Editor's Highlight: Base Excision Repair Variants and Pesticide Exposure Increase Parkinson's Disease Risk.
Exposure to certain pesticides induces oxidative stress and increases Parkinson's disease (PD) risk. Mitochondrial DNA (mtDNA) damage is found in dopaminergic neurons in idiopathic PD and following pesticide exposure in experimental models thereof. Base excision repair (BER) is the major pathway responsible for repairing oxidative DNA damage in cells. Whether single nucleotide polymorphisms (SNPs) in BER genes alone or in combination with pesticide exposure influence PD risk is unknown. We investigated the contributions of functional SNPs in 2 BER genes (APEX1 and OGG1) and mitochondrial dysfunction- or oxidative stress-related pesticide exposure, including paraquat, to PD risk. We also studied the effect of paraquat on levels of mtDNA damage and mitochondrial bioenergetics. 619 PD patients and 854 population-based controls were analyzed for the 2 SNPs, APEX1 rs1130409 and OGG1 rs1052133. Ambient pesticide exposures were assessed with a geographic information system. Individually, or in combination, the BER SNPs did not influence PD risk. Mitochondrial-inhibiting (OR = 1.79, 95% CI [1.32, 2.42]), oxidative stress-inducing pesticides (OR = 1.61, 95% CI [1.22, 2.11]), and paraquat (OR = 1.54, 95% CI [1.23, 1.93]) were associated with PD. Statistical interactions were detected, including for a genetic risk score based on rs1130409 and rs1052133 and oxidative stress inducing pesticides, where highly exposed carriers of both risk genotypes were at the highest risk of PD (OR = 2.21, 95% CI [1.25, 3.86]); similar interactions were estimated for mitochondrial-inhibiting pesticides and paraquat alone. Additionally, paraquat exposure was found to impair mitochondrial respiration and increase mtDNA damage in in vivo and in vitro systems. Our findings provide insight into possible mechanisms involved in increased PD risk due to pesticide exposure in the context of BER genotype variants.


Empagliflozin, an SGLT2 inhibitor for the treatment of type 2 diabetes mellitus: a review of the evidence.
To review available studies of empagliflozin, a sodium glucose co-transporter-2 (SGLT2) inhibitor approved in 2014 by the European Commission and the United States Food and Drug Administration for the treatment of type 2 diabetes mellitus (T2DM). PubMed was searched using the search terms empagliflozin, BI 10773, and BI10773, for entries between January 1, 2000, and December 1, 2014. Reference lists from retrieved articles were searched manually for additional peer-reviewed publications. All publications reporting clinical trials of empagliflozin were eligible for inclusion. Empagliflozin is a new once-daily oral SGLT2 inhibitor with a mechanism of action that is independent of β-cell function and the insulin pathway. Data from a comprehensive phase III clinical trial program have demonstrated its efficacy as monotherapy, as add-on to other glucose-lowering agents, and in different patient populations. In these studies, empagliflozin resulted in improvements in blood glucose levels as well as reductions in body weight and blood pressure. Empagliflozin was well tolerated and was not associated with an increased risk of hypoglycemia versus placebo. The oral antidiabetes agent, empagliflozin, can be used as monotherapy or alongside other glucose-lowering treatments, including insulin, to treat T2DM.


Differential roles for the interferon-inducible IFI16 and AIM2 innate immune sensors for cytosolic DNA in cellular senescence of human fibroblasts.
The IFN-inducible IFI16 and AIM2 proteins act as innate immune sensors for cytosolic double-stranded DNA (dsDNA). On sensing dsDNA, the IFI16 protein induces the expression of IFN-β whereas the AIM2 protein forms an inflammasome, which promotes the secretion of IL-1β. Given that the knockdown of IFI16 expression in human diploid fibroblasts (HDF) delays the onset of cellular senescence, we investigated the potential roles for the IFI16 and AIM2 proteins in cellular senescence. We found that increased IFI16 protein levels in old (vs. young) HDFs were associated with the induction of IFN-β. In contrast, increased levels of the AIM2 protein in the senescent (vs. old) HDFs were associated with increased production of IL-1β. The knockdown of type I IFN-α receptor subunit, which reduced the basal levels of the IFI16 but not of the AIM2, protein delayed the onset of cellular senescence. Accordingly, increased constitutive levels of IFI16 and AIM2 proteins in ataxia telangiectasia mutated (ATM) HDFs were associated with the activation of the IFN signaling and increased levels of IL-1β. The IFN-β treatment of the young HDFs, which induced the expression of IFI16 and AIM2 proteins, activated a DNA damage response and also increased basal levels of IL-1β. Interestingly, the knockdown of AIM2 expression in HDFs increased the basal levels of IFI16 protein and activated the IFN signaling. In contrast, the knockdown of the IFI16 expression in HDFs decreased the basal and dsDNA-induced activation of the IFN signaling. Collectively, our observations show differential roles for the IFI16 and AIM2 proteins in cellular senescence and associated secretory phenotype.


Factors contributing to institutionalization in patients with Huntington's disease.
The objective of this study was to determine which factors are predictive of institutionalization in Huntington's disease. Seven hundred and ninety-nine subjects with 4313 examinations from the Baltimore Huntington's Disease Center were included in the data set; 88 of these patients with an average follow-up time of 9.2 years went from living at home to being institutionalized while being observed in our clinic. We examined demographic, genetic, and clinical variables for a relationship with institutionalization using linear regressions, a Cox proportional hazards model, and χ2 or t tests in certain cases. In our linear models, scores on the Quantified Neurologic Examination (R2=0.203, P<.001), Huntington's disease Activities of Daily Living Scale (R2=0.259, P<.001), and Motor Impairment Score (R2=0.173, P<.001) were found to have the strongest correlation with time until institutionalization. In addition, CAG repeat length (R2=0.248, P<.001) was significantly associated with disease duration at institutionalization, when controlling for age at onset. In the Cox proportional hazards model, scores on the Activities of Daily Living Scale, Mini-Mental State Examination, Quantified Neurologic Examination, and Motor Impairment Score all significantly predicted placement in long-term care. Finally, institutionalized patients were shown to have a higher CAG number and a lower level of educational attainment than patients who avoided institutionalization for at least 15 years after disease onset. Neurologic findings, functional capacity, cognitive impairment, and CAG repeat length are all likely determinants of institutionalization. In contrast with other dementing conditions like Parkinson's and Alzheimer's, psychiatric symptoms were not shown to predict institutionalization in Huntington's disease. This may illustrate the especially debilitating nature of the movement disorder of Huntington's disease in comparison with the other dementias.


Serum depletion induced cancer stem cell-like phenotype due to nitric oxide synthesis in oncogenic HRas transformed cells.
Cancer cells rewire their metabolism and mitochondrial oxidative phosphorylation (OXPHOS) to promote proliferation and maintenance. Cancer cells use multiple adaptive mechanisms in response to a hypo-nutrient environment. However, little is known about how cancer mitochondria are involved in the ability of these cells to adapt to a hypo-nutrient environment. Oncogenic HRas leads to suppression of the mitochondrial oxygen consumption rate (OCR), but oxygen consumption is essential for tumorigenesis. We found that in oncogenic HRas transformed cells, serum depletion reversibly increased the OCR and membrane potential. Serum depletion promoted a cancer stem cell (CSC)-like phenotype, indicated by an increase in CSC markers expression and resistance to anticancer agents. We also found that nitric oxide (NO) synthesis was significantly induced after serum depletion and that NO donors modified the OCR. An NOS inhibitor, SEITU, inhibited the OCR and CSC gene expression. It also reduced anchorage-independent growth by promoting apoptosis. In summary, our data provide new molecular findings that serum depletion induces NO synthesis and promotes mitochondrial OXPHOS, leading to tumor progression and a CSC phenotype. These results suggest that mitochondrial OCR inhibitors can be used as therapy against CSC.


[Uncertainty, relations with agencies and asymmetric information (III)].
In this the third part of the article on uncertainty, agent relationships and asymmetrical information, Doctor Lara passes through Reykiavik and London on her to her final description in Seville. She takes advantage of this analogy to conceptualize the problem of principal and agent, analyzing generic agency relationships pertaining directly with health organizations. When dealing with the interesting question if the problem of principal and agent in health organizations can be lessened, a few equally interesting answers were formulated: allow the professionals to assume the mission and the values of the organization; a well thought out leadership policy; and improvement in information provided; an incentive policy; compatibility of interests between the principal and the agent; ethics and competitive advantage; quality management....


[The crude drugs of special location of Sichuan as viewed from the tribute drugs in the Tang dynasty].
As an important location for drug production in history, the tribute drugs from Sichuan in the Tang dynasty occupied a very large quotas. Hence, the study on Sichuan tribute drugs is conducive to the understanding of the distribution of crude drugs of special location and its rules of evolution in different historical periods, with rather significance for the exploration and utilization of crude drugs. There is a rich resource of crude drugs in Sichuan, exhibiting a conspicuous and zone distribution. According to historical documents of the Tang dynasty, the characteristics of the Sichuan tribute drugs include rich species, extensive distribution, rich special and valuable drugs, and processed drugs. There are over 40 kinds of crude drugs of special location produced in Sichuan. The tribute drugs of the Tang dynasty reveal the evolution of crude drugs of special location. Occupying an important position of crude drugs in the whole country, the crude drugs produced in Sichuan are not necessarily to be all tribute drugs.


A Selected Immunohistochemical Panel Aids in Differential Diagnosis and Prognostic Stratification of Subtypes of High-grade Endometrial Carcinoma: A Clinicopathologic and Immunohistochemical Study at a Single Institution.
This study aimed to investigate whether a selected immunohistochemical panel (estrogen receptor, p53, ARID1A, PPP2R1A, HNF-1β) could contribute to the diagnostic process of high-grade endometrial carcinomas (HG-ECs). We also aimed to analyze the correlation of these immunohistochemical results with several morphologic variables and survival data. After revising the diagnosis of 78 HG-ECs, immunohistochemical analysis was performed for each case. After immunohistochemical analysis, a specific diagnosis of prototypic HG-EC was established in most of the cases that were uncertain due to morphologic ambiguity. In the univariate analysis, older patient age, type II morphology, undifferentiated carcinoma and carcinosarcoma type of histology, altered p53 immunostaining, strong membranous staining of PPP2R1A, presence of lymphovascular invasion in serous carcinoma, and microcystic, elongated, and fragmented-type infiltration pattern in endometrioid carcinoma were significantly related to poor prognosis. In the multivariate analysis, only older patient age and carcinosarcoma or undifferentiated/dedifferentiated carcinoma type histology were found to be significantly poor prognostic factors (P=0.011), whereas advanced FIGO stage and type II histology were found to be correlated with poor prognosis, but did not reach statistical significance. We suggest that immunohistochemistry should be used in the differential diagnosis of HG-ECs, especially those with ambiguous morphology. Markers used in this study made a valuable contribution to the diagnostic process as well as prediction of prognosis.


