Aorta response to secretin in intact and diabetic rats.
The influence of secretin (10(-10) - 10(-8) M), a gastrointestinal hormone, on the relaxing response of rat thoracic aorta rings preconstricted by 40 mM KCl was studied by measuring changes in isometric tension in intact and diabetic animals. Initial contraction of aorta rings was markedly decreased in diabetic state. Secretin administered at two higher doses (10(-9), 10(-8) M) caused significant relaxation. In diabetes, relaxing effect was observed at all three doses of the peptide. These data indicate that secretin relaxes thoracic aorta rings and state of diabetes markedly amplifies the relaxant response to this peptide.


Bridging to family and community support for older adults and the domain management model.
Older adults with chronic illness and functional impairment require continuous and long-term monitoring and supervision. Changes in the health care system make this harder to accomplish. Social work assistance can help identify issues and provide help through counseling, patient advocacy, resource utilization, and education. Tools that help include the Domain Management Model and well-organized team care. This article describes these interventions in a senior primary care practice and community-based screening and education program. Both programs bridge from acute illness care to improved chronic illness management in the community.


Federal legislative support for independent living.
This article details the federal legislative background, development, and outlook, and current activities by the states, in relation to establishing services to help adolescents in foster care prepare for independent living. The authors also specify directions that future legislation should take to expand assistance.


Osteopontin is elevated in patients with mitral annulus calcification independent from classic cardiovascular risk factors.
Osteopontin (OPN) regulates the Ca(++)-deposition in bone and coronary arteries. Elevated OPN were also associated with (aortic) valve calcification in healthy individuals. This study aimed to investigate the association between OPN levels and mitral annulus calcification (MAC) in patients with coronary artery disease (CAD). In this cross-sectional study OPN-levels were measured in 223 non-or ex-smoking patients (160 male, mean age: 61,09 ± 11,02 years; 63 female: mean age: 67,49 ± 7,87 years) with CAD. Plasma OPN levels were measured by ELISA and MAC was evaluated by echocardiography. Forward stepwise logistic regression analysis (likelihood quotient) showed significantly higher OPN-levels in patients with MAC compared to patient without calcified mitral annulus independent from the classic risk factors age and severity of coronary artery disease (CAD). In addition to age and the severity of CAD, the circulating OPN amount was a significant predictor for MAC. This is the first clinical trial which observed increased circulating OPN levels in MAC, suggesting a distinct role of OPN in the process of MAC. Considering the current knowledge about OPN it is more likely that OPN does not promote but counteracts valve calcification and therefore is elevated in course of a calcification processes.


In vivo virtual histology intravascular ultrasound correlates of risk factors for sudden coronary death in men: results from the prospective, multi-centre virtual histology intravascular ultrasound registry.
We hypothesized a relationship between virtual histology intravascular ultrasound (VH-IVUS) findings and risk factors histopathologically associated with sudden coronary death (SCD) in men: cigarette smoking and an increased total cholesterol-to-high-density lipoprotein cholesterol (HDL-C) ratio (TC/HDL > 5). We assessed volumetric VH-IVUS parameters in a consecutive series of 473 male patients: fibrous, fibro-fatty, dense calcium (DC), necrotic core (NC), and a calculated NC/DC ratio. Patients' age was 61 ± 11 years, with 27% smokers and 69% having a lipid disorder. The NC/DC ratio was the only VH-IVUS parameter related to both TC/HDL ratio (r = 0.18, P= 0.0008) and low-density lipoprotein cholesterol levels (r = 0.17, P= 0.002); had a negative correlation with HDL-C levels (r = -0.11, P= 0.03); and was higher for smokers [median 1.98 (1.35-3.18)] vs. non-smokers [median 1.70 (1.23-2.53), P= 0.006]. An NC/DC value >3 was the threshold that best identified smokers and/or patients presenting TC/HDL >5 (odds ratio 3.0, 95% CI 1.7-4.9, P= 0.0001), and receiver-operator curves showed the superiority of the NC/DC ratio [area under curve (AUC) 0.64, P < 0.0001] over %DC (AUC 0.58, P= 0.006) or %NC (AUC 0.51, P= 0.43) to identify these patients. The ratio of NC to calcification detected by VH-IVUS in diseased coronary segments is related to known risk factors for SCD and, thus, may be associated with a worse prognosis.


Autonomic dysfunction predicts mortality in patients with multiple organ dysfunction syndrome of different age groups.
Multiple organ dysfunction syndrome (MODS) is the sequential failure of several organ systems after a trigger event, like sepsis or cardiogenic shock. Mortality rate is high, up to 70%. Autonomic dysfunction may substantially contribute to the development of MODS. Our study aimed to characterize a) the spectrum of autonomic dysfunction of critically ill MODS patients; b) whether autonomic dysfunction is different in patients receiving sedation, mechanical ventilation, or catecholamines; c) the age dependency of autonomic dysfunction in MODS; and d) whether autonomic dysfunction predicts mortality in MODS. Prospective cohort study. Twelve-bed medical intensive care unit in a university center. Ninety consecutively admitted score-defined MODS patients. Assessment of heart rate variability, baroreflex sensitivity, and chemoreflex sensitivity as markers of autonomic dysfunction. The patients were followed for 28-day mortality. Baroreflex sensitivity, chemoreflex sensitivity, and almost all indexes of heart rate variability were attenuated in comparison to normal range data. There was no association between the assessed heart rate variability variables, baroreflex sensitivity or chemoreflex sensitivity, and the presence of sedation or catecholamine therapy. Except for frequency-domain variables, pNN50 (percentage of differences of successive RR intervals differing >50 msecs) and rMSSD (root mean square of successive difference of N-N intervals), none of the measured variables were related to the presence of mechanical ventilation. Age dependency was detected for baroreflex sensitivity but not for heart rate variability indexes or chemoreflex sensitivity (across ages 24-96 yrs). lnVLF predicted 28-day mortality best in the entire cohort of patients and in a subgroup of patients with cardiogenic-triggered MODS. Autonomic function of MODS patients is blunted, and this attenuation has prognostic implications. The extensive influence of MODS on autonomic function overwhelms and masks the well-known age dependency of autonomic function seen in healthy persons.


[Rehabilitation of war injuries of the upper extremities with peripheral nerve lesions at the and Praxis Clinic for physical medicine and rehabilitation in Sarajevo].
The war injuries are the most frequent multiple, with the difficult distructions of the tissue and the lesions of the peripheral nerves. By the injuries, the lesions of the nerv system represent the delicate problem for the physical medicine and the rehabilitation during the siege of Sarajevo (1992-1995), in the injuries with the lesions of the peripheral nerves, if they are not treated in the frame of the multiple las urgent, they are postponed the operative treatments longer than three months. This is from the aspect of the successfullness, prognostically and therapeutically, the limiting moment. The successfullness of the operative treatment of the lesions of the peripheral nerves significantly depends on that whether the operation was performed incide tree months. The more difficult consequences in these injuries can appear if simultaneously with the taking care of war injuries does not perform adequately surgical treatment adn the in time physical therapy trough the sufficient long time period. On the four year sample was analyzed the success fullness of the rehabilitation of the injuries of the upper extremities with the lesions of the peripheral nerves. Clinically, uniform, we valorized the success fullness of the treatment of all the patients by the marks from 0.5. In the complete sample the excellent success of the rehabilitation we confirmed in more than a half of patients. The better results show the group of the operated patients, in which is performed neuropathia or neurolisis. The capability of the regeneration of the injured peripheral nerves offers the real possibility for extraordinary recovery also in the most difficult injuries, then the preventions of the significant number of the consequences.


Clinical trials.
Clinical trials are a relatively underused form of investigation in family medicine. This paper presents an overview of those considering conducting or assessing a clinical trial. A bibliography for further reading is also provided. Topics covered include aspects of: the development of a protocol; design such as randomization and binding; the special role of non-drug studies in family medicine; measurement--especially combating bias; analysis--particularly the management of drop-outs; and, the problem of generalizability.


Dicoumarol toxicity in cattle associated with ingestion of silage containing sweet vernal grass (Anthoxanthum odoratum).
A diagnosis of dicoumarol toxicity in a herd of Friesian cattle was made following investigation of the deaths of three mature cows and eleven yearling heifers. Affected stock had been fed wrapped, bailed silage containing approximately 90% sweet vernal grass (Anthoxanthum odoratum). Sweet vernal grass contains coumarin, which can be converted to dicoumarol, a vitamin K antagonist, through the action of moulds. Most deaths were preceded by lethargy, severe anaemia and subcutaneous and internal haemorrhage. Dicoumarol toxicosis was suspected based on clinical signs, necropsy findings and prolonged prothrombin and activated partial thromboplastin times. Dicoumarol analysis of blood from affected animals and silage confirmed the diagnosis. Activated partial thromboplastin time Haemoglobin Packed cell volume Prothrombin time Red cell count


Predictive Mixing for Density Functional Theory (and Other Fixed-Point Problems).
Density functional theory calculations use a significant fraction of current supercomputing time. The resources required scale with the problem size, the internal workings of the code, and the number of iterations to convergence, with the latter being controlled by what is called "mixing". This paper describes a new approach to handling trust regions within these and other fixed-point problems. Rather than adjusting the trust region based upon improvement, the prior steps are used to estimate what the parameters and trust regions should be, effectively estimating the optimal Polyak step from the prior history. Detailed results are shown for eight structures using both the "good" and "bad" multisecant versions as well as the Anderson method and a hybrid approach, all with the same predictive method. Additional comparisons are made for 36 cases with a fixed algorithm greed. The predictive method works well independent of which method is used for the candidate step, and it is capable of adapting to different problem types particularly when coupled with the hybrid approach.


