Temporally limited role of substantia nigra-central amygdala connections in surprise-induced enhancement of learning.
Prediction error plays an important role in modern associative learning theories. For example, the omission of an expected event (surprise) can enhance attention to cues that accompany those omissions, such that subsequent new learning about those cues is more rapid. Many studies from our laboratories have demonstrated that circuitry that includes the amygdala central nucleus (CeA), the cholinergic neurons in the substantia innominata/nucleus basalis region and their innervation of the posterior parietal cortex is critical for this surprise-induced enhancement of attention in learning. We recently showed that midbrain dopamine neurons, known to code prediction error, are also important for surprise-induced enhancement of learning through their interaction with CeA. The present study examined whether in rats the communication between the substantia nigra pars compacta (SNc) and CeA is critical only at the time of surprise, for example to detect prediction error information, or is also needed to maintain and later express that information as enhanced learning. All animals received unilateral CeA lesions and unilateral cannula implants targeting the SNc located contralateral to the lesioned CeA. As the SNc-CeA connections are mainly ipsilateral, inactivating SNc contralateral to the lesioned CeA provided transient blockage of SNc and CeA communication. The results show that SNc-CeA communication is critical for processing prediction error information at the time of surprise, but neither SNc nor SNc-CeA communication is necessary to express that information as enhanced learning later.


Hazards of lung biopsy in asbestos workers.
An investigation into the problem of the frequency and hazards of lung biopsy in asbestos workers was performed in two ways. The first study was into the frequency of lung biopsy among 2907 long term asbestos insulation workers in 1981-3 and the second was into the frequency of fatal complications of lung biopsy in 168 deaths from asbestosis among 2271 consecutive deaths of asbestos insulation workers 1967-76. Only 25 (0.9%) of the 2907 asbestos insulation workers reported having had either an open lung biopsy, a needle biopsy, or a transbronchial biopsy. Seven (24%) of these men suffered difficulties as a result of the biopsy. Lung biopsies had been performed on 14 of the 168 workers who died of asbestosis. Three (21%) of these 14 patients had died within 30 days of biopsy as a direct result of the procedure. In most cases there is no need for lung biopsy to establish a diagnosis of asbestosis; generally, it may be defined by history of exposure, clinical and radiological findings, and other well established non-invasive diagnostic procedures. Certainly, legal and compensation recommendation for biopsy should be considered with the possibility of death in mind. If biopsy is performed precautions should be taken, including adequate observation in hospital.


Memories of pain after burn injury--the patient's experience.
Pain after burns is a major clinical problem and researchers continue to report that burn pain remains undertreated. Adequate pain management could contribute to the prevention of posttraumatic stress disorder and can give a growing sense of patients' self-confidence and strength. Freedom from pain might be unrealistic, but the objective should be to reduce pain as much as possible. The purpose of this study was to describe burn patients' experiences and memories of pain during burn care and to acquire a deeper understanding of how patients cope with the experience. The study method was qualitative and interviews were conducted with 12 adult burn patients (eight men and four women) 6 to 12 months postburn (mean = 7 months). The mean burn size for the group was 10.6% mean of TBSA and the mean stay in hospital was 16 days. The interviews were analyzed using Kvales' method for structuring analysis. The patients' experiences and memories of pain during the trajectory of care were clearly described by the informants during the interviews. Four themes were identified for pain: becoming aware of pain, allowing oneself to feel pain, different pain experiences, and fragile body surface. Four themes were identified for coping: pragmatic coping, allowing someone to care for you, carrying the pain, and perspectives on the trauma. Both good and bad memories were recorded during the care trajectory, and it is evident that the patient has to carry the pain experience by themselves to a large extent.


Climate change and health: Why should India be concerned?
Overwhelming evidence shows that climate change presents growing threats to public health security - from extreme weather-related disasters to wider spread of such vector-borne diseases as malaria and dengue. The impacts of climate on human health will not be evenly distributed around the world. The Third Assessment Report (Intergovernmental Panel on Climate Change-2001) concluded that vulnerability to climate change is a function of exposure, sensitivity, and adaptive capacity. Developing country populations, particularly in small island states, arid and high mountain zones, and in densely populated coastal areas are considered to be particularly vulnerable. India is a large developing country, with the Great Himalayas, the world's third largest ice mass in the north, 7500 km long, and densely populated coast line in the south. Nearly 700 million of her over one billion population living in rural areas directly depends on climate-sensitive sectors (agriculture, forests, and fisheries) and natural resources (such as water, biodiversity, mangroves, coastal zones, grasslands) for their subsistence and livelihoods. Heat wave, floods (land and coastal), and draughts occur commonly. Malaria, malnutrition, and diarrhea are major public health problems. Any further increase, as projected in weather-related disasters and related health effects, may cripple the already inadequate public health infrastructure in the country. Hence, there is an urgent need to respond to the situation. Response options to protect health from effects of climate change include mitigation as well as adaptation. Both can complement each other and together can significantly reduce the risks of climate change.


Prevalence of chronic medical conditions among inmates in the Texas prison system.
Given the rapid growth and aging of the US prison population in recent years, the disease profile and health care needs of inmates portend to have far-reaching public health implications. Although numerous studies have examined infectious disease prevalence and treatment in incarcerated populations, little is known about the prevalence of non-infectious chronic medical conditions in US prison populations. The purpose of this study was to estimate the prevalence of selected non-infectious chronic medical conditions among inmates in the Texas prison system. The study population consisted of the total census of inmates who were incarcerated in the Texas Department of Criminal Justice for any duration from September 1, 2006 through August 31, 2007 (N=234,031). Information on medical diagnoses was obtained from a system-wide electronic medical record system. Overall crude prevalence estimates for the selected conditions were as follows: hypertension, 18.8%; asthma, 5.4%; diabetes, 4.2%; ischemic heart disease, 1.7%; chronic obstructive pulmonary disease, 0.96%; and cerebrovascular disease, 0.23%. Nearly one quarter (24.5%) of the study population had at least one of the selected conditions. Except for asthma, crude prevalence estimates of the selected conditions increased monotonically with age. Nearly two thirds (64.6%) of inmates who were >or=55 years of age had at least one of the selected conditions. Except for diabetes, crude prevalence estimates for the selected conditions were lower among Hispanic inmates than among non-Hispanic White inmates and African American inmates. Although age-standardized prevalence estimates for the selected conditions did not appear to exceed age-standardized estimates from the US general population, a large number of inmates were affected by one or more of these conditions. As the prison population continues to grow and to age, the burden of these conditions on correctional and community health care systems can be expected to increase.


Perioperative erythropoietin protects the CNS against ischemic lesions in patients after open heart surgery.
The aim of this study was to establish erythropoietin as a protective factor against brain ischemia during open heart surgery. A total of 36 consecutive patients scheduled for revascularization heart surgery were included in the study. Of the patients 18 received 3 intravenous doses of recombinant human erythropoietin (rHuEpo, 24,000 IU) and 18 patients received a placebo. Magnetic resonance imaging (MRI) to detect new brain ischemic lesions was performed. Additionally, S100A, S100B, neuron-specific enolase A and B (NSE-A and B) and the concentration of antibodies against N‑methyl-D-aspartate receptors (NMDAR) to identify new neurological complications were determined. Patients who received rHuEpo showed no postoperative ischemic changes in the brain on MRI images. In the control group 5 (27.8 %) new ischemic lesions were found. The NMDAR antibody concentration, S100A, S100B and NSE showed no significant differences between the groups for new cerebral ischemia. High levels of lactate before and after external aortic compression (p = 0.022 and p = 0.048, respectively) and duration of operation could predict new ischemic lesions (p = 0.009). The addition of rHuEpo reduced the formation of lesions detectable by MRI in the brain and could be used clinically as neuroprotection in cardiac surgery.


Outcome of intra-arterial thrombolysis in patients with diabetes and acute lower limb ischemia: a propensity score adjusted analysis.
The presence of diabetes mellitus is rarely addressed in acute lower limb ischaemia (ALLI). The aim of this study was to evaluate the outcome of local intra-arterial thrombolysis for ALLI in patients with diabetes mellitus (DM). Outcome of all thrombolytic events performed in an endovascular first-strategy centre during a 13-year period between 2001 and 2013 in patients with ALLI were followed to January 2017. A propensity score adjusted analysis was performed to evaluate results in patients with (n = 83) versus without (n = 316) DM. Patients with DM were younger (p = 0.001), more often women (p = 0.014), more often had renal insufficiency (p = 0.041), foot ulcers (p < 0.001), and thrombosis (p = 0.032) than the patients without DM. At presentation, patients with DM had a lower degree of ischemia judged by the Rutherford classification, compared to those without DM (p = 0.023). None of the 83 diabetic patients had a popliteal artery aneurysm, compared to 25 (7.9%) of the 316 patients without DM (p = 0.008). The amount of tPA administered to patients with DM was higher than to patients without DM (p = 0.03). In the propensity score adjusted analysis, patients with DM had a higher rate of major amputation at 1 (OR 2.52; 95% CI 1.22-5.20) and 3 years (OR 2.52; 95% CI 1.26-5.04), and a lower amputation-free survival at 3 years (OR 0.46; 95% CI 0.25-0.85), than those without DM. Patients with DM presenting with ALLI differ in clinical characteristics, presentation, and aetiology compared to patients with DM, and have a higher rate of major amputation and lower amputation-free survival rate after intra-arterial thrombolysis.


Brain derived neurotropic factor in first-episode psychosis.
There is much interest, derived from current neurochemical, genetic, and therapeutic research, in the role of brain neurotrophins in schizophrenia. Neurotrophins play key roles in neuronal development and differentiation (i.e., promoting dendritogenesis and synaptogenesis), and in orchestrating the neuronal response to stress/noxious stimuli. Additionally, neurotrophins are modulators across monominergic (dopamine and serotonin), gabaergic and cholinergic systems. These roles focus on important areas of the etiopathophysiology of schizophrenia. Clinical studies show reductions in brain-derived neurotrophic factor (BDNF) and nerve growth factor (NFG) in schizophrenic patients as compared to normal control subjects, as well as differences in patients receiving first-generation antipsychotics (FGAs) or second-generation antipsychotics (SGAs). We now report on BDNF levels in subjects with first-episode psychosis in comparison with normal healthy controls. Compared to normal controls (N=14; 290.5+/-38.81 pg/ml), first-episode psychotic patients showed significant reduction (N=15; 135+/-21.77 pg/ml; P=0.001; f=12.873) in plasma BDNF. Additionally, plasma BDNF levels showed a significant negative correlation (N=13' r=0.584, P=0.0362) only with positive symptom scores at base line and no significant correlations were found with any of the cognitive performance test battery or motor function test scores. Low BDNF levels at the onset of psychosis suggest that it may contribute to the pathogenesis of schizophrenia and/or perhaps could be a helpful neurobiological marker for possible early treatment intervention.