ZmABA2, an interacting protein of ZmMPK5, is involved in abscisic acid biosynthesis and functions.
In maize (Zea mays), the mitogen-activated protein kinase ZmMPK5 has been shown to be involved in abscisic acid (ABA)-induced antioxidant defence and to enhance the tolerance of plants to drought, salt stress and oxidative stress. However, the underlying molecular mechanisms are poorly understood. Here, using ZmMPK5 as bait in yeast two-hybrid screening, a protein interacting with ZmMPK5 named ZmABA2, which belongs to a member of the short-chain dehydrogenase/reductase family, was identified. Pull-down assay and bimolecular fluorescence complementation analysis and co-immunoprecipitation test confirmed that ZmMPK5 interacts with ZmABA2 in vitro and in vivo. Phosphorylation of Ser173 in ZmABA2 by ZmMPK5 was shown to increase the activity of ZmABA2 and the protein stability. Various abiotic stimuli induced the expression of ZmABA2 in leaves of maize plants. Pharmacological, biochemical and molecular biology and genetic analyses showed that both ZmMPK5 and ZmABA2 coordinately regulate the content of ABA. Overexpression of ZmABA2 in tobacco plants was found to elevate the content of ABA, regulate seed germination and root growth under drought and salt stress and enhance the tolerance of tobacco plants to drought and salt stress. These results suggest that ZmABA2 is a direct target of ZmMPK5 and is involved in ABA biosynthesis and functions.


Creation and Evaluation of a Clinical Informatics Match: Initial Findings.
Clinical Informatics (CI) fellowship programs utilize the Electronic Residency Application Service (ERAS) to gather applications but until recently used an American Medical Informatics Association (AMIA) member-developed, simultaneous offer-acceptance process to match fellowship applicants to programs. In 2021, program directors collaborated with the AMIA to develop a new match to improve the process. Describe the results of the first 2 years of the match and address opportunities for improvement. We obtained applicant data for fellowship applicants in 2021 and 2022 from the ERAS and match data for the same years from the AMIA. We analyzed our data using descriptive statistics. There were 159 unique applicants over the 2-year period. Applicants submitted 2,178 applications with a median of 10 per applicant (interquartile range [IQR] 3-20). One hundred and four applicants (65.4%) participated in the match and ranked a median of seven programs (2-12). Forty-two programs in 2021 and 47 programs in 2022 offered a combined total 153 positions in the match. Participating programs ranked a median of eight applicants per year (IQR 5-11). Of participating applicants, 95 (91.3%) successfully matched and of those 66 (69.5%) received their top choice. Thirty-two programs (76.2%) matched at least one candidate in 2021 and 33 programs (70.2%) matched at least one candidate in 2022. In both years, 24 programs filled all available slots (57.1% in 2021 and 51.1% in 2022). Applicants were extremely successful in the new match, which successfully addressed most of the challenges of the simultaneous offer-acceptance process identified by program directors. However, applicant attrition resulted in a quarter of programs going unmatched. Although many programs still filled slots outside the match, fellowship slots may remain unfilled while the CI practice pathway remains open.


Caffeine junkie: an unprecedented glutathione S-transferase-dependent oxygenase required for caffeine degradation by Pseudomonas putida CBB5.
Caffeine and other N-methylated xanthines are natural products found in many foods, beverages, and pharmaceuticals. Therefore, it is not surprising that bacteria have evolved to live on caffeine as a sole carbon and nitrogen source. The caffeine degradation pathway of Pseudomonas putida CBB5 utilizes an unprecedented glutathione-S-transferase-dependent Rieske oxygenase for demethylation of 7-methylxanthine to xanthine, the final step in caffeine N-demethylation. The gene coding this function is unusual, in that the iron-sulfur and non-heme iron domains that compose the normally functional Rieske oxygenase (RO) are encoded by separate proteins. The non-heme iron domain is located in the monooxygenase, ndmC, while the Rieske [2Fe-2S] domain is fused to the RO reductase gene, ndmD. This fusion, however, does not interfere with the interaction of the reductase with N1- and N3-demethylase RO oxygenases, which are involved in the initial reactions of caffeine degradation. We demonstrate that the N7-demethylation reaction absolutely requires a unique, tightly bound protein complex composed of NdmC, NdmD, and NdmE, a novel glutathione-S-transferase (GST). NdmE is proposed to function as a noncatalytic subunit that serves a structural role in the complexation of the oxygenase (NdmC) and Rieske domains (NdmD). Genome analyses found this gene organization of a split RO and GST gene cluster to occur more broadly, implying a larger function for RO-GST protein partners.


Physician preference motivates the use of anti-tumor necrosis factor therapy independent of clinical disease activity.
Physician preference has previously been shown to be an important determinant of prescription patterns, independent of patient-specific factors. We evaluated whether physician preference was important in the decision to select anti-tumor necrosis factor (anti-TNF) therapy rather than nonbiologic disease-modifying antirheumatic drugs (DMARDs) among rheumatoid arthritis (RA) patients initiating a new RA medication. Using data from the Consortium of Rheumatology Researchers of North America, we identified RA patients who had never taken biologics initiating either anti-TNF therapy or a DMARD in 2001-2008. Physician preference for the use of anti-TNF agents was calculated using data from the preceding calendar year for each physician's other RA patients. Multivariable logistic regression with generalized estimating equations accounted for clustering of patients within the physician practice and evaluated the relationship between physician preference and receipt of anti-TNF therapy, controlling for patient-related factors and disease activity using the Clinical Disease Activity Index. We identified 1,532 RA patients initiating anti-TNF therapy or a DMARD. In models adjusting for tender and swollen joint counts and global disease activity, physician preference for the use of anti-TNF therapy was an independent predictor of receipt of these agents. Patients of physicians in the highest and middle tertiles of physician preference had a 2.50 (95% confidence interval [95% CI] 1.76-3.56) and 1.70 (95% CI 1.22-2.39) greater likelihood of receiving anti-TNF medications, respectively. Physician preference is an important determinant of patients' receipt of anti-TNF therapy and may be useful to examine in future studies of RA treatment patterns, costs, and medication safety.


Comparative analysis of various root active promoters by evaluation of GUS expression in transgenic Arabidopsis.
To prepare various root active promoters for expressing transgenes and prevent gene silencing caused by the repeated use of the same promoter, the expression characteristics of various root active promoters were comparatively evaluated using GUS as a reporter gene. The high-affinity potassium transporter (HKT1;1), the Shaker family potassium ion channel (SKOR), the Shaker family inward rectifying potassium channel (AKT1), the major facilitator superfamily protein (MFS1), and the senescence associated gene 14 (SAG14) promoter from Arabidopsis (Arabidopsis thaliana) were used, and for comparison, four additional constitutive or green tissue specific promoters in the expression vectors were also employed. As the Gateway cloning technology provided by Invitrogen can offer high efficiency and cloning reliability, and easy manipulation of fusion constructs in vitro, our expression vectors are based on binary (destination) vectors compatible with this cloning technique. These destination vectors are also advantageous for stable expression of the transgene, as the heat shock protein terminator is utilized. The AtHKT1;1, SKOR, AKT1, MFS1 and SAG14 promoters were all active in roots but showed slightly different tissue specificities: AtHKT1;1, SKOR, and MFS1 were dominantly active in vascular bundle tissue, while AtHKT1;1 and MFS1- but not SKOR, AKT1, and SAG14-were active in root tips. SKOR showed the strongest root-specificity, and SAG14 showed the highest activity among the five root active promoters. The activity of MFS was developmentally regulated. These destination vectors are now available to express multiple transgenes in transgenic plants, especially in roots.


Recombination of dispersed repeated DNA sequences in yeast.
Yeast transformation can be used to insert new sequence arrangements into a variety of chromosomal locations by homologous recombination. These newly inserted sequences can recombine with similar sequences located on other chromosomes. In these events, information is duplicated without being lost at the site from which it is derived. Similar mechanisms might be utilized by cells to provide new functions during development or differentiation.


Prosodic comprehension and expression in schizophrenia.
Schizophrenics, manics, depressives and normal subjects (15 in each group) were tested for their ability to understand and express the prosodic quality of speech. Sentences in which one word was stressed (stress prosody comprehension) or in which a particular emotion was conveyed (emotional prosody comprehension) were taped and played to subjects to test their comprehension. Subjects were then asked to read out a list of sentences either stressing a nominated word (stress prosody expression) or conveying a nominated emotion (emotional prosody expression), and their efforts were rated by a panel of normal raters. The main results were 1) that schizophrenics were significantly inferior to the normal group, but equivalent to manics and depressives on emotional prosody comprehension; 2) schizophrenics were significantly inferior to all other groups on emotional prosody expression; and 3) all four groups were equivalently proficient on stress prosody comprehension and expression. The results are interpreted as lending support for the idea that there is an underlying right hemisphere dysfunction in schizophrenia.


Chemopreventive and Chemotherapeutic Effects of Fish Oil derived Omega-3 Polyunsaturated Fatty Acids on Colon Carcinogenesis.
Colorectal cancer is the third most common cause of cancer related death in the world. Multiple lines of evidence suggest that there is an association between consumption of dietary fat and colon cancer risk. Not only the amount but also the type and the ratio of fatty acids comprising dietary fats consumed have been implicated in the etiology and pathogenesis of colon cancer. Omega-3 (n-3) polyunsaturated fatty acids (PUFAs), such as docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA), have been known to inhibit development of colon cancer by downregulating the expression of genes involved in colon carcinogenesis and also by altering the membrane lipid composition. Data from laboratory, epidemiological, and clinical studies substantiate the beneficial role of n-3 PUFAs in preventing colitis and subsequent development of colon cancer. In addition, recent studies suggest that some n-3 PUFAs can be effective as an adjuvant with chemotherapeutic agents and other natural anticancer compounds in the management of colon cancer. In this review, we discuss chemopreventive and therapeutic effects of fish oil derived long chain n-3 PUFAs, particularly EPA and DHA, with focus on synergetic effects of which they exert when combined with chemotherapeutic agents and other natural compounds.