Endometrial ablation complicated by fatal hyponatremic encephalopathy.
To investigate the occurrence of hyponatremic encephalopathy and the effects of therapy for symptomatic hyponatremia on outcome in women undergoing endometrial ablation. Consultations from university-affiliated and community hospitals from June 1991 to June 1992. Four generally healthy women who underwent elective endometrial ablation for dysfunctional bleeding and developed hyponatremic encephalopathy. Three patients were promptly treated with hypertonic (514 mmol/L) sodium chloride such that the serum sodium level increased from 102 to 123 mmol/L within 24 hours. The fourth patient was not treated until after suffering grand mal seizures followed by respiratory arrest. Of the four patients who developed hyponatremic encephalopathy, the diagnosis was established before respiratory arrest occurred in three. The operative procedure was terminated and all three were aggressively treated with intravenous hypertonic sodium chloride, which raised the serum sodium level to modestly hyponatremic levels (120 to 130 mmol/L). All three completely recovered without sequelae. The fourth patient suffered respiratory arrest before therapy could be initiated. The patient remained comatose, and central diabetes mellitus and insipidus developed. She never regained consciousness and died after several days. Autopsy revealed cerebral edema and tonsillar herniation. The mean (+/- SD) preoperative serum sodium level was 138 +/- 1 mmol/L; at the time of diagnosis of hyponatremia, it was 107 +/- 13 mmol/L. In two patients, hyponatremic encephalopathy was diagnosed intraoperatively because of tremulousness and either hypothermia or hypoxemia. In the other two patients, the diagnosis was made postoperatively because of headache, nausea, emesis, and in one of these patients, respiratory arrest. Women undergoing elective endometrial ablation can develop severe symptomatic hyponatremia, which can be fatal. The presence of symptoms suggesting hypo-osmolality should lead to immediate measurement of plasma sodium level. If hyponatremia with hypo-osmolality is present, early and appropriate therapy for the hyponatremia should be instituted before respiratory insufficiency occurs.


Host regulation of lysogenic decision in bacteriophage lambda: transmembrane modulation of FtsH (HflB), the cII degrading protease, by HflKC (HflA).
The cII gene product of bacteriophage lambda is unstable and required for the establishment of lysogenization. Its intracellular amount is important for the decision between lytic growth and lysogenization. Two genetic loci of Escherichia coli are crucial for these commitments of infecting lambda genome. One of them, hflA encodes the HflKC membrane protein complex, which has been believed to be a protease degrading the cII protein. However, both its absence and overproduction stabilized cII in vivo and the proposed serine protease-like sequence motif in HflC was dispensable for the lysogenization control. Moreover, the HflKC protein was found to reside on the periplasmic side of the plasma membrane. In contrast, the other host gene, ftsH (hflB) encoding an integral membrane ATPase/protease, is positively required for degradation of cII, since loss of its function stabilized cII and its overexpression accelerated the cII degradation. In vitro, purified FtsH catalyzed ATP-dependent proteolysis of cII and HflKC antagonized the FtsH action. These results, together with our previous finding that FtsH and HflKC form a complex, suggest that FtsH is the cII degrading protease and HflKC is a modulator of the FtsH function. We propose that this transmembrane modulation differentiates the FtsH actions to different substrate proteins such as the membrane-bound SecY protein and the cytosolic cII protein. This study necessitates a revision of the prevailing view about the host control over lambda lysogenic decision.


High-resolution peptide separations using nano-LC at ultra-high pressure.
We report on the optimization of nano-LC gradient separations of proteomic samples that vary in complexity. The gradient performance limits were visualized by kinetic plots depicting the gradient time needed to achieve a certain peak capacity, while using the maximum system pressure of 80 MPa. The selection of the optimal particle size/column length combination and corresponding gradient steepness was based on scouting the performance of 75 μm id capillary columns packed with 2, 3, and 5 μm fully porous silica C18 particles. At optimal gradient conditions, peak capacities up to 500 can be obtained within a 120 min gradient using 2 μm particle-packed capillary columns. Separations of proteomic samples including a cytochrome c tryptic digest, a bovine serum albumin tryptic digest, a six protein mix digest, and an Escherichia coli digest are demonstrated while operating at the kinetic-performance limit, i.e. using 2-μm packed columns, adjusting the column length and scaling the gradient steepness according to sample complexity. Finally, good run-to-run retention time stability (RSD values below 0.18%) was demonstrated applying ultra-high pressure conditions.


Finite Element Simulation and Sensitivity Analysis of the Cohesive Parameters for Delamination Modeling in Power Electronics Packages.
Delamination is a critical failure mode in power electronics packages that can significantly impact their reliability and performance, due to the large amounts of electrical power managed by the most recent devices which induce remarkable thermomechanical loads. The finite element (FE) simulation of this phenomenon is very challenging for the identification of the appropriate modeling tools and their subsequent calibration. In this study, we present an advanced FE modeling approach for delamination, together with fundamental guidelines to calibrate it. Considering a reference power electronics package subjected to thermomechanical loads, FE simulations with a global-local approach are proposed, also including the implementation of a bi-linear cohesive zone model (CZM) to simulate the complex interfacial behavior between the different layers of the package. A parametric study and sensitivity analysis is presented, exploring the effects of individual CZM variables on the delamination behavior, identifying the most crucial ones and accurately describing their underlying functioning. Then, this work gives valuable insights and guidelines related to advanced and aware FE simulations of delamination in power electronics packages, useful for the design and optimization of these devices to mitigate their vulnerability to thermomechanical loads.


Hypophysectomy for palliation of breast cancer with especial reference to surgical technique.
In a series of 19 patients who had hypophysectomy for palliation of cancer of the breast, all had striking relief of pain. Objective evidence of remission of disease was observed in ten patients. The survival time after operation averaged 19.8 months for patients who had remission and 2.1 months for patients who did not. Results were better in patients who had had cancer for a long time before operation than in those who had had the disease a relatively short time. Also it was noted that results were better in patients who had had preoperative response to endocrine therapy than in those who had not. Hence these factors may be considered in selection of patients for hypophysectomy.


[Degree of trait anxiety in a sample of athletes of the Puerto Rican delegation to the XII Pan American Games held in Winnipeg '99].
This study was conducted in an attempt to determine the degree of anxiety generated by athletes of the Puerto Rican delegation to the XII Pan American Games held in Winnipeng'99 to develop support programs that contribute to the enhancement of the athletes' mental skills and the development of sports, psychology, in particular. The sample consisted of 55 athletes that participated in the abovementioned Pan American Games. The athletes represented 16 sorts and made up 40% of the Puerto Rican athletes officially registered to participate. The Sport Competition Anxiety Test-Form A was used to detect trait anxiety during competition. An analysis of the variables was conducted utilizing t-tests to determine whether there were significant (p < or = 0.05) between the quantitative variables under study. The results indicated that the athletes of the Puerto Rican delegation that participated in the XII Pan American Games have a degree of trait anxiety that needs professional attention.


Salt-induced nephropathy in obese spontaneously hypertensive rats via paradoxical activation of the mineralocorticoid receptor: role of oxidative stress.
Aldosterone is implicated in the pathogenesis of proteinuria and chronic kidney disease. We previously demonstrated the contribution of elevated serum aldosterone in the early nephropathy of SHR/NDmcr-cp (SHR/cp), a rat model of metabolic syndrome. In the present study, we investigated the effect of salt loading on renal damage in SHR/cps and explored the underlying mechanisms. SHR/cps fed a high-sodium diet for 4 weeks developed severe hypertension, massive proteinuria, and advanced renal lesions. High salt also worsened glomerular podocyte impairment. Surprisingly, selective mineralocorticoid receptor (MR) antagonist eplerenone dramatically ameliorated the salt-induced proteinuria and renal injury in SHR/cps. Although salt loading reduced circulating aldosterone, it increased nuclear MR and expression of aldosterone effector kinase Sgk1 in the kidney. Gene expressions of transforming growth factor-beta1 and plasminogen activator inhibitor-1 were also enhanced in the kidneys of salt-loaded SHR/cps, and eplerenone completely inhibited these injury markers. To clarify the discrepancy between decreased aldosterone and enhanced MR signaling by salt, we further investigated the role of oxidative stress, a putative key factor mediating salt-induced tissue damage. Interestingly, antioxidant Tempol attenuated the salt-evoked MR upregulation and Sgk1 induction and alleviated proteinuria and renal histological abnormalities, suggesting the involvement of oxidative stress in salt-induced MR activation. MR activation by salt was not attributed to increased serum corticosterone or reduced 11beta-hydroxysteroid dehydrogenase type 2 activity. In conclusion, sodium loading exacerbated proteinuria and renal injury in metabolic syndrome rats. Salt reduced circulating aldosterone but caused renal MR activation at least partially via induction of oxidative stress, and eplerenone effectively improved the nephropathy.


Mutant gene-induced disorders of structure, function and thyroglobulin synthesis in congenital goitre (cog/cog) in mice.
We have investigated thyroid structure and function in mice homozygous for the chromosome 15 mutation, congenital goitre (cog). Abnormal thyroidal hypertrophy and reduced iodine uptake in cog/cog mice were observed as early as day 18 of gestation, corresponding to the onset of thyroid function. Growth continued unabated in mutants throughout the 10-month period of observation. By 2 months of age, thyroid cell hypertrophy obliterated nearly all follicular lumina in cog/cog glands and by 10 months mean mutant thyroid mass exceeded that of age-matched littermates. Twenty-fold serum concentrations of thyrotrophin were significantly increased at all ages examined. While wild type (+/+) and heterozygote (+/cog) mice are indistinguishable from each other, thyroids of homozygote mutants (cog/cog) and the +/cog type are easily discernible from thyroids of the +/+ type by microscopic and thyroglobulin (Tg) analyses. Thyrofollicular cells of both cog/cog and +/cog genotypes contain large vesicles of accumulated, nonglycosylated proteinaceous material not observed in cells from +/+ mice. Autoradiography showed 125I was incorporated only into Tg within recognizable follicular lumina of thyroids from +/cog mice. Serum concentrations of tri-iodothyronine are depressed during development in cog/cog mice. Serum concentrations of thyroxine are depressed during postnatal development but increase progressively to normal concentrations by 10 months of age. Our analyses indicate that full size Tg is produced in thyroid cells from cog/cog mice, though in a greatly reduced quantity, and that Tgs which are several sizes smaller than normal are also produced in both homozygote and heterozygote thyroids.(ABSTRACT TRUNCATED AT 250 WORDS)


Pregnancy outcome in women requiring chronic hemodialysis.
Chronic renal failure is strongly associated with poor pregnancy outcome. Women dependent upon hemodialysis before conception rarely achieve a successful live birth. Pregnancies in three women requiring chronic hemodialysis before conception and throughout gestation resulted in three live births, of whom one survived the neonatal period. The risks of dialysis in pregnancy (hemodialysis and peritoneal dialysis) are significant to both the mother and fetus. Polyhydramnios appears to be a major complication in women dependent upon chronic hemodialysis.