Drosophila p53 integrates the antagonism between autophagy and apoptosis in response to stress.
The tumor suppressor TP53/p53 is a known regulator of apoptosis and macroautophagy/autophagy. However, the molecular mechanism by which TP53 regulates 2 apparently incompatible processes remains unknown. We found that Drosophila lacking p53 displayed impaired autophagic flux, higher caspase activation and mortality in response to oxidative stress compared with wild-type flies. Moreover, autophagy and apoptosis were differentially regulated by the p53 (p53B) and ΔNp53 (p53A) isoforms: while the former induced autophagy in differentiated neurons, which protected against cell death, the latter inhibited autophagy by activating the caspases Dronc, Drice, and Dcp-1. Our results demonstrate that the differential use of p53 isoforms combined with the antagonism between apoptosis and autophagy ensures the generation of an appropriate p53 biological response to stress.


Prenatal rapamycin results in early and late behavioral abnormalities in wildtype C57BL/6 mice.
Mammalian target of rapamycin (mTOR) signaling has been shown to be deregulated in a number of genetic, neurodevelopmental disorders including Tuberous Sclerosis Complex, Neurofibromatosis, Fragile X, and Rett syndromes. As a result, mTOR inhibitors, such as rapamycin and its analogs, offer potential therapeutic avenues for these disorders. Some of these disorders-such as Tuberous Sclerosis Complex-can be diagnosed prenatally. Thus, prenatal administration of these inhibitors could potentially prevent the development of the devastating symptoms associated with these disorders. To assess the possible detrimental effects of prenatal rapamycin treatment, we evaluated both early and late behavioral effects of a single rapamycin treatment at embryonic day 16.5 in wildtype C57Bl/6 mice. This treatment adversely impacted early developmental milestones as well as motor function in adult animals. Rapamycin also resulted in anxiety-like behaviors during both early development and adulthood but did not affect adult social behaviors. Together, these results indicate that a single, prenatal rapamycin treatment not only adversely affects early postnatal development but also results in long lasting negative effects, persisting into adulthood. These findings are of importance in considering prenatal administration of rapamycin and related drugs in the treatment of patients with neurogenetic, neurodevelopmental disorders.


[Diagnosis and surgical treatment of Mirizzi's syndrome].
Retrospective analysis was done concerning the course of cholelithic disease, the results of laboratory, ultrasonographic, radiological, endoscopical methods of investigation, conducted in 52 patients with different forms of Mirizzi syndrome. The method and extent of surgical correction of the disease are recommended.


The management of hypertension in the overweight and obese patient: is weight reduction sufficient?
The management of hypertension in the overweight and obese patient is a frequently encountered but under investigated clinical problem. The conventional management of such patients involves weight reduction with dietary therapy or a combined approach with dietary and anti-obesity drug therapy. However, long-term weight reduction, which is necessary to sustain blood pressure (BP) control, is not feasible in over 80% of patients. Anti-obesity therapy with orlistat has inconsistent effects on BP and may benefit only patients who have uncontrolled or non-medicated hypertension. Anti-obesity therapy with sibutramine may be associated with a modest worsening of BP control. Consequently, antihypertensive drug therapy is often required to supplement a weight reduction programme, and also in patients with severe hypertension or hypertension-associated end-organ damage. Treatment with a thiazide diuretic should be considered as first-line antihypertensive drug therapy in overweight and obese patients. ACE inhibitors or non-dihydropyridine calcium channel antagonists are reasonable alternatives where clinically indicated, or they can be used in combination with a thiazide diuretic if treatment with the diuretic alone is insufficient. If such treatment is inadequate for BP control, the addition or substitution of an alpha- or beta-adrenoceptor antagonist may be considered, although the latter can be associated with weight gain. Concurrent disease is an important determinant of first-line and supplementary antihypertensive drug therapy. Additional studies are needed to determine the long-term (>1 year) efficacy and safety of antihypertensive and anti-obesity management strategies in the overweight and obese hypertensive patient.


Survival of human enteroviruses in the Hawaiian ocean environment: evidence for virus-inactivating microorganisms.
The stability of certain human enteroviruses in the Hawaiian ocean environment was examined. The present data indicated that the time for 90% reduction of poliovirus type 1 at 24 +/- 1 degree C in seawater samples obtained from different sites in Hawaii ranged from 24 to 48 h, and complete inactivation occurred within 72 to 96 h. The accumulated evidence also strongly indicated that a virus-inactivating agent(s) of a microbiological nature was present in both clean and sewage-polluted seawaters, but not in fresh, mountain stream waters. The antiviral activity was lost when the seawater samples were subjected to boiling, autoclaving, or filtration through a 0.22- or 0.45-micrometer, but not a 1.0-micrometer, membrane filter. That the antiviral activity of the seawater was related to the growth activities of microorganisms was corroborated by the observed effects of added nutrients, a lower temperature of incubation, and the presence of certain antibiotics. Other enteric viruses, such as coxsackie virus B-4 and echo virus-7, were also shown to be similarly inactivated in seawater.


Quantitative measurements of HER2 and phospho-HER2 expression: correlation with pathologic response to neoadjuvant chemotherapy and trastuzumab.
Preoperative therapy with chemotherapy and the HER2-targeted monoclonal antibody trastuzumab is valuable for patients with large or locally advanced HER2-positive (HER2+) breast cancers but traditional methods of measuring HER2 expression do not accurately stratify patients for likelihood of response. Quantitative immunofluorescent approaches have the potential to provide a mathematically continuous measure of HER2. Here we seek to determine whether quantitative measurement of HER2 or phospho-HER2 correlates with likelihood of response to trastuzumab- containing neoadjuvant therapy. We evaluated core biopsy samples from 27 HER2+ breast cancer patients enrolled in a preoperative clinical trial using trastuzumab, nab-paclitaxel and carboplatin combination therapy (BrUOG BR-211B (NCT00617942)). Tumor core biopsies were taken before initiation of treatment and 9-13 days after patients received "run-in" doses of either single agent trastuzumab or nab-paclitaxel. The AQUA method of quantitative immunofluorescence was used for analysis of in situ protein expression. Patients then received 18 weeks of treatment, followed by surgery to assess pathologic response to the neoadjuvant regimen. A HER2 score of 2111 by AQUA analysis has been shown to be equivalent to HER2 3+ by immunohistochemical staining in previous studies. Of 20 evaluable patients, 10 cases who achieved a pathologic complete response (pathCR) with neoadjuvant treatment had a mean HER2 level of 10251 compared with 4766 in the patients without pathCR (p = 0.0021). Measurement of phospho-HER2 showed no difference in pathCR vs non-pathCR groups. In 9 patients who had HER2 levels repeated after a single treatment with trastuzumab there was no evidence of a reduction in the HER2 or phospho-HER2 levels following that exposure. High levels of HER2 are associated with achievement of a pathCR in the preoperative setting, while levels of Phospho-HER2 were not predictive of response. This data suggests that accurate measurement of HER2 may help determine the likelihood of response in the pre-surgical setting. Further validation in larger cohorts is required, but this pilot data shows the feasibility of this approach.


Cell carriers as the next generation of cell therapy for cartilage repair: a review of the matrix-induced autologous chondrocyte implantation procedure.
Since the first patient was implanted with autologous cultured chondrocytes more than 20 years ago, new variations of cell therapies for cartilage repair have appeared. Autologous chondrocyte implantation, a first-generation cell therapy, uses suspended autologous cultured chondrocytes in combination with a periosteal patch. Collagen-covered autologous cultured chondrocyte implantation, a second-generation cell therapy, uses suspended cultured chondrocytes with a collagen type I/III membrane. Today's demand for transarthroscopic procedures has resulted in the development of third-generation cell therapies that deliver autologous cultured chondrocytes using cell carriers or cell-seeded scaffolds. To review the current evidence of the matrix-induced autologous chondrocyte implantation procedure, the most widely used carrier system to date. Also discussed are the characteristics of type I/III collagen membranes, behavior of cells associated with the membrane, surgical technique, rehabilitation, clinical outcomes, and quality of repair tissue. Systematic review. Relevant publications were identified by searching Medline from its inception (1949) to December 2007; peer-reviewed publications of preclinical and clinical cell behavior, manufacturing process, surgical technique, and rehabilitation protocols were identified. Preclinical and clinical studies were included if they contained primary data and used a type I/III collagen membrane. Data from these studies demonstrate that patients treated with matrix-induced autologous chondrocyte implantation have an overall improvement in clinical outcomes. Reduced visual analog scale pain levels (range, 1.7-5.32 points) and improvements in the modified Cincinnati (range, 3.8-34.2 points), Lysholm-Gillquist (range, 23.09-47.6 points), Tegner-Lysholm (range, 1.39-3.9 points), and International Knee Documentation Classification scale (P <.05) were observed. Patients had good-quality (hyaline-like) repair tissue as assessed by arthroscopic evaluation (including International Cartilage Repair Society score), magnetic resonance imaging, and histology, as well as a low incidence of postoperative complications. The findings suggest that matrix-induced autologous chondrocyte implantation is a promising third-generation cell therapy for the repair of symptomatic, full-thickness articular cartilage defects.


Cigarette smoking and p16INK4α gene promoter hypermethylation in non-small cell lung carcinoma patients: a meta-analysis.
Aberrant methylation of promoter DNA and transcriptional repression of specific tumor suppressor genes play an important role in carcinogenesis. Recently, many studies have investigated the association between cigarette smoking and p16(INK4α) gene hypermethylation in lung cancer, but could not reach a unanimous conclusion. Nineteen cross-sectional studies on the association between cigarette smoking and p16(INK4α) methylation in surgically resected tumor tissues from non-small cell lung carcinoma (NSCLC) patients were identified in PubMed database until June 2011. For each study, a 2×2 cross-table was extracted. In total, 2,037 smoker and 765 nonsmoker patients were pooled with a fixed-effects model weighting for the inverse of the variance. Overall, the frequency of p16(INK4α) hypermethylation was higher in NSCLC patients with smoking habits than that in non-smoking patients (OR = 2.25, 95% CI = 1.81-2.80). The positive association between cigarette smoking and p16(INK4α) hypermethylation was similar in adenocarcinoma and squamous-cell carcinoma. In the stratified analyses, the association was stronger in Asian patients and in the studies with larger sample sizes. Cigarette smoking is positively correlated to p16(INK4α) gene hypermethylation in NSCLC patients.