Eating behavior in relation to prevalence of overweight among Japanese men.
Obesity is an important risk factor for lifestyle diseases. There has been much interest in the role of eating behavior in the development of obesity, but findings from population-based epidemiologic studies are limited and inconsistent. The objective of our cross-sectional study was to examine the association between eating behavior and prevalence of overweight. Subjects were 290 men of two municipal offices in northeastern Kyusyu, Japan. A positive association was found between overweight and some eating behaviors; multivariate-adjusted odds ratios (95% confidence intervals) for overweight were 4.33 (2.46-7.64), 2.29 (1.22-4.32), and 2.01 (1.06-3.80) for the behaviors of eating quickly, eating until full and eating fatty foods, respectively. Moreover, the prevalence of overweight was further increased when these eating behaviors were combined; multivariate odds ratios (95% confidence intervals) of overweight for men with 1, 2 and ≥3 versus no high-risk eating behaviors were 2.66 (1.27-5.56), 4.32 (1.87-9.97) and 7.32 (3.01-17.84), respectively (pp<0.0001 for trend). Our findings suggest that eating quickly, eating until full and eating fatty foods are related to overweight in Japanese men.


Respiratory insufficiency in a patient with preeclamsia and hypothyroidism developed after general anesthesia for emergency cesarean section: A case report.
Anesthesia and surgery in a patients with undiagnosed or untreated hypothyroidism can carries the risk of potential complications such as prolonged unconsciousness, respiratory insufficiency, hypotension, hyponatremia, congestive heart failure, and even coma. A 33-year-old gravida was admitted at 36 weeks gestation with hypertension, proteinuria, generalized edema, and intrauterine fetal growth retardation. She had thyroidectomy for thyroid mass six years ago. Because of doubt of uterine abruption, emergency cesarean section was performed without result of thyroid function test. Preeclamsia and mild hypothyroidism show similar symptoms and it can make difficult to diagnose hypothyroidism. We experienced delayed recovery and respiratory insufficiency in a patient with preeclamsia and undetected hypothyroidism during emergence from general anesthesia. She was fully recovered after ventilatory care in intensive care unit.


Are cholinesterase inhibitors effective in the management of the behavioral and psychological symptoms of dementia in Alzheimer's disease? A systematic review of randomized, placebo-controlled trials of donepezil, rivastigmine and galantamine.
The study aim was to conduct a systematic review of the evidence from randomized, placebo controlled trials related to the efficacy of donepezil, rivastigmine and galantamine in the treatment of behavioral and psychological symptoms of Alzheimer's disease. Electronic database searches of MEDLINE, EMBASE and the Cochrane Central Register of Controlled Trials were carried out using multiple search terms. Articles included were original publications of randomized, placebo-controlled trials of monotherapy of donepezil, rivastigmine or galantamine that reported a behavioral outcome measure. 14 studies were identified that matched inclusion criteria. Nine were of donepezil, three of galantamine and two of rivastigmine. Median study treatment length was 24 weeks (range 12-170). Most studies used the Neuropsychiatric Inventory as a behavioral outcome measure although three used specific scales for either agitation or apathy. Four studies were specifically designed to assess behavioral outcomes whilst in the majority of studies behavioral outcomes were only secondary measures. Three studies found statistically significant, albeit modest, differences in the change of NPI total score between drug and placebo. The interpretation of the results of many studies is limited by methodological considerations, including generally low NPI scores at baseline and the investigation of behavioral and psychological symptoms of dementia (BPSD) as secondary outcomes. The evidence base regarding the efficacy of cholinesterase inhibitors in BPSD is limited, in part due to methodological considerations. In the absence of alternative safe and effective management options, the use of cholinesterase inhibitors is an appropriate pharmacological strategy for the management of BPSD in Alzheimer's disease.


Human NK cells licensed by killer Ig receptor genes have an altered cytokine program that modifies CD4+ T cell function.
NK cells are innate immune cells known for their cytolytic activities toward tumors and infections. They are capable of expressing diverse killer Ig-like receptors (KIRs), and KIRs are implicated in susceptibility to Crohn's disease (CD), a chronic intestinal inflammatory disease. However, the cellular mechanism of this genetic contribution is unknown. In this study, we show that the "licensing" of NK cells, determined by the presence of KIR2DL3 and homozygous HLA-C1 in host genome, results in their cytokine reprogramming, which permits them to promote CD4(+) T cell activation and Th17 differentiation ex vivo. Microfluidic analysis of thousands of NK single cells and bulk secretions established that licensed NK cells are more polarized to proinflammatory cytokine production than unlicensed NK cells, including production of IFN-γ, TNF-α, CCL-5, and MIP-1β. Cytokines produced by licensed NK augmented CD4(+) T cell proliferation and IL-17A/IL-22 production. Ab blocking indicated a primary role for IFN-γ, TNF-α, and IL-6 in the augmented T cell-proliferative response. In conclusion, NK licensing mediated by KIR2DL2/3 and HLA-C1 elicits a novel NK cytokine program that activates and induces proinflammatory CD4(+) T cells, thereby providing a potential biologic mechanism for KIR-associated susceptibility to CD and other chronic inflammatory diseases.


Empiric treatment in hospitalized community-acquired pneumonia. Impact on mortality, length of stay and re-admission.
To evaluate adherence to guidelines when choosing an empirical treatment and its impact upon the prognosis of community-acquired pneumonia (CAP). A prospective multicentre study was conducted in 425 CAP patients hospitalized on ward. Initial empirical treatment was classified as adhering or not to Spanish guidelines. Adherent treatment was defined as an initial antimicrobial regimen consisting of beta-lactams plus macrolides, beta-lactam monotherapy and quinolones. Non-adherent treatments included macrolide monotherapy and other regimens. Initial severity was graded according to pneumonia severity index (PSI). The end point variables were mortality, length of stay (LOS) and re-admission at 30 days. Overall 30-day mortality was 8.2%, the mean LOS was 8+/-5 days, and the global re-admission rate was 7.6%. Adherence to guidelines was 76.5%, and in most cases the empirical treatment consisted of beta-lactam and macrolide in combination (57.4%). Logistic regression analysis showed that other regimens were associated with higher mortality OR=3 (1.2-7.3), after adjusting for PSI and admitting hospital. Beta-lactam monotherapy was an independent risk factor for re-admission. LOS was independently associated with admitting hospital and not with antibiotics. A high adherence to CAP treatment guidelines was found, though with considerable variability in the empirical antibiotic treatment among hospitals. Non-adherent other regimens were associated with greater mortality. Beta-lactam monotherapy was associated with an increased re-admission rate.


A molecular basis for differential developmental anomalies in Axenfeld-Rieger syndrome.
Pitx2, a bicoid-like homeodomain transcription factor and Dlx2 are two transcriptional markers observed during early tooth development. PITX2 binds to bicoid and bicoid-like elements in the Dlx2 promoter and activates this promoter 30-fold in Chinese hamster ovary cells. Mutations in PITX2 associated with Axenfeld-Rieger syndrome (ARS) provided the first link of this homeodomain transcription factor to tooth development. We are investigating the molecular basis of developmental anomalies associated with human PITX2 mutations. A phenotypically less severe ARS mutant (without tooth anomalies), PITX2 R84W, has a similar DNA binding specificity compared to wild-type PITX2 and transactivates the Dlx2 promoter. This mutation is associated with iris hypoplasia (IH); in contrast a Rieger syndrome mutation, PITX2 T68P, which presents clinically with the full spectrum of developmental anomalies (including tooth anomalies), is unable to transactivate the Dlx2 promoter. Since Dlx2 expression is required for tooth and craniofacial development the lack of tooth anomalies in the patient with IH may be due to the residual activity of this mutant in activating the Dlx2 promoter. We demonstrate that PITX2 phosphorylation increases PITX2 and PITX2 R84W DNA binding. The PITX2 T68P ARS mutation occurs at a protein kinase C phosphorylation site in the homeodomain. Surprisingly, phosphorylation of PITX2 T68P is increased compared to wild-type PITX2 but has little effect on its DNA binding activity. Altogether these data suggest a molecular mechanism for tooth development involving Dlx2 gene expression in ARS patients.


Multiple alignment tensors from a denatured protein.
The structural content of the denatured state has yet to be fully characterized. In recent years, large residual dipolar couplings (RDCs) from denatured proteins have been observed under alignment conditions produced by bicelles and strained polyacrylamide gels. In this report, we describe efforts to extend our picture of the residual structure in denatured nuclease by measuring RDCs with multiple alignment tensors. Backbone amide 15N-1H RDCs were collected from 4 M urea for a total of eight RDC data sets. The RDCs were analyzed by singular value decomposition (SVD) to determine the number of independent alignment tensors present in the data. On the basis of the resultant singular values and propagated error estimates, it is clear that there are at least three independent alignment tensors. These three independent RDC datasets can be reconstituted as orthogonal linear combinations, (OLC)-RDC datasets, of the eight actually recorded. The first, second, and third OLC-RDC datasets are highly robust to the removal of any single experimental RDC dataset, establishing the presence of three independent alignment tensors, sampled well above the level of experimental uncertainty. The observation that the RDC data span three or more dimensions of the five-dimensional parameter space demonstrates that the ensemble average structure of denatured nuclease must be asymmetric with respect to these three orthogonal principal axes, which is not inconsistent with earlier work demonstrating that it has a nativelike topology.


G protein signaling modulator-3: a leukocyte regulator of inflammation in health and disease.
G protein signaling modulator-3 (GPSM3), also known as G18 or AGS4, is a member of a family of proteins containing one or more copies of a small regulatory motif known as the GoLoco (or GPR) motif. GPSM3 interacts directly with Gα and Gβ subunits of heterotrimeric G proteins to regulate downstream intracellular signals initiated by G protein coupled receptors (GPCRs) that are activated via binding to their cognate ligands. GPSM3 has a selective tissue distribution and is highly expressed in immune system cells; genome-wide association studies (GWAS) have recently revealed that single nucleotide polymorphisms (SNPs) in GPSM3 are associated with chronic inflammatory diseases. This review highlights the current knowledge of GPSM3 function in normal and pathologic immune-mediated conditions.


Physiological noise and signal-to-noise ratio in fMRI with multi-channel array coils.
Sensitivity in BOLD fMRI is characterized by the signal to noise ratio (SNR) of the time-series (tSNR), which contains fluctuations from thermal and physiological noise sources. Alteration of an acquisition parameter can affect the tSNR differently depending on the relative magnitude of the physiological and thermal noise, therefore knowledge of this ratio is essential for optimizing fMRI acquisitions. In this study, we compare image and time-series SNR from array coils at 3T with and without parallel imaging (GRAPPA) as a function of image resolution and acceleration. We use the "absolute unit" SNR method of Kellman and McVeigh to calculate the image SNR (SNR(0)) in a way that renders it comparable to tSNR, allowing determination of the thermal to physiological noise ratio, and the pseudo-multiple replica method to quantify the image noise alterations due to the GRAPPA reconstruction. The Kruger and Glover noise model, in which the physiological noise standard deviation is proportional to signal strength, was found to hold for the accelerated and non-accelerated array coil data. Thermal noise dominated the EPI time-series for medium to large voxel sizes for single-channel and 12-channel head coil configurations, but physiological noise dominated the 32-channel array acquisition even at 1 mm × 1mm × 3 mm resolution. At higher acceleration factors, image SNR is reduced and the time-series becomes increasingly thermal noise dominant. However, the tSNR reduction is smaller than the reduction in image SNR due to the presence of physiological noise.


