The First Paenibacillus larvae Bacteriophage Endolysin (PlyPl23) with High Potential to Control American Foulbrood.
Endolysins, which are peptidoglycan-degrading enzymes expressed during the terminal stage of the reproduction cycle of bacteriophages, have great potential to control Gram-positive pathogens. This work describes the characterization of a novel endolysin (PlyPl23) encoded on the genome of Paenibacillus larvae phage phiIBB_Pl23 with high potential to control American foulbrood. This bacterial disease, caused by P. larvae, is widespread in North America and Europe and causes important economic losses in apiculture. The restriction to antibiotic residues in honey imposed by the EU legislation hinders its therapeutic use to combat American foulbrood and enforces the development of alternative antimicrobial methods. The new endolysin described herein has an N-acetylmuramoyl-L-alanine amidase catalytic domain and exhibits a broad-spectrum activity against common P. larvae genotypes. Moreover, the enzyme displays high antimicrobial activity in a range of pH that matches environmental conditions (pH between 5.0 and 7.0), showing its feasible application in the field. At pH 7.0, a concentration of 0.2 μM of enzyme was enough to lyse 104 CFU.mL-1 of P. larvae in no more than 2 h. The presence of sucrose and of the substances present in the larvae gut content did not affect the enzyme activity. Interestingly, an increase of activity was observed when PlyPl23 was previously incubated in royal jelly. Furthermore, in vivo safety evaluation assays demonstrated that this enzyme is not toxic to the bee larvae. The present work describes for the first time an endolysin encoded in a P. larvae phage that presents high potential to integrate a commercial product to control the problematic American foulbrood.


Effect of hyperthyroidism and its treatment on bone mineral content.
Patients with hyperthyroidism may develop osteopenia associated with fractures; however, there has been no general agreement on the incidence of osteopenia in hyperthyroidism or the recovery of the mineral loss after treatment of hyperthyroidism. We conducted a longitudinal prospective study on the effect of hyperthyroidism and its treatment on bone mineral content (BMC) using photon absorptiometry. We observed that both young and older hyperthyroid patients showed a significantly decreased baseline BMC compared with age- and sex-matched controls. We also observed a slight recovery of BMC in hyperthyroid patients at the two-year interval after a euthyroid state had been achieved. However, the BMC was still much lower than that of controls, and we did not find any significant restoration of BMC following "cure" of hyperthyroidism.


An approach to incorporate individual personality in modeling fish dispersal across in-stream barriers.
Animal personalities are an important factor that affects the dispersal of animals. In the context of aquatic species, dispersal modeling needs to consider that most freshwater ecosystems are highly fragmented by barriers reducing longitudinal connectivity. Previous research has incorporated such barriers into dispersal models under the neutral assumption that all migrating animals attempt to ascend at all times. Modeling dispersal of animals that do not perform trophic or reproductive migrations will be more realistic if it includes assumptions of which individuals attempt to overcome a barrier. We aimed to introduce personality into predictive modeling of whether a nonmigratory invasive freshwater fish (the round goby, Neogobius melanostomus) will disperse across an in-stream barrier. To that end, we experimentally assayed the personalities of 259 individuals from invasion fronts and established round goby populations. Based on the population differences in boldness, asociability, and activity, we defined a priori thresholds with bolder, more asocial, and more active individuals having a higher likelihood of ascent. We then combined the personality thresholds with swimming speed data from the literature and in situ measurements of flow velocities in the barrier. The resulting binary logistic regression model revealed probabilities of crossing a barrier which depended not only on water flow and fish swimming speed but also on animal personalities. We conclude that risk assessment through predictive dispersal modeling across fragmented landscapes can be advanced by including personality traits as parameters. The inclusion of behavior into modeling the spread of invasive species can help to improve the accuracy of risk assessments.


Role of calcium-activated chloride current in regulating pulmonary vasomotor tone.
Many agonists induce vasoconstriction by releasing intracellularly stored Ca2+ and promoting Ca2+ influx. Activation of Ca(2+)-activated Cl- (ClCa) channels may be a critical mechanism by which a rise in intracellular free Ca2+ concentration ([Ca2+]i) causes membrane depolarization that serves to sustain the elevated [Ca2+]i and maintain vascular tone. In this study the biophysical and pharmacological properties of ClCa currents [ICKCa] were characterized in rat pulmonary artery (PA) smooth muscle cells, and their relationship to the regulation of pulmonary vascular tone was determined. When K+ currents were eliminated by using Cs(+)-containing internal solution, depolarization elicited an inward Ca2+ current followed by a time-dependent outward Cl- current that reversed near Cl- equilibrium potential. Repolarizing voltage steps produced a large inward tail Cl- current that also reversed at a potential close to Cl- equilibrium potential. Replacement of extracellular Ca2+ with Ba2+ significantly augmented the Ca2+ current but abolished the Cl- currents. The Cl- channel blocker niflumic acid (10-50 microM) diminished the time-dependent outward Cl- current and the inward tail Cl- current, decreased serotonin-induced membrane depolarization, and inhibited agonist-induced PA contraction. In the absence of extracellular Ca2+, cyclopiazonic acid, which releases Ca2+ from sarcoplasmic reticulum, elicited an inward Cl- current at a holding potential of -70 mV. These results indicate that rat PA myocytes possess ClCa channels that are activated by depolarization-induced Ca2+ influx and agonist-induced Ca2+ release. This Cl- current contributes to agonist-induced pulmonary vasoconstriction via membrane depolarization.


Synthesis of NiMo/La-Al2O3 powders for efficient catalytic transesterification of triglyceride with the high yield of 95.2.
Enhancement of the transesterification efficiency of triglyceride has come under heated study in biodiesel-making industry. In this research, the NiMo/La-Al2O3 nanopowders have been prepared for producing biodiesel efficiently. The screening test showed that the NiMo/La-Al2O3 catalyst has the best catalytic activity for triglyceride transesterification. Besides, the process parameters including reaction temperature, time, oil-to-alcohol ratio and catalyst loading etc., also have been investigated for optimization of the transesterification process. The results showed that with 5 wt% of catalyst loading, and oil to methanol molar ratio of 1:9, the conversion yield of triglyceride could be up to 95.2% within 120 min at 160°C. The NiMo/La-Al2O3 catalyst has the outstanding recycle property, which proved that the prepared NiMo/La-Al2O3 powders can be suitable for biodiesels' production.


