INCREASED PROBABILITY OF EXTINCTION DUE TO DECREASED GENETIC EFFECTIVE POPULATION SIZE: EXPERIMENTAL POPULATIONS OF CLARKIA PULCHELLA.
We established replicated experimental populations of the annual plant Clarkia pulchella to evaluate the existence of a causal relationship between loss of genetic variation and population survival probability. Two treatments differing in the relatedness of the founders, and thus in the genetic effective population size (Ne ), were maintained as isolated populations in a natural environment. After three generations, the low Ne treatment had significantly lower germination and survival rates than did the high Ne treatment. These lower germination and survival rates led to decreased mean fitness in the low Ne populations: estimated mean fitness in the low Ne populations was only 21% of the estimated mean fitness in the high Ne populations. This inbreeding depression led to a reduction in population survival: at the conclusion of the experiment, 75% of the high Ne populations were still extant, whereas only 31% of the low Ne populations had survived. Decreased genetic effective population size, which leads to both inbreeding and the loss of alleles by genetic drift, increased the probability of population extinction over that expected from demographic and environmental stochasticity alone. This demonstrates that the genetic effective population size can strongly affect the probability of population persistence.


Voltage dependent calcium channels in adrenal glomerulosa cells and in insulin producing cells.
We have examined the structure and function of Ca2+ channels in excitable endocrine cell types, in rat adrenal glomerulosa cells and in two insulin producing cell types, the rat pancreatic beta cell and the INS-1 cell line. In previous studies on glomerulosa cells, we observed low (T-type) and high threshold (L-type) voltage dependent Ca2+ currents in addition to a K+ induced inward rectifying Ca2+ current (Igl). beta cells are known to exhibit T-, L- and N-type currents. We have now found that INS-1 cells also show low threshold (T-type) and high threshold Ca2+ currents. The latter was further resolved by organic inhibitors into L-type and P/Q-type currents and no Igl was detected. The expression of the pore-forming alpha 1 subunit of voltage dependent Ca2+ channels was studied by means of reverse transcription-polymerase chain reaction (RT-PCR), followed by restriction enzyme mapping and/or sequencing. Both in glomerulosa and pancreatic beta cells, the neuroendocrine (D) class of the alpha 1 subunit, known to be responsible for L-type current, represents the majority of the PCR product. Comparable amounts of the neuroendocrine (D) and the neuronal A-type alpha 1 subunits dominate the message in INS-1 cells. Different characteristics of Ca2+ currents in these cell types is discussed in view of the channel repertoire.


Hereditary angioedema: Linking complement regulation to the coagulation system.
Congenital deficiency of C1 inhibitor, the main inhibitor of the classic complement system pathway, leads to paroxysmal angioedema (hereditary angioedema) that can be debilitating or life-threatening for affected patients. In the past few years many new insights on the pathogenesis of angioedema formation in the presence of low levels of C1 inhibitor has been accumulated. There is a central role for bradykinin that is released upon activation of the kallikrein-kinin system that is insufficiently controlled by adequate levels of C1 inhibitor. As C1 inhibitor also possesses a central regulatory role of other plasma systems, including the contact activation system of coagulation and the plasminogen-plasmin system that governs endogenous fibrinolysis, it is interesting to observe the effects of C1 inhibitor deficiency on activation of these systems and relevance for hemostasis in vivo and thrombo-embolic disease. Interestingly, and despite significant activation of these pathways, C1 inhibitor deficiency is not at all associated with a hemorrhagic tendency or prothrombotic state. New therapeutic options for treatment of C1 inhibitor efficiency have become available in recent years, including various forms of C1 inhibitor concentrate. Restoration of C1 inhibitor levels in patients with hereditary angioedema has not resulted in thrombotic complications or any other relevant disorder associated with the hemostatic system.


A case of invasive pulmonary aspergillosis during treatment for acute exacerbation of interstitial lung disease.
Prolonged immunosuppressive therapy is a risk factor for invasive pulmonary aspergillosis. We report a case of a 79-yearold man who underwent immunosuppressive therapy with methylprednisolone and cyclosporine for an acute exacerbation of interstitial lung disease. Ten days after initiation of immunosuppressive therapy, the patient reported night sweats and purulent sputum, and chest computed tomography scan revealed consolidation. He was diagnosed with invasive pulmonary aspergillosis, and required vasopressor support with oxygen therapy. After the administration of voriconazole and the modulation of immunosuppressive therapy, his condition improved. Short-term immunosuppressive therapy can also induce invasive pulmonary aspergillosis.


Phenotypic profiles and functional genomics in Alzheimer's disease and in dementia with a vascular component.
Alzheimer's disease (AD) and dementia with vascular component (DVC) are the most prevalent forms of dementia. Both clinical entities share many similarities, but they differ in major phenotypic and genotypic profiles as revealed by structural and functional genomics studies. Comparative phenotypic studies have identified significant differences in 25% of more than 100 parametric variables, including anthropometry, cardiovascular function, aortic atherosclerosis, brain atrophy, blood pressure, blood biochemistry, hematology, thyroid function, folate and vitamin B12 levels, brain hemodynamics and lymphocyte markers. The phenotypic profile of patients with DVC differs from that of AD patients in the following: anthropometric values (weight, height); cardiovascular function (ECG, heart rate); blood pressure; lipid metabolism (HDL-CHO, TGs); uric acid metabolism; peripheral calcium homeostasis; liver function (GOT, GPT, GGT); alkaline phosphatase; lactate dehydrogenase; red and white blood cells; regional brain atrophy (left temporal region, inter-hippocampal distance); and left anterior blood flow velocity. Functional genomics studies incorporating APOE-related changes in biological markers extended the difference between AD and DVC up to 57%. Brain perfusion studies show a severe brain hypoperfusion in dementia associated with enlarged age-dependent arterial perfusion times. Structural genomics studies with AD-related genes, including APP, MAPT, APOE, PS1, PS2, A2M, ACE, AGT, cFOS and PRNP genes, demonstrate different genetic profiles in AD and DVC, with an absolute genetic variation rate ranging from 30% to 80%, depending upon genes and genetic clusters. Single gene analysis identifies relative genetic variations ranging from 0% to 5%. The relative polymorphic variation in genetic clusters integrated by two, three or four genes associated with AD ranges from 1% to 3%. The main phenotypic differences between AD and DVC are genotype-dependent, especially in AD, probably indicating that different genomic factors are determinant for the expression of dementia symptoms which might be accelerated or induced by environmental and/or cerebrovascular factors.


Mutational analysis of the donor substrate binding site of the ribosomal peptidyltransferase center.
Previous experiments have shown that the top of helix 90 of 23S rRNA is highly important for the ribosomal peptidyltransferase activity and might be part of the donor (P) site. Developing on these studies, mutations in the 23S rRNA at the highly conserved positions G2505, G2582, and G2583 were investigated. None of the mutations affected assembly, subunit association, or the capacity of tRNA binding to A and P sites. A "selective transpeptidation assay" revealed that the mutations specifically impaired peptide bond formation. Results with a modified "fragment" assay using the minimal donor substrate pA-fMet are consistent with a model where the nucleotides psiGG2582 form a binding pocket for C75 of the tRNA.


Using motives for smoking to distinguish between different college student smoker typologies.
Relatively little is known about how to categorize different types of smokers, especially occasional smokers. Because of the prevalence of occasional smoking among college students, the current study aimed to gain an understanding of the different typologies of smokers on campus. To accomplish this, a latent class analysis (LCA) was conducted using 17 motives for smoking items (N = 327). The LCA revealed that four smoker groups were present: (1) addicted smokers who endorsed smoking due to pleasure and habit/addiction; (2) stress smokers, who endorsed smoking to relax, to reduce levels of stress, and to regulate mood; (3) social smokers, who endorsed smoking because of social factors such as to fit in or because friends smoke; and (4) nonendorsing smokers, who had a low endorsement for all the items. An additional LCA with covariates revealed that age of initiation, current smoking patterns, smoker self-classification, and quit likelihood differentiated these groups of smokers whereas current age and alcohol use did not. These typologies should be considered when designing interventions for occasional smokers.


Bone resorption in chronic otitis media: the role of the osteoclast.
Most of the pathology associated with cholesteatoma is the result of osteoclast-mediated bone resorption in the middle ear. Cytokines, prostaglandins, nitric oxide, neurotransmitters and growth factors are associated with chronic inflammation and have been implicated in cholesteatoma-induced bone resorption. Although many different factors are known to regulate osteoclast activation, there is a final common pathway of osteoclast differentiation and resorption. Recent advances in molecular techniques and the increasing availability of genetically altered mice have provided valuable insight into the molecular mechanism of osteoclastic bone resorption. This review focuses on osteoclast biology, lessons from genetically altered mice, and their contribution to our understanding of cholesteatoma-induced bone resorption.


Beyond Nursing Staff Levels: The Association of Nursing Home Quality and the Five-Star Quality Rating System's New Staffing Measures.
Recently, the Centers for Medicare & Medicaid Services (CMS) introduced staffing measures related to staffing variability and turnover in the Nursing Home (NH) Care Compare Five-Star Quality Rating System. While the consensus within the literature is that reduced variability and turnover are associated with higher NH quality of care and life, no existing studies have evaluated the relationship between CMS's newly introduced staffing measures and quality. This study uses regression analysis to estimate the relationship between 13 quality measures (used in Care Compare) and CMS's new staffing measures (i.e., weekend nursing staff levels, total nursing and registered nurse staff turnover, and administrator turnover) as well as a measure of daily staffing variation recently introduced in the literature called the coefficient of variation. Regressions analysis finds strong evidence of an association between quality and these staffing measures, though some measures (e.g., nursing staff turnover) are highly correlated and may be duplicative.


Efficacy of intraarticular hyaluronic acid injection through a lateral approach under fluoroscopic control for advanced hip osteoarthritis.
Hyaluronic acid (HA) is used for intraarticular treatment of hip osteoarthritis (OA). The objective of this study was to determine the efficacy of intraarticular injection of HA through a lateral approach under fluoroscopic control for advanced hip OA. The study included 21 patients with advanced hip OA. All patients received 2.5 ml HA injection once a week for 3 weeks by lateral approach under fluoroscopic control. Disability (Lequesne index), pain scores (visual analog scale-VAS) and analgesic use of patients were assessed before treatment and 1, 3 and 6 months after the treatment. Lequesne index and VAS pain scores measured 1, 3 and 6 months after treatment were significantly lower compared to baseline scores (p<0.001). Although analgesic use was significantly reduced 1 and 3 months after treatment compared to baseline (p<0.05), no difference was determined in analgesic use at the 6th month (p>0.05). No side effect was observed. In conclusion, intraarticular HA injection through a lateral approach under fluoroscopic control was shown to be a safe and effective method for patients with advanced hip OA.