Concentrations of lead in maternal blood, cord blood, and breast milk.
Lead concentrations in maternal blood, umbilical cord blood, and breast milk from 114 women who were not occupationally exposed to lead were determined by graphite atomic absorption spectrophotometry. The mean concentrations of lead in maternal blood, umbilical cord blood and breast milk were 0.7, 0.55, and 0.23 mumol/l, respectively. A significant correlation was observed between maternal and umbilical cord blood (r = 0.63). A lower correlation was noted between maternal blood and breast milk (r = 0.29). These results suggest that lead freely crosses the placental barrier from mother to fetus and the transfer of this heavy metal from maternal tissues to breast milk is possible, but the metabolic mechanisms are more complicated. In addition, a longitudinal study was conducted of concentrations of lead in breast milk in nine lactating women. Results suggested no significant change in the content of lead in breast milk during early lactation.


Evaluation of leukocyte differential counts on the QBC centrifugal hematology analyzer according to NCCLS standard H20-T.
A performance study of the QBC centrifugal hematology system was conducted according to reference and analytical procedures defined in NCCLS Tentative Standard for Leukocyte Differential Counting, H20-T. A complex mathematical transformation and analysis of variance (ANOVA) were applied to the test data. This statistical technique is intended to segregate and emphasize differences in samples and methods. The comparative study of leukocyte differential counts show that QBC is equivalent in performance to the NCCLS reference method when the latter counts are grouped according to the WBC subpopulations reported by QBC. Within these grouped subpopulations, QBC counts were more precise than manual reference counts and otherwise comparable in terms of results. Similarly, QBC compared favorably with the reference method in its clinical sensitivity to abnormals, based on between-method versus within-method studies of hospital specimens.


LiaR-independent pathways to daptomycin resistance in Enterococcus faecalis reveal a multilayer defense against cell envelope antibiotics.
The lipopeptide antibiotic daptomycin (DAP) is a key drug against serious enterococcal infections, but the emergence of resistance in the clinical setting is a major concern. The LiaFSR system plays a prominent role in the development of DAP resistance (DAP-R) in enterococci, and blocking this stress response system has been proposed as a novel therapeutic strategy. In this work, we identify LiaR-independent pathways in Enterococcus faecalis that regulate cell membrane adaptation in response to antibiotics. We adapted E. faecalis OG1RF (a laboratory strain) and S613TM (a clinical strain) lacking liaR to increasing concentrations of DAP, leading to the development of DAP-R and elevated MICs to bacitracin and ceftriaxone. Whole genome sequencing identified changes in the YxdJK two-component regulatory system and a putative fatty acid kinase (dak) in both DAP-R strains. Deletion of the gene encoding the YxdJ response regulator in both the DAP-R mutant and wild-type OG1RF decreased MICs to DAP, even when a functional LiaFSR system was present. Mutations in dak were associated with slower growth, decreased membrane fluidity and alterations of cell morphology. These findings suggest that overlapping stress response pathways can provide protection against antimicrobial peptides in E. faecalis at a significant cost in bacterial fitness.


Estimating exposure to polycyclic aromatic hydrocarbons: a comparison of survey, biological monitoring, and geographic information system-based methods.
Our objective was to compare polycyclic aromatic hydrocarbon (PAH) exposure estimates based on survey, biological monitoring, and geographic information system (GIS) methods. The 304 participants in this study supplied a urine sample and completed questionnaires about exposure to potential PAH sources. We assayed urine samples for 1-hydroxypyrene-O-glucuronide (1-OHPG), the major metabolite of pyrene, a common PAH. We used a GIS to estimate traffic exhaust exposure using vehicle count data at the residence and workplace. The five subjects who reported smoking during the 48-hour period had median 1-OHPG concentrations 10-fold that of nonsmokers (1.6 versus 0.16 pmol/mL; P = 0.01). Among nonsmokers, those who reported eating grilled, roasted, or broiled meat had significantly higher 1-OHPG concentrations than those who did not reported eating meat prepared by these methods (0.25 versus 0.06 pmol/mL; P = 0.02). Nonsmokers who reported traveling on roads for > or =3 hours during the 48-hour period also had significantly higher 1-OHPG levels than those who traveled <3 hours (0.23 versus 0.11 pmol/mL; P = 0.03). 1-OHPG levels were also correlated with hours of secondhand smoke exposure among nonsmokers (P = 0.04). In this study, 1-OHPG urine concentrations were not associated with self-reported exposures to cooking smoke, wood burning, or traffic levels near the home or to traffic density or urban/rural status determined using a GIS. Self-reported indicators of residential proximity to high traffic volume were, however, associated with GIS traffic density measures.


Streptococcus gordonii utilizes several distinct gene functions to recruit Porphyromonas gingivalis into a mixed community.
Dental plaque biofilm formation proceeds through a developmental pathway initiated by the attachment of pioneer organisms, such as Streptococcus gordonii, to tooth surfaces. Through a variety of synergistic interactions, pioneer organisms facilitate the colonization of later arrivals including Porphyromonas gingivalis, a potential periodontal pathogen. We have investigated genes of S. gordonii required to support a heterotypic biofilm community with P. gingivalis. By screening a plasmid integration library of S. gordonii, genes were identified that are crucial for the accumulation of planktonic P. gingivalis cells into a multispecies biofilm. These genes were further investigated by specific mutation and complementation analyses. The biofilm-associated genes can be grouped into broad categories based on putative function as follows: (i) intercellular or intracellular signalling (cbe and spxB), (ii) cell wall integrity and maintenance of adhesive proteins (murE, msrA and atf), (iii) extracellular capsule biosynthesis (pgsA and atf), and (iv) physiology (gdhA, ccmA and ntpB). In addition, a gene for a hypothetical protein was identified. Biofilm visualization and quantification by confocal microscopy confirmed the role of these genes in the maturation of the multispecies community, including biofilm architectural development. The results suggest that S. gordonii governs the development of heterotypic oral biofilms through multiple genetic pathways.


Radiographic evaluation of alveolar ridge preservation using a chitosan/polyvinyl alcohol nanofibrous matrix: A randomized clinical study.
The objective of this randomized clinical trial (RCT) was to assess the effectiveness of electrospun chitosan/polyvinyl alcohol (CS/PVA) nanofibrous scaffolds in preserving the alveolar ridge and enhancing bone remodeling following tooth extraction when compared to a control group. In this split RCT, 24 human alveolar sockets were randomly assigned to two groups, with 12 sockets receiving CS/PVA nanofibrous scaffold grafts (test group) and 12 left to heal by secondary intention as the control group. Cone-beam computed tomography (CBCT) was performed at two different time points: immediately after extraction (T0) and 4 months post-extraction (T4). After 4 months, linear vertical and horizontal radiographic changes and bone density of extraction sockets were assessed in both the test and control groups. The RCT included 12 patients (4 male and 8 female) with a mean age of 24 ± 3.37 years. The test group had a significantly lower mean vertical resorption vs the control group, with a mean difference of 1.1 mm (P < 0.05). Similarly, the control group's mean horizontal bone resorption was -2.01 ± 1.04 mm, while the test group had a significantly lower mean of -0.69 ± 0.41 mm, resulting in a mean difference of 1.35 mm (P < 0.05). Furthermore, the study group exhibited a significant increase in bone density (722.03 ± 131.17 HU) after 4 months compared to the control group (448.73 ± 93.23 HU). In conclusion, we demonstrated within the limitations of this study that CS/PVA nanofibrous scaffold significantly limited alveolar bone resorption horizontally and vertically and enhanced bone density in alveolar sockets after 4 months when compared to results in the control group (TCTR20230526005).


Epstein barr virus encodes miRNAs to assist host immune escape.
Epstein-barr virus (EBV) is a definite tumorigenic virus, which can form life-long latency in the host, which is difficult to be recognized and completely eliminated by the immune system. It is closely related to the occurrence and development of nasopharyngeal cancer, gastric cancer and various types of lymphoma. At present, a total of 44 Epstein-barr virus-encoded microRNAs (EBV miRNAs) have been found. In response to the immune system of the body, EBV miRNAs can inhibit the expression and presentation of viral antigens, inhibit immune activation and immunotoxicity, assisting host cells to escape from immunity, and providing conditions for further immortalized tumorigenesis of the host cells.


Anatomy of stroke injury predicts gains from therapy.
Many therapies are emerging that aim to improve motor function in people with stroke. Identifying key biological substrates needed for treatment gains would help to predict treatment effects and to maximize treatment impact. The current study addressed the hypothesis that behavioral gains from therapy targeting distal upper extremity are predicted by the structural integrity of key motor system white matter tracts. Twenty-three subjects with chronic left-sided stroke underwent robotic therapy targeting the distal right upper extremity. MRI was obtained at baseline and used to outline the infarct. For each subject, the degree to which stroke injured each of 4 descending white matter tracts (from the primary motor cortex, supplementary motor area, dorsal premotor cortex, and ventral premotor cortex, respectively) was determined. Correlations between tract-specific injury and behavioral gains from therapy were then examined. Numerous examples were found whereby tract-specific injury predicted treatment gains. The strongest correlations pertained to stroke injury to tracts descending from the primary motor cortex and dorsal premotor cortex. Infarct volume and baseline behavior were weak predictors of treatment gains. Extent of injury to specific motor tracts predicts behavioral gains from treatment in subjects with chronic stroke. This supports a role for these tracts in mediating treatment effects and reinforces the importance of lesion location in stroke. Tract-specific injury was stronger than infarct volume or baseline clinical status at predicting gains, identifies subjects with sufficient biological substrate to improve from therapy, and so might be useful as an entry criterion in repair-based trials.