Opinion and Special Article: The Need for Specialized Training in Women's Neurology.
Women's Neurology is an emerging subspecialty that focuses on neurologic disorders across a woman's lifetime. This new domain recognizes that both health and disease are directly affected by hormonal and reproductive changes throughout the life span. This field includes neurologic diseases with a higher prevalence in women and diseases that require specialized management during pregnancy, postpartum period, lactation, and menopause. A survey was sent to US neurology residency program directors to understand the state of training in the area. Their responses highlighted an urgent need for additional education in this field for neurology residents. In this study, we discuss the educational gaps in this area, the clinical benefits of a women's neurology discipline, and the instructional gaps in this area and provide practical recommendations for training programs in women's neurology using 2 innovative fellowship programs.


Mouse and rat plasma renin concentration and gene expression in (mRen2)27 transgenic rats.
The (mRen2)27 transgenic rat [TGR(mRen2)27] is said to have low plasma levels of active renin. We used a direct radioimmunoassay (RIA) for mouse submaxillary renin, as well as an indirect enzyme-kinetic assay based on the generation of angiotensin I with modification of the pH optimum, to measure rat and mouse plasma renin activity (PRA), plasma renin concentration (PRC), and plasma prorenin in TGR before and after lisinopril. The relationship between rat PRC and % rat kidney extract was steepest at pH 6.0 and flat at pH 8.5, whereas the relationship between mouse PRC and purified mouse renin was steepest at pH 8.5 and flat at pH 6.0. Mouse PRC was highly correlated with direct RIA measurements (r = 0.93). PRA before lisinopril was little influenced by pH, whereas the increase with lisinopril was greatest at pH 6.5. PRC before lisinopril was fourfold higher at pH 8.5 compared with that at pH 6.0. Lisinopril increased both PRC values but reversed the pH dependency. Prorenin was fourfold higher at pH 8.5 compared with that at pH 6.0 and decreased slightly with lisinopril. Renal renin concentration was higher at pH 6.0 than at pH 8.5. With lisinopril, renal renin concentration increased at both pH values. Mouse PRC was not changed by lisinopril. Ribonuclease protection assay showed both rat and mouse renin gene expression in the kidney, which increased with lisinopril. Thus TGR have circulating active rat and mouse renin and prorenin. The notion that TGR are a "low renin" model should be revised.


Brain age predicts mortality.
Age-associated disease and disability are placing a growing burden on society. However, ageing does not affect people uniformly. Hence, markers of the underlying biological ageing process are needed to help identify people at increased risk of age-associated physical and cognitive impairments and ultimately, death. Here, we present such a biomarker, 'brain-predicted age', derived using structural neuroimaging. Brain-predicted age was calculated using machine-learning analysis, trained on neuroimaging data from a large healthy reference sample (N=2001), then tested in the Lothian Birth Cohort 1936 (N=669), to determine relationships with age-associated functional measures and mortality. Having a brain-predicted age indicative of an older-appearing brain was associated with: weaker grip strength, poorer lung function, slower walking speed, lower fluid intelligence, higher allostatic load and increased mortality risk. Furthermore, while combining brain-predicted age with grey matter and cerebrospinal fluid volumes (themselves strong predictors) not did improve mortality risk prediction, the combination of brain-predicted age and DNA-methylation-predicted age did. This indicates that neuroimaging and epigenetics measures of ageing can provide complementary data regarding health outcomes. Our study introduces a clinically-relevant neuroimaging ageing biomarker and demonstrates that combining distinct measurements of biological ageing further helps to determine risk of age-related deterioration and death.


Detection of high levels of heparin binding growth factor-1 (acidic fibroblast growth factor) in inflammatory arthritic joints.
The synovium from patients with rheumatoid arthritis (RA) and LEW/N rats with streptococcal cell wall (SCW) arthritis, an experimental model resembling RA, is characterized by massive proliferation of synovial connective tissues and invasive destruction of periarticular bone and cartilage. Since heparin binding growth factor (HBGF)-1, the precursor of acidic fibroblast growth factor (FGF), is a potent angiogenic polypeptide and mitogen for mesenchymal cells, we sought evidence that it was involved in the synovial pathology of RA and SCW arthritis. HBGF-1 mRNA was detected in RA synovium using the polymerase chain reaction technique, and its product was immunolocalized intracellularly in both RA and osteoarthritis (OA) synovium. HBGF-1 staining was more extensive and intense in synovium of RA patients than OA and correlated with the extent and intensity of synovial mononuclear cell infiltration. HBGF-1 staining also correlated with c-Fos protein staining. In SCW arthritis, HBGF-1 immunostaining was noted in bone marrow, bone, cartilage, synovium, ligamentous and tendinous structures, as well as various dermal structures and developed early in both T-cell competent and incompetent rats. Persistent high level immunostaining of HBGF-1 was only noted in T-cell competent rats like the disease process in general. These observations implicate HBGF-1 in a multitude of biological functions in inflammatory joint diseases.


[Circadian rhythm and encephalic vascular disease: a correlative study with risk factors].
The human biological mechanisms show a predictable clinical variability in time, which has allowed a deeper reevaluation of present-day medical practices, regarding the circadian rhythm (CR) and the mechanisms that produce the supported variations in all biological levels. We have made a study aiming to relate the CR and onset of the neurological clinic situation due to the encephalic vascular lesion, correlating with modifying risk factors. Fifty three patients were studied, 50,94% female (n=27) and 49,50% male (n=26), at average age 66.4 years old. Four intervals of six hours each (0-6; 6-12; 12-18; 18-24) were used to analyze the frequency of the ictus and the incidence in each interval. We found an incidence of 6(11.32%) patients in the 0-6 hs interval; 21 (39.62%) patients in the 6-12 hs interval; 10(18.86%) patients in the 12-18 hs interval; 16 (30.18%) patients in the 18-24 interval. A correlative study with the risk factors has shown that arterial hypertension [(81.25%)] and smoking habit [ (56.25)] were predominant during the 18-24 hs interval, while sedentary [11(52.38%)] stress [11(52.38%)] diabetes [(47.61%)] hyperlipidemia [8 (38.09%)] and alcoholism [8 (38.09%] were predominant during the 6-12 hs interval; and cardiac diseases in the 12-18hs interval.


Functional versus folding landscapes: the same yet different.
Protein functional landscapes are characterized by a modest number of states compared with the folding landscapes, allowing brute force sampling of these states for smaller proteins using computer simulations. On the other hand, because the functional landscape topographies are complicated, the native state dynamics are often difficult to interpret. Nevertheless, a number of experimental and computational techniques have recently emerged that are designed to reveal the essential features of the native landscape, such as the hierarchical organization of conformational substates. These studies also shed light on the mechanisms of protein function, for example, explaining how chemical energy is transduced in molecular motors. Overall, interpreting experimental results in the light of the functional landscape paradigm considerably enhances the understanding of complex biomolecular processes.