Neuron-like spiking and bursting in Josephson junctions: A review.
The superconducting Josephson junction shows spiking and bursting behaviors, which have similarities with neuronal spiking and bursting. This phenomenon had been observed long ago by some researchers; however, they overlooked the biological similarity of this particular dynamical feature and never attempted to interpret it from the perspective of neuronal dynamics. In recent times, the origin of such a strange property of the superconducting junction has been explained and such neuronal functional behavior has also been observed in superconducting nanowires. The history of this research is briefly reviewed here with illustrations from studies of two junction models and their dynamical interpretation in the sense of biological bursting.


A misinterpreted psychoanalyst: Herbert Silberer and his theory of symbol-formation.
The primary aim of this article is to give a more detailed exposition of the cultural, personal, and theoretical contexts in which the Viennese psychoanalyst, Herbert Silberer's theories were born. When assessing the broader picture that this approach offers, it can be concluded that Silberer was an innovative thinker who inspired several of his contemporaries. Recognized in many respects by the society and scholars of this time, he represented quite a different viewpoint that was significantly influenced by several forms of Western esoteric thinking. Yet his main aim was to contribute to the field of psychoanalysis and develop a theory in which rationalistic psychoanalytic interpretations were combined with nonreductive approaches to mystical experiences. Silberer's name is frequently mentioned in a specific context in which his tragic suicide is emphasized rather than his innovations. Upon evaluating the materials recording Silberer's private life, it seems very likely that his suicide was not triggered by the criticism of Freud alone. Silberer's family affairs, his relationship with his father, and his financial and professional struggles could have all contributed to his tragic decision. This paper contends that Silberer's oeuvre deserves greater attention and must be evaluated based upon its own merit.


[Ursodeoxycholic acid in the treatment of chronic cholestatic liver disease. Documented delay in disease progress inspires hope].
The effect of ursodeoxycholic acid (UDCA) in primary biliary cirrhosis (PBC) has been investigated in a large number of randomized controlled studies. Meta-analyses of these studies have not proved a beneficial effect on survival. However, these analyses have also comprised patients with end-stage PBC as well as patients with very early PBC. A prolonged survival in such groups of patients is not expected to be demonstrated in short-term studies. Modified meta-analyses are underway. A number of beneficial effects on the disease process and complications still make an optimistic view of the drug in PBC justifiable. Limited experience suggests a beneficial effect of UDCA also in other cholestatic diseases.


Tracking Fish Lifetime Exposure to Mercury Using Eye Lenses.
Mercury (Hg) uptake in fish is affected by diet, growth, and environmental factors such as primary productivity or oxygen regimes. Traditionally, fish Hg exposure is assessed using muscle tissue or whole fish, reflecting both loss and uptake processes that result in Hg bioaccumulation over entire lifetimes. Tracking changes in Hg exposure of an individual fish chronologically throughout its lifetime can provide novel insights into the processes that affect Hg bioaccumulation. Here we use eye lenses to determine Hg uptake at an annual scale for individual fish. We assess the widely distributed benthic round goby (Neogobius melanostomus) from the Baltic Sea, Lake Erie, and the St. Lawrence River. We aged layers of the eye lens using proportional relationships between otolith length at age and eye lens radius for each individual fish. Mercury concentrations were quantified using laser ablation inductively coupled plasma mass spectrometry. The eye lens Hg content revealed that Hg exposure increased with age in Lake Erie and the Baltic Sea but decreased with age in the St. Lawrence River, a trend not detected using muscle tissues. This novel methodology for measuring Hg concentration over time with eye lens chronology holds promise for quantifying how global change processes like increasing hypoxia affect the exposure of fish to Hg.


Cannabis Use and the Risk of Cardiovascular Diseases: A Mendelian Randomization Study.
Several observational studies have shown that cannabis use has negative effects on the cardiovascular system, but the causality of this relationship has not been confirmed. The aim of the current study was to estimate the effects of genetically determined cannabis use on risk of cardiovascular diseases. Ten single-nucleotide polymorphisms related to cannabis use were employed as instruments to estimate the association between genetically determined cannabis use and risk of cardiovascular diseases using a two-sample Mendelian randomization (MR) method. Summary statistics data on exposure and outcomes were obtained from different genome-wide association meta-analysis studies. The results of this MR analysis showed no causal effects of cannabis use on the risk of several common cardiovascular diseases, including coronary artery disease, myocardial infarction, stroke and ischemic stroke subtypes, atrial fibrillation (AF), and heart failure. Various sensitivity analyses yielded similar results, and no heterogeneity and directional pleiotropy were observed. After adjusting for tobacco use and body mass index, multivariable MR analysis suggested a causal effect of cannabis use on small vessel stroke (SVS) [odds ratio (OR) 1.17; 95% CI 1.02-1.35; p = 0.03] and AF (OR 1.06; 95% CI 1.01-1.10; p = 0.01), respectively. This two-sample MR study did not demonstrate a causal effect of genetic predisposition to cannabis use on several common cardiovascular outcomes. After adjusting for tobacco use and body mass index, the multivariable MR analysis suggested a detrimental effect of cannabis use on the risk of SVS and AF, respectively.


Sb102- and Sb22- found in [K(18-crown-6)]6[Sb10] [Sb4{Mo(CO)3}2]·2en: two missing family members.
Reaction of KMnSb and C7H8Mo(CO)3 produced Sb102- and Sb22- found in [K(18-crown-6)]6[Sb10][Sb4{Mo(CO)3}2]·2en (1·2en) adding two new family members for antimony polyanions. The unusually short Sb-Sb bond in the Sb2 subunits of 1 reveals the formation of the second doubly bonded diatomic anion between heavy elements following Bi22-, and the first example of such doubly bonded anions stabilized by metal carbonyl groups.


[Outpatient echographic follow-up of patients with Fraley's syndrome].
The follow-up of the degree and evolution of the upper hydrocalicosis and of the parenchymatous changes in patients with Fraley's syndrome is very important. All authors agree that these patients should be followed up periodically. 20 patients with Fraley's syndrome were followed up and the authors conclude that the echographic examination is a very reliable method for prophylactic medical examinations since it is informative, harmless and economically advantageous.


Surgical management of pulmonary hydatid cysts in children in KwaZulu-Natal Province, South Africa.
Pulmonary hydatid cyst disease is still a major problem in countries like South Africa (SA), where livestock farming is common. Hydatidosis has a variable clinical course depending on the size, location and complications of the cyst. For pulmonary cysts of any size, surgery remains the gold standard for treatment, with lung conservation surgery being the ideal. To describe the epidemiology, clinical presentation, surgical management, and surgical outcomes of paediatric pulmonary hydatid disease in children referred to the Department of Cardiothoracic Surgery at Inkosi Albert Luthuli Central Hospital (IALCH) in KwaZulu-Natal Province, SA. We retrospectively reviewed and analysed the medical records of 38 children between the ages of 0 and 18 years with pulmonary hydatid cysts, who were referred to the Department of Cardiothoracic Surgery at IALCH and underwent surgical management between 1 January 2008 and 31 December 2018. The medical records were evaluated for patients' demographics, clinical evaluation, surgical management strategies and surgical outcomes. Of the 38 patients, 60.5% were male, with a mean age of 6.5 years. More than two-thirds of the patients (68.4%) were from the Eastern Cape and 31.6% were from KwaZulu-Natal. The majority of the cysts (84.2%; n=32) were classified as large (5 - 9 cm in diameter) and giant (≥10 cm in diameter). Forty-eight surgical procedures were performed and lung preservation surgery by enucleation and capitonnage was preferred. Some patients developed postoperative complications such as prolonged air leaks from bronchopleural fistula (18.8%; n=9) and 88.9% (n=8) of these patients healed with chest tube drainage and physiotherapy. Lung resection was only required in 4% (n=2) of the patients. The mean (standard deviation) number of days spent in hospital was 7 (4) days. All children survived with no recurrences. Conservative surgical procedures such as enucleation of the cysts of any size are possible, safe, reliable and reproducible.


Transcranial direct current and transcranial magnetic stimulations for chronic pain.
Chronic pain is debilitating and difficult to treat with pharmacotherapeutics alone. Consequently, exploring alternative treatment methods for chronic pain is essential. Noninvasive brain stimulation techniques, such as transcranial direct current stimulation (tDCS) and transcranial magnetic stimulation (TMS) are increasingly being investigated for their neuropharmacological effects in the treatment of chronic pain. This review aims to examine and evaluate the present state of evidence regarding the use of tDCS and TMS in the treatment of chronic pain. Despite conflicting evidence in the early literature, evidence from recent rigorous research supports the use of tDCS and TMS in treating chronic pain conditions. For both tDCS and TMS, standardized stimulation parameters have been identified with the recommendation for repeated maintenance stimulation to ensure that the analgesic effect is sustained beyond discontinuation of therapy. Due to a lack of defined stimulation protocols, early findings on the efficacy of tDCS and TMS are mixed. Although the application of tDCS and TMS as pain relief approaches is still in its early stages, the introduction of standardized stimulation protocols is paving the way for more robust and informed research.


Genetic analysis and molecular mapping of a nuclear recessive male sterility gene, ms91(t), in rice.
Mutations that result in plant male sterility provide means not only to probe reproductive development but also to facilitate commercial heterosis application and hybrid seed production. In this study, we report a novel male sterility gene, ms91(t), in a spontaneous mutant line (SH38) from a Chinese rice cultivar (Oryza sativa subsp. japonica 'Jijing14'). The sterility of SH38 was studied by examining its progenies derived from crosses with 6 japonica cultivars. Corresponding F2 populations were obtained by selfing each of the 6 F1s and a backcross population was produced by crossing SH38 to the F1 of SH38 x C18. Our results revealed that SH38 has normal agronomic traits but produces no pollen grains. Segregations of male-sterile and male-fertile progenies in the F2 and backcross populations fit well with ratios of 3:1 and 1:1, respectively, indicating that ms91(t) is a single recessive gene. Amplified fragment length polymorphism (AFLP) analysis of SH38 and Jijing14 plants showed the presence of a unique band in SH38. Simple sequence repeat (SSR) analysis of the bulked and individual progenies of the F2 population of SH38 x C18 showed linkage of ms91(t) with the SSR marker RM5853 on chromosome 1. Subsequently, ms91(t) was fine-mapped to the interval between markers RM7075 (3.75 cM) and RM5638 (3.57 cM). Our results would facilitate the isolation of ms91(t) and male sterility in heterosis application.