Linkage and association of the mitochondrial aspartate/glutamate carrier SLC25A12 gene with autism.
Autism/autistic disorder (MIM number 209850) is a complex, largely genetic psychiatric disorder. The authors recently mapped a susceptibility locus for autism to chromosome region 2q24-q33 (MIM number 606053). In the present study, genes across the 2q24-q33 interval were analyzed to identify an autism susceptibility gene in this region. Mutation screening of positional candidate genes was performed in two stages. The first stage involved identifying, in unrelated subjects showing linkage to 2q24-q33, genetic variants in exons and flanking sequence within candidate genes and comparing the frequency of the variants between autistic and unrelated nonautistic subjects. Two single nucleotide polymorphisms (SNPs) that showed evidence for divergent distribution between autistic and nonautistic subjects were identified, both within SLC25A12, a gene encoding the mitochondrial aspartate/glutamate carrier (AGC1). In the second stage, the two SNPs in SLC25A12 were further genotyped in 411 autistic families, and linkage and association tests were carried out in the 197 informative families. Linkage and association were observed between autistic disorder and the two SNPs, rs2056202 and rs2292813, found in SLC25A12. Using either a single affected subject per family or all affected subjects, evidence for excess transmission was found by the Transmission Disequilibrium Test for rs2056202, rs2292813, and a two-locus G*G haplotype. Similar results were observed using TRANSMIT for the analyses. Evidence for linkage was supported by linkage analysis with the two SNPs, with a maximal multipoint nonparametric linkage score of 1.57 and a maximal multipoint heterogeneity lod score of 2.11. Genotype relative risk could be estimated to be between 2.4 and 4.8 for persons homozygous at these loci. A strong association of autism with SNPs within the SLC25A12 gene was demonstrated. Further studies are needed to confirm this association and to decipher any potential etiological role of AGC1 in autism.


Nucleotide sequence analysis of the long terminal repeat of human T-cell leukemia virus type II.
The nucleotide sequence of the human T-cell leukemia virus type II (HTLV-II) long terminal repeat (LTR) and its surrounding regions were determined. Our results show the following structural features: (i) the LTR is 763 base pairs (bp) in length and consists of 314 +/- 1 bp of region U3, 248 +/- 1 bp of region R, and 201 bp of region U5; (ii) the terminal nucleotides in the LTR form an inverted repeat of T-G.....C-A; (iii) 6-bp direct repeats of cellular sequences flanking the provirus were present; and (iv) the putative functional signals for initiation or termination of viral RNA synthesis were identified. Comparison of the HTLV-II LTR sequence with that previously published for adult T-cell leukemia virus (ATLV; HTLV-I) shows that the LTRs are distinct. Some small regions are conserved between HTLV-II and ATLV, involving sequences important for transcription and a sequence of 21 nucleotides repeated three times in U3. This 21-bp repeat may be important in regulating viral transcription in lymphoid cells.


Turbo Processing for Speech Recognition.
Speech recognition is a classic example of a human/machine interface, typifying many of the difficulties and opportunities of human/machine interaction. In this paper, speech recognition is used as an example of applying turbo processing principles to the general problem of human/machine interface. Speech recognizers frequently involve a model representing phonemic information at a local level, followed by a language model representing information at a nonlocal level. This structure is analogous to the local (e.g., equalizer) and nonlocal (e.g., error correction decoding) elements common in digital communications. Drawing from the analogy of turbo processing for digital communications, turbo speech processing iteratively feeds back the output of the language model to be used as prior probabilities for the phonemic model. This analogy is developed here, and the performance of this turbo model is characterized by using an artificial language model. Using turbo processing, the relative error rate improves significantly, especially in high-noise settings.


Precise Prediction of Calpain Cleavage Sites and Their Aberrance Caused by Mutations in Cancer.
As a widespread post-translational modification of proteins, calpain-mediated cleavage regulates a broad range of cellular processes, including proliferation, differentiation, cytoskeletal reorganization, and apoptosis. The identification of proteins that undergo calpain cleavage in a site-specific manner is the necessary foundation for understanding the exact molecular mechanisms and regulatory roles of calpain-mediated cleavage. In contrast with time-consuming and labor-intensive experimental methods, computational approaches for detecting calpain cleavage sites have attracted wide attention due to their efficiency and convenience. In this study, we established a novel computational tool named DeepCalpain (http://deepcalpain.cancerbio.info/) for predicting the potential calpain cleavage sites by adopting deep neural network and the particle swarm optimization algorithm. Through critical evaluation and comparison, DeepCalpain exhibited superior performance against other existing tools. Meanwhile, we found that protein interactions could enrich the calpain-substrate regulatory relationship. Since calpain-mediated cleavage was critical for cancer development and progression, we comprehensively analyzed the calpain cleavage associated mutations across 11 cancers with the help of DeepCalpain, which demonstrated that the calpain-mediated cleavage events were affected by mutations and heavily implicated in the regulation of cancer cells. These prediction and analysis results might provide helpful information to reveal the regulatory mechanism of calpain cleavage in biological pathways and different cancer types, which might open new avenues for the diagnosis and treatment of cancers.


Dynamic wrinkling of freely floating smectic films.
We demonstrate spontaneous wrinkling as a transient dynamical pattern in thin freely floating smectic liquid-crystalline films. The peculiarity of such films is that, while behaving liquid-like with respect to flow in the film plane, they cannot quickly expand their thickness because that requires stacking of additional smectic layers. At short time scales, they therefore behave like quasi-incompressible membranes, very different from soap films. Smectic films can develop a transient undulation instability or form bulges in response to lateral compression. Optical experiments with freely floating bubbles on parabolic flights and in ground lab experiments are reported. The characteristic wavelengths of the wrinkles are in the submillimeter range. We demonstrate the dynamic nature of the pattern formation mechanism and develop a basic model that explains the physical mechanism for the wavelength selection and wrinkle orientation.


Study on the usefulness of APR scores from the viewpoint of proinflammatory cytokines.
Delayed diagnosis and treatment of newborn infection adversely impact outcomes. Clinical laboratory parameters have aimed to obtain the most correct and prompt diagnosis and treatment of this disease. This study simultaneously observed changes over time in APR as well as proinflammatory cytokines and anti-proinflammatory cytokines and aims to clarify usefulness of APR scores. We evaluated the usefulness of acute phase reactants (APR) in 46 newborns whose serum up to age 7 days had been stored, with comparison of three types (Group I: infection 15, Group F: fetal inflammatory response syndrome 17, and Group C: control 14) of APR-based scores, those of C-reactive protein (CRP), alpha1-acid glycoprotein (AGP), and haptoglobin (Hp), with proinflammatory cytokine levels. APR scores for CRP, AGP, and Hp and the levels of the proinflammatory cytokines IL-1β, IL-6, IL-8, IL-10, and TNFα were determined. The cytokine levels started to increase from age 0 days and then decreased rapidly. The three APR scores, CRP, AG, and Hp, were elevated at age 0 days and then gradually decreased in infection (Group I) and fetal inflammatory response syndrome (Group F). The duration of antibiotic administration according to APR scores was significantly shorter in Group F than in Group I. This study demonstrated APR scores to be more useful for deciding whether antibiotics should be discontinued than proinflammatory cytokine levels.


In vivo cytochrome and alternative pathway respiration in leaves of Arabidopsis thaliana plants with altered alternative oxidase under different light conditions.
The in vivo activity of the alternative pathway (ν(alt)) has been studied using the oxygen isotope fractionation method in leaves of Arabidopsis thaliana modified for the expression of the AtAOX1a gene by anti-sense (AS-12) or overexpression (XX-2). Under non-stressful conditions, ν(alt) was similar in all plant lines regardless of its different alternative pathway capacities (V(alt)). Total leaf respiration (V(t)) and V(alt) were directly related to growth light conditions while electron partitioning between the cytochrome pathway (CP) and alternative pathway (AP) was unchanged by light levels. Interestingly, the AP functioned at full capacity in anti-sense plants under both growth light conditions. The role of the AP in response to a high light stress induced by short-term high light treatment (HLT) was also studied. In wild type and XX-2, both CP and AP rates increased proportionally after HLT while in AS-12, where the AP was unable to increase its rate, the CP accommodated all the increase in respiration. The results obtained under high light stress suggest that flexibility in the response of the mitochondrial electron transport chain is involved in sustaining photosynthetic rates in response to this stress while the saturated AP in AS-12 plants may contribute to the observed increase in photoinhibition.


Disability indicators and WHO programme: "health for all by the year 2000".
The WHO worldwide policy "Health for all by the year 2000" has been adapted into "Targets for health for all by the year 2000" for Europe. The issue of the elderly is addressed by the slogan "Add life to years", the issue of the disabled by "Better opportunities for disabled persons". To support the monitoring process towards attainment of the regional targets, a list of regional indicators was proposed. These indicators are either parts of the national health information systems already, or are developed ad hoc through sample surveys. The indicators in care of the elderly and disabled are reviewed with regard to previous developments of data base on consequences of diseases, malformations and injuries.


Survival Probability, Weibull Characteristics, Stress Distribution, and Fractographic Analysis of Polymer-Infiltrated Ceramic Network Restorations Cemented on a Chairside Titanium Base: An In Vitro and In Silico Study.
Different techniques are available to manufacture polymer-infiltrated ceramic restorations cemented on a chairside titanium base. To compare the influence of these techniques in the mechanical response, 75 implant-supported crowns were divided in three groups: CME (crown cemented on a mesostructure), a two-piece prosthetic solution consisting of a crown and hybrid abutment; MC (monolithic crown), a one-piece prosthetic solution consisting of a crown; and MP (monolithic crown with perforation), a one-piece prosthetic solution consisting of a crown with a screw access hole. All specimens were stepwise fatigued (50 N in each 20,000 cycles until 1200 N and 350,000 cycles). The failed crowns were inspected under scanning electron microscopy. The finite element method was applied to analyze mechanical behavior under 300 N axial load. Log-Rank (p = 0.17) and Wilcoxon (p = 0.11) tests revealed similar survival probability at 300 and 900 N. Higher stress concentration was observed in the crowns' emergence profiles. The MP and CME techniques showed similar survival and can be applied to manufacture an implant-supported crown. In all groups, the stress concentration associated with fractographic analysis suggests that the region of the emergence profile should always be evaluated due to the high prevalence of failures in this area.