Anti-metastatic properties of the leaves of Eriobotrya japonica.
The leaves of Eriobotrya japonica Lindl. have been widely used as a traditional medicine for the treatment of many diseases including gastroenteric disorders, diabetes mellitus, chronic bronchitis and asthma. In the present study, the anti-metastatic action of the EtOAc fraction of the leaves of E. japonica (LEJ) was investigated. LEJ showed potent inhibitory effects on MMP-2 and MMP-9 activities and expressions via down-regulation of NF-κB translocation to the nucleus in B16F10 cells. In addition, the cell migration and invasion were down-regulated by LEJ. LEJ also significantly suppressed lung metastasis in vivo. Moreover, we isolated the compounds ursolic acid and 2α-hydroxyursolic acid from LEJ and both compounds also significantly suppressed MMP-2 and MMP-9 activities, indicating that they are the active components of LEJ. The present results demonstrate that LEJ may be used as valuable antimetastatic agent for the treatment of cancer metastasis.


Drosophila p53 is a structural and functional homolog of the tumor suppressor p53.
The importance of p53 in carcinogenesis stems from its central role in inducing cell cycle arrest or apoptosis in response to cellular stresses. We have identified a Drosophila homolog of p53 ("Dmp53"). Like mammalian p53, Dmp53 binds specifically to human p53 binding sites, and overexpression of Dmp53 induces apoptosis. Importantly, inhibition of Dmp53 function renders cells resistant to X ray-induced apoptosis, suggesting that Dmp53 is required for the apoptotic response to DNA damage. Unlike mammalian p53, Dmp53 appears unable to induce a G1 cell cycle block when overexpressed, and inhibition of Dmp53 activity does not affect X ray-induced cell cycle arrest. These data reveal an ancestral proapoptotic function for p53 and identify Drosophila as an ideal model system for elucidating the p53 apoptotic pathway(s) induced by DNA damage.


[Changes in blood pressure in the pulmonary artery in myocardial infarct].
A polygraphic study involved 122 patients with acute myocardial infarction mostly of transmural nature. In 42 cases using parallel catheterization, barograms of the right portion of the heart and pulmonary artery were recorded. The above non-invasive techniques were tested in relation to their suitability for the indirect determination of the pulmonary circulation pressure. Mathematically substantiated formula are presented designed for the indirect measurement of right atrium and pulmonary pressure. The localization of the primary infarction in the left ventricle anterior wall, recurrent necrosis of the cardiac muscle, attendant essential hypertension and diabetes mellitus are shown to be associated with a more pronounced pulmonary hypertension. An attempt is made to elicit the mechanisms of pulmonary hypertension by correlating the phasic structures of the systole of the right and left ventricles.


Improvements in absorbed dose measurements for external radiation therapy using ferrous dosimeter gel and MR imaging (FeMRI).
A ferrous gel, based on ferrous (Fe) sulphate and agarose, was used with a clinical magnetic resonance imaging (MRI) scanner to obtain relative dose distribution data from therapeutic photon and electron beams. The FeMRI gel was scanned using a new MRI acquisition protocol optimized for T1 measurements. Thorough comparisons with silicon semiconductor detector and ionization chamber measurements, as well as with Monte Carlo calculations, were performed in order to quantify the improvements obtained using FeMRI for dose estimations. Most of the relative doses measured with FeMRI were within 2% of the doses measured with other methods. The larger discrepancies (2-4%) found at shallow depths are discussed. The uncertainty in relative dose measurements using FeMRI was significantly improved compared with previously reported results (5-10%, one standard deviation, 1 SD), and is today between 1.6% and 3.3% (depending on dose level, 2 SD). This corresponds to an improvement in the minimum detectable dose (3 SD above background) from approximately 2 Gy to better than 0.6 Gy. The results obtained in this study emphasize the importance of obtaining basic FeMRI dose data before the method is extended to complicated treatment regimes.


Quantification, identification and comparison of oligopeptides on five tea categories with different fermentation degree by Kjeldahl method and ultra-high performance liquid chromatography coupled with quadrupole-orbitrap ultra-high resolution mass spectrometry.
Peptides with different lengths or amino sequences could have specific tastes or bio-activities. So far, either the quantity or pattern differences of peptide among various of teas were unknown. Here, firstly, we developed a method for tea oligopeptide quantification and made comparison of their contents. Secondly, we applied ultra-high performance liquid chromatography coupled with quadrupole-orbitrap ultra-high resolution mass spectrometry (UHPLC-Quadrupole-Orbitrap-UHRMS) to sequence oligopeptides. As a result, the total amount of oligopeptides in white tea and dark tea were higher, followed by black tea and green tea, finally with oolong tea. It suggested that withering which undergoes with endogenous protease and post-fermented that undergoes with a participation of exotic micro-organisms were key in oligopeptide enrichment. Thirdly, a total of 902 abundant identified peptides, most of which were tripeptide, tetrapeptide, pentapeptide, and hexapeptide were screened against several existing peptide databases. There were a series of taste peptides and bio-active peptides existing.


Glucometabolic characteristics and higher vascular complication risk in Korean patients with type 2 diabetes with non-albumin proteinuria.
We investigated the clinical relevance of non-albumin proteinuria (NAP) in Korean patients with type 2 diabetes (T2D). We enrolled 883 T2D patients who had both their urinary albumin-to-creatinine ratio (uACR) and protein-to-creatinine ratio (uPCR) measured. We classified the patients into non-proteinuria (NP; uPCR <150 mg/g and uACR <30 mg/g), isolated NAP (iNAP; uPCR ≥150 mg/g and uACR <30 mg/g), and albuminuria (uACR ≥30 mg/g) groups. The associations between uPCR, uACR, and several indices of glucose metabolism were investigated. The glucometabolic pathophysiology of iNAP (96 [10.9%]) group was more associated with a decrease in homeostatic model assessment (HOMA)-beta value (aOR 1.89 [95% CI, 1.21-2,96]) than with an increase in HOMA-insulin resistance (aOR 1.29 [95% CI, 0.83-2.01]). uPCR ≥150 mg/g was also found to have more consistent and stronger association with vascular complications than uACR ≥30 mg/g (aOR 1.44 [95% CI, 1.03-2.02] vs. 1.26 [95% CI, 0.89-1.79]). The nephropathy of iNAP may be mainly attributed to decreased beta cell function. Furthermore, uPCR might be a more sensitive urinary biomarker than uACR for the detection of vascular complications in T2D patients.


Accessing restoration margins--a multidisciplinary approach.
Dentists frequently balance periodontal, restorative, and esthetic concerns that will provide adequate tooth length, ensure periodontal health, and provide biological predictability. This report discusses the current and classical literature and provides a multidisciplinary approach of predictably "accessing" the restoration margins to facilitate long-term gingival health and tooth survival.


Radar-echo tracking by use of invariant moments.
A number of techniques to track rainfall patterns by use of radar observations have been developed over the years. We present a method for radar-echo tracking based on Hu invariant moments. The method has been tried on several sequences of test images, and the derived displacement fields were in good agreement with the real motions of the tested objects. For the real data obtained from the conventional meteorological radar in Legionowo the method occasionally failed when changes in the radar echo between observations were too large.


Puerarin prevents isoprenaline-induced myocardial fibrosis in mice by reduction of myocardial TGF-β1 expression.
It has been reported that soy isoflavones could significantly increase peroxisome proliferator-activated receptor α/γ gene expressions, while the activation of peroxisome proliferator-activated receptor α/γ may attenuate myocardial fibrosis. Puerarin is the main isoflavone isolated from the root of the wild leguminous creeper Pueraria lobata (Willd) Ohwi, so we thought that puerarin could inhibit myocardial fibrotic formation. A mouse myocardial fibrotic model was induced by hypodermic injection of isoprenaline when these mice were simultaneously treated with puerarin 600 and 1200 mg/kg by gavage for 40 days, respectively. The results showed that puerarin could significantly improve myocardial fibrosis and decrease the collagen accumulation, collagen volume fraction, hydroxyproline content in myocardial tissue and cardiac weight index. The results from reverse transcription polymerase chain reaction indicated that the messenger RNA (mRNA) expression of transforming growth factor-β1 in myocardial tissue was decreased, while the mRNA expressions of peroxisome proliferator-activated receptor α/γ were increased, in the puerarin groups as compared with the model group. Importantly, puerarin could significantly decrease the protein expressions of transforming growth factor-β1 and nuclear factor-κB in myocardial tissue. These results suggested that puerarin could prevent isoprenaline-induced myocardial fibrosis in mice, and its mechanisms might be related to reduction of transforming growth factor-β1 expression via activation of peroxisome proliferator-activated receptor α/γ and subsequent inhibition of nuclear factor-κB in myocardial tissue.


Development of the retinofugal pathway in birds and mammals: evidence for a common 'timetable'.
We have compared changes in axon numbers in the developing optic nerves of eight homeotherms (seven mammals and one bird) using data from the available literature and our own material. The proportion of axons lost during development is smaller in the chick (35%) than in mammals (54-74%). The relative magnitude of this loss does not correlate with the extent of the binocular visual field or the size of the retinofugal ipsilateral projection. The timing of developmental events in the retinofugal pathway was compared as a proportion of the period between conception and eye opening (percentage of the 'caecal period', CP). In eutherian mammals, retinal ganglion cells and their target neurons are generated between 30 and 49% of CP (a duration of 19% of CP). In homeotherms, a phase of rapid axon generation begins around 38% of CP and the peak number of axons is reached at about 56% of CP (a duration of 18% of CP). A phase of rapid axon loss begins thereafter, and in most species it ends at about 74% of CP (a duration of 18% of CP), the rate of rapid axons loss being about half the rate of rapid axon generation. The similarity in relative timing (homochrony), like the similarity in the relative magnitude of the axon loss, suggests that cell generation and loss in the retinofugal pathway are influenced by a mechanism common to all homeotherms. We propose that in homeotherms each cohort of retinal ganglion cells is numerically matched with a group of target cells that is at an appropriate stage of maturation ('temporal matching' hypothesis). About twice as many ganglion cells are produced in each cohort than are needed, and their survival is determined by natural selection.


KDM6B Counteracts EZH2-Mediated Suppression of IGFBP5 to Confer Resistance to PI3K/AKT Inhibitor Treatment in Breast Cancer.
Despite showing promise against PIK3CA-mutant breast cancers in preclinical studies, PI3K/AKT pathway inhibitors demonstrate limited clinical efficacy as monotherapy. Here, we found that histone H3K27me3 demethylase KDM6B-targeted IGFBP5 expression provides a protective mechanism for PI3K/AKT inhibitor-induced apoptosis in breast cancer cells. We found that overexpression of KDM6B and IGFBP5 in luminal breast cancer are positively associated with poorer disease outcomes. Mechanistically, KDM6B promotes IGFBP5 expression by antagonizing EZH2-mediated repression, and pharmacologic inhibition of KDM6B augments apoptotic response to PI3K/AKT inhibitor treatment. Moreover, the IGFBP5 expression is upregulated upon acquired resistance to the PI3K inhibitor GDC-0941, which is associated with an epigenetic switch from H3K27me3 to H3K27Ac at the IGFBP5 gene promoter. Intriguingly, GDC-0941-resistant breast cancer cells remained sensitive to KDM6B or IGFBP5 inhibition, indicating the dependency on the KDM6B-IGFBP5 axis to confer the survival advantage in GDC-0941-resistant cells. Our study reveals an epigenetic mechanism associated with resistance to targeted therapy and demonstrates that therapeutic targeting of KDM6B-mediated IGFBP5 expression may provide a useful approach to mitigate both intrinsic and acquired resistance to the PI3K inhibitor in breast cancer. Mol Cancer Ther; 17(9); 1973-83. ©2018 AACR.