Vascular island skin-flap tolerance to warm ischemia: an analysis by perfusion fluorometry.
Fiberoptic perfusion fluorometry and assessment of ultimate viability were used to analyze the tolerance to warm ischemia of rat vascular island skin flaps. Both acute flaps and flaps raised 24 hours previously and then reraised were subjected to 0 (control), 6, 8, 10, and 12 hours of vascular pedicle clamping. Following clamp release, serial fluorometry documented the progressive delay in effective reflow resulting from extended periods of ischemia. Fluorometry, furthermore , suggested that flaps constructed 24 hours previously had an improved hemodynamic status with a significantly reduced period of poor reflow following clamp release. The improved hemodynamics were associated with increased viability, confirming the increased tolerance of 24-hour-old flaps to warm ischemia.


The syngeneic mixed leukocyte reaction: the genetic requirements for the recognition of self resemble the requirements for the recognition of antigen in association with self.
We have used cells from inbred strain 2 and strain 13 guinea pigs in order to define further the role of Ia antigens in the syngeneic mixed leukocyte reaction (MLR). The guinea pig syngeneic MLR resembled the autologous MLR in man in that it demonstrated both memory and specificity. The Ia antigens appeared to be the proliferative stimuli in that the primary stimulator cell was an Ia-positive adherent peritoneal exudate cell (PEC) and the reaction could be specifically inhibited by anti-Ia sera directed to the stimulator cell. We also demonstrated the existence of two (2 x 13)F1 T cell populations that were capable of reacting to one or the other parental PEC in the absence of any known exogenous antigen. These results suggest that the syngeneic MLR may represent T cell activation mediated through a receptor for self Ia.


[Gymnic art of Iccus of Taranto in the contest of Magna Grecia].
Iccus of Taranto was a famous athlete living in the V century B.C. and participating in Olympic competitions, of which was a winner in the LXXXVII games. He was quoted by Plato for the accuracy in the preparation of his athletic body: before competitions he avoided sexual relationships and took a lot of care to diet, so that he is considered the founder of athletic dietology. His thought was influenced by pytagoreans, mainly by Alcmaeon of Crotone: for this he taught the importance of the harmony of body and mind, the frugality in eating, the self control. An appropriate but frugal diet for athletes was known as meal of Iccus and used before races.


Outcomes in a digital weight management intervention with one-on-one health coaching.
Some guidelines state that in-person weight management interventions are more efficacious than those delivered digitally. However, digital programs are more scalable and accessible. We hypothesized that one-on-one health coaching via app-based video chat would simulate an in-person experience and help achieve outcomes comparable to those of in-person interventions. A 12-month digital weight management intervention was provided to overweight or obese adults recruited from a large technology company. One-on-one health coaching sessions were offered during a 24-week intensive phase as well as subsequent maintenance phase. Focused on sustainable changes in activity and diet, the intervention incorporates SMART goals, in-app food and activity logs, Fitbit integration, as well as optional sleep and stress modules. Self-Determination Theory and the Transtheoretical Model are incorporated to drive behavior change. Multilevel mixed-effects models were used to analyze weight changes retrospectively. Six hundred eighty-three participants reported 29,051 weights. At 12 months, mean percent changes in body weight were -7.2% and -7.6% for overweight and obese groups, respectively. A weight change of -5% is commonly targeted for in-person weight management interventions. Observed weight loss exceeded this target by 2.2% (95% CI, 0.7% to 3.8%; P < .01) for the overweight group and 2.6% (95% CI, 1.4% to 3.9%; P < .01) for the obese group. Further research is needed with randomization to in-person or digital interventions. Though limited by an observational, retrospective design, preliminary results suggest that some digital weight management programs with one-on-one coaching may achieve outcomes comparable to those of robust, in-person interventions.


Fibroblasts from the oral masticatory and lining mucosa have different gene expression profiles and proliferation rates.
To compare the gene expression profiles and proliferation rates of fibroblasts from the oral lining and masticatory mucosae. Primary human fibroblasts were retrieved from the posterior masticatory hard palate and the lining alveolar mucosa of five individuals. The gene expression profile was evaluated using total RNA sequencing. The proliferation rate was determined colorimetrically. Substantial differences in specific gene groups and pathways were observed between fibroblasts from the two tissues. Significantly enriched gene ontology processes were focused on the extracellular components. Lining mucosa fibroblasts exhibited significantly higher expression of the principal structural collagens, cranial neural crest markers, and homeobox genes associated with positional memory. Masticatory mucosa fibroblasts showed greater expression of genes related to transforming growth factor-β signalling, which may be associated with fibrosis. In addition, they expressed higher levels of the EP2 prostaglandin E2 receptor and Toll-like receptor 1. Finally, masticatory mucosa fibroblasts exhibited a 10%-30% higher proliferation rate. Fibroblasts from the lining and masticatory oral mucosae are phenotypically heterogeneous, presenting distinct gene expression profiles and proliferation rates. These features may contribute to their specific physiological functions and have relevance for potential therapeutic applications.


Effectiveness of mRNA Booster Vaccination Against Mild, Moderate, and Severe COVID-19 Caused by the Omicron Variant in a Large, Population-Based, Norwegian Cohort.
Understanding how booster vaccination can prevent moderate and severe illness without hospitalization is crucial to evaluate the full advantage of mRNA boosters. We followed 85 801 participants (aged 31-81 years) in 2 large population-based cohorts during the Omicron BA.1/2 wave. Information on home testing, PCR testing, and symptoms of coronavirus disease 2019 (COVID-19) was extracted from biweekly questionnaires covering the period 12 January 2022 to 7 April 2022. Vaccination status and data on previous SARS-CoV-2 infection were obtained from national registries. Cox regression was used to estimate the effectiveness of booster vaccination compared to receipt of 2-dose primary series >130 days previously. The effectiveness of booster vaccination increased with increasing severity of COVID-19 and decreased with time since booster vaccination. The effectiveness against severe COVID-19 was reduced from 80.9% shortly after booster vaccination to 63.4% in the period >90 days after vaccination. There was hardly any effect against mild COVID-19. The effectiveness tended to be lower among subjects aged ≥60 years than those aged <50 years. This is the first population-based study to evaluate booster effectiveness against self-reported mild, moderate, and severe COVID-19. Our findings contribute valuable information on duration of protection and thus timing of additional booster vaccinations.


The glenoid notch and its relation to the shape of the glenoid cavity of the scapula.
The prevalence of a notch in the anterior margin of the glenoid cavity of 236 scapulae (118 female, 118 male) was investigated. The notch was found in 129 scapulae (55%) and gave rise to a pear-shaped cavity. In 107 scapulae (45%) the notch was absent, the shape of the cavity being oval. No sex difference was found in the prevalence of the notch. If a distinct notch exists, the glenoid labrum is not attached to bone at the notch and is therefore liable to be sheared off (Bankart lesion).


[Reoperation following correction of partial defects of the endocardial cushion (indication, incidence, results)].
In the period between 1955 and 1982, 297 patients underwent surgical correction of a partial AV-canal. Closure of the ostium primum defect was performed either by direct suture or by patch (prosthetic material or pericardium). Only in cases with severe mitral incompetence was the cleft in the anterior leaflet of the mitral valve surgically treated. At an average of 6 years (range: 3 months - 22 years) after the initial procedure 21 patients (7.8 per cent) underwent reoperation. In 20 patients reoperation was necessary for hemodynamic reasons (recurrence of ASD: n = 8; severe AV-valve regurgitation: n = 3; or both: n = 9). One patient with moderate mitral valve incompetence suffered from severe "patch-hemolysis" due to direction of the blood-jet towards the prosthetic patch. Residual or recurrent atrial septal defects were closed by using a patch in cases with previous direct suture (39 patients - 8 reoperations) or by reinsertion or enlargement of the present patch (258 patients - 13 reoperations). AV-valve incompetence could be treated in all cases but two with reconstructive methods. In two patients implantation of a prosthetic valve was necessary. In the single case with "patch-hemolysis" the previous prosthetic patch was replaced by a pericardial one, together with a suture of the mitral cleft 3 months after operation. Mortality of reoperation was 14.5 per cent (3 early deaths). Major complications in the surviving patients did not occur, with one exception: one patient with postoperative total AV-block received a permanent pacemaker.


Genomic cloning of human Echinococcus granulosus DNA: isolation of recombinant plasmids and their use as genetic markers in strain characterization.
A small, size selected (0.5-5.0 Kbp) genomic DNA library has been constructed in the bacterial plasmid pAT153 using DNA extracted from a human isolate (Kenyan origin) of the hydatid disease organism, Echinococcus granulosus. A panel of taeniid cestode DNAs has been used in conjunction with hybridization and restriction-enzyme analysis to identify in the library two recombinant plasmids with Echinococcus-specific inserts and a single recombinant plasmid (coded pEG18) with a DNA fragment unique for E. granulosus. These, and other recombinant plasmids with E. granulosus DNA inserts, have been used in restriction endonuclease, Southern transfer and hybridization analysis to independently and reproducibly discriminate between the UK horse and sheep strains of E. granulosus; these probes may well prove of value for characterizing isolates from other endemic areas. The feasibility of using a cloned DNA fragment as the basis of a simple field test for distinguishing eggs of E. granulosus from those of other taeniid cestodes is discussed. The recombinant insert (approximately 2.3 Kbp in size) of pEG18 has a low copy number - estimated at approximately 26 - and it may not be sufficiently sensitive for practical use as a DNA probe in the identification of small numbers of E. granulosus eggs by molecular hybridization.


Dopamine Transporter Availability in Early Parkinson's Disease is Dependent on Sunlight Exposure.
Preliminary studies suggested seasonality of dopaminergic functioning, but it is unknown whether dopamine transporter (DAT) expression in humans is also dependent on the seasons. We, therefore, investigated seasonal and sunlight-dependent effects on DAT availability in early Parkinson's disease (PD) patients and healthy controls. DAT single-photon emission computed tomography scans (n = 730) were gathered from the Parkinson's Progression Marker Initiative (PPMI) database. We used global horizontal irradiance (GHI) as proxy for sun exposure/month and assessed associations between striatal DAT availability and season (autumn/winter versus spring/summer), GHI and latitude of the PPMI site. In PD patients, DAT availability in the left caudate nucleus was higher in spring/summer (B [standard error (SE)] = 0.05 [0.02], P = 0.03) and positively associated with higher sun exposure (B [SE] = 0.59 [0.22] × 10-3 , P = 0.007). Latitude (in degrees north) of the PPMI site was negatively associated with DAT availability in both PD and healthy controls. Striatal DAT availability may be influenced by daylight exposure. © 2023 The Authors. Movement Disorders published by Wiley Periodicals LLC on behalf of International Parkinson and Movement Disorder Society.