Assent observed: children's involvement in leukemia treatment and research discussions.
Assent is a critical issue in pediatric ethics that has not been well-studied. We examined the role of older children in discussing a recent diagnosis of acute leukemia and treatment options, including participation in a randomized, clinical trial (RCT). Our sample was drawn from data collected as part of a larger, National Cancer Institute-funded study in which we observed, tape-recorded, and transcribed the informed consent conference (ICC) for children with leukemia at 6 institutions. We analyzed the data for the child's presence and role, examined 5 discussion categories relevant to assent in the context of pediatric cancer treatment/research, conducted parent interviews, and administered clinician questionnaires to examine these issues. We report a qualitative analysis of 14 cases in which the child was present for the ICC. Clinicians varied with regard to whom they directed specific discussions when patients were present. Questions asked by the children predominantly related to details of the disease and treatment, with few questions about the RCT. Clinicians and parents attribute responsibility for decision-making about participation in RCTs in a variety of ways. In conferences that included the child-patient, parents asked significantly fewer questions than in other ICCs. There is significant variation among ICCs that include the older child. Additional empirical study of assent is warranted, and our data suggest several hypotheses that should be tested in future research.


Probabilistic HIV recency classification-a logistic regression without labeled individual level training data.
Accurate HIV incidence estimation based on individual recent infection status (recent vs long-term infection) is important for monitoring the epidemic, targeting interventions to those at greatest risk of new infection, and evaluating existing programs of prevention and treatment. Starting from 2015, the Population-based HIV Impact Assessment (PHIA) individual-level surveys are implemented in the most-affected countries in sub-Saharan Africa. PHIA is a nationally-representative HIV-focused survey that combines household visits with key questions and cutting-edge technologies such as biomarker tests for HIV antibody and HIV viral load which offer the unique opportunity of distinguishing between recent infection and long-term infection, and providing relevant HIV information by age, gender, and location. In this article, we propose a semi-supervised logistic regression model for estimating individual level HIV recency status. It incorporates information from multiple data sources - the PHIA survey where the true HIV recency status is unknown, and the cohort studies provided in the literature where the relationship between HIV recency status and the covariates are presented in the form of a contingency table. It also utilizes the national level HIV incidence estimates from the epidemiology model. Applying the proposed model to Malawi PHIA data, we demonstrate that our approach is more accurate for the individual level estimation and more appropriate for estimating HIV recency rates at aggregated levels than the current practice - the binary classification tree (BCT).


Prediction of Potential Associations Between MicroRNA and Disease Based on Bayesian Probabilistic Matrix Factorization Model.
The association between microRNAs (miRNAs) and diseases is significant to understand the development and progression of many human diseases. Given the cost and complexity of biological experiments, the computational method for predicting the potential association between miRNAs and disease will be an effective complement. In this article, we have developed a model (microRNA and disease based on Bayesian probabilistic matrix factorization, MDBPMF) based on a fully Bayesian treatment of the probabilistic matrix factorization to find potential associations between miRNAs and diseases by using the HMDDv2.0 database, which contains proven miRNA-disease associations. We show that Bayesian probabilistic matrix factorization models can be efficiently trained using Markov chain Monte Carlo methods by applying them to the HMDDv2.0 database. MDBPMF achieves reliable prediction with an average area under receiver operating characteristic curve of 0.8755 for eight complex diseases based on fivefold cross-validation, which indeed outperforms the state-of-the-art method. In addition, a case study of lung cancer further verifies the utility of our method.


Flood protection and endogenous sorting of households: the role of credit constraints.
Human migration is increasingly seen as a promising climate change adaptation and flood risk reduction strategy. The purpose of this paper is to investigate how spatial differences in flood risk, due to differences in flood protection, reduce the mobility of vulnerable households through a credit constraint mechanism. Using an equilibrium model with two households types and endogenous sorting, we show how spatial differences in flood protection lead to clustering of vulnerable households in a risky region, in a real-world setting of common United States (US) flood zones. We find clustering effects of some size for flood zones with return periods of less than 30 years.


Testing the Validity of Taxonic Schizotypy Using Genetic and Environmental Risk Variables.
Meehl regarded schizotypy as a categorial liability for schizophrenia that is the product of genes, environment, and gene-environment interactions. We sought to test whether schizophrenia-related genotypes and environmental risk factors predict membership in classes defined by taxometric analyses of positive (cognitive-perceptual), negative (interpersonal), and disorganized schizotypy. Participants (n = 500) completed the Schizotypal Personality Questionnaire (SPQ) and provided information on the following risk factors: cannabis use, pregnancy and obstetric complications, social adjustment, and family history of psychosis. Saliva samples were obtained so that the frequency of single-nucleotide polymorphism (SNP) alleles associated with risk for developing schizophrenia could be determined. Genotyped SNPs were rs1625579 (MIR137), rs7004633 (MMP16), rs7914558 (CNNM2), and rs12966547 (CCDC68). Sets of SPQ items were subject to multiple coherent cut kinetic (CCK) analyses, including mean-above-minus-below-a-cut, maximum covariance, maximum eigenvalue, and latent modes analyses. CCK analyses indicated latent taxonicity of schizotypy across the 3 item sets. The cognitive-perceptual class had a base rate of 25%, and membership was predicted by the rs7004633 SNP (odds ratio = 2.33, 95% confidence interval = 1.15-4.72 in adjusted analyses). Poor social adjustment predicted memberships in the interpersonal (16%) and disorganized (21%) classes. Classes were found not to be mutually exclusive. Schizotypy is taxonic and schizotypy class membership is predicted by genetic and environmental factors that predict schizophrenia. The findings hold the promise that a more complete understanding of schizotypy as a schizophrenia liability state will come from investigation of other genes and environmental factors associated with schizophrenia.


Preparation and microstructure of nanocrystalline Ni-Mo films.
Ni-Mo films were prepared by electrodeposition technique with citric acid as a complexing agent. The influence of the main technical parameters such as the concentration of Na2MoO4 x 2H2O, the current density, the bath temperature and pH on the component content in the Ni-Mo films were investigated by energy dispersive spectrometer (EDS), the microstructure and surface morphology of Ni-Mo films were characterized by employing X-ray diffractometer (XRD) and scanning electron microscope (SEM). The results showed that the excellent Ni-Mo films with 35.51 wt% Mo was obtained when the concentration of Na2MoO4 x 2H2O was 15 g/L, the current density was 8 A/dm2, the bath temperature was 30 degrees C and the pH was 5.0. The mircostructure of the Ni-Mo films were nanocrystalline and the average size of grain was about 20.12 nm by calculating using Scherrer Equation.


AIDS and intravenous drug use: a growing menace.
The AIDS virus is transmitted through the exchange of bodily fluids between an infected person and an uninfected person, and the sharing of needles by IV drug users has been one of the primary means of transmission. The incidence of infection by this method has increased in the world, and in the inner cities of the United States has been a particular problem. Infected drug users also transmit the disease to their sexual partners and to their unborn children. Educational efforts have been directed at changing the behavior of drug users to reduce their sharing of needles, and some cities have tried handing out free needles. Such efforts, of course, run counter to the admonition not to use drugs at all, but health professionals believe changing behavior is the most important way of attacking the disease. While transmission of the HIV virus through IV drug abuse remains a significant problem, there is evidence that the educational efforts have been working and that, in some areas at least, behaviors have been changed and the risk has been reduced.


Two-photon absorption spectra of fluorescent isomorphic DNA base analogs.
Fluorescent DNA base analogs and intrinsic fluorophores are gaining importance for multiphoton microscopy and imaging, however, their quantitative nonlinear excitation properties have been poorly documented. Here we present the two-photon absorption (2PA) spectra of 2-aminopurine (2AP), 7-methyl guanosine (7MG), isoxanthopterin (IXP), 6-methyl isoxanthopterin (6MI), as well as L-tryptophan (L-trp) and 3-methylindole (3MI) in aqueous solution and some organic solvents measured in the wavelength range 550 - 810 nm using femtosecond two-photon excited fluorescence (2PEF) and nonlinear transmission (NLT) methods. The peak 2PA cross section values range from 0.1 GM (1 GM = 10-50 cm4 s photon-1) for 2AP to 2.0 GM for IXP and 7MG. Assuming typical excitation conditions for a scanning 2PEF microscope, we estimate a maximum image frame rate of ~175 frames per second (FPS).


Purification and structural determination of hematopoietic stem cell-stimulating monoacetyldiglycerides from Cervus nippon (deer antler).
A mixture of monoacetyldiglycerides was newly isolated from the chloroform extract of antlers of Cervus nippon, guided by the hematopoietic stimulation of stem cells. The structures of monoacetyldiglycerides were determined by various spectroscopic methods: FAB MS, CID tandem MS, and 1D and 2D NMR. A mixture of at least nine inseparable sn-3-monoacetyldiglycerides was identified: 1 [C(39)H(72)O(6) (C16 : 0/C18 : 1)], 2 [C(39)H(72)O(6) (C18 : 1/C16 : 0)], 3 [C(39)H(70)O(6) (C16 : 0/C18 : 2)], 4 [C(39)H(70)O(6) (C18 : 2/C16 : 0)], 5 [C(41)H(74)O(6) (C18 : 0/C18 : 2), 6 [C(41)H(74)O(6) (C18 : 2/C18 : 0)], 7 [C(41)H(74)O(6) (C18 : 1/C18 : 1)], 8 [C(43)H(74)O(6) (C18 : 0/C20 : 4)], and 9 [C(43)H(74)O(6) (C20 : 4/C18 : 0)]. Among these nine monoacetyldiglycerides in deer antlers, compound 3 was one of the major compounds and was efficiently synthesized from glycerol. Spectral data of synthetic monoacetyldiglyceride 3 were compared with the corresponding data for the mixture of natural monoacetyldiglycerides. The mixture of natural monoacetyldiglycerides from deer antlers showed potent activity on the hematopoiesis (stimulation index=1.40+/-0.05, p<0.02 at 1 microg/ml), and synthetic monoacetyldiglyceride 3 showed even better activity (stimulation index=1.54+/-0.12, p<0.001, at 1 microg/ml).