Safety and immunogenicity of a mRNA rabies vaccine in healthy adults: an open-label, non-randomised, prospective, first-in-human phase 1 clinical trial.
Vaccines based on mRNA coding for antigens have been shown to be safe and immunogenic in preclinical models. We aimed to report results of the first-in-human proof-of-concept clinical trial in healthy adults of a prophylactic mRNA-based vaccine encoding rabies virus glycoprotein (CV7201). We did an open-label, uncontrolled, prospective, phase 1 clinical trial at one centre in Munich, Germany. Healthy male and female volunteers (aged 18-40 years) with no history of rabies vaccination were sequentially enrolled. They received three doses of CV7201 intradermally or intramuscularly by needle-syringe or one of three needle-free devices. Escalating doses were given to subsequent cohorts, and one cohort received a booster dose after 1 year. The primary endpoint was safety and tolerability. The secondary endpoint was to determine the lowest dose of CV7201 to elicit rabies virus neutralising titres equal to or greater than the WHO-specified protective antibody titre of 0·5 IU/mL. The study is continuing for long-term safety and immunogenicity follow-up. This trial is registered with ClinicalTrials.gov, number NCT02241135. Between Oct 21, 2013, and Jan 11, 2016, we enrolled and vaccinated 101 participants with 306 doses of mRNA (80-640 μg) by needle-syringe (18 intradermally and 24 intramuscularly) or needle-free devices (46 intradermally and 13 intramuscularly). In the 7 days post vaccination, 60 (94%) of 64 intradermally vaccinated participants and 36 (97%) of 37 intramuscularly vaccinated participants reported solicited injection site reactions, and 50 (78%) of 64 intradermally vaccinated participants and 29 (78%) of 37 intramuscularly vaccinated participants reported solicited systemic adverse events, including ten grade 3 events. One unexpected, possibly related, serious adverse reaction that occurred 7 days after a 640 μg intramuscular dose resolved without sequelae. mRNA vaccination by needle-free intradermal or intramuscular device injection induced virus neutralising antibody titres of 0·5 IU/mL or more across dose levels and schedules in 32 (71%) of 45 participants given 80 μg or 160 μg CV7201 doses intradermally and six (46%) of 13 participants given 200 μg or 400 μg CV7201 doses intramuscularly. 1 year later, eight (57%) of 14 participants boosted with an 80 μg needle-free intradermal dose of CV7201 achieved titres of 0·5 IU/mL or more. Conversely, intradermal or intramuscular needle-syringe injection was ineffective, with only one participant (who received 320 μg intradermally) showing a detectable immune response. This first-ever demonstration in human beings shows that a prophylactic mRNA-based candidate vaccine can induce boostable functional antibodies against a viral antigen when administered with a needle-free device, although not when injected by a needle-syringe. The vaccine was generally safe with a reasonable tolerability profile. CureVac AG.


[The influence of environmental factors on the incidence and prevalence of multiple sclerosis].
257 patients with multiple sclerosis (MS) participated in the study in Yaroslavl region. The main epidemiological indices (incidence and prevalence of MS) for the last 25 years were determined. The evaluation of environment pollution has been realized in different districts of the city according to the level of snow covering pollution. Farther in snow-test contents of 38 chemical elements found in industrial waste were determined and summary index of pollution of environment were calculated. Significant direct correlation is revealed between level of environment pollution and increasing of epidemiological characteristics of MS.


Monitoring of the Palladium Concentration in River Water and Sediment at an Acidic Hot Spring Spa Area in Gunma Prefecture.
In an effort to elucidate the deposition pathway of Pd in river sediments, we analyzed the amount of Pd in the river water and sediments of the Yukawa and Yazawa rivers, as well as in the sediments of the Shinaki dam-lake of the Kusatsu hot-spring area, which is located northwest in Gunma Prefecture of Japan. The crystal structures and elemental compositions of the river sediment samples differed significantly before and after neutralization. This was attributed to the lime input, which also affected the Pd abundance ratio obtained by the sequential extraction procedure. Additionally, the low leachability of Pd in the sediment suggested possible difficulties in its recovery. Considering the analysis of the Pd variation in the environments, it was concluded that the Pd content in the sediment of the Shinaki dam-lake was mainly supplied by the Yukawa river water inflowing the surrounding tributary rivers and hot-spring waters before neutralization, rather than the lime input.


Membrane specializations in mammalian lens fiber cells: distribution of square arrays.
Fragments of intact rat, pig and bovine lenses and isolated membranes from bovine lenses were examined by freeze-fracture-etch electron microscopy employing samples ultra-rapidly frozen in the absence of fixatives or cryoprotectants. Complementary replicas of the bovine outer cortex clearly display small patches of square arrays (6.6 nm repeat) as structures distinct from gap junctions in the same membranes. In the outer cortex square arrays appear in single membranes where the extracellular space is not thinned, whereas gap junctions, as in other tissues, only occur where the extracellular space is greatly attenuated. Square arrays in the middle cortex appear as patches of varying size surrounded by smooth membrane fracture faces. Within the undulating membranes of tongue-and-groove interdigitations in the inner cortex and nucleus, the square arrays are extensive and are located specifically on the regions which have convex curvature toward the cytoplasm. Square crystalline regions alternate with non-crystalline regions within each membrane of an undulating pair. Across the extracellular space, crystalline regions are matched with non-crystalline regions, thus making it unlikely that the square array participates in intercellular communication. Because the undulations occur in isolated membranes following urea washing to remove cytoplasmic and extrinsic membrane proteins, the square arrays probably play a crucial role in the formation of the undulations and in the maintenance of membrane curvature. X-ray diffraction experiments show reflections from the crystalline square array for all twenty of our preparations of isolated membranes. These x-ray experiments indicate that the square arrays observed by freeze-fracture are abundant in lens fiber cell membranes.


Poor outcome associated with symptomatic deterioration among help-seeking individuals at risk for psychosis: a naturalistic follow-up study.
It remains debatable whether early intervention for psychosis is capable of meeting the needs of at-risk subjects. The aims of this study were to describe the actual impact of interventions on subjective difficulties and to explore the factors that may be associated with a poor outcome. Participants were help-seeking outpatients at a university hospital who met the Criteria of Prodromal Syndromes. Changes in the symptoms, subjective experience and current insight were assessed using the Scales of Prodromal Symptoms, the Subjective Well-being under Neuroleptics, and the Scale to Assess Unawareness of Mental Disorder, respectively. Global functioning, social functioning and subjective quality of life were evaluated using the Global Assessment of Functioning Scale, the Social Functioning Scale, and the WHO-Quality of Life 26, respectively. These measures were assessed both at baseline and after 1 year. Forty-six patients agreed to participate. Of the 27 patients who completed the reassessment at the follow-up point, 13 patients (48%) showed little improvement in their positive/negative symptoms, subjective well-being or awareness of their symptoms. Additionally, less severe negative symptoms, more severe general symptoms and lower subjective well-being at baseline significantly predicted a deterioration of positive/negative symptoms after 1 year. Our findings suggest that the current strategy for reducing psychosis risk based on positive symptoms should be reappraised.


Lung vascular permeability after reversal of fibronectin deficiency in septic sheep. Correlation with patient studies.
Plasma fibronectin deficiency and opsonic dysfunction exist in critically ill septic surgical, trauma, and burn patients with multiple organ failure. Fibronectin deficiency can be reversed by infusion of fresh plasma cryoprecipitate. The influence of therapy with human cryoprecipitate on lung vascular permeability in septic sheep with plasma fibronectin deficiency following surgery was evaluated. Additionally, selected studies on pulmonary function in septic surgical and trauma patients after infusion of plasma cryoprecipitate were completed. In patients, ventilation-perfusion balance appeared to improve as measured by the multiple inert gas elimination technique. With the lung lymph fistula preparation in fibronectin deficient sheep, infusion of human plasma cryoprecipitate (10 units; 250 ml) delayed the onset and minimized the increase in lung vascular permeability during postoperative Pseudomonas sepsis (5 X 10(9) bacteria, I.V.; 5 X 10(10) bacteria, I.P.). For example, in a first group of sheep, the transvascular protein clearance (TPC) at 2 hrs in septic sheep (n = 4) treated with only saline (volume control) was 20.1 +/- 3.1 ml/hr, compared to 11.23 +/- 0.83 ml/hr in the sheep (n =a 4) treated with fibronectin-rich cryoprecipitate (p less than 0.05). In a second group of sheep, cryoprecipitate depleted of fibronectin by affinity chromatography was used as the control solution. It also did not manifest this protective effect with respect to lung vascular permeability. Thus, at 2 hrs the lymph flow (Qlym) was 30.2 ml/hr and the transvascular protein clearance (TPC) was 18.0 ml/hr in septic sheep given fibronectin-deficient cryoprecipitate. In contrast, in the fibronectin-rich cryoprecipitate treated sheep, the Qlym was 14.8 ml/hr and the TPC was 8.12 ml/hr. It is suggested that fibronectin may influence lung vascular integrity during sepsis following surgery and trauma.


Postoperative Limb-Offset Discrepancy Notably Affects Soft-Tissue Tension in Total Hip Arthroplasty.
Inadequate soft-tissue tension in total hip arthroplasty is regarded as one cause of dislocation or abductor muscle weakness. The purpose of the present study was to assess how the postoperative discrepancy in limb offset (consisting of both femoral offset and acetabular offset) affects soft-tissue tension compared with other factors among patients with unilateral hip disease undergoing total hip arthroplasty. A total of 89 consecutive patients underwent mini-incision total hip arthroplasty involving an anterolateral or posterior approach and with use of a computer navigation system. Soft-tissue tension was measured by applying traction amounting to 40% of body weight with the joint positioned at 0°, 15°, 30°, and 45° of flexion. The separation between the cup and the prosthetic head was measured using the navigation system. The cup-head separation differed significantly for varying angles of flexion (p < 0.001), with the greatest distance noted at 15° of flexion (mean and standard deviation, 11 ± 5 mm). Stepwise multiple regression analysis showed that postoperative limb-offset discrepancy, an anterolateral approach, and preoperative abduction range of motion were correlated with the cup-head separation at 15° of flexion. Postoperative limb-offset discrepancy was negatively correlated with the cup-head separation at 0°, 15°, and 30° of flexion. Postoperative limb-offset discrepancy significantly affected the soft-tissue tension in total hip arthroplasty at varying degrees of flexion. This indicated that it is important to restore normal limb offset without overlengthening to obtain adequate soft-tissue tension in total hip arthroplasty. Therapeutic Level IV. See Instructions for Authors for a complete description of levels of evidence.