A novel GDAP1 mutation 439delA is associated with autosomal recessive CMT disease.
Charcot-Marie-Tooth (CMT) disease is the most common form of inherited motor and sensory neuropathy. Based on neurophysiological and neuropathological criteria CMT has been sub-classified into two main types: demyelinating and axonal. Furthermore, it is genetically heterogeneous with autosomal dominant, autosomal recessive (AR) and X-linked modes of inheritance. Thus far, seven genes have been identified in association with the demyelinating AR-CMT disease. We hereby report our clinical and molecular genetic findings in a consanguineous family with AR-CMT. Two young sisters with AR-CMT and other non-affected family members were clinically and electrophysiologically evaluated and then molecular genetic investigation was carried out in order to identify the pathogenic mutation. Following an initial indication for linkage of the family to the CMT4A locus on chromosome 8, we sequenced the Ganglioside-induced differentiation-associated protein 1 (GDAP1) gene and identified a single nucleotide deletion in exon 3 that is associated with AR-CMT in the family. We identified a novel GDAP1 439delA mutation that is associated with AR-CMT in a consanguineous family of Iranian descent with two affected young girls and a history in other members of the family.


Recent advances in the development of metal complexes as antibacterial agents with metal-specific modes of action.
The mounting burden of antimicrobial resistance (AMR) is one of the most concerning threats to public health worldwide. With low economic incentives and a dwindling supply of new drugs in clinical pipelines, more innovative approaches to novel drug design and development are desperately required. Metal-based compounds are rapidly emerging as an alternative to organic drugs, as they have the ability to kill pathogens via metal-specific modes of action. We herein review recent advances in metal-based antibacterial agents, including metal complexes, metal ions and catalytic metallodrugs. The review concludes with a perspective on the rational design of metal-based antibiotics, and how we can exploit their unique properties to tackle AMR.


Adult respiratory distress syndrome associated with disseminated toxoplasmosis.
Diffuse lung injury may result as a complication of disseminated toxoplasmosis in patients with AIDS. We report the case of an AIDS patient who had multiple organ failure associated with disseminated toxoplasmosis. The patient died because of ventilatory failure, and at autopsy findings in regard to the lungs were consistent with adult respiratory distress syndrome.


Early histologic findings of pulmonary SARS-CoV-2 infection detected in a surgical specimen.
Despite the current pandemic season, reports on pathologic features of coronavirus disease 19 (Covid-19) are exceedingly rare at the present time. Here we describe the pathologic features of early lung involvement by Covid-19 in a surgical sample resected for carcinoma from a patient who developed SARS-CoV-2 infection soon after surgery. The main histologic findings observed were pneumocyte damage, alveolar hemorrhages with clustering of macrophages, prominent and diffuse neutrophilic margination within septal vessels, and interstitial inflammatory infiltrates, mainly represented by CD8+ T lymphocytes. These features are similar to those previously described in SARS-CoV-1 infection. Subtle histologic changes suggestive pulmonary involvement by Covid-19 may be accidentally encountered in routine pathology practice, especially when extensive sampling is performed for histology. These findings should be carefully interpreted in light of the clinical context of the patient and could prompt a pharyngeal swab PCR test to rule out the possibility of SARS-CoV-2 infection in asymptomatic patients.


Application of prophylactic cranial irradiation in limited-stage small-cell lung cancer: which patients could benefit?
To analyze the role of prophylactic cranial irradiation (PCI) on the survival for patients with limited-stage small-cell lung cancer (LS-SCLC). We screened patients from SEER database. Kaplan-Meier analysis and Cox proportional hazard model were used to evaluate factors influencing survival. LS-SCLC patients who receiving PCI were associated with better overall survival (OS; p < 0.001) and cancer-specific survival (CSS; p < 0.001). Multivariable Cox analysis revealed PCI was an independent prognostic factor for OS (p < 0.001) and CSS (p < 0.001). In subgroup analysis, there were no OS and CSS differences between PCI and no PCI groups in black patients and patient with a tumor size <5 cm (all p > 0.05). PCI remains an effective method for most LS-SCLC patients. However, caution should be taken in recommending PCI for black patients and patients with a tumor size <5 cm. Further clinical trials are necessary to validate our results and identify the most suitable patients for PCI in the modern era.


Rearrangement, amplification and overexpression of c-Ha Ras gene in premalignant lesion of Turcot's syndrome.
Ras gene family (c-Ha, Ki, N ras) and c-myc proto-oncogenes were analyzed in seven colonic and three gastric adenomatous polyps obtained from a patient with Turcot's syndrome. The rearrangement and amplification as well as overexpression of c-Ha ras gene in one colonic adenomatous polyp were determined. The amplified c-Ha ras gene in this polyp revealed larger fragments of BamHI, PstI and SacI than those in the normal colonic mucosa from the same patient. But, such abnormalities were not observed in other polyps. No abnormalities of c-Ki ras, c-N ras or c-myc gene were observed in any polyps. These results suggest that the alternations of c-Ha ras gene in this patient may not be responsible for the adenomatous change, but may be related to the transition from adenoma to carcinoma of the colon.


Enabling Flexible Heterostructures for Li-Ion Battery Anodes Based on Nanotube and Liquid-Phase Exfoliated 2D Gallium Chalcogenide Nanosheet Colloidal Solutions.
2D metal chalcogenide (MC) nanosheets (NS) have displayed high capacities as lithium-ion battery (LiB) anodes. Nevertheless, their complicated synthesis routes coupled with low electronic conductivity greatly limit them as promising LiB electrode material. Here, this work reports a facile single-walled carbon nanotube (SWCNT) percolating strategy for efficiently maximizing the electrochemical performances of gallium chalcogenide (GaX, X = S or Se). Multiscaled flexible GaX NS/SWCNT heterostructures with abundant voids for Li+ diffusion are fabricated by embedding the liquid-exfoliated GaX NS matrix within a SWCNT-percolated network; the latter improves the electron transport and ion diffusion kinetics as well as maintains the mechanical flexibility. Consequently, high capacities (i.e., 838 mAh g-1 per gallium (II) sulfide (GaS) NS/SWCNT mass and 1107 mAh g-1 per GaS mass; the latter is close to the theoretical value) and good rate capabilities are achieved, which can be majorly attributed to the alloying processes of disordered Ga formed after the first irreversible GaX conversion reaction, as monitored by in situ X-ray diffraction. The presented approach, colloidal solution processing of SWCNT and liquid-exfoliated MC NS to produce flexible paper-based electrode, could be generalized for wearable energy storage devices with promising performances.


Robust homeostasis of cellular cholesterol is a consequence of endogenous antithetic integral control.
Although cholesterol is essential for cellular viability and proliferation, it is highly toxic in excess. The concentration of cellular cholesterol must therefore be maintained within tight tolerances, and is thought to be subject to a stringent form of homeostasis known as Robust Perfect Adaptation (RPA). While much is known about the cellular signalling interactions involved in cholesterol regulation, the specific chemical reaction network structures that might be responsible for the robust homeostatic regulation of cellular cholesterol have been entirely unclear until now. In particular, the molecular mechanisms responsible for sensing excess whole-cell cholesterol levels have not been identified previously, and no mathematical models to date have been able to capture an integral control implementation that could impose RPA on cellular cholesterol. Here we provide a detailed mathematical description of cholesterol regulation pathways in terms of biochemical reactions, based on an extensive review of experimental and clinical literature. We are able to decompose the associated chemical reaction network structures into several independent subnetworks, one of which is responsible for conferring RPA on several intracellular forms of cholesterol. Remarkably, our analysis reveals that RPA in the cholesterol concentration in the endoplasmic reticulum (ER) is almost certainly due to a well-characterised control strategy known as antithetic integral control which, in this case, involves the high-affinity binding of a multi-molecular transcription factor complex with cholesterol molecules that are excluded from the ER membrane. Our model provides a detailed framework for exploring the necessary biochemical conditions for robust homeostatic control of essential and tightly regulated cellular molecules such as cholesterol.


Biotransformation of glyceryl trinitrate and elevation of cyclic GMP precede glyceryl trinitrate-induced vasodilation.
In this study, we examined glyceryl trinitrate (GTN) biotransformation and cyclic GMP elevation in vascular smooth muscle before onset of GTN-induced relaxation. Isolated rabbit aortic strips (RAS) and strips of bovine pulmonary artery (BPA) and bovine pulmonary vein (BPV) were contracted submaximally and incubated with [3H]GTN. Before onset of GTN-induced vasodilation, the tissues were freeze-clamped and then analyzed for GTN, glyceryl-1,2-dinitrate (1,2-GDN), and glyceryl-1,3-dinitrate (1,3-GDN) and for cyclic GMP. Before onset of relaxation of RAS, BPA, and BPV, there was significant biotransformation of GTN to GDN and significant elevation of cyclic GMP. There was significantly greater biotransformation of GTN and elevation of cyclic GMP by BPV than by BPA incubated with the same concentration of GTN, which was temporally related with the more rapid onset of relaxation induced in BPV than in BPA. These results are consistent with the hypothesis that the magnitude of GTN biotransformation before vasodilation is the important determinant of subsequent tissue relaxation. In GTN biotransformation before vasodilation, there was preferential formation of 1,2-GDN. These data indicate that the mechanism of GTN biotransformation to 1,2-GDN is related to elevation of cyclic GMP and subsequent vasodilation.


T cells and autoimmune kidney disease.
Glomerulonephritis is traditionally considered to result from the invasion of the kidney by autoantibodies and immune complexes from the circulation or following their formation in situ, and by cells of the innate and the adaptive immune system. The inflammatory response leads to the proliferation and dysfunction of cells of the glomerulus, and invasion of the interstitial space with immune cells, resulting in tubular cell malfunction and fibrosis. T cells are critical drivers of autoimmunity and related organ damage, by supporting B-cell differentiation and antibody production or by directly promoting inflammation and cytotoxicity against kidney resident cells. T cells might become activated by autoantigens in the periphery and become polarized to secrete inflammatory cytokines before entering the kidney where they have the opportunity to expand owing to the presence of costimulatory molecules and activating cytokines. Alternatively, naive T cells could enter the kidney where they become activated after encountering autoantigen and expand locally. As not all individuals with a peripheral autoimmune response to kidney antigens develop glomerulonephritis, the contribution of local kidney factors expressed or produced by kidney cells is probably of crucial importance. Improved understanding of the biochemistry and molecular biology of T cells in patients with glomerulonephritis offers unique opportunities for the recognition of treatment targets for autoimmune kidney disease.