Methylenetetrahydrofolate reductase (MTHFR) C677T polymorphism is associated with spinal BMD in 9-year-old children.
The C677T MTHFR polymorphism has been associated with lumbar spine and hip BMD. In older adults, the genetic effect has been reported in women only. However, in younger adults, this influence may only be present in men. This study is the first to investigate associations between the C677T MTHFR polymorphism and bone phenotypes in children. Regression analyses were used to study the relationship between MTHFR genotype and bone phenotypes derived from total body DXA scans in children 9.9 yr of age from the Avon Longitudinal Study of Parents and Children (ALSPAC). A total of 5816 children had both genetic and DXA data for the total body less head region (TBLH) and 3196 for the spine. A strong association was observed between the C677T MTHFR genotype and spine BMD (p < 0.001; 0.10 SD decrease per T allele). There was some evidence that this genetic effect was stronger in boys compared with girls (p = 0.04 for sex interaction). In contrast, there was no association between the C677T MTHFR genotype and TBLH BMD. The association between MTHFR genotype and spine BMD was attenuated particularly in girls by high maternal dietary intakes of vitamin B(6) and folate during pregnancy but not by child dietary intakes at 7 yr. To the extent that these findings reflect known influences of C677T MTHFR genotype on plasma homocysteine levels, our results suggest that the latter is an important regulator of spinal BMD in childhood.


IgG antibodies to Aspergillus fumigatus in cystic fibrosis: a laboratory correlate of disease activity.
Serum was collected from 50 patients with cystic fibrosis, and IgG antibodies to Aspergillus fumigatus were measured by enzyme linked immunosorbent assay (ELISA). In addition, total IgE and Aspergillus specific IgE antibodies were measured in 41 of the 50. A close association was found between pulmonary function and clinical state, and IgG antibodies to Aspergillus. There was no association between pulmonary function or clinical state and IgE antibodies. It is postulated that in patients with cystic fibrosis, Aspergillus fumigatus may contribute to deterioration in pulmonary function by local pathogenicity, or by hypersensitivity mechanisms mediated by IgG.


Return to work after Post-COVID: describing affected employees' perceptions of personal resources, organizational offerings and care pathways.
Most individuals recover from the acute phase of infection with the SARS-CoV-2 virus, however, some encounter prolonged effects, referred to as the Post-COVID syndrome. Evidence exists that such persistent symptoms can significantly impact patients' ability to return to work. This paper gives a comprehensive overview of different care pathways and resources, both personal and external, that aim to support Post-COVID patients during their work-life reintegration process. By describing the current situation of Post-COVID patients pertaining their transition back to the workplace, this paper provides valuable insights into their needs. A quantitative research design was applied using an online questionnaire as an instrument. Participants were recruited via Post-COVID outpatients, rehab facilities, general practitioners, support groups, and other healthcare facilities. The analyses of 184 data sets of Post-COVID affected produced three key findings: (1) The evaluation of different types of personal resources that may lead to a successful return to work found that particularly the individuals' ability to cope with their situation (measured with the FERUS questionnaire), produced significant differences between participants that had returned to work and those that had not been able to return so far (F = 4.913, p = 0.001). (2) In terms of organizational provisions to facilitate successful reintegration into work-life, predominantly structural changes (i.e., modification of the workplace, working hours, and task) were rated as helpful or very helpful on average (meanworkplace 2.55/SD = 0.83, meanworking hours 2.44/SD = 0.80; meantasks 2.55/SD = 0.83), while the remaining offerings (i.e., job coaching or health courses) were rated as less helpful or not helpful at all. (3) No significant correlation was found between different care pathways and a successful return to work. The results of the in-depth descriptive analysis allows to suggests that the level of ability to cope with the Post-COVID syndrome and its associated complaints, as well as the structural adaptation of the workplace to meet the needs and demands of patients better, might be important determinants of a successful return. While the latter might be addressed by employers directly, it might be helpful to integrate training on coping behavior early in care pathways and treatment plans for Post-COVID patients to strengthen their coping abilities aiming to support their successful return to work at an early stage.


Differential sensitivity to 2-deoxy-D-glucose between two pancreatic cell lines correlates with GLUT-1 expression.
To determine whether the differential growth inhibition of pancreatic tumor cells to the glycolytic inhibitor 2-deoxy-D-glucose (2-DG) correlates with uptake, expression of GLUT-1 transporter, and levels of hypoxic-inducible factor-1alpha. Growth inhibition assays with 2-DG, lactic acid analysis, Western blots of GLUT-1, and hypoxic-inducible factor-1alpha were correlated with each other and with uptake and accumulation of radio-labeled 2-DG in 2 pancreatic cell lines. Under normal oxygen tension, we find that the pancreatic cell line MIA PaCa2 (1420) is 7 times more sensitive to 2-DG and equally sensitive to oxamate, as compared with Panc-1 (1469). Lactate levels in both cell types are similarly low, indicating that mitochondria are functioning normally in these cells and that they are not solely dependent on glycolysis for survival under an aerobic microenvironment. Since oxamate does not use glucose transporters for entry into the cell, the equal sensitivity to this drug suggests that the selective growth inhibition of 2-DG in these 2 cell types might be reflective of differential expression of glucose transporters. Indeed, we find that GLUT-1 is more highly expressed in 1420 and that this cell line accumulates 2-DG twice as much as 1469. Additionally, hypoxic-inducible factor, which is known to upregulate the expression of GLUT-1, is found at equally low levels in both cell types. Thus, the increased expression of GLUT-1 in 1420 appears to be independent of hypoxia-inducible factor. Overall, our results suggest that certain pancreatic tumors may be inherently sensitive to 2-DG, even under normal oxygen tension, due to greater intracellular accumulation of this inhibitor. Moreover, if 2-DG shows clinical efficacy, it may be possible to predict which pancreatic tumors would be sensitive to this agent based on their GLUT-1 expression profile and their increased uptake of 2-fluoro-deoxy-D-glucose currently used to image tumors via PET scanning.


Key and recollection vector effects on heteroassociative memory performance.
Most associative memory work has concentrated on autoassociative memories (AAMs). These associative processors provide reduced noise and error correction in their output data. We will consider heteroassociative memories (HAMs), which are needed to provide decisions on the class of the input data and inferences for subsequent processing. We derive new equations for the storage capacity and noise performance of HAMs, emphasize how they differ from those derived for AAMs, suggest new performance measures to be used, and show how different recollection vector encodings can improve HAM performance.