Fluoro-gold and pentamidine inhibit the in vitro and in vivo release of dopamine in the striatum of rat.
Fluoro-Gold (FG), first developed as an antifungal/antiparasitic agent, is now also used extensively as a retrograde tracer in histological studies of nervous tissue. The fact that FG is taken up by dopamine (DA) terminals before its retrograde transport to DA cell bodies implies a presynaptic interaction, though the biochemical target(s) and mechanism(s) are unknown. To further elucidate, FG and another aromatic diamidine, pentamidine, were tested on [3H]DA release and uptake in vitro from striatal slices and synaptosomes. Neither compound affected [3H]DA uptake in synaptosomes and slices, and neither inhibited DA efflux mediated through reversal of DA uptake mechanisms. NMDA-mediated glutamate-evoked DA release was completely inhibited by either FG (IC50 approximately 3 microM) or pentamidine (IC50 approximately 1 microM), and 20 mM K(+)-evoked DA release was inhibited by similar concentrations but only to 60% of control. Arginine (up to 500 microM) and spermidine (200 microM) failed to reverse 33 microM FG inhibition of either the spontaneous or the glutamate-evoked DA release, indicating that FG inhibition of release was not necessarily via blockade of either nitric oxide generation or spermidine binding to NMDA receptors. Interestingly, FG (33 microM) and pentamidine (10 microM) inhibited 1 and 5 microM D-methamphetamine (METH)-evoked [3H]DA release to approximately 50% of control, and in striatal synaptosomes, FG (33 microM) and pentamidine (10 microM) inhibited 5 microM METH- and 1.25 mM Ca(++)-evoked DA release. Additionally, in vivo brain microdialysis supported the in vitro results; 100 microM FG in the microdialysis buffer inhibited 70% of the increase in extracellular DA in the striatum produced by 2.5 mg/kg METH.(ABSTRACT TRUNCATED AT 250 WORDS)


Hodgkin's disease: diagnostic accuracy of fine needle aspiration; a report based on 62 consecutive cases.
We report on our series of 62 cases occurring between January 1977 and December 1990, which were diagnosed as Hodgkin's disease by fine needle aspiration (FNA) samples. The overall accuracy of the cytological diagnosis was high, with only four incorrect diagnoses and a positive predictive value of 93.5%. The value of FNA as a first level diagnostic technique in the screening of lymphadenopathies is discussed, as well as the limitations and pitfalls of the cytological diagnosis.


Diffuse pulmonary interstitial fibrosis. Evidence of humoral antibody mediated pathogenesis.
Immunochemical studies of post-mortem lung tissue from a patient suffering from diffuse pulmonary interstitial fibrosis were conducted. Identifiable deposition of IgG on the alveolar septa was noted by immunoperoxidase and immunofluorescein methods. The antemortem serum obtained from the patient showed high affinity immunologic binding to normal human lung tissue. An antibody-antigen reaction was also detectable by microcomplement fixation procedure with buffer extracts of normal lung tissue, while normal pooled serum showed no significant reaction. These findings, in addition to the gross, light microscopic, and electron microscopic observations, strongly suggest that a humoral antibody mediated reaction is one of the underlying pathogenetic mechanisms eliciting the morphologically and clinically observable manifestations of diffuse interstitial pulmonary fibrosis.


Oxygen tension regulates the stability of insulin receptor substrate-1 (IRS-1) through caspase-mediated cleavage.
The insulin and insulin-like growth factor-1 (IGF-1) receptors mediate signaling for energy uptake and growth through insulin receptor substrates (IRSs), which interact with these receptors as well as with downstream effectors. Oxygen is essential not only for ATP production through oxidative phosphorylation but also for many cellular processes, particularly those involved in energy homeostasis. The oxygen tension in vivo is significantly lower than that in the air and can vary widely depending on the tissue as well as on perfusion and oxygen consumption. How oxygen tension affects IRSs and their functions is poorly understood. Our findings indicate that transient hypoxia (1% oxygen) leads to caspase-mediated cleavage of IRS-1 without inducing cell death. The IRS-1 protein level rebounds rapidly upon return to normoxia. Protein tyrosine phosphatases (PTPs) appear to be important for the IRS-1 cleavage because tyrosine phosphorylation of the insulin receptor was decreased in hypoxia and IRS-1 cleavage could be blocked either with H(2)O(2) or with vanadate, each of which inhibits PTPs. Activity of Akt, a downstream effector of insulin and IGF-1 signaling that is known to suppress caspase activation, was suppressed in hypoxia. Overexpression of dominant-negative Akt led to IRS-1 cleavage even in normoxia, and overexpression of constitutively active Akt partially suppressed IRS-1 cleavage in hypoxia, suggesting that hypoxia-mediated suppression of Akt may induce caspase-mediated IRS-1 cleavage. In conclusion, our study elucidates a mechanism by which insulin and IGF-1 signaling can be matched to the oxygen level that is available to support growth and energy metabolism.


Morphological study of Streptococcus mutans and two extracellular polysaccharide mutants.
Two extracellular polysaccharide mutants of Streptococcus mutans GS-5 were obtained and examined. The mutants were distinguished by colonial morphology and by growth on and adherence to hard surfaces. A technique was devised which allowed these bacteria to be studied as they appeared when grown on a hard surface in liquid medium which contained sucrose. Negative stains, replicas, and scanning electron micrography clearly revealed differences in cellular aggregation due to the various extracellular polysaccharides produced. Comparison of sections of the adherent parent strain (GS-5) with those of the nonadherent mutant (GS-511) allowed the extracellular polysaccharide(s) responsible for adhesion to be visually localized.


Microstructural characterization of degradation in a high-chromium alloy in the sugar industry.
A new high (24%)-chromium alloy was experimentally manufactured by combining the particular chemical composition of various materials with the use of thermal treatments to produce pump impellers for the sugar industry. The alloy metal was tested in situ in a sugar medium and then evaluated by electrochemical measurements; the two former procedures were associated with a nontraditional method to assess the susceptibility and resistance to corrosion. Potentiostatic and atomic absorption measurements were carried out to characterize the degradation and damage morphology by scanning electron microscopy (SEM) observations. The trials proved a good performance for the new alloy in the sugar medium in spite of nonsignificant localized damages involving erosion-corrosion mechanisms and surface microcrackings of the material.


The microcystin composition of the cyanobacterium Planktothrix agardhii changes toward a more toxic variant with increasing light intensity.
The cyanobacterium Planktothrix agardhii, which is dominant in many shallow eutrophic lakes, can produce hepatotoxic microcystins. Currently, more than 70 different microcystin variants have been described, which differ in toxicity. In this study, the effect of photon irradiance on the production of different microcystin variants by P. agardhii was investigated using light-limited turbidostats. Both the amount of the mRNA transcript of the mcyA gene and the total microcystin production rate increased with photon irradiance up to 60 micromol m(-2) s(-1), but they started to decrease with irradiance greater than 100 micromol m(-2) s(-1). The cellular content of total microcystin remained constant, independent of the irradiance. However, of the two main microcystin variants detected in P. agardhii, the microcystin-DeRR content decreased twofold with increased photon irradiance, whereas the microcystin-DeLR content increased threefold. Since microcystin-DeLR is considerably more toxic than microcystin-DeRR, this implies that P. agardhii becomes more toxic at high light intensities.


A comparison of the Cambridge Automated Neuropsychological Test Battery (CANTAB) with "traditional" neuropsychological testing instruments.
The Cambridge Neuropsychological Test Automated Battery (CANTAB) is frequently used in research protocols and increasingly in clinical practice. Despite the frequency of its use, important aspects of its measurement validity have yet to be established in healthy adults. Two hundred and fifty-five individuals completed the CANTAB and traditional neuropsychological tests commonly used in clinical practice, including selected subtests from the Wechsler Adult Intelligence Scale, Controlled Oral Word Association Test, Animal Naming, Trail Making Tests A and B, the Stroop test, and the Green Story Recall test. Results showed that CANTAB subtests were modestly correlated with traditional subtests. Correlations between CANTAB subtests and traditional subtests were less consistent when age and education were controlled for. In conclusion, the CANTAB shows modest associations with traditional neuropsychological test measures.


Analysis of lncRNA sequences: FAM3D-AS1, LINC01230, LINC01315 and LINC01468 in endometrial cancer.
The analysis of long non-coding RNA (lncRNA) in endometrial cancer is a novel field of science. Although numerous lncRNA sequences have been identified until today, their correlation with endometrial cancer is still undetermined. The aim of this study was to analyze the expression of four lncRNA sequences: FAM3D-AS1, LINC01230, LINC01315 and LINC01468 and to investigate their significance in endometrial cancer. LncRNA sequences were investigated in paraffin blocks (tumor tissue and non-malignant endometrial tissue in archival postoperative specimens) in endometrial cancer patients (Cases, n = 120) and in cancer-free controls (n = 80) using real-time PCR assay. This study revealed a lower expression of LINC01468 in endometrial cancer patients than in controls. Both LINC01468 and FAM3D-AS1 were positively correlated with Body Mass Index (BMI) in cancer-free controls. LncRNA LINC01468 may be a protective factor in development of endometrial cancer.


Maturation of the mouse oocyte-cumulus cell complex: stimulation by lectins.
We have used carbohydrate-binding proteins, or lectins, as tools to investigate the physiological phenomena associated with the preovulatory maturation of the oocyte-cumulus cell complex. Certain lectins are mitogens, and since other mitogenic agents such as growth factors are known to stimulate meiotic maturation and cumulus expansion, we tested the ability of lectins to provoke these physiological responses. Cumulus cell-enclosed oocytes (CEO) from primed mice were maintained in meiotic arrest in vitro with dibutyryl cyclic adenosine 3',5'-monophosphate (dbcAMP) and treated with one of eleven different lectins. With the exception of pokeweed mitogen (PWM), all of the mitogenic lectins tested were able to induce germinal vesicle breakdown (GVB) in meiotically arrested oocytes, and this action required the presence of the somatic cumulus cells; in fact, either there was no effect or maturation was suppressed when cumulus cell-free oocytes (denuded oocytes; DO) were treated with lectins. None of the nonmitogenic lectins stimulated meiotic maturation in either CEO or DO. The mitogenic lectin concanavalin A (Con A) also induced maturation in CEO when meiotic arrest was maintained with hypoxanthine, guanosine, or 3-isobutyl-1-methylxanthine. The kinetics of spontaneous oocyte maturation in inhibitor-free medium were not altered by Con A. Only the mitogenic lectins that induced meiotic maturation stimulated cumulus expansion, with Con A the most active lectin. The actions of Con A on the maturation of the oocyte-cumulus cell complex were inhibited by methyl-alpha-D-mannopyranoside as predicted by its sugar-binding specificity. These results demonstrate that (1) lectins can stimulate maturation of the mouse oocyte-cumulus cell complex; (2) mitogenicity is associated with the positive activity of the lectins; and (3) cumulus cells mediate the stimulatory action of lectins on oocyte maturation, while inhibition of GVB occurs at the oocyte level. These data support the idea that common signals mediate the mitogenic and maturation-inducing actions of lectins.


MicroRNAs: a new piece in the paediatric cardiovascular disease puzzle.
Cardiovascular diseases in children comprise a large public health problem. The major goals of paediatric cardiologists and paediatric cardiovascular researchers are to identify the cause(s) of these diseases to improve treatment and preventive protocols. Recent studies show the involvement of microRNAs (miRs) in different aspects of heart development, function, and disease. Therefore, miR-based research in paediatric cardiovascular disorders is crucial for a better understanding of the underlying pathogenesis of the disease, and unravelling novel, efficient, preventive, and therapeutic means. The ultimate goal of such research is to secure normal cardiac development and hence decrease disabilities, improve clinical outcomes, and decrease the morbidity and mortality among children. This review focuses on the role of miRs in different paediatric cardiovascular conditions in an effort to encourage miR-based research in paediatric cardiovascular disorders.


