM-1 levels than healthy controls, and plasma sTREM-1 levels were correlated with disease activity measures, suggesting that plasma sTREM-1 could play a role in the inflammatory process associated with TNF-α, and that it may be a useful disease activity marker in RA.


The Kleisin Subunits of Cohesin are Involved in the Fate Determination of Embryonic Stem Cells.
As a potential candidate to generate an everlasting cell source to treat various diseases, embryonic stem cells are regarded as a promising therapeutic tool in the regenerative medicine field. Cohesin, a multi-functional complex that controls various cellular activities, plays roles not only in organizing chromosome dynamics but also in controlling transcriptional activities related to self-renewal and differentiation of stem cells. Here, we report a novel role of the α-kleisin subunits of cohesin (RAD21 and REC8) in the maintenance of the balance between these two stem-cell processes. By knocking down REC8, RAD21, or the non-kleisin cohesin subunit SMC3 in mouse embryonic stem cells, we show that reduction in cohesin level impairs their self-renewal. Interestingly, the transcriptomic analysis revealed that knocking down each cohesin subunit enables the differentiation of embryonic stem cells into specific lineages. Specifically, embryonic stem cells in which cohesin subunit RAD21 were knocked down differentiated into cells expressing neural alongside germline lineage markers. Thus, we conclude that cohesin appears to control the fate determination of embryonic stem cells.


Preventing kidney failure: primary care physicians must intervene earlier.
Mild chronic kidney disease often goes unnoticed until a substantial loss of renal function has occurred. Given the increasing incidence of chronic kidney disease, primary care physicians play a critical role in the early evaluation and intervention of patients at risk. This article discusses the key steps, with emphasis on patients with mild disease due to diabetes or hypertension.