Evaluation of dextromethorphan and carbetapentane as anticonvulsants and N-methyl-D-aspartic acid antagonists in mice.
Two antitussives, dextromethorphan and carbetapentane, which have been reported to bind to a common binding site in brain tissue and produce anticonvulsant effects in rats, were evaluated for their anticonvulsant effects against maximal electroshock-induced seizures, for their neurological impairing effects on the horizontal screen test, and their protective effects against N-methyl-D-aspartic acid (NMDA)-induced lethality in mice. Both compounds protected animals against maximal electroshock-induced seizures in a dose-related fashion after either intraperitoneal or oral administration. The neurologically impairing doses were approximately 1.5 times the anticonvulsant doses. As a function of dose, dextromethorphan, but not carbetapentane, protected mice from NMDA-induced lethality. Since carbetapentane had an anticonvulsant action without protecting against NMDA-induced lethality, these data support the hypothesis that dextromethorphan and carbetapentane may have a common anticonvulsant action separate from the phencyclidine-like, NMDA-antagonist action which only dextromethorphan exhibits.


Substrate replenishment and byproduct removal improve yeast cell-free protein synthesis.
Cell-free protein synthesis (CFPS) platforms are now considered a powerful tool for synthesizing a variety of proteins at scales from pL to 100 L with accelerated process development pipelines. We previously reported the advancement of a novel yeast-based CFPS platform. Here, we studied factors that cause termination of yeast CFPS batch reactions. Specifically, we characterized the substrate and byproduct concentrations in batch, fed-batch, and semi-continuous reaction formats through high-performance liquid chromatography (HPLC) and chemical assays. We discovered that creatine phosphate, the secondary energy substrate, and nucleoside triphosphates were rapidly degraded during batch CFPS, causing a significant drop in the reaction's energy charge (E.C.) and eventual termination of protein synthesis. As a consequence of consuming creatine phosphate, inorganic phosphate accumulated as a toxic byproduct. Additionally, we measured amino acid concentrations and found that aspartic acid was rapidly consumed. By adopting a semi-continuous reaction format, where passive diffusion enables substrate replenishment and byproduct removal, we achieved over a 70% increase in active superfolder green fluorescent protein (sfGFP) as compared with the batch system. This study identifies targets for the future improvement of the batch yeast CFPS reaction. Moreover, it outlines a detailed, generalized method to characterize and improve other CFPS platforms.


Toward a liberal education in medicine.
The central thesis of Abraham Flexner's analyses of North American and European medical education was that the university is essential to the provision of a medical education. The authors invoke the spirit of Flexner to envision further contributions of the university at large to undergraduate medical education. Medical curricula now include elements of a variety of other disciplines that are better represented in other parts of the university. Most schools, however, even those closely affiliated with a comprehensive university, do not take full advantage of these resources, nor do they offer sufficient opportunities for students to pursue individualized interests and learning goals. Medical school now plays a different role in the education of physicians than it did a century ago-it remains the definitive, but is no longer the ultimate, stage in a continuum involving college, professional, postgraduate, and continuing education. The authors explore the medical school years as an opportunity for a liberal education in medicine. Beyond the assurance of competence in core knowledge, skills, and perspectives, this model places more emphasis on nurturing students' intellectual curiosity about phenomena of illness and disease, their understanding of the human condition, and their exploration of the many other disciplines related to medicine and the life sciences. A richer, broader education can be achieved through more flexible and individualized paths to the MD and facilitated by realizing medical schools' full academic citizenship in the university.


Occurrence of a furin-like prohormone processing enzyme in Aplysia neuroendocrine bag cells.
1. Strong evidence is accumulating that the endoproteases which process prohormones at dibasic residue cleavage sites are members of a subtilisin-related class of proteases. 2. Using the polymerase chain reaction (PCR), we have isolated and characterized an Aplysia californica neuroendocrine bag cell cDNA product (270 base pairs) that encodes a sequence which is highly homologous to the subtilisin-related class of processing proteases that includes yeast Kex2, human/mouse/Drosophila furin, human PC2, and mouse PC1/PC3 and PC2. 3. The characterized cDNA PCR product showed the highest degree of residue identity with the furin-related group of proteins (human/mouse furin 71%; Drosophila furin 63%). 4. These results establish that Aplysia contain a subtilisin-like gene and suggest that the expression of this gene may play a role in processing Aplysia precursor proteins in the bag cells and likely also in the exocrine atrial gland. 5. Furthermore, the Aplysia nucleotide sequence results, together with available sequence information from human, mouse, and Drosophila furins, provide reasonable evidence that the furin-like enzymes may represent a separate subclass of the subtilisin-like processing enzymes.


Computed analysis of three-dimensional cone-beam computed tomography angiography for determination of tumor-feeding vessels during chemoembolization of liver tumor: a pilot study.
The purpose of this study was to evaluate computed analysis of three-dimensional (3D) cone-beam computed tomography angiography (CTA) of the liver for determination of subsegmental tumor-feeding vessels (FVs). Eighteen consecutive patients underwent transarterial chemoembolization (TACE) from January to October 2008 for 25 liver tumors (15 hepatocellular carcinomas [HCCs] and 10 neuroendocrine metastases). Anteroposterior projection angiogram (two-dimensional [2D]) and 3D cone-beam CTA images were acquired by injection of the common hepatic artery. Retrospectively, FVs were independently identified by three radiology technologists using a software package (S) that automatically determines FVs by analysis of 3D images. Subsequently, three interventional radiologists (IRs) independently identified FVs by reviewing the 2D images followed by examination of the 3D images. Finally, the "ground truth" for the number and location of FVs was obtained by consensus among the IRs, who were allowed to use any imaging-including 2D, 3D, and all oblique or selective angiograms-for such determination. Sensitivities, durations, and degrees of agreement for review of 2D, 3D, and S results were evaluated. Sensitivity of 3D (73%) was higher than 2D (64%) images for identification of FVs (P = 0.036). The sensitivity of S (93%) was higher than 2D (P = 0.02) and 3D (P = 0.005) imaging. The duration for review of 3D imaging was longer than that for 2D imaging (187 vs. 94 s, P = 0.0001) or for S (135 s, P = 0.0001). The degree of agreement between the IRs using 2D and 3D imaging were 54% and 62%, respectively, whereas it was 82% between the three radiology technologists using S. These preliminary data show that computed determination of FVs is both accurate and sensitive.


Generation of two isogenic iPS cell lines (IRFMNi002-A and IRFMNi002-B) from a patient affected by Focal Segmental Glomerulosclerosis carrying a heterozygous c.565G>A mutation in PAX2 gene.
Focal Segmental Glomerulosclerosis (FSGS) is the typical renal histologic lesion in familial steroid-resistant nephrotic syndrome, for which there is currently no treatment. Dysfunction of the glomerular podocyte, a specialized cell that forms the glomerular filtration barrier, is central in the pathogenesis of FSGS. Here, we reported the generation of two isogenic iPS cell lines from a patient affected by FSGS, carrying the c.565G > A mutation in the PAX2 gene. The iPS cell lines we generated expressed pluripotency markers at the mRNA and protein levels and differentiated into all three germ layers. These iPSCs will be instrumental in understanding FSGS pathogenesis.


Comparative rates of desensitization of beta-adrenergic receptors by the beta-adrenergic receptor kinase and the cyclic AMP-dependent protein kinase.
Three separate processes may contribute to rapid beta-adrenergic receptor desensitization: functional uncoupling from the stimulatory guanine nucleotide-binding protein Gs, mediated by phosphorylation of the receptors by two distinct kinases, the specific beta-adrenergic receptor kinase (beta ARK) and the cyclic AMP-dependent protein kinase A (PKA), as well as a spatial uncoupling via sequestration of the receptors away from the cell surface. To evaluate the relative importance and potential role of the various processes in different physiological situations, a kinetic analysis of these three mechanisms was performed in permeabilized A431 epidermoid carcinoma cells. To allow a separate analysis of each mechanism, inhibitors of the various desensitization mechanisms were used: heparin to inhibit beta ARK, the PKA inhibitor peptide PKI to inhibit PKA, and concanavalin A treatment to prevent sequestration. Isoproterenol-induced phosphorylation of beta 2 receptors in these cells by beta ARK occurred with a t1/2 of less than 20 sec, whereas phosphorylation by PKA had a t1/2 of about 2 min. Similarly, beta ARK-mediated desensitization of the receptors proceeded with a t1/2 of less than 15 sec, and PKA-mediated desensitization with a t1/2 of about 3.5 min. Maximal desensitization mediated by the two kinases corresponded to a reduction of the signal-transduction capacity of the receptor/adenylyl cyclase system by about 60% in the case of beta ARK and by about 40% in the case of PKA. Receptor sequestration was much slower (t1/2 of about 10 min) and involved no more than 30% of the cell surface receptors. It is concluded that beta ARK-mediated phosphorylation is the most rapid and quantitatively most important factor contributing to the rapid desensitization. This rapidity of the beta ARK-mediated mechanism makes it particularly well suited to regulate beta-adrenergic receptor function in rapidly changing environments such as the synaptic cleft.


Activity-based assay for human mono-ADP-ribosyltransferases ARTD7/PARP15 and ARTD10/PARP10 aimed at screening and profiling inhibitors.
Poly(ADP-ribose) polymerases (PARPs) or diphtheria toxin like ADP-ribosyl transferases (ARTDs) are enzymes that catalyze the covalent modification of proteins by attachment of ADP-ribose units to the target amino acid residues or to the growing chain of ADP-ribose. A subclass of the ARTD superfamily consists of mono-ADP-ribosyl transferases that are thought to modify themselves and other substrate proteins by covalently adding only a single ADP-ribose moiety to the target. Many of the ARTD enzymes are either established or potential drug targets and a functional activity assay for them will be a valuable tool to identify selective inhibitors for each enzyme. Existing assays are not directly applicable for screening of inhibitors due to the different nature of the reaction and different target molecules. We modified and applied a fluorescence-based assay previously described for PARP1/ARTD1 and tankyrase/ARTD5 for screening of PARP10/ARTD10 and PARP15/ARTD7 inhibitors. The assay measures the amount of NAD(+) present after chemically converting it to a fluorescent analog. We demonstrate that by using an excess of a recombinant acceptor protein the performance of the activity-based assay is excellent for screening of compound libraries. The assay is homogenous and cost effective, making it possible to test relatively large compound libraries. This method can be used to screen inhibitors of mono-ARTDs and profile inhibitors of the enzyme class. The assay was optimized for ARTD10 and ARTD7, but it can be directly applied to other mono-ARTDs of the ARTD superfamily. Profiling of known ARTD inhibitors against ARTD10 and ARTD7 in a validatory screening identified the best inhibitors with submicromolar potencies. Only few of the tested ARTD inhibitors were potent, implicating that there is a need to screen new compound scaffolds. This is needed to create small molecules that could serve as biological probes and potential starting points for drug discovery projects against mono-ARTDs.