The predictive value of lactate levels in vaginal fluid on the latent period in pregnant women with preterm premature rupture of membranes.
The aim of the present study was to investigate the relationship between lactate level in vaginal fluid and the latent phase of labour in pregnancies complicated by preterm premature rupture of membranes (PPROM). Seventy pregnant women with PPROM during 28-34 weeks' gestation were selected for this prospective observational study. All subjects underwent a pelvic examination involving the insertion of a vaginal speculum, and lactate levels were measured in vaginal fluid samples. The relationship between the lactate levels in the vaginal fluid and the latent phase of the labour was analysed using a logistic regression test. Of the patients, 48 (68.6%) had a latent period of 48 h or less, and 22 patients (31.4%) had a latent period longer than 48 h. The median lactate level was 3.81 mmol/L in patients with a latent period ≤ 48 h, and 3.36 mmol/L in patients with a latent period > 48 h. The lactate level in vaginal fluid was not found to be distinctive in the differentiation of patients according to the duration of the latent phase (receiver operating characteristic or ROC: 0.509; 95% confidence interval or CI: 0.361-0.657; p = 0.904). There was no significant correlation between the lactate level in the vaginal fluid and the transition from the latent phase to the active phase of labour in pregnancies complicated by PPROM.


Does cyclic 3' ,5'-adenosine monophosphate act as second messenger in a voltage-dependent response to 5-hydroxytryptamine in Aplysia?
1 The possibility that cyclic adenosine 3',5'-monophosphate (cyclic AMP) mediates a voltage-dependent inward current elicited by 5-hydroxytryptamine (5-HT) in RB and LB cells of the abdominal ganglion of Aplysia was tested. 2 Intracellular injection of cyclic AMP elicited an inward current with a similar time course, potential dependence and ionic sensitivity as the response to 5-HT. 3 Intracellular injection of guanylyl imidodiphosphate (GMP-PNP), which activated adenylate cyclase, neither mimicked nor enhanced the 5-HT-evoked current. On the contrary, it reduced the current. 4 The phosphodiesterase inhibitors, Ro20-1724, isobutyl methylxanthine (IBMX) and theophylline, each antagonized the voltage-dependent response to 5-HT. To varying degrees they each induced an inward current. 5 The adenylate cyclase antagonist, dithiobisnitrobenzoic acic (DTNB), had no effect on the response to 5-HT when applied either intracellularly or extracellularly. Intracellular injection of the phosphodiesterase activator imidazole also had no effect. 6 Tubocurarine and neostigmine did not reduce the voltage-dependent inward current evoked by 5-HT; methysergide elicited an inward current. 7 Although the observations that cyclic AMP and 5-HT can evoke similar voltage-dependent inward currents in RB and LB neurones of Aplysia might suggest a second messenger role for the cyclic nucleotide, the pharmacological data are inconsistent with this hypothesis.


Identification of Key Genes and Pathways Involved in Circulating Tumor Cells in Colorectal Cancer.
Characterization of the features associated with circulating tumor cells (CTCs) is one of major interests for predicting clinical outcome of colorectal cancer (CRC) patients. However, the molecular features of CTCs remain largely unclear. For identification of key genes and pathways, GSE31023, contained CTCs from six metastatic CRC patients and three controls, was retrieved for differentially expressed gene (DEG) analysis. Protein-protein interaction networks of DEGs were constructed. Hub genes from the network were prognostic analyzed, as well as the association with tumor-infiltrating immune cells. 1353 DEGs were identified between the CTC and control groups, with 403 genes upregulated and 950 downregulated. 32 pathways were significantly enriched in KEGG, with ribosome pathway as top. The top 10 hub genes were included, including eukaryotic translation elongation factor 2 (EEF2), ribosomal protein S2 (RPS2), ribosomal protein S5 (RPS5), ribosomal protein L3 (RPL3), ribosomal protein S3 (RPS3), ribosomal protein S14 (RPS14), ribosomal protein SA (RPSA), eukaryotic translation elongation factor 1 alpha 1 (EEF1A1), ribosomal protein S15a (RPS15A), and ribosomal protein L4 (RPL4). The correlation between CD4+ T cells and RPS14 (correlation = -0.5) was the highest in colon cancer while CD8+ T and RPS2 (correlation = -0.53) was the highest in rectal cancer. This study identified potential role of ribosome pathway in CTC, providing further insightful therapeutic targets and biomarkers for CRC.


Experiments with hierarchical concept-based search.
Many digital libraries use hierarchical indexing schema, such as MeSH to enable concept based search in the retrieval phase. However, improving or outperforming the traditional full text search isn't trivial. We present an extensive set of experiments using a hierarchical concept based search retrieval method, applied in addition to several baselines, within the Vaidruya search and retrieval framework. Concept Based Search applied in addition to a low baseline is outperforming significantly, especially when queried on concepts in the third level and using disjunction within the hierarchical trees.


[Primary epiretinal gliosis (author's transl)].
Analyzing the findings of 45 patients with primary epiretinal gliosis in 51 otherwise entirely healthy eyes, the clinical features of the disease are elaborated. The disorder is characterized by the occurance of a glial membrane covering the retina in the area of the posterior pole and leading to a distorsion of the posterior pole and leading to a distorsion of the subjacent and surrounding tissues. Most frequently, the foveal region is involved, but the foveola is often spared. With increasing distance from the center the frequency of involvement decreases. Only in very rare cases the membrane extends beyond a distance of 5,5 mm from the center of the retina. The density of the membrane varies from case to case. Average visual acuity of the eyes evaluated in this study was 0,45. In no case it was worse than 0,05. The incidence of the disease increased with both increasing myopia and hyperopia. The average age of the patients was 55 years. There was no predominance of either men or women. The glia cells which compose the membrane originate from the innermost layers of the retina, from where they extend on the retinal surface through a defect in the inner limiting membrane. The reason for the formation of the defects is not yet understood. It is also unknown if the presence of such defects is absolutely necessary for the development of this condition.


A bent Laue analyser crystal for Rayleigh-to-Compton computed tomography.
A new optical system to perform tomography based on the Rayleigh-to-Compton (RC) method with high spatial and spectral resolution is presented. The RC technique allows the effective atomic number of a sample to be measured and finds application in bone mineral densitometry in medicine. It is particularly useful for the characterization of the distribution of biological materials which do not exhibit distinctive diffraction peaks. The system is based on the separation of the elastic line from the spectrum that is scattered by the sample by means of a bent Laue analyser crystal, and the subsequent independent detection of the elastic and inelastic parts of the spectrum with two large-area scintillation counters. The high energy resolution permits operation at low momentum transfer, where the RC method has its best contrast-to-noise ratio for low-Z materials. The geometrical and spectral requirements in terms of the incident beam and the conical analyser crystal are discussed. A first-generation tomographic imaging system (pencil beam, scanned sample) as implemented at the ESRF Compton-Scattering Station ID15B is described. A high-resolution tomographic reconstruction of a bone sample is presented.


Internal medicine residents' perceptions of the balance between service and education in their night-call activities.
To quantify the workload of residents on night call and to determine the residents' perceptions of the balance between service and education in their night-call activities. Twenty-two internal medicine residents--nine first-year residents and 13 senior residents (i.e., in their second, third, or fourth year)--kept logs of their night-call activities for two periods of 16 days each in 1991-92, at a 772-bed teaching hospital affiliated with Wright State University School of Medicine. The residents used the following five-point scale to rate each activity: 1, strictly service; 3, even balance; 5, strictly education. They also recorded the total times spent on various activities. Data were analyzed by using the independent and paired t-tests. Ninety-eight percent of the logs were returned. The first-year and senior residents did not differ significantly in times spent on logged events. Both the first-year and the senior residents perceived many night-call activities to be weighted toward education, especially those involving evaluation of acutely ill patients being considered for hospital admission (overall mean rating of 3.3). There was no striking difference between the first-year and senior residents' perceptions of the balance between service and education in their activities. The residents' perceptions of the relationship between service and education indicate that there was an even balance between the two. The educational aspects of night-call duties can and should remain an integral part of residents' training.


Computer model of antiepileptic effects mediated by alterations in GABA(A)-mediated inhibition.
Results from a computer model of a thalamic network predict that agents augmenting GABA(A)-mediated inhibition in the reticular thalamic (RE) nucleus will be antiepileptic or desynchronizing. This provides support for the hypothesis that antiepileptics like benzodiazepines may exert their effects through an isolated increase of inhibition in the RE nucleus. When desynchronized, the model thalamocortical neurons showed a decreased probability of firing a low threshold spike, a decreased secondary inhibitory postsynaptic potential and a higher frequency of oscillations. The transition to desynchrony was also accompanied by an increased frequency in the firing of the model RE neurons.


Resonantly pumped high efficiency Ho:GdTaO4 laser.
We demonstrate a continuous-wave 2.1-µm laser with a new Ho:GdTaO4 crystal pumped by a 1940.3-nm Tm fiber laser at room temperature. The maximum output power of 11.2 W at 2068.39 nm was achieved, corresponding to a slope efficiency of 72.9%. Moreover, the beam quality factor was measured to be about 1.4 at the maximum output level.