Uptake of direct acting antiviral therapies for the treatment of hepatitis C virus among people who inject drugs in a universal health-care system.
Direct acting antiviral (DAA) treatment can reduce hepatitis C virus (HCV) infection incidence and mortality, although large scale uptake of these treatments is necessary to achieve those reductions. Targeting people who inject drugs (PWID) will be crucial to achieve the necessary reductions. Previously, treatment uptake has been very low (1-2%) among PWID, but these new DAAs have been subsidised in Australia since March 2016. This study describes treatment uptake among a sample of PWID in Australia and examines sociodemographic and drug use variables associated with treatment uptake. An Australian national cross-sectional sample (n = 817) of regular PWID drawn from a convenience sentinel sample aged ≥18 years. Information on demographics, drug use, HCV antibody testing, RNA testing, recent DAA treatment initiation and DAA prescriber settings was collected via an interviewer-administered survey. There were 291 HCV RNA positive participants, of who one-third reported initiating DAA treatment since the introduction of subsidised DAA medications. Current opioid substitution treatment was significantly associated with increased treatment uptake (adjusted odds ratio 2.20, 95% confidence interval 1.31, 3.68) while those with unstable housing were less likely to initiate treatment (adjusted odds ratio 0.42, 95% confidence interval 0.22, 0.82). This study found that DAA treatment uptake has increased among current PWID, a priority population for treatment. Efforts to maintain this uptake through linking current PWID with treatment providers through harm reduction services such as opioid substitution therapy, needle and syringe programs, and outreach services are required to meet incidence and mortality reduction targets.


In Doxorubicin-Adapted Hodgkin Lymphoma Cells, Acquiring Multidrug Resistance and Improved Immunosuppressive Abilities, Doxorubicin Activity Was Enhanced by Chloroquine and GW4869.
Classical Hodgkin lymphoma (cHL) is a highly curable disease (70-80%), even though long-term toxicities, drug resistance, and predicting clinical responses to therapy are major challenges in cHL treatment. To solve these problems, we characterized two cHL cell lines with acquired resistance to doxorubicin, KM-H2dx and HDLM-2dx (HRSdx), generated from KM-H2 and HDLM-2 cells, respectively. HRSdx cells developed cross-resistance to vinblastine, bendamustin, cisplatin, dacarbazine, gemcitabine, brentuximab vedotin (BV), and γ-radiation. Both HDLM-2 and HDLM-2dx cells had intrinsic resistance to BV but not to the drug MMAE. HDLM-2dx acquired cross-resistance to caelyx. HRSdx cells had in common decreased CD71, CD80, CD54, cyt-ROS, HLA-DR, DDR1, and CD44; increased Bcl-2, CD58, COX2, CD26, CCR5, and invasive capability; increased CCL5, TARC, PGE2, and TGF-β; and the capability of hijacking monocytes. In HRSdx cells less sensitive to DNA damage and oxidative stress, the efflux drug transporters MDR1 and MRP1 were not up-regulated, and doxorubicin accumulated in the cytoplasm rather than in the nucleus. Both the autophagy inhibitor chloroquine and extracellular vesicle (EV) release inhibitor GW4869 enhanced doxorubicin activity and counteracted doxorubicin resistance. In conclusion, this study identifies common modulated antigens in HRSdx cells, the associated cross-resistance patterns, and new potential therapeutic options to enhance doxorubicin activity and overcome resistance.


Congestive Heart Failure at the Department of Medicine: Demographic and Clinical Characteristics.
Congestive heart failure (CHF) with reduced ejection fraction (HFrEF) or with preserved ejection fraction (HFpEF) is a common diagnosis in patients hospitalized in the department of internal medicine. Recently, the therapeutic regimens were updated, as the sodium-glucose cotransporter-2 (SGLT2) inhibitors became an integral part of the therapeutic regimen for either HFrEF or HFpEF. To define the demographic and clinical characteristics of CHF patients hospitalized in the department of medicine. We conducted a retrospective cohort study that included all patients hospitalized in the departments of medicine at the Rabin Medical Center, Israel, between 2016 and 2019. Demographic and clinical background, in-hospital procedures, discharge regimens, and outcome parameters were evaluated according to HFrEF/HFpEF. The cohort included 4458 patients. The majority (97%) presented with a preexisting diagnosis, whereas HF was an active condition in only half of them. The rates of HFrEF/HFpEF were equal. In most cases, the trigger of the exacerbation could not be determined; however, infection was the most common cause. There were basic differences in the demography, clinical aspects, and therapeutic regimens at discharge between HFrEF and HFpEF. Both conditions were associated with high in hospital mortality (8%) and re-admissions rates (30 days [20%], 90 days [35%]) without any difference between them. HFrEF/HFpEF patients differed by demographics and co-morbidities. They were equally represented among patients admitted to medical wards and had similar prognosis. For both diagnoses, hospitalization should be considered for updating therapeutic regimens, especially with SGLT2 inhibitors.


Post-traumatic left ventricular false aneurysm.
Most false aneurysms of the heart represent contained ventricular free wall ruptures after myocardial infarction. Post-traumatic aneurysms also may follow penetrating or non-penetrating trauma to the chest. Regardless of the origin of the false aneurysm there is a propensity for aneurysm rupture. We report a patient who developed a false aneurysm of her left ventricle that developed post-motor vehicle accident. Her orthopedic problems were the clinical problems identified and after a hospital admission of 10 days she was discharged home. Four weeks later she died suddenly from anterior left ventricle false aneurysm rupture and tamponaide. Patients with significant chest wall trauma should be assessed for cardiac pathology prior to discharge. Presentation may be delayed and be overshadowed by more evident pathology. Trauma-related aneurysms may cause sudden death, and this may occur some later time after the trauma. Attributing the cause of death to the trauma, which may be remote, is important for the forensic investigator to remember.


Plasticity in morphogenetic expression in plant.
This article discusses the plasticity of morphogenetic programmes in plants. It outlines the numerous ways in which the flowering programme can be accelerated or modified into partial flower formation and describes the differentiation of naked ovules on anthers in intact plants. The generation of embryos and flowers from cultured cells is then described and some of the properties of hypohaploids outlined. It is concluded that in vitro culture provides a useful facility for manipulating plant development and that the interaction between the nucleus and the cytoplasm may exemplify and explain the process of developmental plasticity.