Traditional Uses, Pharmacological Activities, and Phytochemical Analysis of Diospyros mespiliformis Hochst. ex. A. DC (Ebenaceae): A Review.
Diospyros mespiliformis Hochst. ex. A. DC is widely distributed throughout Africa and around the world. It is utilized ethnobotanically to treat fevers, wounds, malaria, diabetes mellitus, and other diseases. This review aims to provide an exhaustive overview of the traditional uses, pharmacology, and phytochemical analysis of D. mespiliformis, with the objective of identifying its therapeutic potential for further research. Scientific resources, including Google Scholar, Science Direct, Web of Science, Pub Med, and Scopus, were used to find pertinent data on D. mespiliformis. Secondary metabolites tentatively identified from this species were primarily terpenoids, naphthoquinones, phenolics, and coumarins. D. mespiliformis has been reported to demonstrate pharmacological activities, including antimicrobial, antiproliferative, antiparasitic, antioxidant, anti-inflammatory, antiviral, anti-hypersensitivity, and antidiabetic properties. The phytochemicals and extracts from D. mespiliformis have been reported to have some pharmacological effects in in vivo studies and were not toxic to the animal models that were utilized. The D. mespiliformis information reported in this review provides researchers with a comprehensive summary of the current research status of this medicinal plant and a guide for further investigation.


[Therapeutic options for androgenetic alopecia].
Androgenetic alopecia (AGA) is the most common type of hair loss. There are different treatment possibilities for the prevention of and as a remedy for AGA. The broad spectrum of possibilities varies from doing nothing to hair transplantation. Treating AGA is difficult because it is a natural, ongoing and personal process. The progression of AGA can be slowed down by medication. If one desires a full head of hair and immediate results, however, a hairpiece could be an option. A permanent result is only attainable by hair transplantation. In terms of cost effectiveness over the short term (1 year) and intermediate term (10 years), medication is the best option. Hair transplantation, however, remains the best solution for a permanent result over the long term (more than 10 years). Given the advantages of partial longitudinal follicle unit transplantation (PL-FUT) in comparison with traditional hair transplantation techniques, our preferences go out to PL-FUT and medicinal treatment.


Hydroxychloroquine in the primary thrombosis prophylaxis of antiphospholipid antibody positive patients without systemic autoimmune disease.
Objective The objective of this study was to determine the efficacy of hydroxychloroquine (HCQ) in the primary thrombosis prevention of antiphospholipid antibody (aPL)-positive patients with no other systemic autoimmune diseases. Methods Under the auspices of Antiphospholipid Syndrome Alliance for Clinical Trials and International Networking, a multicenter, international, randomized controlled trial (RCT) was initiated, in which persistently aPL-positive but thrombosis-free patients without systemic autoimmune diseases were randomized to receive HCQ or no treatment in addition to their standard regimen. The primary objective was the efficacy of HCQ in preventing the first thrombosis. The secondary objectives were the thrombosis incidence rate, and the effects of HCQ on aPL profile and mortality rate. Patients were risk-stratified based on antiplatelet agent use. The goal was to follow patients every 6 months for 5 years. Results We recruited 20 persistently aPL-positive patients (female: 19, mean age: 46.6 ± 9.9 years, and baseline antiplatelet medication: 14); 9/20 were randomized to HCQ. During the mean follow-up of 1.7 years, no patients developed thrombosis or a serious adverse event. The study was terminated early due to the low recruitment rate, exacerbated by the prolonged manufacturing shortage and significant price increase of HCQ in the United States. Conclusion Given that a small number of patients with a relatively short follow-up were enrolled in our RCT, and no patients developed thrombosis, we cannot accurately assess the effectiveness of HCQ for primary thrombosis prevention in persistently aPL-positive patients with no other systemic autoimmune diseases. Our experience suggests that conducting an international RCT, especially without pharmaceutical support, is an extremely challenging undertaking.


A Plague of Their Own: Injury Incidence Remains Elevated in the 2021 Major League Baseball Season Compared to Pre-COVID-19 Seasons.
Significant increases in injuries were observed in the 2020 Major League Baseball (MLB) season; these were attributed to an increased acute to chronic workload due to the interrupted preseason and compressed season during the coronavirus disease of 2019 (COVID-19) pandemic. In 2021, the MLB resumed its regular schedule. The purpose of this study was to determine the injury incidence and epidemiology of the 2021 MLB season compared to the injury incidence in the 2020 season and pre-COVID-19 seasons. The hypothesis was that, with the return to normal preseason training, injury incidence in 2021 would return to pre-COVID-19 rates. Additionally, it was hypothesized that injury list (IL) placements at midseason 2021 would be decreased relative to 2020 due to the uninterrupted preseason yet increased at full season 2021 due to increased overall workload from 2020. Descriptive epidemiology study. The MLB transactions database was searched for players placed on the IL between 2018 and 2021. Injuries were categorized by body part and player position. Incidence per 1000 athlete-exposures was calculated for the pre-COVID-19 (2018-2019), 2020, and 2021 seasons. The z test for proportions was used to determine significant differences between injury incidences. The injury incidence rate by midseason 2021 (9.32) compared to 2020 (8.66) was not significantly different (p=0.234). At full season 2021, injury incidence rate (8.69) was significantly higher than pre-COVID-19 seasons (5.13, p<0.001), but not 2020 (p=0.952). When comparing full season 2021 to 2020, increased foot/ankle (0.50 vs 0.14, respectively, p<0.001) and miscellaneous (1.92 vs 0.68, respectively, p<0.001) injuries were observed. The overall injury incidence in 2021 was significantly higher than pre-COVID-19 seasons, and no significant difference was observed between both mid- and full season 2021 and 2020, refuting the hypotheses. This signifies that injury incidence remained elevated in the 2021 season despite resumption of preseason training and a regular season. 3.


Automatic external defibrillators used by emergency medical technicians. A controlled clinical trial.
In a randomized controlled clinical trial, the effectiveness of emergency medical technician (EMT) use of automatic external defibrillators (AEDs) was compared with EMT use of standard defibrillators for patients in cardiac arrest. A total of 321 cardiac arrest patients were treated during the study: 116 were treated by EMTs using the AED (AUTO group), 158 were treated by EMTs using the standard defibrillators (standard group), and 47 were treated by EMTs using the standard defibrillator when they were assigned to use the AED. There was no significant differences in hospital admission or discharge rates between the AUTO group (54% admitted, 28% discharged) and the standard group (52% admitted, 23% discharged) for patients in ventricular fibrillation (VF), for patients in non-VF rhythms, or for all patients combined. The only significant difference observed was in the time from power ON to first shock: 1.1 minutes average AUTO group and 2.0 minutes average standard group. The treatment groups did not differ significantly in sensitivity for VF (78% AED, 76% standard), specificity for non-VF rhythms (100% AED, 95% standard), or rates of defibrillation to a non-VF rhythm (62% AED, 57% standard). We conclude that in clinical outcomes and device performance, AEDs are comparable with standard defibrillators and should be considered an acceptable alternative. Automatic external defibrillators appear to have advantages over standard defibrillators in training, skill retention, and faster operation. Such devices can make early defibrillation available for a much larger portion of the population. They are a major innovation for the prehospital care of cardiac arrest patients.


Lectin-like oxidized low-density lipoprotein receptor-1 (LOX-1), a relevant target for diabetic vasculopathy?
Mechanisms through which major risk factors accelerate diabetic angiopathy include low density lipoprotein (LDL) oxidation and advanced glycation end products (AGEs) formation. Lectin-like oxidized LDL receptor (LOX-1) is a newly identified vascular receptor for oxidized LDL (oxLDL) and AGEs. LOX-1 is up-regulated in vascular endothelium of diabetic animals and thus may be relevant to the development and progression of human diabetic vasculopathy. The mechanisms responsible for LOX-1 induction in diabetes remain unclear but appear to involve metabolic and inflammatory stimuli relevant to diabetes. Such factors may impact on LOX-1-mediated pro-atherogenic events, including endothelial dysfunction and plaque destabilization. Previous studies have shown that drugs commonly used in the treatment of type 2 diabetic patients, including statins and antidiabetic agents, inhibit endothelial LOX-1 expression. This review summarizes recent advances related to the role of LOX-1 in macrovascular diseases, its regulation by some derangements commonly found in diabetic patients and its modulation by vasculoprotective drugs.


Amino acids at the site of V kappa-J kappa recombination not encoded by germline sequences.
Murine V kappa-J kappa recombination is characterized by a maintenance of size at the site of recombination and the use of nucleic acids found only in germline sequences. This is in contrast to heavy chain VH-D-JH assembly where random nucleotides are added at the recombination sites to produce considerable size variation, even though the heptamer/nonomer recombination sequences are identical in both kappa and heavy chain genes. We have examined the origin of an unusual amino acid, Ile, found at the site of V kappa-J kappa recombination in antigalactan antibodies, by sequence analysis of the corresponding rearranged and germline genes. Results indicate that the Ile codon can be generated by use of a single nucleotide 3' of the V kappa segment in combination with the second and third nucleotides of the first codon of J kappa 5 or J kappa 4. However, several antigalactan antibodies express Ile in combination with J kappa 2. An Ile codon cannot be generated by recombination in any reading frame between germline V kappa and J kappa 2 segments. These results suggest that the origin of the Ile codon in lines using J kappa 2 may represent a novel even in murine light chain assembly, possibly similar to the de novo addition of nucleotides observed in heavy chain gene recombination.


Scaling of near-wall flows in quasi-two-dimensional turbulent channels.
The law of the wall and the log law rule the near-wall mean velocity profile of three-dimensional turbulent flows. These well-known laws, which are validated by legions of experiments and simulations, may be universal. Here, using a soap-film channel, we report the first experimental test of these laws in quasi-two-dimensional turbulent channel flows under two disparate turbulent spectra. We find that despite the differences with three-dimensional flows, the laws prevail, albeit with notable distinctions: the two parameters of the log law are markedly distinct from their three-dimensional counterpart; further, one parameter (the von Kármán constant) is independent of the spectrum whereas the other (the offset of the log law) depends on the spectrum. Our results suggest that the classical theory of scaling in wall-bounded turbulence is incomplete wherein a key missing element is the link with the turbulent spectrum.