Anti-HBc IgM and anti-delta screening by EIA method.
The clinical value of an enzyme-linked immunosorbent assay (ELISA) for the detection of anti-HBc IgM was evaluated by testing 202 sera from acute viral hepatitis B (AVHB), hepatitis B (HB), chronic hepatitis (CAH), chronic liver disease (CLD), cirrhosis, primary hepatoma, HBsAg carrier, acute viral hepatitis A (AVHA), hepatitis A (HA), non-A, non-B (NANB) hepatitis and miscellaneous conditions other than hepatic disease, and 19 additional various hepatic disease cases were examined for anti-delta. In clinical situations the accurate diagnosis of HB is not always possible and the differential diagnosis seems to be very important especially in making decisions of treatment and estimation of prognosis. In overall cases the highest positive rate of anti-HBc IgM was found in AVHB as shown as 74.3% (26/35) comparing to other conditions in which the positive rate was extremely low (2.1%). The anti-HBc IgM appeared to be highly specific to AVHB (83.9%) as compared to the other. The positive rate of HBsAg was high in AVHB, CAH and HBsAg carrier (100.0%) followed by CLD, cirrhosis and HB (up to 70.8%). The ALT activities and ALPalb fractions were significantly high in AVHB (p less than 0.005). The correlation between the positivity of anti-HBc IgM and highly abnormal ALT appeared be high. AVHB was confined mostly to 10-20 age group and the male to female ratio was about 6 to 1. Subgroup of AVHB II with positive anti-HBc IgM appeared to have a greater chance being positive for HBsAg and ALPalb. The S/N ratio of anti-HBc IgM was as high as 20 which was unique to AVHB.(ABSTRACT TRUNCATED AT 250 WORDS)


New noninvasive methodology for real-time monitoring of lipid flip.
A new methodology for the detection of lipid flip was developed. This methodology relies on the quenching of the fluorescence of the cascade-blue-labeled lipid through complex formation with a membrane-impermeable cyclen-tetranaphthalenethiourea synthetic receptor for this dye. The high affinity of the receptor to cascade-blue label allows the use of micromolar concentrations of this receptor during the experiment. At these low concentrations, the receptor does not interfere with the membrane integrity and, therefore, renders this new methodology less invasive to the model and cell membranes than commonly utilized 7-nitro-1,2,3-benzoxadiazol-4-yl (NBD)-dithionite methodology. Unlike with the NBD-dithionite assay, where the fluorescence quenching of the NBD group is achieved through its chemical modification, this new assay relies on the noncovalent interactions between cascade-blue label and the receptor. Therefore, the quenching can be reverted by either competitive displacement of the lipid-attached label with a water-soluble substrate or by enzymatic degradation of the receptor leading to the label release and fluorescence dequenching. We demonstrate that this new methodology is suitable for the study of lipid flip in both model spherical bilayer membranes and in-vitro experiments.


Development and characterization of the replicon system of Japanese encephalitis live vaccine virus SA14-14-2.
Viral self-replicating sub-genomic replicons represent a powerful tool for studying viral genome replication, antiviral screening and chimeric vaccine development. Many kinds of flavivirus replicons have been developed with broad applications. The replicon system of JEV live vaccine strain SA14-14-2 was successfully developed in this study. Two kinds of replicons that express enhanced green fluorescent protein (EGFP) and Renilla luciferase (R.luc) were constructed under the control of SP6 promoter, respectively. Robust EGFP and R.luc signals could be detected in the replicon-transfected BHK-21 cells. Furthermore, the potential effects of selected amino acids in the C-terminal of envelope protein on replication were characterized using the replicon system. Our results provide a useful platform not only for the study of JEV replication, but also for antiviral screening and chimeric vaccine development.


Retinal nerve fiber layer thickness in children <18 years by spectral-domain optical coherence tomography.
To establish a normative database for retinal nerve fiber layer (RNFL) thickness in children<18 years by spectral-domain optical coherence tomography (SD-OCT). Children <18 years, who were consecutively enrolled from July-August 2011 in this cross-sectional hospital study, underwent Cirrus SD-OCT. Recorded demographics and clinical characteristics included age, sex, axial length, spherical equivalent, intraocular pressure (by Perkins tonometry) and cup disc ratio. The RNFL thickness full circle (RNFL-FC), superior, (RNFL-S), inferior (RNFL-I), temporal (RNFL-T), and nasal (RNFL-N) and the influence of clinical and demographic factors on the RNFL thickness were assessed using linear regression analysis. 148 eyes of 74 children were analyzed (females: males = 37: 37) with a mean age of 10 ± 3.4 years (range 4-17 years) and a mean spherical equivalent of -0.6 ± 1.2 dioptres. The mean RNFL-FC was 94 ± 10.9 and 93 ± 10.6 µm in the right and left eyes, respectively, with maximum thickness found in the superior quadrant. The RNFL-FC and RNFL-S decreased by 1.9 µm for every 1 dioptre increase in myopic shift, p < 0.001, and by 1.9 µm for every mm increase in axial length, p < 0.001. There was no effect of age on RNFL thickness in females or males. The RNFL thickness in children was influenced most by axial length and refractive error while it was independent of age. The normative data from this study could serve as reference for further studies on pediatric glaucoma using newer imaging devices.


Clinical outcomes by supplemental oxygen use in remdesivir-treated, hospitalised adults with COVID-19.
Clinical trials show different effects of remdesivir on clinical outcomes relative to COVID-19 severity at hospital admission; in Europe, there are few real-world data. A multicentre, multinational retrospective cohort study in adult patients hospitalised with PCR-confirmed COVID-19 was conducted to understand remdesivir clinical use in different countries and to describe outcomes for patients receiving remdesivir stratified by oxygen use. Primary endpoints were all-cause mortality at day 28 and hospitalisation duration. Patients were categorised by baseline disease severity: no supplemental oxygen (NSO); low flow oxygen ≤ 6 litres (l)/minute (LFO); high flow oxygen > 6 l/minute (HFO). Four hundred and forty-eight (448) patients (72 [16.1%] HFO; 295 [65.8%] LFO; 81 (18.1%] NSO) were included; median age was 65 years and 64% were male. Mortality was higher in patients on HFO (rate 23.6%) compared to LFO (10.2%; p = 0.001) or NSO (6.2%; p = 0.002). Duration of hospitalisation was longer in patients on HFO (13 days) compared to LFO (9 days; p = 0.003) and NSO (9 days; p = 0.021). Patients who initiated remdesivir ≥ 2 days compared to within a day of hospitalisation had a 4.2 times higher risk of death, irrespective of age, sex, comorbidities, and oxygen support at baseline. Requirement for mechanical ventilation/ECMO and readmission within 28 days of discharge was similar across groups. Remdesivir use and outcomes differed by country. A higher mortality rate and duration of hospitalisation was seen in remdesivir-treated COVID-19 patients on HFO compared to LFO and NSO. Initiation of remdesivir upon admission as opposed to delayed initiation has a mortality benefit. NCT04847622.


Using protein-based motifs to stabilize peptides.
While the use of synthetically derived novel inhibitor peptides as a source of new therapeutics for medicine remains incredibly promising, there is a major problem with implementing this technology, as many synthetic peptides have proven to be unstable and are degraded by peptidases in the host cell. In this study, we have investigated methods by which peptides can be stabilized using protein-based motifs in order to prevent the action of peptidases. Using an in vivo approach our laboratory developed to screen for synthetic peptides which can inhibit the growth of Escherichia coli, we found that protecting the amino or carboxyl terminus of the peptides via fusion to the very stable Rop protein, or the incorporation of two proline residues, increased the frequency at which potent inhibitor peptides could be isolated. Using an in vitro degradation assay in which extracts from several different cell types were tested, we demonstrated that peptides stabilized with multiple proline residues were more resistant to degradation than peptides stabilized by amidation or acetylation, two approaches that are routinely utilized to improve the stability of peptide drugs.


Long-term survival and virulence of Mycobacterium leprae in amoebal cysts.
Leprosy is a curable neglected disease of humans caused by Mycobacterium leprae that affects the skin and peripheral nerves and manifests clinically in various forms ranging from self-resolving, tuberculoid leprosy to lepromatous leprosy having significant pathology with ensuing disfiguration disability and social stigma. Despite the global success of multi-drug therapy (MDT), incidences of clinical leprosy have been observed in individuals with no apparent exposure to other cases, suggestive of possible non-human sources of the bacteria. In this study we show that common free-living amoebae (FLA) can phagocytose M. leprae, and allow the bacillus to remain viable for up to 8 months within amoebic cysts. Viable bacilli were extracted from separate encysted cocultures comprising three common Acanthamoeba spp.: A. lenticulata, A. castellanii, and A. polyphaga and two strains of Hartmannella vermiformis. Trophozoites of these common FLA take up M. leprae by phagocytosis. M. leprae from infected trophozoites induced to encyst for long-term storage of the bacilli emerged viable by assessment of membrane integrity. The majority (80%) of mice that were injected with bacilli extracted from 35 day cocultures of encysted/excysted A. castellanii and A. polyphaga showed lesion development that was similar to mice challenged with fresh M. leprae from passage mice albeit at a slower initial rate. Mice challenged with coculture-extracted bacilli showed evidence of acid-fast bacteria and positive PCR signal for M. leprae. These data support the conclusion that M. leprae can remain viable long-term in environmentally ubiquitous FLA and retain virulence as assessed in the nu/nu mouse model. Additionally, this work supports the idea that M. leprae might be sustained in the environment between hosts in FLA and such residence in FLA may provide a macrophage-like niche contributing to the higher-than-expected rate of leprosy transmission despite a significant decrease in human reservoirs due to MDT.


The investigation of spin-orbit effect for the F((2)P)+HD reaction.
In this paper, we employ the time-dependent quantum wave packet method to study the reaction of F((2)P(3/2), (2)P(1/2)) with HD on the Alexander-Stark-Werner potential energy surface. The reaction probabilities and total integral cross sections of the spin-orbit ground and excited states for the two possible products of the system are calculated. Because the reaction channel of the excited spin-orbit state is closed at the resonance energy, the resonance feature does not appear in the reaction probabilities and cross section for the F((2)P(1/2))+HD(v=j=0)-->HF+D reaction, in contrast with that found for the ground spin--orbit state. We also compare the average cross sections of the two possible products with the experimental measurement. The resonance peak in the present average cross section for the HF+D product is slightly larger than the experimental result, but much smaller than that of the single-state calculations on the potential energy surface of Stark and Werner. It seems that the spin--orbit coupling would play a relatively important role in this reaction. Moreover, the isotope effects of the ground and excited spin--orbit states and the reactivity of the two product channels from the excited spin--orbit state are presented.