Lipozyme RM IM-catalyzed acidolysis of Cinnamomum camphora seed oil with oleic acid to produce human milk fat substitutes enriched in medium-chain fatty acids.
In the present study, a human milk fat substitute (HMFS) enriched in medium-chain fatty acids (MCFAs) was synthesized through acidolysis reaction from Cinnamomum camphora seed oil (CCSO) with oleic acid in a solvent-free system. A commercial immobilized lipase, Lipozyme RM IM, from Rhizomucor miehei, was facilitated as a biocatalyst. Effects of different reaction conditions, including substrate molar ratio, enzyme concentration, reaction temperature, and reaction time were investigated using response surface methodology (RSM) to obtain the optimal oleic acid incorporation. After optimization, results showed that the maximal incorporation of oleic acid into HMFS was 59.68%. Compared with CCSO, medium-chain fatty acids at the sn-2 position of HMFS accounted for >70%, whereas oleic acid was occupied predominantly at the sn-1,3 position (78.69%). Meanwhile, triacylglycerol (TAG) components of OCO (23.93%), CCO (14.94%), LaCO (13.58%), OLaO (12.66%), and OOO (11.13%) were determined as the major TAG species in HMFS. The final optimal reaction conditions were carried out as follows: substrate molar ratio (oleic acid/CCSO), 5:1; enzyme concentration, 12.5% (w/w total reactants); reaction temperature, 60 °C; and reaction time, 28 h. The reusability of Lipozyme RM IM in the acidolysis reaction was also evaluated, and it was found that it could be reused up to 9 times without significant loss of activities. Urea inclusion method was used to separate and purify the synthetic product. As the ratio of HMFS/urea increased to 1:2, the acid value lowered to the minimum. In a scale-up experiment, the contents of TAG and total tocopherols in HMFS (modified CCSO) were 77.28% and 12.27 mg/100 g, respectively. All of the physicochemical indices of purified product were within food standards. Therefore, such a MCFA-enriched HMFS produced by using the acidolysis method might have potential application in the infant formula industry.


Housing conditions and microbial environment do not affect the efficacy of vaccines for treatment of opioid use disorders in mice and rats.
Vaccines offer a promising prophylactic and therapeutic intervention to counteract opioid use disorders (OUD) and fatal overdoses. Vaccines generate opioid-specific antibodies that bind the target opioid, reducing drug distribution to the brain and preventing drug-induced behavioral and pharmacological effects. Due to their selectivity, anti-opioid vaccines can be administered in combination with FDA-approved medications. Because patients with OUD or other substance use disorders may be affected by other multifactorial co-morbidities, such as infection or depression, it is important to test whether vaccine efficacy is modified by factors that may impact individual innate or adaptive immunity. To that end, this study tested whether housing conditions would affect the efficacy of two lead vaccine formulations targeting oxycodone and fentanyl in male mice and rats, and further analyzed whether differences in the gastrointestinal (GI) microbiome would be correlated with either vaccine efficacy or housing conditions. Results showed that housing mice and rats in either conventional (non-controlled) or specific pathogen-free (SPF, sterile barrier maintained) environment did not affect vaccine-induced antibody responses against oxycodone and fentanyl, nor their efficacy against oxycodone- and fentanyl-induced antinociception, respiratory depression, and bradycardia. Differences in the GI microbiome detected via 16S rRNA gene sequencing were related to the housing environment. This study supports use of anti-opioid vaccines in clinical populations that may display deficits in microbiome function.


Clinical characteristics of asymptomatic and symptomatic patients with mild COVID-19.
Detailed knowledge on the prevalence of asymptomatic cases of coronavirus disease 2019 (COVID-19) and the clinical characteristics of mild COVID-19 is essential for effective control of the COVID-19 pandemic. We determined the prevalence of asymptomatic cases of COVID-19 and characterized the symptoms of patients with mild COVID-19. Study participants were recruited from a community facility designated for the isolation of patients without moderate-to-severe symptoms of COVID-19 in South Korea. The prevalence of asymptomatic patients at admission and the detailed symptoms of mild COVID-19 were evaluated through a questionnaire-based survey. Diagnosis of COVID-19 was confirmed by real-time RT-PCR. Of the 213 individuals with COVID-19, 41 (19.2%) were asymptomatic until admission. Among the remaining patients with mild COVID-19, the most common symptom was cough (40.1%; 69/172), followed by hyposmia (39.5%; 68/172) and sputum (39.5%; 68/172). Of the 68 individuals with hyposmia, 61 (90%) had accompanying symptoms such as hypogeusia, nasal congestion or rhinorrhoea. Fever (>37.5°C) was only observed in 20 (11.6%) individuals. As much as one-fifth of individuals with COVID-19 remained asymptomatic from exposure to admission. Hyposmia was quite frequent among individuals with mild COVID-19, but fever was not. Social distancing should be strongly implemented to prevent disease transmission from asymptomatic individuals or those with mild and inconspicuous symptoms.


Grazoprevir/elbasvir combination therapy for HCV infection.
Interferon-free regimens combine different second-wave direct-acting antiviral agents (DAAs), which target the main viral proteins involved in the replication cycle of hepatitis C virus (HCV): NS3/4A protease inhibitors (simeprevir or paritaprevir boosted by ritonavir), NS5B nucleos(t)idic (sofosbuvir) and nonnucleos(t)idic (dasabuvir) polymerase inhibitors, NS5A replication complex inhibitors (daclatasvir, ledipasvir, elbasvir, velpatasvir). Combinations of two or three DAAs, given for 8-24 weeks reach sustained virology response (SVR) rates greater than 90% with good tolerance. SVR rates and safety are similar in clinical trials and in real life, usually higher than 95% in the per-protocol analysis. Next-generation DAAs are now expected. To be competitive, these new combinations need to prove their added value regarding the pill burden, the reduced duration of treatment, the drug-drug interaction profile and safety. Zepatier is a fixed-dose combination product coformulating MK-5172 [grazoprevir (GZR), 100 mg QD] and MK-8742 [elbasvir or (EBR) 50 mg QD]: it combines highly potent inhibitors of the HCV NS3/4A protease and NS5A replication complex, respectively. This review provides a summary of the main evidence available for the use of GZR/EBR and highlights the strength of this combination.


Twenty Years of Progress on Maternal and Child Health in the Philippines: An Equity Lens.
This article assesses trends and inequalities in maternal and child health in the Philippines between 1993 and 2013, using 6 national household surveys, and also compares the Philippines' performance to 15 other Asia-Pacific countries. Thirteen indicators of child health outcomes and maternal and child health interventions are examined. Two measures of inequality are used: the absolute difference between the poorest and wealthiest quintile, and the concentration index. Coverage of all indicators has improved, both on average and among the poorest quintile; however, increases are very small for child health interventions (especially immunization coverage). By the first measure of inequality, all indicators show narrowing inequalities. By the second measure, inequality has fallen only for maternal health interventions. Compared with other 15 other developing Asia-Pacific countries, the Philippines performs among the best on the child health outcomes examined and above average on maternal health interventions (except family planning), but only at or below average on child health interventions.


Superoxide generation via the NR2B-NMDAR/RasGRF1/NOX2 pathway promotes dendritogenesis.
N-methyl-D-aspartate receptors (NMDARs) that contain the NR2A and NR2B subunits play a critical role in neuronal plasticity and dendritogenesis. Gain-and-loss-of function studies indicate that NR2B, but not NR2A, promotes dendritic branching. Accumulating evidence indicates that stimulation of NMDARs activates NADPH oxidase (NOX2), thereby generating superoxide. However, the molecular underpinnings of this process are not understood. RasGRF1, a guanine nucleotide exchange factor, is key for several forms of neuronal plasticity and interacts directly with the tail of NR2B. We investigated whether the NR2B-NMDAR/RasGRF1 pathway regulates the activity of NOX2 and whether superoxide production is required for dendritogenesis. We measured superoxide production in developing primary cultures of hippocampal neurons from 3 to 25 days in vitro (DIV) with the probe dihydroethidium (dHE). We found the highest dHE levels at early and intermediate developmental stages (3-15 DIV), when the NR2B-NMDAR expression is abundant. During these early/intermediate developmental stages, but not in mature neurons (>15 DIV), NMDAR activity is required for superoxide production. We also found that disrupting the NR2B-RasGRF1 interaction led to reduced dHE fluorescence intensity and moreover inhibited dendritic branching in hippocampal neurons. Together, our data indicate that superoxide production is induced by the NR2B-NMDARs/RasGRF1/NOX2 pathway and promotes dendritogenesis.


Phototoxic effect of photodynamic therapy on lung cancer cells grown as a monolayer and three dimensional multicellular spheroids.
Photodynamic therapy (PDT) is a minimally invasive therapeutic modality for the treatment of neoplastic and non-neoplastic diseases. The photochemical interaction of light, photosensitizer (PS) and molecular oxygen produces singlet oxygen which induces cell death. The effectiveness of zinc sulfophthalocyanine (ZnPcSmix ) has been shown on A549 monolayers and not on MCTS. The objective of this study was to investigate whether the same pattern of cell death would be induced in lung cancer cells (A549) grown as monolayer cells compared to three dimensional multicellular tumor spheroids (MCTS) using the same laser parameters. Commercially purchased A549 cells used in this study were cultured as monolayers and as MCTS. ZnPcSmix at different concentrations [0, 5, 10, 20, and 40 µM] was used and activated at a wavelength of 680 nm with 5 J/cm2. Lysosomal and mitochondrial damage after PDT and reactive oxygen species (ROS) production were determined by fluorescent microscopy. Changes in cellular responses were evaluated using cell morphology, viability, proliferation, cytotoxicity, and cell death analysis. Cells exposed to neither laser light nor PS, showed no changes in cell morphology, proliferation, cytotoxicity, and ROS production. However, cells treated with light activated ZnPcSmix resulted in a significant production of ROS and a dose dependant decrease in viability and proliferation as well as an increase in cell membrane damage in both monolayer and MCTSs. ZnPcSmix PS used in this study induced damage to vital organelles and was effective in inducing apoptosis in lung cancer cells grown as a monolayer and MTCS through ROS production.


A Whole-Mount Approach for Accurate Quantitative and Spatial Assessment of Fetal Oocyte Dynamics in Mice.
Depletion of oocytes from the embryonic ovary is a key feature of mammalian oogenesis; however, the rational and molecular bases for this phenomenon remain poorly understood. Presently in the field, the most systematic analysis used to understand the effect of a given molecular pathway on fetal oocyte attrition is to count the number of oocytes in ovaries at different stages of development. This analysis is commonly done using a sampling method based on sectioning of the ovary, a technique that includes many laborious steps culminating in an inaccurate estimate of oocyte number contained within that ovary. This inability to generate data that are directly comparable between labs hinders the field and raises questions about the timing and rate of oocyte depletion. Therefore, we set out to implement a robust method that can be easily used by most research laboratories to study the dynamics of oogenesis during fetal mouse ovary development in both normal and experimental conditions. Here we describe an approach to accurately count the total number of oocytes in embryonic ovaries. This method is based on whole-mount immunofluorescence, tissue clearing with sucrose and ScaleA2 reagent, and automatic detection and counting of germ cells in intact ovaries using confocal microscopy and three-dimensional software analyses. We demonstrate the power of the method by assessing variation of fetal oocyte numbers between left and right ovaries and between litters of mice. Finally, we anticipate that the method could be adopted to the analysis of substages of meiotic prophase I and ovarian somatic cells.