Steady-state and time-resolved spectroscopy of F420 extracted from methanogen cells and its utility as a marker for fecal contamination.
Methanogenic bacteria, which are common inhabitants of the animal digestive tract, contain the fluorescent compound F420 (coenzyme 420), a 7,8-didemethyl-8-hydroxy-5-deazariboflavin chromophore. F420 was characterized as an initial step in determining if this compound would be useful as a fluorescent marker for the detection of fecal and ingesta contamination. Using a single anion exchange chromatographic process, F420 was separated from other cell components of a Methanobrevibacter sp. cell culture. The extent of separation was determined spectroscopically. To aid in the development of possible techniques for the detection of fecal contamination using F420 as a marker, further spectroscopic investigation of F420 was conducted using steady-state and time-resolved fluorescence methods. The fluorescence lifetime of F420 in an elution buffer of pH 7.5 was found to be 4.2 ns. At higher pH values, the fluorescence decay, F(t), was best described by a sum of two exponentials: at pH 13, F(t) = 0.31 exp(-t/4.20 ns) + 0.69 exp(-t/1.79 ns). Further investigation using front-faced fluorescence techniques has shown that emission from F420 can be collected efficiently from samples of methanogen cell cultures as well as from fecal material.


[Acute toxicity of ginger essential oil to Drosophila melanogaster].
Drosophila melanogaster adults were fed with different concentration ginger essential oil, and time-dosage-mortality model was used to simulate their cumulative mortality within 96 h. The results indicated that higher concentrations of ginger essential oil had high acute toxicity to D. melanogaster adults. Seventy-two hours after treated with 0.5% ginger essential oil, the cumulative mortality of the adults was 88% for female and 79% for male, while with 1% and 10% ginger essential oil, the cumulative mortality of both female and male adults was up to 100%. The LC50 of ginger essential oil decreased gradually with increasing time. Tween-20 as an emulsifying agent also had high toxicity to D. melanogaster adults, especially to male adults, which was more obvious in high concentrations.


The Epidemiology of Smoking in Older Adults: A National Cohort Study.
Older smokers account for the greatest tobacco-related morbidity and mortality in the USA, while quitting smoking remains the single most effective preventive health intervention for reducing the risk of smoking-related illness. Yet, knowledge about patterns of smoking and smoking cessation in older adults is lacking. Assess trends in prevalence of cigarette smoking between 1998 and 2018 and identify patterns and predictors of smoking cessation in US older adults. Retrospective cohort study PARTICIPANTS: Individuals aged 55+ enrolled in the nationally representative Health and Retirement Study, 1998-2018 MAIN MEASURES: Current smoking was assessed with the question: "Do you smoke cigarettes now?" Quitting smoking was defined as having at least two consecutive waves (between 2 and 4 years) in which participants who were current smokers in 1998 reported they were not currently smoking in subsequent waves. Age-adjusted smoking prevalence decreased from 15.9% in 1998 (95% confidence interval (CI) 15.2, 16.7) to 11.2% in 2018 (95% CI 10.4, 12.1). Among 2187 current smokers in 1998 (mean age 64, 56% female), 56% of those living to age 90 had a sustained period of smoking cessation. Smoking less than 10 cigarettes/day was strongly associated with an increased likelihood of quitting smoking (subdistribution hazard ratio 2.3; 95% CI 1.9, 2.8), compared to those who smoked more than 20 cigarettes/day. Smoking prevalence among older persons has declined and substantial numbers of older smokers succeed in quitting smoking for a sustained period. These findings highlight the need for continued aggressive efforts at tobacco cessation among older persons.


The development of new chemotherapeutic agents.
There is a great need for new drugs to treat the increasing numbers of patients with disseminated cancers. Many of the known anticancer drugs, including the taxoids, docetaxel (Taxotere) and paclitaxel (Taxol), are derived from natural products and there will be many more active compounds to be discovered amongst the 300000 plant species available for evaluation. Cytotoxic agents may kill cancer cells by a variety of means. Thymidilate synthase inhibitors such as LY231514 and raltitrexed (Tomudex) have shown activity against a variety of solid tumours including colorectal cancer. The antifolate, trimetrexate, and the nucleoside analogue, gemcitabine, have also shown anticancer activity. Amongst the topoisomerase I inhibitors, CPT-11 is being developed for use against colorectal cancer. Non-cytotoxic agents which interfere with processes such as angiogenesis may also have a role in future treatments for disseminated cancers.


FOXP1 Expression in Normal and Neoplastic Erythroid and Myeloid Cells.
FOXP1 protein was firstly analyzed in normal tissues, and afterwards in different tumor tissues, mainly carcinoma and lymphoma. In B-cell malignancies, its role was well explored; its expression was shown to be connected with disease prognosis in certain B-non Hodgkin lymphomas. In this study, 16 bone marrow trephine samples from patients with no hematopoietic malignancies and 10 samples from peripheral blood of healthy individuals were immunostained with anti-FOXP1 antibody. Positive cells in bone marrows were not only lymphocytes, but also cells that are immunohistochemically positive for glycophorin C or myeloperoxidase. Peripheral blood samples showed no other positive cells, but small round lymphocytes. Additionally 60 samples from patients with myeloid lineage neoplasms were analyzed. 25 samples from patients with myelodysplastic syndrome (MDS) and 35 patients with myeloproliferative disease (MPD) were double immunostained with anti-FOXP1/anti-glycophorin C and anti-FOXP1/anti-myeloperoxidase antibodies. FOXP1 was found to be expressed in 22 cases of MDS and in none of MPD cases. Its expression in MDS was observed mostly in myeloperoxidase positive cells in contrast to gylcophorin C positive cells. Only two cases revealed both myeloperoxidase positive cells and gylcophorin C positive cells expressing FOXP1 transcription factor. Our results show that FOXP1 is present in normal cells of erythroid and myeloid linages and thus suggest its possible role in development of all hematopoetic cells as well as possible involvement in neoplasm development of myeloid disorders.


Climate change and overfishing increase neurotoxicant in marine predators.
More than three billion people rely on seafood for nutrition. However, fish are the predominant source of human exposure to methylmercury (MeHg), a potent neurotoxic substance. In the United States, 82% of population-wide exposure to MeHg is from the consumption of marine seafood and almost 40% is from fresh and canned tuna alone1. Around 80% of the inorganic mercury (Hg) that is emitted to the atmosphere from natural and human sources is deposited in the ocean2, where some is converted by microorganisms to MeHg. In predatory fish, environmental MeHg concentrations are amplified by a million times or more. Human exposure to MeHg has been associated with long-term neurocognitive deficits in children that persist into adulthood, with global costs to society that exceed US$20 billion3. The first global treaty on reductions in anthropogenic Hg emissions (the Minamata Convention on Mercury) entered into force in 2017. However, effects of ongoing changes in marine ecosystems on bioaccumulation of MeHg in marine predators that are frequently consumed by humans (for example, tuna, cod and swordfish) have not been considered when setting global policy targets. Here we use more than 30 years of data and ecosystem modelling to show that MeHg concentrations in Atlantic cod (Gadus morhua) increased by up to 23% between the 1970s and 2000s as a result of dietary shifts initiated by overfishing. Our model also predicts an estimated 56% increase in tissue MeHg concentrations in Atlantic bluefin tuna (Thunnus thynnus) due to increases in seawater temperature between a low point in 1969 and recent peak levels-which is consistent with 2017 observations. This estimated increase in tissue MeHg exceeds the modelled 22% reduction that was achieved in the late 1990s and 2000s as a result of decreased seawater MeHg concentrations. The recently reported plateau in global anthropogenic Hg emissions4 suggests that ocean warming and fisheries management programmes will be major drivers of future MeHg concentrations in marine predators.


A global approach to identify novel broad-spectrum antibacterial targets among proteins of unknown function.
Attempted allelic replacement of 144 Streptococcus pneumoniae open reading frames of previously uncharacterized function led to the identification of 36 genes essential for growth under laboratory conditions. Of these, 14 genes (obg, spoIIIJ2, trmU, yacA, yacM, ydiC, ydiE, yjbN, yneS, yphC, ysxC, ytaG, yloI and yxeH4) were also essential in Staphylococcus aureus and Haemophilus influenzae or Escherichia coli, 2 genes (yrrK and ydiB) were only essential in H. influenzae as well as S. pneumoniae and 8 genes were necessary for growth of S.pneumoniae and S. aureus and did not have a homolog in H. influenzae(murD2, ykqC, ylqF, yqeH, ytgP, yybQ) or were not essential in that organism (yqeL, yhcT). The proteins encoded by these genes could represent good targets for novel antibiotics covering different therapeutic profiles. The putative functions of some of these essential proteins, inferred by bioinformatic analysis, are presented. Four mutants, with deletions of loci not essential for in vitro growth, were found to be severely attenuated in a murine respiratory tract infection model, suggesting that not all targets for antibacterial therapeutics are revealed by simple in vitro essentiality testing. The results of our experiments together with those collated from previously reported studies including Bacillus subtilis, E. coli and Mycoplasma sp. demonstrate that gene conservation amongst bacteria does not necessarily indicate that essentiality in one organism can be extrapolated to others. Moreover, this study demonstrates that different experimental procedures can produce apparently contradictory results.


NNVA: Neural Network Assisted Visual Analysis of Yeast Cell Polarization Simulation.
Complex computational models are often designed to simulate real-world physical phenomena in many scientific disciplines. However, these simulation models tend to be computationally very expensive and involve a large number of simulation input parameters, which need to be analyzed and properly calibrated before the models can be applied for real scientific studies. We propose a visual analysis system to facilitate interactive exploratory analysis of high-dimensional input parameter space for a complex yeast cell polarization simulation. The proposed system can assist the computational biologists, who designed the simulation model, to visually calibrate the input parameters by modifying the parameter values and immediately visualizing the predicted simulation outcome without having the need to run the original expensive simulation for every instance. Our proposed visual analysis system is driven by a trained neural network-based surrogate model as the backend analysis framework. In this work, we demonstrate the advantage of using neural networks as surrogate models for visual analysis by incorporating some of the recent advances in the field of uncertainty quantification, interpretability and explainability of neural network-based models. We utilize the trained network to perform interactive parameter sensitivity analysis of the original simulation as well as recommend optimal parameter configurations using the activation maximization framework of neural networks. We also facilitate detail analysis of the trained network to extract useful insights about the simulation model, learned by the network, during the training process. We performed two case studies, and discovered multiple new parameter configurations, which can trigger high cell polarization results in the original simulation model. We evaluated our results by comparing with the original simulation model outcomes as well as the findings from previous parameter analysis performed by our experts.