Masitinib for the treatment of Alzheimer's disease.
The actual standard treatment for mild-to-moderately severe Alzheimer's disease only attacks its symptoms. Masitinib is a potent and selective phenylaminothiazole-type tyrosine kinase inhibitor which is currently in Phase III studies for the treatment of Alzheimer's disease (AD) with the aim of modifying its evolution and with multiple pharmacological targets such as inhibition of mast cells activity, inhibition of microglia activation, modulation of Aβ and Tau protein signaling pathway and prevention of synaptic damage. Here, we review the preclinical and clinical studies that investigated the administration of masitinib treatment in monotherapy in AD. All research studies revealed positive effects concerning the cognitive functions in AD and generally with good safety and tolerability.


Wearable Smartwatch Facilitated Remote Health Management for Patients Undergoing Transcatheter Aortic Valve Replacement.
Background In the recent decades, the development of novel digital health technologies enables doctors to monitor ECG and vital signs remotely. But the data on applying the noninvasive wearable smartwatch on patients with transcatheter aortic valve replacement (TAVR) are unknown. Methods and Results We performed a prospective, observational cohort study to evaluate the feasibility of a novel, virtual, and remote health care strategy for patients with TAVR discharged to home with smart wearable devices. A total of 100 consecutive patients with severe aortic stenosis who underwent elective transfemoral TAVR were enrolled and received the Huawei smartwatch at least 1 day before TAVR. Vital signs, including heart rate, rhythm, oxygen saturation, and activity, were continuously recorded. Single-lead ECG was recorded twice per day in the week following TAVR discharge and at least 2 days a week for the subsequent month after TAVR discharge. A designated heart team member provided remote health care with the data from the smartwatch when the patient had a need. Thirty-eight cardiac events were reported in 34 patients after discharge, with most of the events (76.0%) detected and confirmed by the smartwatch. Six patients were advised and readmitted to the hospital for arrhythmia events detected by the smartwatch, of whom 4 patients received pacemaker implantation. The remaining 28 (82.4%) patients received telemedicine monitoring instead of face-to-face clinical visits, and 3 of them received new medication treatment under the online guidance of doctors. New-onset left branch bundle block was found in 48 patients, with transient characteristics, and recovered spontaneously in 30 patients, and new-onset atrial fibrillation was detected in 4 patients. There were no significant differences in the average weekly heart rates or the ratio of abnormal or low oxygen saturation when compared with the baseline. The average daily steps increased over time significantly (baseline, 870±1353 steps; first week, 1986±2406 steps; second week, 2707±2716 steps; third week, 3059±3036 steps; fourth week, 3678±3485 steps, P<0.001). Conclusions Smartwatches can facilitate remote health care for patients discharged to home after undergoing TAVR and enable a novel remote follow-up strategy. The majority of cardiac clinical events that occurred within 30-day follow-up were detected by the smartwatch, mainly because of the record of conduction abnormality. Registration URL: https://www.clinicaltrials.gov; Unique identifier: NCT04454177.


Diagnosis of acute foetal distress does not preclude banking of umbilical cord blood units.
Clinical diagnosis of acute foetal distress (AFD) is based on several parameters such as abnormal foetal heart rate (FHR) pattern and/or meconium liquid staining (MLS). Standards for cord blood (CB) banking indicate that AFD should be considered as exclusion criteria for CB collection, but precise guidelines on how to proceed with CB collection in the presence of AFD signs during labour are not available. We evaluated whether the presence of FHR abnormality and/or MLS during labour 1) reduced the CB collection activity; 2) were associated with the infant's acidaemia or asphyxia and 3) deteriorated the biological characteristics of CB units. Thirty-three units of CB were evaluated for biological parameters, gas values and newborn's Apgar score. The results were compared with a control group of 33 consecutive units previously banked. No differences were observed between the two groups and all but one newborn showed normal Apgar score and absence of metabolic acidaemia. The results showed that 1) AFD reduced the CB collection activity by 10% each year; 2) the majority of CB units collected in the presence of abnormal FHR and/or meconium have biological characteristics eligible for banking; 3) FHR alterations or meconium in the presence of normal gas analysis do not represent certain diagnosis of AFD.


[Homicides without clues in children].
From 1990 to 1995, 274 deaths of infants (age range 7 days-6 years) were investigated in Münster, where there were no indications of suspicious circumstances. In 167 cases a legal autopsy was carried out and revealed 5 homicides. In 107 cases where an autopsy was not ordered by the prosecutor 3 homicides were discovered. A similar frequency of homicides in both groups indicates that the criteria used to form the basis of a decision by the prosecutor to release the body are insufficient. On average one out of 30 cases was found to be a homicide as the result of the autopsy where the initial assumption was death from natural causes.


Implementation of the Critical Care Pain Observation Tool increases the frequency of pain assessment for noncommunicative ICU patients.
Pain is a common stressor for ICU patients, necessitating routine assessment. For patients who are unable to communicate, self-report tools are unsuitable, and the use of an observational tool is required to assess pain appropriately. The Critical Care Pain Observation Tool (CPOT) is the most reliable tool currently available to assess pain in these patients. We investigated whether the implementation of the CPOT in one Australian ICU could increase frequency of appropriate pain assessments, and if this would affect the administration of analgesia and sedation. In this before and after study, we first performed a retrospective chart audit on 441 adult ICU patient charts, over 49 days. Data collected included frequency and type of pain assessments, sedation and analgesia administered, communication and CAM-ICU status, and bedside nurse-perceived pain. During the implementation phase, new policy and guideline documents were released, and ICU charts were redesigned to incorporate the CPOT. All nursing staff attended an education session on pain assessment and correct use of the CPOT. The chart audit was repeated, capturing 344 charts over 43 days. Mean total assessments in 24 hours increased from 7.2 to 7.9 for communicative, 3.0 to 8.9 for non-communicative, and 5.1 to 9.1 for transitioning patients. For non-communicative patients there was a significant increase in observational assessments including the CPOT (1.7 to 8.3), and a decrease in inappropriate use of self-report tools (1.3 to 0.2). We also observed significant increases in administration of paracetamol, opiates, propofol, patient-controlled analgesia, modified-release opiates, and neuropathic pain agents. Implementation of the CPOT using standardised education and resources led to increased frequency of pain assessment, particularly for non-communicative patients. Appropriate observational assessments were also more frequently used for these patients. Analgesic administration generally increased, as did the use of propofol.


T-cell receptor variable beta genes show differential expression in CD4 and CD8 T cells.
Studies in transgenic and inbred strains of mice have shown that the critical molecular interactions controlling positive selection involve major histocompatibility complex (MHC), T-cell receptor (TCR), and CD4 or CD8 coreceptor molecules. Correlations have been established between MHC gene products and the percentage of CD4 or CD8 T cells that express specific variable (V) beta-gene products as part of the alpha beta heterodimer. These studies have important implications regarding potential mechanisms of HLA-linked autoimmune diseases in humans. If similar interactions are required for positive selection in humans, one would predict that the TCR repertoire expressed by mature, peripheral blood CD4 and CD8 T cells would vary. To test this hypothesis the expression of specific TCR V beta-region genes by CD4 and CD8 T cells from healthy individuals was compared using both triple-color flow cytometry and polymerase chain reaction based experimental approaches. The results show that the TCR repertoire does vary as a function of CD4 and CD8 T-cell subsets. Among unrelated individuals certain V beta genes were consistently overrepresented in the CD4 population (V beta-5.1, -6.7a, and -18); some were skewed to the CD8 population (V beta-14) while others showed variable patterns (V beta-12 and -17). Deletion of entire V beta gene families was not observed suggesting that this is a rare event in humans. Attempts to correlate the expressed TCR repertoire in humans with HLA alleles will require consideration of these differences in expression as a function of subset.


Hue Selectivity in Human Visual Cortex Revealed by Functional Magnetic Resonance Imaging.
The variability of color-selective neurons in human visual cortex is considered more diverse than cone-opponent mechanisms. We addressed this issue by deriving histograms of hue-selective voxels measured using fMRI with a novel stimulation paradigm, where the stimulus hue changed continuously. Despite the large between-subject difference in hue-selective histograms, individual voxels exhibited selectivity for intermediate hues, such as purple, cyan, and orange, in addition to those along cone-opponent axes. In order to rule the possibility out that the selectivity for intermediate hues emerged through spatial summation of activities of neurons selectively responding to cone-opponent signals, we further tested hue-selective adaptations in intermediate directions of cone-opponent axes, by measuring responses to 4 diagonal hues during concurrent adaptation to 1 of the 4 hues. The selective and unidirectional reduction in response to the adapted hue lends supports to our argument that cortical neurons respond selectively to intermediate hues.


Toward the use of mixed microbial cultures for the biological production of adipic and levulinic acid.
Biological synthesis of high added-value compounds like adipic acid (AA), levulinic acid (LA), or polyhydroxybutyrate (PHB) using pure culture has been separately reported. However, pure culture requires sterile conditions and the use of specific carbon sources resulting in high operating costs. Different alternatives based on the use of mixed microbial cultures (MMC) have been explored to resolve this problem. MMC have been widely reported for the production of PHB, but scarcely reported for LA production and never for AA synthesis. This work presents a novel strategy for the co-production of AA LA, and PHB using MMC. The strategy consists in selecting an MMC producer of AA, LA and PHB from an inoculum obtained from a wastewater treatment plant, which is then subjected to the feast and famine culture strategy in a sequential batch reactor, coupled with a batch reactor step to enhance the accumulation of AA and LA. The results showed that the MMC could produce a 16 ± 2, 23 ± 1 and 5 ± %1 (g compound/g volatile solids) of AA, LA and PHB, respectively, using a non-fermented residual biomass rich in pentose, namely synthetic hemicellulose hydrolysate (SHH) as the carbon source. These results contribute to generating future research to better understand and optimise the biosynthesis of these compounds by MMC.