Assessment of corneal topography indices after collagen crosslinking for keratoconus.
Riboflavin UVA (collagen crosslinking [CXL]) treatment for keratoconus is a method for stabilizing or improving corneal properties. Our aim was to evaluate changes in corneal topography indices after CXL. A total of 38 eyes of 25 patients (mean age 29.36 ± 9.7 years) were treated with CXL technique. The follow-up period was 36 months. Slit-lamp examination, visual acuity tests, and pachymetry measurements were performed; in order to obtain the following numerical data, corneal topography (TMS-4; Tomey) was used: simulated keratometry (K1, K2), CYL (cylinder value), SAI (surface asymmetry index), IAI (irregular astigmatism index), SRI (surface regulatory index), ACP (average corneal power), and CEI (corneal eccentricity index). Treatments were performed with In-Pro CCL-Lix (Germany) equipment. The values of mean K1 (p = 0.82), cylinder (p = 0.73), SAI (p = 0.99), IAI (p = 0.82), SRI (p = 0.73), ACP (p = 0.73), K2 (p = 0.8), and CEI (p = 0.99) showed no significant changes at the end of the examination period. Uncorrected visual acuity remained unchanged (p = 0.32). However, best-corrected visual acuity changed from 0.21 ± 0.17 to 0.12 ± 0.11 in logMAR scale, but this deviation was not considered significant (p = 0.08). No change was observed in intraocular pressure (p = 0.79). Cornea remained clear and during follow-up period no corneal thinning was observed (p = 0.78). Our results suggest that corneal collagen CXL can be a successful treatment at certain stages of keratoconus, offering a way to stop progression in keratometric indices.


Colonic Anastomotic Leak Model in Swine.
Anastomotic leaks are serious postoperative complications. Current experimental studies designed to investigate leaks are based on acute intraoperative dehiscence of the anastomosis. Clinically, however, AL usually happens later in the post-operative course. Presented here is a clinically relevant colonic anastomotic leak model in swine. 17 Yorkshire pigs were divided into 2 groups: the control group (n=6) and the experimental group (n=11). An enterotomy was performed on the descending colon and an end-to-end handsewn anastomosis was created in the groups. The proximal and distal ends of the suture were exteriorized and tied to a plastic tube. Subsequently, the suture was cut and pulled to induce breakdown of the anastomosis in the experimental group 3-4 hours postoperatively. Study endpoints included behavioral changes, clinical assessment, laboratory indicators, and macroscopic indicators of leakage. Leaks were successfully created in 8/11 of the experimental group animals and confirmed through exploratory relaparotomy. Seven of the experimental pigs showed complete anastomotic breakdown and one showed partial rupture. Fecal peritonitis and enteric spillage were observed macroscopically within the abdomen of the experimental pigs, confirming the presence of a leak. The remaining (3/11) experimental pigs did not experience those findings due to either a tamponade/containment by the abdominal wall or surrounding organs. Statistical significance (p<0.05) was achieved between the experimental and control cohorts for laboratory and clinical indicators including fever, leukocytosis, and decreased blood potassium. This animal model generated postoperative induced leak in approximately three-quarters (8/11) of experimental pigs, allowing control over the time of leak onset to simulate clinical settings.


Transcriptional repression of DNA repair genes is a hallmark and a cause of cellular senescence.
Cellular senescence response is (i) activated by numerous stresses, (ii) is characterized by a stable proliferation arrest, and (iii) by a set of specific features. Timely regulated senescence is thought to be beneficial, whereas chronic senescence such as during normal or premature aging is deleterious as it favors most, if not all, age-related diseases. In this study, using in-house or publicly available microarray analyses of transcriptomes of senescent cells, as well as analyses of the level of expression of several DNA repair genes by RT-qPCR and immunoblot, we show that repression of DNA repair gene expression is associated with cellular senescence. This repression is mediated by the RB/E2F pathway and it may play a causal role in senescence induction, as single DNA repair gene repression by siRNA induced features of premature senescence. Importantly, activating RB independently of direct DNA damage also results in repression of DNA repair genes and in the subsequent induction of DNA damage and senescence. The dogma is that DNA damage observed during cellular senescence is directly provoked by DNA lesions following genotoxic attack (UV, IR, and ROS) or by induction of replicative stress upon oncogenic activation. Our in vitro results support a largely unsuspected contribution of the loss of DNA repair gene expression in the induction and the accumulation of the DNA damage observed in most, if not all, kinds of cellular senescence, and thus in the induction of cellular senescence. Further demonstration using in vivo models will help to generalize our findings.


Cefazolin susceptibility of coagulase-negative staphylococci (CoNS) causing late-onset neonatal bacteraemia.
CoNS bacteraemia causes significant neonatal morbidity. Previous work has suggested that β-lactam antibiotics vary in their binding affinity to PBP2a (produced by the mecA gene) present in most CoNS. We evaluated cefazolin MICs for CoNS isolated in an Australian neonatal ICU (NICU) and correlated them with isolate genotype and phenotype. Significant blood isolates from 2009 to 2017 were speciated and underwent broth microdilution testing for cefazolin, cefoxitin, oxacillin and flucloxacillin. Correlation with mecA presence and PBP2a expression was evaluated. A selection of Staphylococcus capitis isolates underwent WGS. The CoNS (n = 99) isolates were confirmed as S. capitis (n = 57), Staphylococcus epidermidis (n = 32), Staphylococcus haemolyticus (n = 2) and Staphylococcus warneri (n = 8). The MIC of cefazolin was ≤2 mg/L for 30% of isolates and 75% had an MIC of ≤8 mg/L (MIC90 = 16 mg/L). This contrasted with MIC90s of cefoxitin, oxacillin and flucloxacillin, which were all ≥32 mg/L. WGS found a number of S. capitis isolates closely related to the globally established NRCS-A clone. CoNS displayed distinctly lower MIC values of cefazolin than of other agents tested. MIC variation may be related to binding affinity of PBP2a or regulation of expression of mecA by mecR1-mecI functional genes. Further, NRCS-A S. capitis strains were present in this Australian NICU before and after the unit underwent physical relocation, which raised questions about a common environmental source. It is considered justified to conduct a randomized clinical trial that assesses cefazolin versus vancomycin for management of late-onset neonatal sepsis.


A to I editing in disease is not fake news.
Adenosine deaminases acting on RNA (ADARs) are zinc-containing enzymes that deaminate adenosine bases to inosines within dsRNA regions in transcripts. In short, structured dsRNA hairpins individual adenosine bases may be targeted specifically and edited with up to one hundred percent efficiency, leading to the production of alternative protein variants. However, the majority of editing events occur within longer stretches of dsRNA formed by pairing of repetitive sequences. Here, many different adenosine bases are potential targets but editing efficiency is usually much lower. Recent work shows that ADAR-mediated RNA editing is also required to prevent aberrant activation of antiviral innate immune sensors that detect viral dsRNA in the cytoplasm. Missense mutations in the ADAR1 RNA editing enzyme cause a fatal auto-inflammatory disease, Aicardi-Goutières syndrome (AGS) in affected children. In addition RNA editing by ADARs has been observed to increase in many cancers and also can contribute to vascular disease. Thus the role of RNA editing in the progression of various diseases can no longer be ignored. The ability of ADARs to alter the sequence of RNAs has also been used to artificially target model RNAs in vitro and in cells for RNA editing. Potentially this approach may be used to repair genetic defects and to alter genetic information at the RNA level. In this review we focus on the role of ADARs in disease development and progression and on their potential use to artificially modify RNAs in a targeted manner.