[Various individual features of human adaptation to altitude].
Two groups of young healthy men--natives of lowlands who for one year lived and worked in chronic hypoxia (Group 1 at an altitude of 1680 m with PO2 = 120 mm Hg and Group 2 at an altitude of 3650 with PO2 = 90 mm Hg) were examined. It was found that after this prolonged exposure the subjects showed a higher sensitivity of the respiration system to hypoxia, an enhanced lung ventilation and circulation, a lower gas exchange and physical work capacity. The concentration of lactic acid at rest in the Group 2 subjects was 47% higher than in the Group 1 subjects. The lactate/pyruvate ratio in the Group 2 subjects increased by 46% thus indicating an enhanced rate of anaerobic processes. A higher deficiency of buffer bases, a lower concentration of bicarbonates in blood at rest and during exercise tests of the Group 2 subjects pointed to metabolic acidosis. The subjects with a higher rate of anaerobic metabolism in a low PO2 environment displayed a diminished sensitivity of the hypoxic stimulation of respiration, an increased tolerance to the very low PAO2 and a reduced work capacity in chronic hypoxia.


A Mixed Methods Approach to Evaluate Partnerships and Implementation of the Massachusetts Prevention and Wellness Trust Fund.
Background: Strong partnerships are critical to integrate evidence-based prevention interventions within clinical and community-based settings, offering multilevel and sustainable solutions to complex health issues. As part of Massachusetts' 2012 health reform, The Prevention and Wellness Trust Fund (PWTF) funded nine local partnerships throughout the state to address hypertension, pediatric asthma, falls among older adults, and tobacco use. The initiative was designed to improve health outcomes through prevention and disease management strategies and reduce healthcare costs. Purpose: Describe the mixed-methods study design for investigating PWTF implementation. Methods: The Consolidated Framework for Implementation Research guided the development of this evaluation. First, the study team conducted semi-structured qualitative interviews with leaders from each of nine partnerships to document partnership development and function, intervention adaptation and delivery, and the influence of contextual factors on implementation. The interview findings were used to develop a quantitative survey to assess the implementation experiences of 172 staff from clinical and community-based settings and a social network analysis to assess changes in the relationships among 72 PWTF partner organizations. The quantitative survey data on ratings of perceived implementation success were used to purposively select 24 staff for interviews to explore the most successful experiences of implementing evidence-based interventions for each of the four conditions. Conclusions: This mixed-methods approach for evaluation of implementation of evidence-based prevention interventions by PWTF partnerships can help decision-makers set future priorities for implementing and assessing clinical-community partnerships focused on prevention.


Transbilayer inhibition of protein kinase C by the lipophosphoglycan from Leishmania donovani.
Lipophosphoglycan (LPG), the predominant molecule on the surface of the parasite Leishmania donovani, has previously been shown to be a potent inhibitor of protein kinase C (PKC) isolated from rat brain. The mechanism by which LPG inhibits PKC was further investigated in this study. LPG was found to inhibit the PKC alpha-catalyzed phosphorylation of histone in assays using large unilamellar vesicles composed of 1-palmitoyl, 2-oleoyl phosphatidylserine and 1-palmitoyl, 2-oleoyl phosphatidylcholine either with or without 1% 1,2 diolein added. The results also indicated that while PKC binding to sucrose-loaded vesicles was not substantially reduced in the presence of LPG at concentrations of 1-2%, the activity of membrane-bound PKC was inhibited by 70%. This inhibition of the membrane-bound form of PKC is not a consequence of reduced substrate availability to the membrane. However, Km shifted from approximately 31 +/- 4 microM to 105 +/- 26 microM in the presence of 5% LPG. LPG caused PKC to bind to membranes without inducing a conformational change as revealed by the lack of an increased susceptibility to trypsin. An LPG fragment containing only one repeating disaccharide unit was not as effective as the entire LPG molecule or of larger fragments in inhibiting the membrane-bound form of the enzyme. The shorter fragments were also less potent in raising the bilayer to hexagonal phase transition temperature of a model membrane. LPG is also able to inhibit the membrane-bound form of PKC alpha from the inner monolayer of large unilamellar vesicles, the opposite monolayer to which the enzyme binds in our assay. Inhibition is likely a result of alterations in the physical properties of the membrane. To our knowledge, this is the first example of a membrane additive that can inhibit the membrane-bound form of PKC in the presence of other lipid cofactors.


Analysis of Copy Number Variations in Patients with Autism Using Cytogenetic and MLPA Techniques: Report of 16p13.1p13.3 and 10q26.3 Duplications.
Autism is a common neuropsychiatric disorder affecting 1 in 68 children. Copy number variations (CNVs) are known to be major contributors of autism spectrum disorder (ASD). There are different whole genome or targeted techniques to identify CNVs in the patients including karyotyping, multiplex ligation-dependent probe amplification (MLPA) and array CGH. In this study, we used karyotyping and MLPA to detect CNVs in 50 Iranian patients with autism. GTG banding and 4 different MLPA kits (2 subtelomeric and 2 autism kits) were utilized. To elevate our detection rate, we selected the sporadic patients who had additional clinical features including intellectual disability, seizure, attention deficit hyperactivity disorder, and abnormal head circumference. Two out of 50 patients (4%) showed microscopic chromosome abnormalities and 5 out of 50 (10%) demonstrated copy number gains or losses using MLPA kits. Including one overlapping result between karyotype and MLPA techniques, our overall detection rate was 6 out of 50 (12%). Three out of 6 CNVs were de novo and three others were paternally inherited. Two of CNVs detected by karyotyping and MLPA tests were 16p13.1q13.3 and 10q26.3 duplications, respectively. For these two CNVs genotype and phenotype of the patients were compared with other studies. Although the pathogenicity of cytogenetic results was certain, most of MLPA results needed to be better refined using other more accurate techniques such as array CGH. Our findings suggest that it might be possible to obtain some useful information using MLPA technique but it cannot be used as a single diagnostic tool for the autism.


The effect of the shape of gold core-mesoporous silica shell nanoparticles on the cellular behavior and tumor spheroid penetration.
Size, surface charge, and shape have a huge influence on the behavior, cellular uptake, and cytotoxic profile of nanoparticles. Herein, gold core and silica shell based nanoparticles (Au-MSSs) with spherical or rod-like shape were produced, in order to disclose the effect of the shape of nanomaterials on the cellular uptake, reactive oxygen species (ROS) production, 3D tumor spheroid penetration and cytotoxicity towards cancer cells. The Au-MSS spheres induced greater reduction of the ROS content of cancer cells and also displayed a more homogeneous distribution and penetration in 3D tumor spheroids. However, the Au-MSS rods show enhanced cellular migration and uptake in 2D cell cultures, which results in a higher drug delivering capacity. Furthermore, the Au-MSS rods displayed an enhanced cellular cytotoxicity upon laser irradiation (808 nm, 1.7 W cm-2, 10 min), where less than 10% of cells remained viable. In addition, both Au-MSSs have the potential to be used as imaging agents, which further expands their applicability as theranostic agents in the biomedical area. In summary, the obtained results show that the shape of Au-MSSs is crucial for their biological performance, which will have a great influence on their therapeutic outcome. Therefore, our findings may provide useful information for the development and design of new drug delivery systems towards a more effective therapy.


Methane production from acetate and associated methane fluxes from anoxic coastal sediments.
The apparent microbial conversion of acetate to methane ranges seasonally from 0.7 to 88 micromoles per liter of whole wet sediment per hour in the top 5 centimeters of methane-producing sediments underlying sulfate-reducing sediments in Cape Lookout Bight, North Carolina. The associated methane flux across the sediment-water interface into overlying waters exhibits the same seasonal pattern. Significant methane production from acetate is observed only in sulfate-depleted sediments.


The odorant-sensitive adenylate cyclase of olfactory receptor cells. Differential stimulation by distinct classes of odorants.
We have characterized odorant-stimulated adenylate cyclase activity in isolated chemosensory cilia prepared from frog and rat olfactory epithelium. Cilia from both species exhibit high levels of adenylate cyclase activity. Basal activity is stimulated approximately 2-fold by GTP and approximately 5-fold by guanosine 5'-(3-O-thio)triphosphate and forskolin. Odorants augment enzyme activity 30-65% above the basal level in a tissue-specific and GTP-dependent manner. Calcium reduces GTP-stimulated activity with a 50% effective concentration at 10 microM. Odorants vary in their influence upon olfactory adenylate cyclase activity. Most fruity, floral, minty, and herbaceous odorants stimulate the enzyme. 3,7-Dimethyl-2,6-octadienenitrile (citralva), menthone, D-carvone, L-carvone, and 2-isobutyl-3-methoxypyrazine display similar potencies in activating the adenylate cyclase upto concentrations of 100 microM. Putrid odorants, such as isovaleric acid, triethylamine, pyridine, thiazole, and methoxypyrazine, and odorous chemical solvents, do not stimulate enzyme activity. In homologous series of pyrazine, thiazole, and pyridine odorants, compounds with the longest hydrocarbon side chains are best able to enhance enzyme activity. The failure of certain odorants to affect adenylate cyclase activity suggests that additional transduction mechanisms besides the formation of cAMP are involved in olfaction.


Atrial fibrillation in patients with end-stage renal disease on hemodialysis: Magnitude of the problem and new approach to oral anticoagulation.
Atrial fibrillation (AF) is a frequent comorbid condition in patients with end-stage renal disease on hemodialysis (HD) with a prevalence of up to 27%. The incidence rate of stroke in AF patients on HD is approximately 5%. The AF-associated risk of stroke is a major clinical challenge because current evidence for anticoagulation in HD patients with AF is based on observational data. Results from these observational studies is largely contradictory because they do not show a clear benefit of vitamin K antagonists over no treatment in terms of stroke prevention, and they show an increased risk of hemorrhage associated with anticoagulation treatment in HD patients. HD patients were not included in randomized trials of the direct oral anticoagulants (DOACs), and therefore there is no evidence to support efficacy and safety of DOACs compared to vitamin K antagonists in HD patients. The pharmacological characteristics of DOACs are of particular interest in the HD setting. The factor Xa inhibitors rivaroxaban, apixaban, and edoxaban are not predominantly eliminated via the kidneys. The thrombin inhibitor dabigatran is 80% eliminated via the kidneys but is dialyzable due to its low protein binding. In this narrative review, we examine the current state of evidence regarding the prevalence of AF in patients on HD, the associated risk of stroke, and the efficacy and safety of anticoagulation for stroke prevention in the HD setting. Further, based on the pharmacokinetic properties of DOACs, we discuss their potential use in patients on HD and ongoing randomized trials.