Variations in the vestibular cortical bone of permanent canine teeth in orthodontic patients. a comparative study: linear tomography vs. cbct (3d accuitomo).
The aim of this study was to compare the results of measuring the height of the vestibular cortical bone of canine teeth by linear tomography (LT) and 3-D Accuitomo cone beam computed tomography (CBCT) before and after aligning dental arches by orthodontic treatment. LT and CBCT were performed before and after orthodontic alignment on 12 canines in three patients undergoing orthodontic treatment, and the height of the canine vestibular cortical bones measured in mm. Measurements were taken by double-blinded operators. The mean variation in height of the vestibular cortical bone with orthodontic treatment was - 0,33 mm} 0.233 standard error using CBCT and -0,08mm } 0.55 standard error using LT. Analysis of variance (ANOVA) was performed to compare the techniques, the patients and upper and lower canines. No significant difference was found for any of the cases. Using LT to evaluate vestibular crest cortical bone in canines is comparable in efficiency to using CBCT. Height in millimeters is less in LT because image resolution is lower and when it is very thin it is not appreciable by this method.


Efficacy of a single-dose regimen of inactivated whole-cell oral cholera vaccine: results from 2 years of follow-up of a randomised trial.
A single-dose regimen of inactivated whole-cell oral cholera vaccine (OCV) is attractive because it reduces logistical challenges for vaccination and could enable more people to be vaccinated. Previously, we reported the efficacy of a single dose of an OCV vaccine during the 6 months following dosing. Herein, we report the results of 2 years of follow-up. In this placebo-controlled, double-blind trial done in Dhaka, Bangladesh, individuals aged 1 year or older with no history of receipt of OCV were randomly assigned to receive a single dose of inactivated OCV or oral placebo. The primary endpoint was a confirmed episode of non-bloody diarrhoea for which the onset was at least 7 days after dosing and a faecal culture was positive for Vibrio cholerae O1 or O139. Passive surveillance for diarrhoea was done in 13 hospitals or major clinics located in or near the study area for 2 years after the last administered dose. We assessed the protective efficacy of the OCV against culture-confirmed cholera occurring 7-730 days after dosing with both crude and multivariable per-protocol analyses. This trial is registered at ClinicalTrials.gov, number NCT02027207. Between Jan 10, 2014, and Feb 4, 2014, 205 513 people were randomly assigned to receive either vaccine or placebo, of whom 204 700 (102 552 vaccine recipients and 102 148 placebo recipients) were included in the per-protocol analysis. 287 first episodes of cholera (109 among vaccine recipients and 178 among placebo recipients) were detected during the 2-year follow-up; 138 of these episodes (46 in vaccine recipients and 92 in placebo recipients) were associated with severe dehydration. The overall incidence rates of initial cholera episodes were 0·22 (95% CI 0·18 to 0·27) per 100 000 person-days in vaccine recipients versus 0·36 (0·31 to 0·42) per 100 000 person-days in placebo recipients (adjusted protective efficacy 39%, 95% CI 23 to 52). The overall incidence of severe cholera was 0·09 (0·07 to 0·12) per 100 000 person-days versus 0·19 (0·15 to 0·23; adjusted protective efficacy 50%, 29 to 65). Vaccine protective efficacy was 52% (8 to 75) against all cholera episodes and 71% (27 to 88) against severe cholera episodes in participants aged 5 years to younger than 15 years. For participants aged 15 years or older, vaccine protective efficacy was 59% (42 to 71) against all cholera episodes and 59% (35 to 74) against severe cholera. The protection in the older age groups was sustained throughout the 2-year follow-up. In participants younger than 5 years, the vaccine did not show protection against either all cholera episodes (protective efficacy -13%, -68 to 25) or severe cholera episodes (-44%, -220 to 35). A single dose of the inactivated whole-cell OCV offered protection to older children and adults that was sustained for at least 2 years. The absence of protection of young children might reflect a lesser degree of pre-existing natural immunity in this age group. Bill & Melinda Gates Foundation to the International Vaccine Institute.


TdP Incidence in Methoxamine-Sensitized Rabbit Model Is Reduced With Age but Not Influenced by Hypercholesterolemia.
Metabolic syndrome is associated with hypercholesterolemia, cardiac remodeling, and increased susceptibility to ventricular arrhythmias. Effects of diet-induced hypercholesterolemia on susceptibility to torsades de pointes arrhythmias (TdP) together with potential indicators of arrhythmic risk were investigated in three experimental groups of Carlsson's rabbit model: (1) young rabbits (YC, young control, age 12-16 weeks), older rabbits (AC, adult control, age 20-24 weeks), and older age-matched cholesterol-fed rabbits (CH, cholesterol, age 20-24 weeks). TdP was induced by α-adrenergic stimulation by methoxamine and IKr block in 83% of YC rabbits, 18% of AC rabbits, and 21% of CH rabbits. High incidence of TdP was associated with high incidence of single (SEB) and multiple ectopic beats (MEB), but the QTc prolongation and short-term variability (STV) were similar in all three groups. In TdP-susceptible rabbits, STV was significantly higher compared with arrhythmia-free rabbits but not with rabbits with other than TdP arrhythmias (SEB, MEB). Amplitude-aware permutation entropy analysis of baseline ECG could identify arrhythmia-resistant animals with high sensitivity and specificity. The data indicate that the TdP susceptibility in methoxamine-sensitized rabbits is affected by the age of rabbits but probably not by hypercholesterolemia. Entropy analysis could potentially stratify the arrhythmic risk and identify the low-risk individuals.


Acetic acid used for the elimination of Pseudomonas aeruginosa from burn and soft tissue wounds.
Acetic acid was used topically at concentrations of between 0.5% and 5% to eliminate Pseudomonas aeruginosa from the burn wounds or soft tissue wounds of 16 patients. In-vitro studies indicated the susceptibility of P. aeruginosa to acetic acid; all strains exhibited a minimum inhibitory concentration of 2 per cent. P. aeruginosa was eliminated from the wounds of 14 of the 16 patients within two weeks of treatment. Acetic acid was shown to be an inexpensive and efficient agent for the elimination of P. aeruginosa from burn and soft tissue wounds.


Intracellular calcium handling heterogeneities in intact guinea pig hearts.
Regional heterogeneities of ventricular repolarizing currents and their role in arrhythmogenesis have received much attention; however, relatively little is known regarding heterogeneities of intracellular calcium handling. Because repolarization properties and contractile function are heterogeneous from base to apex of the intact heart, we hypothesize that calcium handling is also heterogeneous from base to apex. To test this hypothesis, we developed a novel ratiometric optical mapping system capable of measuring calcium fluorescence of indo-1 at two separate wavelengths from 256 sites simultaneously. With the use of intact Langendorff-perfused guinea pig hearts, ratiometric calcium transients were recorded under normal conditions and during administration of known inotropic agents. Ratiometric calcium transients were insensitive to changes in excitation light intensity and fluorescence over time. Under control conditions, calcium transient amplitude near the apex was significantly larger (60%, P < 0.01) compared with the base. In contrast, calcium transient duration was significantly longer (7.5%, P < 0.03) near the base compared with the apex. During isoproterenol (0.05 microM) and verapamil (2.5 microM) administration, ratiometric calcium transients accurately reflected changes in contractile function, and, the direction of base-to-apex heterogeneities remained unchanged compared with control. Ratiometric optical mapping techniques can be used to accurately quantify heterogeneities of calcium handling in the intact heart. Significant heterogeneities of calcium release and sequestration exist from base to apex of the intact heart. These heterogeneities are consistent with base-to-apex heterogeneities of contraction observed in the intact heart and may play a role in arrhythmogenesis under abnormal conditions.


Revealing the Chemical Nature of Functional Groups on Graphene Oxide by Integrating Potentiometric Titration and Ab Initio Calculations.
A new graphene oxide (GO) model with reasonable functional group types and distribution modes was proposed by integrating potentiometric titrations and ab initio calculations. Due to the complex synthesis mechanism, the atomic structure of GO has been controversial for a long time. Here, we use density functional theory calculations to mimic the oxidation process, and a series of GO fragments (GOFs) were deduced. A new pKa calculation method (RCDPKA) developed specifically in this work was further used to predict pKa values of the fragments. Then, we performed potentiometric titrations on four different GO samples to confirm the existence of these GOFs and determine the content of functional groups. Interestingly, different GO samples present the same pKa values in titration, and the results are consistent with the predicted ones. Based on the evidence from titration and calculation, prominent correlations between functional groups could be found. Groups at the edges are mainly double-interactive carboxyls (pKa1 ≈ 3.4, pKa2 ≈ 5.7) and double-adjacent phenolic hydroxyls (pKa1 ≈ 8.8, pKa2 ≈ 12.1), while groups on the plane are mainly collocated epoxies and hydroxyls (pKa1 ≈ 11.1, pKa2 ≈ 13.8) on both sides of the plane with a meta-positional hydrogen bond interaction. These findings were further validated by multiple characterizations and GO modifications. These results not only stimulate a fundamental understanding of the GO structure but also provide a quantitative analysis method for functional groups on GO.


Advanced practice in neurocritical care: an innovative orientation and competency model.
The advanced registered nurse practitioner (ARNP) began in the 1960s as an alternative provider to meet the demands of an escalating healthcare resource deficit. As the role evolved and ARNPs demonstrated safe and effective care, these providers began to appear in critical care settings. It is believed that in the specialty of Neurocritical Care, about half the providers are ARNPs. Hiring and training practitioners for this complex environment is daunting. At the University of Florida & Shands Jacksonville, an innovative orientation and competency model for ARNPs hired for the newly opened Neurocritical Care unit was developed and implemented. The program contains a roadmap for knowledge base and skill acquisition as well as competency training and maintenance. Experience with appropriate hiring and screening standards, internally developed training tools, and identification of necessary advanced classes are discussed. This model may be used as a guideline for Neurocritical Care ARNP training as well as adapted for all other critical care settings.