[Obstructive ischemic involvement of the anterior descending coronary artery in asymmetrical septal form of hypertrophic cardiomyopathy].
The significance of necrosis and fibrosis of the interventricular septum in hypertrophic cardiomyopathy (HCM) is controversial. The purpose of this study was then to evaluate the clinical impact of left anterior descending artery (LAD) disease in HCM. Among 158 patients presenting with HCM, 6 (3.79%) had LAD disease. Mean age was 65.16 years (52 to 70), 4 were men and 4 had the obstructive form of HCM. All patients were submitted to complete clinical and laboratory evaluation. Mean time of the diagnosis was 141 months (ranging from 78 to 182) for HCM and 42 months (ranging from one day to 106 months) for LAD disease. Five patients had unstable angina and one had myocardial infarction. LAD disease (60 to 100% coronary narrowing) was present in all patients; one patient had single vessel disease and 5 multivessel disease. During follow-up, 3 patients had coronary artery bypass grafting (CABG), one with associated septal myectomy; one was submitted to coronary angioplasty and 2 were submitted to medical treatment. During a follow-up period ranging from 76 to 124 months after LAD disease diagnosis, one patient died. At the end of the study, a reduction of the thickness of the interventricular septum from 1.53 to 1.40 cm was observed and left ventricular outflow pressure gradient decreased from 56 to 16 mmHg. Left ventricular diastolic diameter increased from 4.55 to 4.85 cm and systolic diameter from 2.83 to 3.13 cm. Left atrium diameter was unchanged. LAD disease is well tolerated in the asymmetrical form of HCM and may contribute to septal fibrosis, improving cardiac function. It does not represent an adverse factor in the evolution of HCM.


Research Review: What have we learned about adolescent substance use?
Adolescence is a critical biological, psychological, and social developmental stage involving heightened risk for substance use and associated adverse consequences. This review, synthesizing emerging findings on this complex topic, is intended to inform research and clinical care focused on adolescents. Literature searches were conducted using PubMed, yielding a cross-section of observational and interventional studies focused on adolescent substance use. Findings were organized and categorized to cover key areas of epidemiology, neurobiology, prevention, and treatment. Adolescent substance-related attitudes and use patterns have evolved over time, informed by adult and peer behaviors, public policy, media messaging, substance availability, and other variables. A number of risk and resiliency factors contribute to individual differences in substance use and related consequences. Advances in observational techniques have provided enhanced understanding of adolescent brain development and its implications for substance use. Prevention efforts have yielded mixed results, and while a number of adolescent-targeted evidence-based treatments for substance use disorders have been developed, effect sizes are generally modest, indicating the need for further research to enhance prevention and treatment outcomes. Substance use in adolescence is heterogeneous, ranging from normative to pathological, and can lead to significant acute and long-term morbidity and mortality. Understanding risk and resiliency factors, underlying neurobiology, and optimal developmentally sensitive interventions is critical in addressing substance-associated problems in adolescence.


A comparison of phosphorus speciation and potential bioavailability in feed and feces of different dairy herds using 31p nuclear magnetic resonance spectroscopy.
An experiment was conducted to examine how potential phosphorus (P) bioavailability (inferred from speciation) differs in feed and feces collected in spring from four dairy herds representing different management systems: (i) total confinement with cows fed total mixed ration (TMR), (ii) total confinement with TMR plus P mineral supplement, (iii) a hybrid of confinement with TMR and pastoral grazing, and (iv) predominantly grazing with supplemental grains. A treatment was included that air dried feces to simulate conditions after dung deposition. Wet chemical techniques and solution (31)P nuclear magnetic resonance spectroscopy ((31)P-NMR) were used to identify P concentrations and compounds present in water (a surrogate for P in overland flow), dilute acid (0.012 M HCl, an estimate of P utilization by cattle), or NaOH-EDTA (a solution that maximizes the organic P extraction) extracts of feed and feces. In general, P concentration in feces paralleled P in feed. Air drying feces decreased water-extractable P by 13 to 61% largely due to a decrease in orthophosphate, whereas NaOH-EDTA-extractable P increased by 18 to 48%. Analysis of dilute HCl was unsuccessful due to orthophosphate precipitation when pH was adjusted to 12 for (31)P-NMR. In water extracts, more P was in bioavailable diester-P forms, undetectable by colorimetry, than in NaOH-EDTA extracts. In feed, orthophosphate dominated (46-70%), but myo-IHP varied with feed (<10% in forage samples but 43% in a TMR sample). The proportion of myo-IHP decreased in feces compared with feed via mineralization but decreased less in systems with a greater proportion of available P input (e.g., orthophosphate and phospholipids). Feed and drying effect the concentrations and forms of P in feces and their potential impact on soil and water quality. Although bioavailable P in feces from pasture-based and confined systems can be similar in spring, dung-P is distributed on a lower kg P ha(-1) rate in grazing systems. The best method to mitigate P loss from feces is to decrease P in feed.


Ineffectiveness of intravenous ascorbic acid as an acidifying agent in man.
The acidifying effect of intravenous (IV) ascorbic acid was studied in seven healthy adult volunteers. After obtaining baseline urine and blood samples, 2-g IV doses of ascorbic acid were administered to each subject during a 20-minute period. Venous blood samples were obtained at times 0.5, 1, and 2 hours, and urine was collected at times 0.5, 1, 2, and 3 hours. Our results show that venous blood pH, plasma bicarbonate concentration, urine PCO2, and urine bicarbonate excretion did not change significantly during the study period. Urinary titratable acidity, ammonium excretion, and net hydrogen ion excretion decreased, and urinary pH actually showed a significant rise at two hours. We therefore conclude that IV ascorbic acid administered in recommended doses does not effectively acidify urine.


High level expression of DNA polymerases from herpesviruses.
The DNA polymerase genes of human cytomegalovirus (HCMV) and varicella-zoster virus (VZV) were inserted separately into the polyhedrin gene of Autographa californica nuclear polyhedrosis virus (AcNPV) by cotransfection of Spodoptera frugiperda (SF9) cells with baculovirus transfer vectors carrying the genes and AcNPV infectious DNA. Infection of SF9 cells with the recombinant viruses resulted in expression from the polyhedrin promoter of proteins of the expected Mrs. These proteins possessed DNA polymerase activities similar to that of the enzymes induced by the respective herpesvirus in infected cells, and were identified as HCMV and VZV DNA polymerase using inhibitors and specific antisera reactive with each enzyme.