Population Control, Family Planning, and Maternal Health Networks in the 1960s/70s: Diary of an International Consultant.
Over the past decade historians have explored the rise of the mid-twentieth-century population/family planning movement on both the international and the local levels. This article bridges the gap between these studies by exploring the work diaries of Dr. Adaline Pendleton ("Penny") Satterthwaite, a midlevel technical advisor who traveled to over two dozen countries for the Population Council from 1965 to 1974. Penny's diaries draw our attention to a diverse network of advocates who mediated between international population activists, state actors, and local communities while also acting as conduits for the transnational spread of strategies and resources. Her experiences also provide evidence of the coercive practices, gendered tensions, and political conflicts shaping the movement while illustrating the resistance and engagement of local actors, the existence of health- and women-centered approaches even during the high period of population control, and the many structural and social barriers shaping family planning projects in practice.


Ileal digestibility of amino acids of unheated and autoclaved pea protein concentrate in broilers.
The effects of autoclaving 2 varieties of micronized (fine grinding) pea protein concentrate (PPC) on the ileal digestibility (ID) of CP and amino acids (AA) were studied in broilers. There was a control diet based on fermented soybean meal (FSBM) and 4 extra diets in which the FSBM was substituted on a CP basis by PPC from 2 different pea cultivars (PPC-1 and PPC-2), either unheated or autoclaved. Chicks were fed a common diet from 1 to 17 d of age and, then, their respective experimental diets from 18 to 21 d of age. Each treatment was replicated 6 times. Autoclaving reduced trypsin inhibitor activity (TIA) but had little effect on the saponin content of the PPC. The apparent ID (AID) of CP was similar for the FSBM and the unheated PPC and lower for both than for the autoclaved PPC. Autoclaving improved (P < 0.001) the AID of CP (87.6 vs. 82.2%) and most indispensable AA (e.g., 92.1 vs. 88.8% for Lys and 83.6 vs. 76.5% for Thr) of the PPC. The improvement in CP and AA digestibility with autoclaving varied with the PPC used and was consistent with the reduction in TIA observed (9.4 to 2.8 mg/g for PPC-1 vs. 9.1 to 5.3 mg/g for PPC-2). The standardized ID (SID) of most indispensable AA was similar for the FSBM and the PPC-2 and higher for both than for the PPC-1 (P < 0.05). For Lys, the lowest SID value was observed for the FSBM and the highest for the PPC-2 either unheated or autoclaved. It is concluded that the ID of the AA of the PPC improved with heating and was in general higher for the autoclaved PPC than for the FSBM. Consequently, heat processed PPC is a good alternative to FSBM and unheated PPC in starter diets for broilers.


Long-term, ambulatory, subcutaneous insulin infusion versus multiple daily injections in brittle diabetic patients.
We compared the blood glucose control of four intact and eight kidney recipient, metabolically unstable, ketosis-prone, insulin-dependent diabetic patients under two different regimens: (a) intensive conventional treatment with two to four insulin injections daily (48 patient-months) and (B) subcutaneous, portable insulin delivery system (IDS) (54 patient-months). Both regimens included frequent home blood glucose and 24-h urine glucose determinations and daily telephone follow-up to maximize compliance with treatment. Analyzed as a group the fasting blood glucose for intact patients (A: 172 +/- 13 mg/dl; B: 141 +/- 12, P less than 0.02) and the nonfasting blood glucose for kidney recipient patients (A: 165 +/- 10; B: 138 +/- 5, P less than 0.01) were significantly lower during treatment with the IDS than with multiple injections. Six out of 12 patients (2/4 intact and 4/8 kidney recipient patients) showed significant and consistent improvement of blood glucose concentrations. Four showed marginal and inconsistent improvement. Two patients (one intact and one kidney recipient) improved on the IDS but maintained the improvement when changed back to conventional treatment. The 24-h urine glucose, maximal glucose excursions, number of blood glucoses less than or equal to 40 mg/dl, and glycosylated hemoglobin decreased significantly in some patients on the pump. We conclude that subcutaneous, portable insulin delivery devices can significantly improve the metabolic control of some ambulatory, unstable diabetic patients during long-term treatment beyond that obtained with intensive, multiple-injection, conventional treatment. Normalization of the metabolic control, however, is not obtained. These infusion systems still pose several problems during ambulatory use, which could have serious consequences in patients less compliant and/or followed less closely than ours.


Treatment cost development of patients undergoing remission induction chemotherapy: a pharmacoeconomic analysis before and after introduction of posaconazole prophylaxis.
Prior clinical trials have demonstrated efficacy and effectiveness of posaconazole in the prophylaxis of invasive fungal diseases in high-risk patients. Controversy exists about the cost-effectiveness of this approach. We performed an analysis comparing the direct costs of posaconazole prophylaxis against polyene mouthwash (thrush) prophylaxis in patients with acute myelogenous leukaemia (AML). Data of AML patients receiving remission-induction chemotherapy were extracted from the CoCoNut (Cologne Cohort of Neutropenic Patients) database to compare hospital costs of patients before (2003-2005) and after (2006-2008) introduction of posaconazole prophylaxis. Treatment on general ward, intensive care unit (ICU), mechanical ventilation, diagnostic procedures, and all anti-infectives were calculated. Patient groups were well matched according to age, gender and duration of neutropenia. The mean costs per patient in the posaconazole group (n = 76) and the polyene mouthwash group (n = 81) were €21 040 (95% confidence interval (CI): €18 204-€23 876) and €23 169 (95% CI: €19 402-€26 937) per patient. Antifungal treatment costs were €4580 (95% CI: €3678-€5482) and €4019 (95% CI: €2825-€5214). Duration on the ICU was 2582 (95% CI: 984.1-4181.7) and 5517 (95% CI: 2206-8827.3) min. In our hospital, primary antifungal prophylaxis by posaconazole was cost-effective. There was a trend towards cost savings, which was primarily caused by a shorter overall length of stay and the less frequent ICU treatment.


Mapping of the tuberous sclerosis genes.
Tuberous sclerosis is a dominantly inherited genetic disorder, with a high frequency of new mutations, which has been shown through genetic linkage studies to be genetically heterogeneous. In this paper we summarize recent progress in linkage studies which indicate that there is one TSC gene on chromosome 9q34 which accounts for approximately 40% of cases and another on chromosome 16p13.3 which accounts for approximately 50% of cases. We discuss the map position of these TSC genes on chromosome 9q34 and 16p13.3. We discuss physical mapping data in these two regions. We postulate on the pathogenesis of TSC lesions and on the cause of the high frequency of new mutations. We present a strategy to progress from the mapping of the TSC genes to isolation of these genes.


Lifetime prevalence, correlates and health consequences of gender-based violence victimisation and perpetration among men and women in Somalia.
Humanitarian emergencies increase the risk of gender-based violence (GBV). We estimated the prevalence of GBV victimisation and perpetration among women and men in urban settings across Somalia, which has faced decades of war and natural disasters that have resulted in massive population displacements. A population-based survey was conducted in 14 urban areas across Somalia between December 2014 and November 2015. A total of 2376 women and 2257 men participated in the survey. One in five men (22.2%, 95% CI 20.5 to 23.9) and one in seven (15.5%; 95% CI 14.1 to 17.0) women reported physical or sexual violence victimisation during childhood. Among women, 35.6% (95% CI 33.4 to 37.9) reported adult lifetime experiences of physical or sexual intimate partner violence (IPV) and 16.5% (95% CI 15.1 to 18.1) reported adult lifetime experience of physical or sexual non-partner violence (NPV). Almost one-third of men (31.2%; 95% CI 29.4 to 33.1) reported victimisation as an adult, the majority of which was physical violence. Twenty-two per cent (21.7%; 95% CI 19.5 to 24.1) of men reported lifetime sexual or physical IPV perpetration and 8.1% (95% CI 7.1 to 9.3) reported lifetime sexual or physical NPV perpetration. Minority clan membership, displacement, exposure to parental violence and violence during childhood were common correlates of IPV and NPV victimisation and perpetration among women and men. Victimisation and perpetration were also strongly associated with recent depression and experiences of miscarriage or stillbirth. GBV is prevalent and spans all regions of Somalia. Programmes that support nurturing environments for children and provide health and psychosocial support for women and men are critical to prevent and respond to GBV.


Posttranslational Regulation of IL-23 Production Distinguishes the Innate Immune Responses to Live Toxigenic versus Heat-Inactivated Vibrio cholerae.
Vibrio cholerae infection provides long-lasting protective immunity, while oral, inactivated cholera vaccines (OCV) result in more-limited protection. To identify characteristics of the innate immune response that may distinguish natural V. cholerae infection from OCV, we stimulated differentiated, macrophage-like THP-1 cells with live versus heat-inactivated V. cholerae with and without endogenous or exogenous cholera holotoxin (CT). Interleukin 23A gene (IL23A) expression was higher in cells exposed to live V. cholerae than in cells exposed to inactivated organisms (mean change, 38-fold; 95% confidence interval [95% CI], 4.0 to 42; P < 0.01). IL-23 secretion was also higher in cells exposed to live V. cholerae than in cells exposed to inactivated V. cholerae (mean change, 5.6-fold; 95% CI, 4.4 to 11; P < 0.001). This increase in IL-23 secretion was more marked than for other key innate immune cytokines (e.g., IL-1β and IL-6) and dependent on exposure to the combination of both live V. cholerae and CT. While IL-23 secretion was reduced following stimulation with either heat-inactivated wild-type V. cholerae or a live isogenic ctxAB mutant of V. cholerae, the addition of exogenous CT restored IL-23 secretion in combination with the live isogenic ctxAB mutant V. cholerae, but not when it was paired with stimulation by heat-inactivated V. cholerae The posttranslational regulation of IL-23 under these conditions was dependent on the activity of the cysteine protease cathepsin B. In humans, IL-23 promotes the differentiation of Th17 cells to T follicular helper cells, which maintain and support long-term memory B cell generation after infection. Based on these findings, the stimulation of IL-23 production may be a determinant of protective immunity following V. cholerae infection.IMPORTANCE An episode of cholera provides better protection against reinfection than oral cholera vaccines, and the reasons for this are still under study. To better understand this, we compared the immune responses of human cells exposed to live Vibrio cholerae with those of cells exposed to heat-killed V. cholerae (similar to the contents of oral cholera vaccines). We also compared the effects of active cholera toxin and the inactive cholera toxin B subunit (which is included in some cholera vaccines). One key immune signaling molecule, IL-23, was uniquely produced in response to the combination of live bacteria and active cholera holotoxin. Stimulation with V. cholerae that did not produce the active toxin or was killed did not produce an IL-23 response. The stimulation of IL-23 production by cholera toxin-producing V. cholerae may be important in conferring long-term immunity after cholera.