Disrupted-In-Schizophrenia 1 regulates integration of newly generated neurons in the adult brain.
Adult neurogenesis occurs throughout life in discrete regions of the adult mammalian brain. Little is known about the mechanism governing the sequential developmental process that leads to integration of new neurons from adult neural stem cells into the existing circuitry. Here, we investigated roles of Disrupted-In-Schizophrenia 1 (DISC1), a schizophrenia susceptibility gene, in adult hippocampal neurogenesis. Unexpectedly, downregulation of DISC1 leads to accelerated neuronal integration, resulting in aberrant morphological development and mispositioning of new dentate granule cells in a cell-autonomous fashion. Functionally, newborn neurons with DISC1 knockdown exhibit enhanced excitability and accelerated dendritic development and synapse formation. Furthermore, DISC1 cooperates with its binding partner NDEL1 in regulating adult neurogenesis. Taken together, our study identifies DISC1 as a key regulator that orchestrates the tempo of functional neuronal integration in the adult brain and demonstrates essential roles of a susceptibility gene for major mental illness in neuronal development, including adult neurogenesis.


CPIELA: Computational Prediction of Plant Protein-Protein Interactions by Ensemble Learning Approach From Protein Sequences and Evolutionary Information.
Identification and characterization of plant protein-protein interactions (PPIs) are critical in elucidating the functions of proteins and molecular mechanisms in a plant cell. Although experimentally validated plant PPIs data have become increasingly available in diverse plant species, the high-throughput techniques are usually expensive and labor-intensive. With the incredibly valuable plant PPIs data accumulating in public databases, it is progressively important to propose computational approaches to facilitate the identification of possible PPIs. In this article, we propose an effective framework for predicting plant PPIs by combining the position-specific scoring matrix (PSSM), local optimal-oriented pattern (LOOP), and ensemble rotation forest (ROF) model. Specifically, the plant protein sequence is firstly transformed into the PSSM, in which the protein evolutionary information is perfectly preserved. Then, the local textural descriptor LOOP is employed to extract texture variation features from PSSM. Finally, the ROF classifier is adopted to infer the potential plant PPIs. The performance of CPIELA is evaluated via cross-validation on three plant PPIs datasets: Arabidopsis thaliana, Zea mays, and Oryza sativa. The experimental results demonstrate that the CPIELA method achieved the high average prediction accuracies of 98.63%, 98.09%, and 94.02%, respectively. To further verify the high performance of CPIELA, we also compared it with the other state-of-the-art methods on three gold standard datasets. The experimental results illustrate that CPIELA is efficient and reliable for predicting plant PPIs. It is anticipated that the CPIELA approach could become a useful tool for facilitating the identification of possible plant PPIs.


A small molecule species specifically inhibits Fusarium myosin I.
Fusarium head blight (FHB) caused by Fusarium graminearum is a devastating disease of cereal crops worldwide. Recently, a novel fungicide JS399-19 has been launched into the marketplace to manage FHB. It is compelling that JS399-19 shows highly inhibitory activity towards some Fusarium species, but not to other fungi, indicating that it is an environmentally compatible fungicide. To explore the mode of action of this species-specific compound, we conducted a whole-genome transcript profiling together with genetic and biochemical assays, and discovered that JS399-19 targets the myosin I of F. graminearum (FgMyo1). FgMyo1 is essential for F. graminearum growth. A point mutation S217L or E420K in FgMyo1 is responsible for F. graminearum resistance to JS399-19. In addition, transformation of F. graminearum with the myosin I gene of Magnaporthe grisea, the causal agent of rice blast, also led to JS399-19 resistance. JS399-19 strongly inhibits the ATPase activity of the wild-type FgMyo1, but not the mutated FgMyo1(S217L/E420K) . These results provide us a new insight into the design of species-specific antifungal compounds. Furthermore, our strategy can be applied to identify novel drug targets in various pathogenic organisms.


Coxsackievirus B3-induced myocarditis. Autoimmunity is L3T4+ T helper cell and IL-2 independent in BALB/c mice.
Male BALB/c mice inoculated with 6 X 10(4) plaque-forming units (pfu) coxsackievirus, group B, type 3 (CVB3), develop myocarditis within 7 days. Two cytolytic T lymphocyte (CTL) populations arise in infected animals. One population belongs to the Lyt 2+ T (cytolytic/suppressor) lymphocyte subset and reacts specifically with uninfected heart cells (autoreactive CTLs, ACTLs), whereas the other belongs to the L3T4+ T (helper) lymphocyte subset and reacts with infected targets (virus-specific CTLs, VSCTLs). Although both immune T lymphocyte populations can induce cardiac inflammation in vivo, ACTLs predominantly cause tissue injury. VSCTL generation can be inhibited by either anti-Tac (antibody to the interleukin 2 [IL-2] receptor) or anti-Iad but not by anti-IAk, indicating that this response is probably both IL-2-dependent and Class II (major histocompatibility complex [MHC] antigen restricted. ACTL generation is independent of IL-2, because neither anti-Tac or cyclosporin A inhibit this response.


Development of fluorescent methods for DNA methyltransferase assay.
DNA methylation modified by DNA methyltransferase (MTase) plays an important role in regulating gene transcription, cell growth and proliferation. The aberrant DNA MTase activity may lead to a variety of human diseases including cancers. Therefore, accurate and sensitive detection of DNA MTase activity is crucial to biomedical research, clinical diagnostics and therapy. However, conventional DNA MTase assays often suffer from labor-intensive operations and time-consuming procedures. Alternatively, fluorescent methods have significant advantages of simplicity and high sensitivity, and have been widely applied for DNA MTase assay. In this review, we summarize the recent advances in the development of fluorescent methods for DNA MTase assay. These emerging methods include amplification-free and the amplification-assisted assays. Moreover, we discuss the challenges and future directions of this area.


Vegetarian diets in children: a systematic review.
While the prevalence of children on vegetarian diets is assumed to be on the rise in industrialized countries, there are hardly any representative data available. In general, vegetarian diets are presumed to be healthy; nevertheless, there are concerns as to whether the dietary specifications required during infancy, childhood, and adolescence can be met. Therefore, the objective of this systematic review was to evaluate studies on the dietary intake and the nutritional or health status of vegetarian infants, children, and adolescents. The database MEDLINE was used for literature search. In addition, references of reviews and expert opinions were considered. Inclusion criteria were (1) sufficient dietary information to define vegetarian type diet and (2) characteristics of nutritional or health status. Case reports and studies from non-industrialized countries were excluded. 24 publications from 16 studies published from 1988 to 2013 met our criteria. Study samples covered the age range from 0 to 18 years, and median sample size was 35. Five studies did not include a control group. With regard to biomarkers, anthropometry, and dietary or nutritional intake, the outcomes were diverse. Growth and body weight were generally found within the lower reference range. The intakes of folate, vitamin C, and dietary fiber were relatively high compared to reference values and/or control groups. Low status of vitamin B12 was reported in one study and low status of vitamin D in two studies. Due to the study heterogeneity, the small samples, the bias towards upper social classes, and the scarcity of recent studies, the existing data do not allow us to draw firm conclusions on health benefits or risks of present-day vegetarian type diets on the nutritional or health status of children and adolescents in industrialized countries.


Combination of immunotherapy with targeted therapies in advanced non-small cell lung cancer (NSCLC).
Treatment for advanced non-small cell lung cancer (NSCLC) has been significantly improved in recent years with the incorporation of drugs targeting antiangiogenesis and more specifically genomic alterations such as the EGFR mutations and ALK translocations. However, most patients invariably progress and die. The emergence of immune checkpoint inhibitors targeting the pathways involved in tumor-induced immunosuppression have redefined the management of the disease, achieving significant long-lasting responses with manageable safety profiles, regardless of histology. Still, response rates with immunotherapy are deemed suboptimal. Current efforts are focusing on new potential combination strategies with synergistic antitumor activity, using immune checkpoint blockade as a partner for targeted agents. Herein we discuss the available data on the combined use of immunotherapy, including PD-1/PD-L1 and CTLA-4 inhibitors, with EGFR and ALK inhibitors and comment on the current status of immunotherapy plus antiangiogenic drugs for molecularly unselected advanced NSCLC.


Together stronger: boundary work within an Australian systems-based prevention initiative.
Complexity and systems science are increasingly used to devise interventions to address health and social problems. Boundaries are important in systems thinking, as they bring attention to the power dynamics that guide decision-making around the framing of a situation, and how it is subsequently tackled. Using complexity theory as an analytical frame, this qualitative exploratory study examined boundary interactions between local government and community health organizations during the operationalization of a systems-based initiative to prevent obesity and chronic diseases (Healthy Together Communities-HTCs) in Victoria, Australia. Across two HTC sites, data was generated through semi-structured interviews with 20 key informants, in mid-2015. Template analysis based on properties of complex systems was applied to the data. The dynamics of boundary work are explored using three case illustrations: alignment, boundary spanning and boundary permeability. Alignment was both a process and an outcome of boundary work, and occurred at strategic, operational and individual levels. Boundary spanning was an important mechanism to develop a unified collaborative approach, and ensure that mainstream initiatives reached disadvantaged groups. Finally, some boundaries exhibited different levels of permeability for local government and community health organizations. This influenced how each organization could contribute to HTC interventions in unique, yet complementary ways. The study of boundary work offers potential for understanding the mechanisms that contribute to the nonlinear behaviour of complex systems. The complementarity of partnering organizations, and boundary dynamics should be considered when designing and operationalizing multilevel, complex systems-informed prevention initiatives.


The nuclear uptake and retention of a synthetic progestin in the cardiovascular system of the baboon.
It has long been known that there is a sexual dimorphism in the incidence of coronary heart disease. This observation, together with more recent reports of increased cardiovascular disease associated with the use of oral contraceptives, led to a search for steroid receptors in the cardiovascular system. In this study we examined the nuclear uptake and retention of a synthetic progestin in the cardiovascular system of the baboons. Long term oophorectomized baboons were primed with estradiol benzoate for 3 days before the experiment (50 micrograms/kg, im) and adrenalectomized 2 days before the experiment. On the day of the experiment, the animals were injected under anesthesia with 2.5 micrograms/kg BW [3H]ORG 2058 (16 alpha-ethyl-21-hydroxy-19-nor-[6,7-3H]pregn-4-ene-3,20-dione) or with [3H] ORG 2058 plus a 1000-fold excess of unlabeled progesterone (control). One hour after the injection, the animals were rapidly exsanguinated, and parts of the cardiovascular system were removed and processed for autoradiography. Localization of the synthetic progestin was found in nuclei of between 25-75% of all smooth muscle cells of the media of all arteries examined and to a lesser extent in the nuclei of the fibroblasts and others cells of the adventitia. Localization of the synthetic progestin in the heart was limited to approximately 1% of the myocardial cells and less than 5% of interstitial cell nuclei. The pattern of localization found differs from that for estrogen and androgen and suggests the possible presence of estrogen-independent progesterone receptors in smooth muscle cells of the media of the aorta and coronary arteries.