Concerted charge and energy transfer processes in a highly flexible fullerene-dye system: a mixed quantum-classical study.
Photoinduced excitation energy transfer and accompanying charge separation are elucidated for a supramolecular system of a single fullerene covalently linked to six pyropheophorbide-a dye molecules. Molecular dynamics simulations are performed to gain an atomistic picture of the architecture and the surrounding solvent. Excitation energy transfer among the dye molecules and electron transfer from the excited dyes to the fullerene are described by a mixed quantum-classical version of the Förster rate and the semiclassical Marcus rate, respectively. The mean characteristic time of energy redistribution lies in the range of 10 ps, while electron transfer proceeds within 150 ps. In between, on a 20 to 50 ps time-scale, conformational changes take place in the system. This temporal hierarchy of processes guarantees efficient charge separation, if the structure is exposed to a solvent. The fast energy transfer can adopt the dye excitation to the actual conformation. In this sense, the probability to achieve charge separation is large enough since any dominance of unfavorable conformations that exhibit a large dye-fullerene distance is circumvented. And the slow electron transfer may realize an averaging with respect to different conformations. To confirm the reliability of our computations, ensemble measurements on the charge separation dynamics are simulated and a very good agreement with the experimental data is obtained.


NrichD database: sequence databases enriched with computationally designed protein-like sequences aid in remote homology detection.
NrichD (http://proline.biochem.iisc.ernet.in/NRICHD/) is a database of computationally designed protein-like sequences, augmented into natural sequence databases that can perform hops in protein sequence space to assist in the detection of remote relationships. Establishing protein relationships in the absence of structural evidence or natural 'intermediately related sequences' is a challenging task. Recently, we have demonstrated that the computational design of artificial intermediary sequences/linkers is an effective approach to fill naturally occurring voids in protein sequence space. Through a large-scale assessment we have demonstrated that such sequences can be plugged into commonly employed search databases to improve the performance of routinely used sequence search methods in detecting remote relationships. Since it is anticipated that such data sets will be employed to establish protein relationships, two databases that have already captured these relationships at the structural and functional domain level, namely, the SCOP database and the Pfam database, have been 'enriched' with these artificial intermediary sequences. NrichD database currently contains 3,611,010 artificial sequences that have been generated between 27,882 pairs of families from 374 SCOP folds. The data sets are freely available for download. Additional features include the design of artificial sequences between any two protein families of interest to the user.


Desensitization for renal transplantation: depletion of donor-specific anti-HLA antibodies, preservation of memory antibodies, and clinical risks.
Desensitization protocols reduce donor-specific anti-HLA antibodies (DSA) and enable renal transplantation in patients with a positive complement-dependent cytotoxic cross-match (CDC-CXM). The effect of this treatment on protective antibody and immunoglobulin levels is unknown. Thirteen patients with end-stage renal disease, DSA and positive CDC-CXM underwent desensitization. Sera collected pre- and post-transplantation were analysed for anti-tetanus and anti-pneumococcal antibodies, total immunoglobulin (Ig) levels and IgG subclasses and were compared to healthy controls and contemporaneous renal transplant recipients treated with standard immunosuppression alone. Ten patients developed negative CDC-CXM and enzyme-linked immunosorbent assay (ELISA) and underwent successful transplantation. Eight recipients achieved good graft function without antibody-mediated or late rejection, BK virus or cytomegalovirus infection. One patient had primary non-function due to recurrent oxalosis, and one patient with immediate graft function died from septicaemia. Seven recipients required post-operative transfusion and three developed septicaemia. DSA remained negative by ELISA at 12 months, but were detectable by Luminex(®) . Anti-tetanus and anti-pneumococcal antibodies, total Ig and IgG subclasses were below the normal range but comparable to levels in renal transplant recipients who had not undergone desensitization. Desensitization protocols effectively reduce DSA and allow successful transplantation. Post-operative bleeding and short-term infectious risk is increased. Protective antibody and serum immunoglobulin levels are relatively preserved.


Characterization of a truncated metabotropic glutamate receptor in a primitive metazoan, the parasitic flatworm Schistosoma mansoni.
A novel glutamate-binding protein was identified in Schistosoma mansoni. The protein (SmGBP) is related to metabotropic glutamate receptors from other species and has a predicted glutamate binding site located within a Venus Flytrap module but it lacks the heptahelical transmembrane segment that normally characterizes these receptors. The SmGBP cDNA was cloned, verified by 5' and 3' Rapid Amplification of cDNA Ends (RACE) and shown to be polyadenylated at the 3'end, suggesting the transcript is full-length. The cloned cDNA was subsequently expressed in bacteria and shown to encode a functional glutamate-binding protein. Other studies, using a specific peptide antibody, determined that SmGBP exists in two forms, a monomer of the expected size and a stable but non-covalent dimer. The monomer and dimer are both present in the membrane fraction of S. mansoni and are resistant to extraction with high-salt, alkaline pH and urea, suggesting SmGBP is either an integral membrane protein or a peripheral protein that is tightly associated with the membrane. Surface biotinylation experiments combined with western blot analyses and confocal immunolocalization revealed that SmGBP localized to the surface membranes of adult male schistosomes, especially the dorsal tubercles. In contrast, we detected little or no expression of SmGBP either in the females or larval stages. A comparative quantitative PCR analysis confirmed that the level of SmGBP expression is several-fold higher in male worms than cercariae, and it is barely detectable in adult females. Together, the results identify SmGBP as a new type of schistosome glutamate receptor that is both gender- and stage-specific. The high-level expression of this protein in the male tubercles suggests a possible role in host-parasite interaction.


2-Amino-1,8-naphthyridine Dimer (ANP77), a High-Affinity Binder to the Internal Loops of C/CC and T/CC Sites in Double-Stranded DNA.
Small molecules targeting DNA regions with structural fluctuation are an important class of molecule as chemical probes for studying the role of these structures in biological systems and the development of neurological disorders. The molecule ANP77 we described here, where a three-atom linker connects two 2-amino-1,8-naphthyridines at the C7 position, was found to form stacked structure with protonation of naphthyridine at low pH, and bound to the internal loop consisting of C/CC and T/CC in double-stranded DNA with affinities of 4.8 and 34.4 nM, respectively. Mass spectrometry and isothermal titration calorimetry analyses determined the stoichiometry for the binding as 1:1, and chemical footprinting with permanganate and NMR structural analysis revealed that the T in the T/CC was forced to flip out toward an extrahelical position upon ANP77 binding. Protonated stacked ANP77 interacted with two adjacent cytosines through hydrogen bonding and occupied the position in the duplex by flipping out the C or T opposite CC. Finally, this study demonstrated the potential of ANP77 for binding to the sequences of biological significance with the TG(T/C)CC repeat of the PIG3 promoter and the telomere repeat CCCTAA.