Alteration of homeobox gene expression by N-ras transformation of PA-1 human teratocarcinoma cells.
We used a series of cell clones from a human teratocarcinoma cell line, PA-1, to study the effect of transformation by an activated N-ras oncogene on the expression of genes involved in retinoic acid (RA)-induced differentiation and growth regulation. Recently, it has been shown that expression of human HOX 2 genes is sequentially activated by RA beginning from Hox 2.9 at the 3' end of the HOX 2 cluster (A. Simeone, D. Acampora, L. Arcioni, P. W. Andrews, E. Boncinelli, and F. Mavilio, Nature [London] 346:763-766, 1990). We now report that six different genes of the cluster HOX 1 are sequentially induced by RA in a similar temporal pattern, beginning with genes at the 3' end of the cluster. However, in N-ras-transformed cell clones, RA-induced expression of these homeobox genes is delayed. Hox 1.4 and Hox 1.3, genes abundantly induced in nontransformed clones after 3 days of RA treatment, are expressed in N-ras-transformed cells only after 10 days of RA treatment. At this time, the cells' growth is arrested at very high density, and no differentiated morphologic characteristics are observed. Constitutive expression of a transfected Hox 1.4 gene under the control of a simian virus 40 promotor leads to differentiated cell morphology similar to that of the RA-induced phenotype and restores the growth-inhibitory effects of RA in N-ras-transformed cells. These observations provide evidence that enhanced proliferation in N-ras-transformed cells compromises teratocarcinoma cell differentiation by a mechanism that transiently suppresses homeobox gene induction and implies a central role for homeobox genes in RA-induced cell differentiation. We conclude that stimulation of a putative growth factor signal pathway, associated with ras-induced proliferation, transiently suppresses the induction of transcription factors functionally involved in cell growth and differentiation.


Insight into metastatic oral cancer tissue from novel analyses using FTIR spectroscopy and aperture IR-SNOM.
A novel machine learning algorithm is shown to accurately discriminate between oral squamous cell carcinoma (OSCC) nodal metastases and surrounding lymphoid tissue on the basis of a single metric, the ratio of Fourier transform infrared (FTIR) absorption intensities at 1252 cm-1 and 1285 cm-1. The metric yields discriminating sensitivities, specificities and precision of 98.8 ± 0.1%, 99.89 ± 0.01% and 99.78 ± 0.02% respectively, and an area under receiver operator characteristic (AUC) of 0.9935 ± 0.0006. The delineation of the OSCC and lymphoid tissue revealed by the image formed from the metric is in better agreement with an immunohistochemistry (IHC) stained image than are either of the FTIR images obtained at the individual wavenumbers. Scanning near-field optical microscopy (SNOM) images of the tissue obtained at a number of key wavenumbers, with high spatial resolution, show variations in the chemical structure of the tissue with a feature size down to ∼4 μm. The image formed from the ratio of the SNOM images obtained at 1252 cm-1 and 1285 cm-1 shows more contrast than the SNOM images obtained at these or a number of other individual wavenumbers. The discrimination between the two tissue types is dominated by the contribution from the 1252 cm-1 signal, which is representative of nucleic acids, and this shows the OSCC tissue to be accompanied by two wide arcs of tissue which are particularly low in nucleic acids. Haematoxylin and eosin (H&E) staining shows the tumour core in this specimen to be ∼40 μm wide and the SNOM topography shows that the core centre is raised by ∼1 μm compared to the surrounding tissue. Line profiles of the SNOM signal intensity taken through the highly keratinised core show that the increase in height correlates with an increase in the protein signal. SNOM line profiles show that the nucleic acids signal decreases at the centre of the tumour core between two peaks of higher intensity. All these nucleic acid features are ∼25 μm wide, roughly the width of two cancer cells.


[The role of Cln3 in filamentous growth and invasive growth of Saccharomyces cerevisiae].
Cln3 is one of G1 cyclins in Saccharomyces cerevisiae. In order to study the function of Cln3 in cell cycle and morphogenesis, we constructed a cln3 null mutant and analyzed its phenotype. Our results indicated that the cln3 null mutant was more sensitive to alpha pheromone, and arrested at G1 phase. The hypersensitivity to alpha-pheromone was not suppressed by overexpression of SGV1. The null mutant showed a different phenotype with that of the other two G1 cyclin mutants. The filamentous growth in diploid cells of cln3 mutant was stronger than that in wild type cells, while invasive growth of the haploid cells was partially inhibited. The results suggested that the Cln3 plays a unique function in morphogenesis under a different mechanism with that used by Cln1 and Cln2.


Food-Dependent, Exercise-Induced Anaphylaxis: Diagnosis and Management in the Outpatient Setting.
Food-dependent, exercise-induced anaphylaxis is a disorder in which anaphylaxis develops most predictably during exercise, when exercise takes place within a few hours of ingesting a specific food. IgE to that food should be demonstrable. It is the combination of the food and exercise that precipitates attacks, whereas the food and exercise are each tolerated independently. Recently, it was demonstrated that exercise is not essential for the development of symptoms, and that if enough of the culprit food is ingested, often with additional augmentation factors, such as alcohol or acetylsalicylic acid, symptoms can be induced at rest in the challenge setting. Thus, food-dependent, exercise-induced anaphylaxis appears to be more correctly characterized as a food allergy syndrome in which symptoms develop only in the presence of various augmentation factors, with exercise being the primary one. However, additional factors are not usually present when the patient exercises normally, so ongoing investigation is needed into the physiologic and cellular changes that occur during exercise to facilitate food-induced anaphylaxis.


Evaluation of occupational health service in the wood processing industry--determination of employee satisfaction.
In order to define priorities for improvement of the occupational health service (OHS) in one primary health care unit, employees' satisfaction was evaluated with a postal questionnaire. The questionnaire covered items on service reservation, personal health education received, quality of the OHS, and use of the OHS within six months by the employees. This evaluation was one step of the quality system that was planned in accordance with the ISO 9004-2 standard for continuous quality improvement of OHS. The study population was all of 1,050 employees working for five employers; 546 (52%) employees responded to the inquiry. However, only 377 of them had made use of the OHS within the last six months before the inquiry. The mean age of the responders was 42 years (SD = 9 years) and 76% of the respondents were men. The percentages of employees with a high degree of satisfaction with the quality of the OHS and personal health education in the OHS were 76% and 69%, respectively. However, concerning the quality of the OHS, the proportion of highly satisfied employees was 27% higher (95% CI = 19-35%) among those who had used OHS within six months compared to the others. Furthermore, when the oldest (> 48 years) and the youngest (< 35 years) age groups were compared, the difference between the proportions of highly satisfied employees was 21% (95% CI = 31%). In conclusion, the high differences suggest a close linkage between the employee's satisfaction with the OHS, the use of the OHS and ageing.


Curvature dependent onset of quantum tunneling in subnanometer gaps.
The quantum tunneling in subnanometer gap sizes in gold dimers is studied in order to account for the dependency of the onset of quantum tunneling on the dimer's radius and accordingly the gap wall's curvature, realized in experiments. Several nanodimers both nanowires and nanospheres with various radii and gap sizes are modelled and simulated based on the quantum corrected model, determining the onset of the quantum tunneling. Results show that the onset of quantum tunneling is both dependent on the gap size as well as on the dimer's radius. As larger dimers result in larger effective conductivity volumes, the influence of the quantum tunneling begins in larger gap sizes in larger dimers.


Dose range-finding studies with frovatriptan in the acute treatment of migraine.
To determine the optimum dose of frovatriptan for the acute treatment of migraine. Frovatriptan is a new triptan developed for the acute treatment of migraine. The dose-response characteristics and safety of frovatriptan have been investigated across a broad range of doses from 0.5 to 40 mg. Two randomized, placebo-controlled, double-blind, parallel-group trials, with a total of 1453 patients, were performed to determine the optimal dose of the 5-HT(B/1D)agonist, frovatriptan, for the acute treatment of migraine. The dose ranges studied were 2.5 to 40 mg in the high-dose study and 0.5 to 5 mg in the low-dose study. At 2 hours postdosing for initial moderate or severe headache (International Headache Society grades 2 or 3), there was an approximate two-fold difference in the proportion of patients taking frovatriptan doses of 2.5 to 40 mg with mild or no headache compared to placebo. Frovatriptan doses of 0.5 mg and 1 mg were not more effective than placebo at 2 hours postdose, and 2.5 mg was identified as the lowest effective dose for the relief of migraine and accompanying symptoms. Above 2.5 mg, no dose-response relationship was observed for any efficacy parameters. There was an increase in the incidence of adverse events from 10 mg and above, but the vast majority were rated as mild in severity and did not impact upon tolerability in a significant manner. Frovatriptan was well tolerated throughout the dose range of 0.5 to 40 mg. The 2.5-mg dose confers the optimal balance of efficacy and tolerability for the acute treatment of migraine.


Modified Fullerenes for Efficient Electron Transport Layer-Free Perovskite/Fullerene Blend-Based Solar Cells.
A variety of novel chemically modified fullerenes, showing different electron-accepting capabilities, has been synthesized and used to prepare electron transport layer (ETL)-free solar cells based on perovskite/fullerene blends. In particular, isoxazolino[60] fullerenes are proven to be a good candidate for processing blend films with CH3 NH3 PbI3 and obtaining enhanced power conversion efficiency (PCE) ETL-free perovskite solar cells (PSCs), improving the state-of-the-art PCE (i.e., 14.3 %) for this simplified device architecture. A beneficial effect for pyrazolino and methano[60]fullerene derivatives versus pristine [60]/fullerene is also shown. Furthermore, a clear correlation between the LUMO energy level of the fullerene component and the open circuit voltage of the solar cells is found. Apart from the new knowledge on innovative fullerene derivatives for PSCs, the universality and versatility of perovskite/fullerene blend films to obtain efficient ETL-free PSCs is demonstrated.


Characteristics and health conditions of a group of nursing home patients with mental-physical multimorbidity - the MAPPING study.
Long-term care facilities have partly taken over the traditional asylum function of psychiatric hospitals and house an increasing group of patients with mental-physical multimorbidity (MPM). Little is known about the characteristics, behavior, and care dependency of these patients. This paper aims to describe these aspects. Explorative, descriptive study among patients with MPM without dementia (n = 142), living in 17 geronto-psychiatric nursing home (NH) units across the Netherlands, stratified by those referred from mental healthcare services (MHS) and other healthcare services (OHS). Data collection consisted of chart review, semi-structured interviews, (brief) neuropsychological testing, and self-report questionnaires. Patients referred from MHS (n = 58) and from OHS (n = 84) were compared by descriptive statistics. Despite exclusion of patients with dementia, the majority of participants had cognitive impairment. Prevalence and severity of frontal impairment were high, as well as the number of patients with clinically relevant neuropsychiatric symptoms. MHS patients were younger, had more chronic psychiatric disorders, and more often used antipsychotics. Neuropsychiatric symptoms, domains of care dependency, physical conditions and concomitant medication use differed not significantly between the subgroups. Both groups of patients with MPM showed heterogeneity in various aspects but differed not significantly regarding the consequences of their multimorbidity. In a variety of characteristics, this group seems to be different from other NH patient groups, which requires extra knowledge and skills of the staff. To uncover which knowledge and skills are necessary, the next step should be to investigate the specific care needs of NH patients with MPM without dementia.