From simple receptors to complex multimodal percepts: a first global picture on the mechanisms involved in perceptual binding.
The binding problem in perception is concerned with answering the question how information from millions of sensory receptors, processed by millions of neurons working in parallel, can be merged into a unified percept. Binding in perception reaches from the lowest levels of feature binding up to the levels of multimodal binding of information coming from the different sensor modalities and also from other functional systems. The last 40 years of research have shown that the binding problem cannot be solved easily. Today, it is considered as one of the key questions to brain understanding. To date, various solutions have been suggested to the binding problem including: (1) combination coding, (2) binding by synchrony, (3) population coding, (4) binding by attention, (5) binding by knowledge, expectation, and memory, (6) hardwired vs. on-demand binding, (7) bundling and binding of features, (8) the feature-integration theory of attention, and (9) synchronization through top-down processes. Each of those hypotheses addresses important aspects of binding. However, each of them also suffers from certain weak points and can never give a complete explanation. This article gives a brief overview of the so far suggested solutions of perceptual binding and then shows that those are actually not mutually exclusive but can complement each other. A computationally verified model is presented which shows that, most likely, the different described mechanisms of binding act (1) at different hierarchical levels and (2) in different stages of "perceptual knowledge acquisition." The model furthermore considers and explains a number of inhibitory "filter mechanisms" that suppress the activation of inappropriate or currently irrelevant information.


Focus on sepsis and intensive care.
The Surviving Sepsis Campaign (SSC) Guidelines collate the evidence for managing sepsis. Most of the interventions suggested by the SSC guidelines are very relevant to military critical care, including rapid microbiologic investigation, early antibiotic administration and many aspects of early goal directed therapy. Other interventions may be more difficult to provide in remote theatres of operation where resources may be limited. This article discusses the application of the SSC guidelines to deployed military hospitals, with suggestions as to which interventions are feasible, and which may not be indicated.


A MATLAB package for the EIDORS project to reconstruct two-dimensional EIT images.
The EIDORS (electrical impedance and diffuse optical reconstruction software) project aims to produce a software system for reconstructing images from electrical or diffuse optical data. MATLAB is a software that is used in the EIDORS project for rapid prototyping, graphical user interface construction and image display. We have written a MATLAB package (http://venda.uku.fi/ vauhkon/) which can be used for two-dimensional mesh generation, solving the forward problem and reconstructing and displaying the reconstructed images (resistivity or admittivity). In this paper we briefly describe the mathematical theory on which the codes are based on and also give some examples of the capabilities of the package.


Doxorubicin selected multidrug-resistant small cell lung cancer cell lines characterised by elevated cytoplasmic Ca2+ and resistance modulation by verapamil in absence of P-glycoprotein overexpression.
Sublines from the small cell lung cancer (SCLC) cell lines U1285 and U1690, denoted U1285-100, U1285-250, U1690-40 and U1690-150, were adapted to grow in the continuous presence of 100, 250, 40 and 150 ng ml-1 doxorubicin (Dox), respectively. The Dox resistance was accompanied by cross-resistance to vincristine (Vcr), Vp-16 and for U1285-100 also to cisplatinum. Sublines of U1690-40 and U1285-100, cultured in absence of Dox for 4 months were only partially reversed with respect to Dox resistance. Neither the parental nor the most Dox resistance sublines had detectable levels of mdr 1 RNA but a small fraction of cells in all cell lines stained weakly positive for P-glycoprotein (P-gp). Verapamil (Ver) at 5 microM reversed the Dox resistance completely and partly in the U1690 and U1285 sublines, respectively, but did not increase the cellular accumulation of Dox. The cytoplasmic free Ca2+ concentration (Ca2+i) was close to 100 nM in both parental cell lines but elevated in the U1285-100 and U1690-40 sublines by 21 and 44%, respectively, and in U1285-250 and U1690-150 by 51 and 91%, respectively. The partly reverted sublines still showed significant but smaller elevations in Ca2+i of 10-30%. Ver was without acute or long term effects of Ca2+i in the U1285-100 and U1690-40 sublines. Selection for Dox resistance in SCLC may thus result in atypical multidrug-resistance characterised by absence of P-gp overexpression and atypical cross-resistance. Although Ver did not seem to affect Dox accumulation it may still work as a resistance modulator.(ABSTRACT TRUNCATED AT 250 WORDS)


Adverse effects of rabies pre- and postexposure prophylaxis in 290 health-care-workers exposed to a rabies infected organ donor or transplant recipients.
The recent unfortunate rabies transmissions through solid organ transplants of an infected donor in Germany required the initiation of a vaccination program to protect health care workers (HCWs) with close contact to rabies-infected patients. A systematic follow-up of adverse effects was initiated. Rabies postexposure prophylaxis (PEP) was started in 269 HCWs at four German hospitals. Pre-exposure prophylaxis (PreEP) was administered to 74 HCWs caring for an already diagnosed rabies patient. At each vaccination date, HCWs were interviewed for symptoms possibly representing adverse effects. Adverse effects of PEP and PrePEP were compared. Out of 269 HCWs, 216 were included for the investigation of adverse effects. Of these 216 HCWs, 114 (53%) individuals developed at least one systemic adverse effect. Incidences of tiredness (30.6%), malaise (26.4%), headache (26.9%), dizziness (14.8%), and chills (13.0%) declined in the course of PEP (p < 0.05), whereas incidences of fever (7.4%), paraesthesias (7.9%), arthralgias (1.9%), myalgias (4.2%), nausea (9.3%), diarrheas (2.8%) and vomiting (1.4%) did not. In 11 (5.1%) HCWs PEP was discontinued mostly due to adverse reactions (four suffered strong headaches, two HCWs meningeal irritations, two chills, one paraesthesia, one malaise, and one a rush). Systemic effects of PEP or PreEP did not differ significantly. Despite relatively high incidences of moderate severe adverse reactions rabies PEP is safe. Strong headache, tiredness, dizziness, and paraesthesias are the most important postvaccinal symptoms. Vaccinees suffering from adverse effects of PEP must be strongly encouraged to complete PEP, as it is to date the only protection against fatal rabies.


Determinants of pharmacologic treatment failure in panic disorder.
We systematically assessed reasons for failure of pharmacologic treatment for panic disorder in patients referred to a specialty anxiety and mood disorders clinic and examined possible determinants of treatment-resistant panic disorder. Interview data were obtained from 106 patients with DSM-III-R panic disorder seen in consultation. Data for each of 252 past medication trials included dose, duration of treatment, side effects, outcome, and reason for discontinuation. T tests and chi-square analyses were used to compare demographic and clinical characteristics of patients failing versus responding to adequate trials and those with and without intolerable medication side effects. Of 252 medication trials, 190 used effective antipanic medications, and only 59 (23%) were adequate in dose and duration. The most common reason for treatment failure was intolerable side effects, occurring in 51 (27%) of 190 trials using effective antipanic medications. Patients discontinuing treatment due to adverse effects had higher Hamilton Rating Scale for Anxiety scores and were less likely to have a history of substance abuse. Discontinuation due to side effects was significantly more common with tricyclic antidepressants than with benzodiazepines. True treatment resistance was reported in 14 (24%) of 59 adequate medication trials. Treatment-resistant patients were younger and had a higher lifetime rate of major depression. Although use of ineffective medications or inadequate trials were important factors, the most common reason for treatment failure was side effects, especially with tricyclic antidepressants. True treatment resistance was less common, since few medication trials were adequate in dose and duration, and may be associated with comorbidity.


The multiple functions of plant serine protease inhibitors: defense against herbivores and beyond.
Plant protease inhibitors (PIs) are a diverse group of proteins which have been intensely investigated due to their potential function in protecting plants against herbivorous insects by inhibiting digestive proteases. Although this mechanism has been well documented for a number of single PIs and their target enzymes, whether this mechanism protects plants in nature remains unclear. Moreover, many plants express a number of different PIs and it was unknown if these proteins work synergistically as defenses or if they also have other functions. We recently identified four serine PIs (SPI) of Solanum nigrum and demonstrated that they differ substantially in substrate specificity, accumulation patterns, and their effect against different natural herbivorous insects in field- and glasshouse experiments. These differences suggest that SPIs have at least partially diversified to provide protection against different attackers. Although we could not detect effects on plant development or growth when silencing SPIs, gene- and tissue-specific expression patterns suggest multiple functions in generative tissues, including a possible involvement in development.


Ultrathin Cu-TCPP MOF nanosheets: a new theragnostic nanoplatform with magnetic resonance/near-infrared thermal imaging for synergistic phototherapy of cancers.
Nanostructures based on metal-organic frameworks (MOFs) have promising potential as theragnostic nanoplatforms for phototherapy of cancer cells. However, the MOFs alone are seldom reported to be used as photothermal agents mainly due to their poor near-infrared (NIR) light absorption. Methods: Ultrathin copper-tetrakis (4-carboxyphenyl) porphyrin (Cu-TCPP) MOF nanosheets were prepared by a facile solvothermal route. The photothermal therapy (PTT), photodynamic therapy (PDT), and T1 -weighted magnetic resonance (MR) imaging capabilities and the high biocompatibility of these composite nanosheets were evaluated in vitro as well as in vivo in a mouse tumor model. Results: The ultrathin Cu-TCPP MOF nanosheets exhibited 1) strong NIR absorption because of the d-d energy band transition of Cu2+ and the ultrathin characteristic translating into excellent photothermal performance, 2) ability to produce singlet oxygen because of the inherent characteristic of TCPP, and 3) capability for MR imaging because of the unpaired 3d electrons of copper. Conclusion: Our study demonstrated that the Cu-TCPP MOF nanosheets are a promising phototherapy nanoplatform with the synergistic ability for PTT and PDT of cancer, guided by MR and infrared thermal imaging.