Relationships between chemotaxis, chemotactic modulators, and cyclic nucleotide levels in tumor cells.
Like many freely moving cells, Walker 256 carcinosarcoma cells respond to chemotactic stimuli. Since cyclic nucleotides are involved in the chemotaxis of other cells, we have examined the action of several nucleosides and nucleotides as chemoattractants and as modulators of tumor cell movement. We have also studied the effect of chemoattractants and prostaglandins on intracellular cyclic nucleotide levels and the effect of prostaglandins as modulators of chemotaxis. Of the agents studied, only the cyclic nucleotides and prostaglandins were found to modulate cellular motility. Neither cyclic adenosine 3':5'-monophosphate (cAMP) nor cyclic guanosine 3':5'-monophosphate (cGMP) was a chemoattractant, but cGMP and N6,O2'-dibutyryl cyclic guanosine 3':5'-monophosphate at low concentrations (approximately 10(-10) M to 10(-8) M) enhanced chemotaxis by 80 +/- 15%, and both cAMP and N6,O2-dibutyryl cyclic adenosine 3':5'-monophosphate had an inhibitory effect at concentrations greater than 10(-6) M. Chemotaxis was suppressed by 21 to 100% in media depleted of Ca2+ and/or Mg2+, but in the presence of 10(-8) M cGMP, there was partial recovery of the chemotactic response. In response to chemotactic stimulation, there was a 28 to 60% rise in intracellular cAMP within 30 sec. This returned to basal levels within 2 min. Intracellular cGMP levels became elevated approximately 3- to 3.5-fold after this time. Incubation of cells with prostaglandins A1 and F2 alpha stimulated chemotaxis at lower concentrations (10(-7) and 10(-9) M, respectively) and resulted in elevation of cGMP, while incubation with prostaglandin E2 resulted in inhibition of chemotaxis and a rise in cAMP levels.


A statistical approach to the initial volume problem in Single Particle Analysis by Electron Microscopy.
Cryo Electron Microscopy is a powerful Structural Biology technique, allowing the elucidation of the three-dimensional structure of biological macromolecules. In particular, the structural study of purified macromolecules -often referred as Single Particle Analysis(SPA)- is normally performed through an iterative process that needs a first estimation of the three-dimensional structure that is progressively refined using experimental data. It is well-known the local optimisation nature of this refinement, so that the initial choice of this first structure may substantially change the final result. Computational algorithms aiming to providing this first structure already exist. However, the question is far from settled and more robust algorithms are still needed so that the refinement process can be performed with sufficient guarantees. In this article we present a new algorithm that addresses the initial volume problem in SPA by setting it in a Weighted Least Squares framework and calculating the weights through a statistical approach based on the cumulative density function of different image similarity measures. We show that the new algorithm is significantly more robust than other state-of-the-art algorithms currently in use in the field. The algorithm is available as part of the software suite Xmipp (http://xmipp.cnb.csic.es) and Scipion (http://scipion.cnb.csic.es) under the name "Significant".


Effects of Caloric Intake and Aerobic Activity in Individuals with Prehypertension and Hypertension on Levels of Inflammatory, Adhesion and Prothrombotic Biomarkers-Secondary Analysis of a Randomized Controlled Trial.
Cardiopulmonary fitness and low calorie diets have been shown to reduce inflammation but few studies have been conducted in individuals with elevated blood pressure (BP) in a randomized intervention setting. Thereby, adhesion biomarkers, e.g., soluble intercellular adhesion molecule (sICAM)-3, have not been examined so far. Sixty-eight sedentary prehypertensive and mildly hypertensive individuals (mean age ± SEM: 45 ± 1 years; mean BP: 141/84 ± 1/1 mmHg) were randomized to one of three 12-week intervention groups: cardio training and caloric reduction, cardio training alone, or wait-list control group. Plasma levels of inflammatory, adhesion and prothrombotic biomarkers were assessed. In a second step, intervention groups were combined to one sample and multivariate regression analyses were applied in order to account for exercise and diet behavior changes. There were no significant differences among the intervention groups. In the combined sample, greater caloric reduction was associated with a larger increase of sICAM-3 (p = 0.026) and decrease of C-reactive protein (p = 0.018) as a result of the interventions. More cardio training was associated with increases of sICAM-3 (p = 0.046) as well as interleukin-6 (p = 0.004) and a decrease of tumor necrosis factor- (p = 0.017) levels. Higher BP predicted higher plasminogen activator inhibitor (PAI)-1 (p = 0.001), and greater fitness predicted lower PAI-1 levels (p = 0.006) after the intervention. In prehypertensive and hypertensive patients, plasma levels of the adhesion molecule sICAM-3 and inflammatory biomarkers have different response patterns to cardio training with and without caloric reduction. Such anti-inflammatory and anti-thrombotic effects may have implications for the prevention of atherothrombotic cardiovascular disease among individuals at increased risk.


A quantitative approach to determining disease response during therapy using multiple biologic markers: application to carcinoma of the breast.
This analytical study was undertaken in an effort to develop a model for a quantitative approach to the evaluation of multiple biological marker levels in blood and urine as a means for determining tumor changes during treatment of patients with malignant disease. The potential biologic markers measured in patients with carcinoma of the breast consist of three urinary polyamines (putrescine, spermidine and spermine), three urinary nucleosides (pseudouridine, N2, N2-dimethylguanosine and 1-methylinosine), and plasma carcinoembryonic antigen (CEA). The distribution patterns of the seven markers measured pretreatment and five weeks after initiating therapy were examined by grouping the patients into the three categories of progression, stable, or regression based on their clinical response to treatment. In addition to the individual marker measurements, the pretreatment and posttreatment values of the ratios of the polyamine levels (spermine/putrescine, spermine/spermidine, and spermidine/putrescine) and the nucleoside levels (N2, N2-dimethylguanosine/pseudouridine, 1-methylinosine/pseudouridine, and 1-methylinosine/N2, N2-dimethylguanosine) were also evaluated. In the pretreatment measurements, CEA levels were elevated for 76% of the patients and the three nucleosides were elevated for 36% of the patients and the three nucleosides were elevated for 36% to 37% of the patients. Urinary spermidine and spermine levels were abnormal for 27% and 24%, respectively, while putrescine levels were elevated for 7% of the patients. When all 14 marker measurements and the 12 ratios of these measurements were considered, the multiple regression equation evaluated the treatment results with a multiple correlation coefficient (R = 0.891; P less than 0.100) about 2.4 times higher than with the most sensitive single marker variable, N2, N2-dimethylguanosine/pseudouridine (R = 0.377; P less than 0.05), alone. Stepwise regression analysis revealed that the minimum number of multiple marker measurements and their ratios required to achieve the maximum value of the multiple correlation coefficient (R = 0.653; p = 0.010) was fifteen. These include the pre and posttreatment measurements of CEA, spermine, N2, N2-dimethylguanosine and 1-methylinosine, as well as two ratios of the polyamines and three ratios of the nucleosides in the post-treatment of the polyamines and three ratios of the nucleosides in the post-treatment measurements. These data suggest that the utilization of regression analysis to evaluate the monitoring utility of multiple marker measurements may be of clinical value.


Extra-corporeal membrane oxygenation-associated infections: implication of extra-intestinal pathogenic Escherichia coli clones.
Extra-corporeal membrane oxygenation (ECMO) is a promising life-saving technique for critically ill patients. Bacterial infection is a frequent complication, and Escherichia coli the predominant causative pathogen, but little is known about the characteristics of E. coli strains in these infections. We therefore conducted a retrospective study of 33 E. coli strains responsible for 33 ECMO-related infections, in 30 subjects. Antimicrobial susceptibility, phylotyping, O-typing, clonal relatedness determination and the screening for four virulence factor genes were conducted. Polymicrobial infections were evidenced in 61.6 % of episodes, irrespective of E. coli characteristics. Extra-intestinal pathogenic strains represented the large majority (69.7 %) of all E. coli isolates. Their advantageous genetic background may explain their predominance in this context. The potential for targeted digestive decontamination should be investigated in these patients for whom infectious complications are a heavy burden.


Cluster of mrdA and mrdB genes responsible for the rod shape and mecillinam sensitivity of Escherichia coli.
Two closely linked genes, mrdA and mrdB, located at ca. 14.2 min on the Escherichia coli chromosomal linkage map, seen to be responsible for the normal rod shape and mecillinam sensitivity of E. coli. The product of mrdA was concluded to be penicillin-binding protein 2, because mrdA mutations caused formation of thermosensitive penicillin-binding protein 2. The product of the mrdB gene is unknown. At 42 degrees, C, mutation in either of these genes caused formation of spherical cells and mecillinam resistance. Both mutations was recessive, and complementation, as detected in +-/-+ meroheterodiploids having the wild-type phenotype, provided strong evidence that the two mutations are in different complementation groups. P1 transduction suggested that the most plausible gene order is leuS-mrdA-mrdB-lip. The rodA mutation reported previously seems to be similar to the mrdB murations, but the identities of the two have not yet been proven.


Influence of neighboring reactive particles on diffusion-limited reactions.
Competition between reactive species is commonplace in typical chemical reactions. Specifically the primary reaction between a substrate and its target enzyme may be altered when interactions with secondary species in the system are substantial. We explore this competition phenomenon for diffusion-limited reactions in the presence of neighboring particles through numerical solution of the diffusion equation. As a general model for globular proteins and small molecules, we consider spherical representations of the reactants and neighboring particles; these neighbors vary in local density, size, distribution, and relative distance from the primary target reaction, as well as their surface reactivity. Modulations of these model variables permit inquiry into the influence of excluded volume and competition on the primary reaction due to the presence of neighboring particles. We find that the surface reactivity effect is long-ranged and a strong determinant of reaction kinetics, whereas the excluded volume effect is relatively short-ranged and less influential in comparison. As a consequence, the effect of the excluded volume is only modestly dependent on the neighbor distribution and is approximately additive; this additivity permits a linear approximation to the many-body effect on the reaction kinetics. In contrast, the surface reactivity effect is non-additive, and thus it may require higher-order approximations to describe the reaction kinetics. Our model study has broad implications in the general understanding of competition and local crowding on diffusion-limited chemical reactions.