A combination of polyunsaturated fatty acid, nonribosomal peptide and polyketide biosynthetic machinery is used to assemble the zeamine antibiotics.
The zeamines are a unique group of antibiotics produced by Serratia plymuthica RVH1 that contain variable hybrid peptide-polyketide moieties connected to a common pentaamino-hydroxyalkyl chain. They exhibit potent activity against a broad spectrum of Gram-positive and Gram-negative bacteria. Here we report a combination of targeted gene deletions, high resolution LC-MS(/MS) analyses, in vitro biochemical assays and feeding studies that define the functions of several key zeamine biosynthetic enzymes. The pentaamino-hydroxyalkyl chain is assembled by an iterative multienzyme complex (Zmn10-13) that bears a close resemblance to polyunsaturated fatty acid synthases. Zmn14 was shown to function as an NADH-dependent thioester reductase and is proposed to release a tetraamino-hydroxyalkyl thioester from the acyl carrier protein domain of Zmn10 as an aldehyde. Despite the intrinsic ability of Zmn14 to catalyze further reduction of aldehydes to alcohols, the initially-formed aldehyde intermediate is proposed to undergo preferential transamination to produce zeamine II. In a parallel pathway, hexapeptide-monoketide and hexapeptide-diketide thioesters are generated by a hybrid nonribosomal peptide synthetase-polyketide synthase multienzyme complex (Zmn16-18) and subsequently conjugated to zeamine II by a stand-alone condensing enzyme (Zmn19). Structures for the resulting prezeamines were elucidated using a combination of high resolution LC-MS/MS and 1- and 2-D NMR spectroscopic analyses. The prezeamines are hypothesized to be precursors of the previously-identified zeamines, which are generated by the action of Zmn22, an acylpeptide hydrolase that specifically cleaves the N-terminal pentapeptide of the prezeamines in a post-assembly processing step. Thus, the zeamine antibiotics are assembled by a unique combination of nonribosomal peptide synthetase, type I modular polyketide synthase and polyunsaturated fatty acid synthase-like biosynthetic machinery.


Immunotherapeutics to prevent the replication of Brucella in a treatment failure mouse model.
Outer membrane vesicles (OMVs) from Brucella melitensis and irradiated Brucella neotomae have been shown to be effective vaccines against a B. melitensis challenge in a mouse model. The present study evaluates the efficacy of these two vaccines as immuno-therapeutics in combination with conventional antibiotics against a B. melitensis infection. BALB/c mice chronically infected with B. melitensis were treated for 4 weeks with doxycycline and gentamicin and vaccinated twice during the course of therapy. Antibiotics in sub-therapeutic concentrations were chosen in such a way that the treatment would result in a therapeutic failure in mice. Although no additive effect of vaccines and antibiotics was seen on the clearance of B. melitensis, mice receiving vaccines along with antibiotics exhibited no Brucella replication post-treatment compared to mice treated only with antibiotics. Administration of irradiated B. neotomae along with antibiotics led to higher production of IFN-γ ex vivo by splenocytes upon stimulation with heat inactivated B. melitensis while no such effect was seen by splenocytes from mice vaccinated with OMVs. OMV vaccinated mice developed significantly higher anti-Brucella IgG antibody titers at the end of the treatment compared to the mice that received only antibiotics. The mice that received only vaccines did not show any significant clearance of Brucella from spleens and livers compared to non-treated control mice. This study suggests that incorporating OMVs or irradiated B. neotomae along with conventional antibiotics might be able to improve therapeutic efficacy and control the progression of disease in treatment failure cases.


Prevention of hepatitis B virus reinfection after orthotopic liver transplantation.
We discuss the prevention of hepatitis B virus reinfection following orthotopic liver transplantation. Sixty-eight cases of chronic fulminant hepatitis B, the end stage of liver cirrhosis and liver carcinoma complicated with HBV cirrhosis, were given antiviral drugs pre- and posttransplantation to prevent hepatitis B virus reinfection. Lamivudine was administered to two cases and lamivudine + HBIG to 63 cases. Adefovir + HBIG was administered to three cases. The serum HBV, HBV DNA, liver biopsy immunohistochemistry and clinical examinations were performed. One of two cases given lamivudine developed reinfection with serum HBSAg, HbeAb, HBcAb, HBV DNA, and positive and liver biopsy immunohistochemistry showing HBSAg phenotype. Two of the 63 cases given lamivudine + HBIG developed reinfection with serum HBSAg, HBeAb, HBcAb positive and liver biopsy immunohistochemistry showing HBSAg phenotype. The serum HBV DNA was positive in one of the two cases. Three cases given adefovir developed no reinfection with HBV. Orthotopic liver transplantation is an effective treatment for HBV infection; lamivudine + HBIG or adefovir + HBIG prevent hepatitis B virus reinfection.


Determinants of inpatient costs of angina pectoris, myocardial infarction, and heart failure in a university hospital setting in Turkey.
This study aimed to determine the correlates of in-hospital costs for angina pectoris (AP), myocardial infarction (MI), and heart failure (HF) in a university hospital setting. This is a retrospective cost-of-illness study using data from the records of patients who were admitted with AP, MI, or HF to Dokuz Eylül University Hospital during 2008. Direct medical costs were calculated from the Social Security Institute perspective using a bottom-up approach. Socio-demographic and clinical information was abstracted from patient files. Costs were presented in Turkish lira (TL). A generalized linear model was used in the multivariate analysis. We included 337 in-patients in total in the study. AP was present in 26.4% (n=89), MI was present in 55.8% (n=188), and HF was present in 17.8% (n=60) of patients. MI was the most costly disease (2760 TL), followed by HF (2350 TL) and AP (1881 TL). The largest proportion of the total cost was formed by medical interventions (27.5%), followed by surgery (22.2%). Presence of DM, smoking, diagnosis of MI, HF, need for intensive care, and resulting in death were strong predictors of treatment costs. Both preadmission characteristics of patients (diabetes mellitus, smoking, use of anti-aggregant before admission) and in-patient characteristics (diagnosis, coronary artery bypass grafting, intensive care need, death) predicted the hospital cost of cardiovascular diseases (CVDs) independently. Our results may be used as input for health-economic models and economic evaluations to support the decision-making of reimbursement and the cost-effectiveness of public health interventions in healthcare.


Transcriptomic and proteomic profile of Aspergillus fumigatus on exposure to artemisinin.
Artemisinin, an antimalarial drug, and its derivatives are reported to have antifungal activity against some fungi. We report its antifungal activity against Aspergillus fumigatus (A. fumigatus), a pathogenic filamentous fungus responsible for allergic and invasive aspergillosis in humans, and its synergistic effect in combination with itraconazole (ITC), an available antifungal drug. In order to identify its molecular targets, we further analyzed transcript and proteomic profiles of the fungus on exposure to the artemisinin. In transcriptomic analysis, a total of 745 genes were observed to be modulated on exposure to artemisinin, and some of them were confirmed by real-time polymerase chain reaction analysis. Proteomic profiles of A. fumigatus treated with artemisinin showed modulation of 175 proteins (66 upregulated and 109 downregulated) as compared to the control. Peptide mass fingerprinting led to the identification of 85 proteins-29 upregulated and 56 downregulated, 65 of which were unique proteins. Consistent with earlier reports of molecular mechanisms of artemisinin and that of other antifungal drugs, we believe that oxidative phosphorylation pathway (64 kDa mitochondrial NADH dehydrogenase), cell wall-associated proteins and enzymes (conidial hydrophobin B protein, cell wall phiA protein, extracellular thaumatin domain protein, 1,3-beta-glucanosyltransferase Gel2) and genes involved in ergosterol biosynthesis (ERG6 and coproporphyrinogen III oxidase, HEM13) are potential targets of artemisinin for further investigations.


Expression of β-1,4-galactosyltransferase I in a surgically-induced rat model of knee osteoarthritic synovitis.
There are few reports of a biological role for glycosyltransferases in the infiltration of osteoarthritic synovitis. The aim of this research was to investigate the expression and cellular location of β-1,4-galactosyltransferase I (β-1,4-GalT-I) in a surgically-induced rat model of knee osteoarthritis (OA), and explore the role of β-1,4-GalT-I in the pathogenesis of OA. Male Sprague-Dawley rats were randomly divided into three groups: OA group, sham group and normal group. The model of OA was established in the right knees of rats by anterior cruciate ligament transaction (ACLT) with partial medial meniscectomy. Fibroblast-like synoviocytes (FLSs) obtained from normal rat synovial tissue were cultured. The expression of β-1,4-GalT-I mRNA in the synovial tissue, articular cartilage and FLSs treated with tumor necrosis factor-α (TNF-α) were assayed by real-time PCR. Western-blotting and immunohistochemisty were used to observe the expression of β-1,4-GalT-I at the protein level. Double immunofluorescent staining was used to define the location of the β-1,4-GalT-I with macrophage-like synoviocytes, FLSs, neutrophils, and TNF-α in the OA synovium. The alteration of TNF-α in FLSs which were treated with lipopolysaccharide (LPS) and β-1,4-GalT-I-Ab were detected by enzyme-linked immunosorbent assay (ELISA). The mRNA and protein expression of β-1,4-GalT-I increased in synovial tissue of the OA group compared with the normal and sham groups at two and four weeks after the surgery, however, no significant difference appeared in the articular cartilage. Immunohistochemistry also indicated that the β-1,4-GalT-I expression in OA synovium at four weeks after surgery increased sharply compared with the control group. β-1,4-GalT-I co-localized with macrophage-like synoviocytes, FLSs, neutrophils and TNF-α in rat OA synovitis. Moreover, in vitro β-1,4-GalT-I mRNA in FLSs was affected in a dose- and time-dependent manner in response to TNF-α stimulation. ELISA revealed that the expression of TNF-α was attenuated in FLSs in vitro when treated with anti β-1,4-GalT-I antibody. β-1,4-GalT-I may play an important role in the inflammation process of rat OA synovial tissue which would provide the foundation for further researching into the concrete mechanism of β-1,4-GalT-I in OA synovitis.