Preservation of Mycoplasma strains by freezing in liquid nitrogen and by lyophilization with sucrose.
Procedures for maintaining Mycoplasma strains are described. Liquid nitrogen storage provided an adequate means for keeping stock cultures stable. Over 74 strains of approximately 26 species have been preserved in this way, some for as long as 9 years. Mycoplasma strains can also be recovered satisfactorily from the freeze-dried state when the procedure described includes the use of 12% (v/v) sucrose as an additive. Fifteen strains representing 12 or more species were subjected to a freeze-drying program with and without sucrose added to concentrated cell suspensions in growth medium. Cell counts indicate improved survival with sucrose.


[Inversion polymorphism of the malaria mosquito Anopheles messeae. IX. Cannibalism in larvae as a selection factor].
The genetic consequences of larval cannibalism were studied in mosquito Anopheles messeae Fall. (Diptera. Culicidae) with "south" inversional combinations. It is shown that mosquito with karyotypes XL11(2)R00(3)R00(01, 11) eat the larvae of the species named more actively than mosquito with karyotypes XL00(01)2R00(3)R00(01)3L00(01, 11). Evidently, adaptation of mosquito with "south" genocomplexes was developed in two directions: 1) more effective utilization of food resources in the larvae with the inversion XL0 which are wide-spread at south-west of the species areal; 2) the spread of cannibalism in the larvae of eldest ages with the XL1 inversion which inhabited south-east of the species areal.


Treatment Course of Patients with Exudative Age-Related Macular Degeneration Using Ocular Hypotensives.
To characterize the visual outcomes and the treatment course of patients with exudative age-related macular degeneration (AMD) based on ocular hypotensive use. A matched retrospective cohort study of patients enrolled in Kaiser Permanente Southern California health plan was conducted. Patients taking ocular hypotensives were identified using pharmacy dispensing data and were matched to controls to compare visual acuity, number of anti-VEGF injections, and conversation to secondary anti-VEGF agents in the first year of treatment. Subgroup analysis was performed based on the number of ocular hypotensive agents (0, 1, 2 or 3+ agents) and drug class (aqueous suppressants and prostaglandin analogs). A total of 234 patients patients were examined. Baseline and final visual acuity did not significantly differ between drop users and controls, including on subgroup analysis. The average number of anti-VEGF injections did not differ between drop users and controls (6.1 vs 6.2, p=0.97), nor did the percentage of patients who were switched to a second-line anti-VEGF agent (23.9% vs 17.9%, p=0.26). Subgroup analysis did not reveal significant differences in the number of anti-VEGF injections and the percentage of patients switched to secondary agents, with patients receiving 6 ±1 injections across regardless of the number or class of ocular hypotensive agents used. Patients with concurrent glaucoma and exudative AMD have similar visual outcomes and treatment courses compared to those not taking ocular hypotensives. Although aqueous suppressants have been suggested to prolong anti-VEGF residence time, patients using these agents did not demonstrate visual benefit or a reduced injection burden in this series.


Factors influencing acceptance, adoption and adherence to sentinel node biopsy recommendations in the Australian Melanoma Management Guidelines: a qualitative study using an implementation science framework.
Sentinel node biopsy (SN biopsy) is a surgical procedure used to accurately stage patients with primary melanoma at high risk of recurrence. Although Australian Melanoma Management Guidelines recommend SN biopsy be considered in patients with melanomas > 1 mm thick, SN biopsy rates in Australia are reportedly low. Our objective was to identify factors impacting the acceptance, adoption and adherence to the Australian SN biopsy guideline recommendations. Opinions of Australian key informants including clinicians, representatives from melanoma education and training providers, professional associations and colleges, and melanoma advocacy organisations were collected through semi-structured interviews (n = 29) and from publicly released statements (n = 14 news articles). Data analysis involved inductive and deductive thematic analysis using Flottorp's determinants framework. A complex interplay of contemporary and historical factors was identified as influencing acceptance, adoption and adherence to the SN biopsy guideline recommendations at the individual, guideline, patient, organisational and social levels. Expert and peer opinion leaders have played an important role in facilitating or inhibiting adoption of guideline recommendations, as have financial incentives driven by healthcare-funding policies and non-financial incentives including professional identity and standing. Of critical importance have been the social and knowledge boundaries that exist between different professional groups to whom the guidelines apply (surgeons, dermatologists and primary care practitioners) with adherence to the guideline recommendations having the potential to shift work across professional boundaries, altering a clinician's workflow and revenue. More recently, the emergence of effective immunotherapies and targeted therapies for patients at high risk of recurrence, the emergence of new opinion leaders on the topic (in medical oncology), and patient demands for accurate staging are playing crucial roles in overcoming the resistance to change created by these social and knowledge boundaries. Acceptance and adherence to SN biopsy guideline recommendations in Australia over the past 20 years has involved a process of renegotiation and reframing of the evidence for SN biopsy in melanoma by clinicians from different professional groups and networks. This process has helped to refine the evidence for SN biopsy and our understanding of appropriate adoption. New effective systemic therapies have changed the balance towards accepting guideline recommendations.


How can we remodel practices into medical homes without a blueprint or a bank account?
Medical homes are widely viewed as a solution to the problems with American medical care, despite lack of answers to many important questions. Review of articles from issues of 5 journals devoted to the medical home in 2010 provides few answers to those questions. However, with some exceptions, those answers seem more likely to come from real-life efforts to implement medical homes than from the research literature. In any other industry, that would be the case, especially the key questions about the financial viability of both the transformation of traditional practices and sustainability of the new care model.


Simultaneous reconstruction and registration algorithm for limited view transmission tomography using a multiple cluster approximation to the joint histogram with an anatomical prior.
We develop a novel simultaneous reconstruction and registration algorithm for limited view transmission tomography. We derive a cost function using Bayesian probability theory, and propose a similarity metric based on the explicit modeling of the joint histogram as a sum of bivariate clusters. The resulting algorithm shows a robust mitigation of the data insufficiency problem in limited view tomography. To our knowledge, our work represents the first attempt to incorporate non-registered, multimodal anatomical priors into limited view transmission tomography by using joint histogram based similarity measures.


A Retrospective 10-Year Analysis of Water Absorbent Bead Ingestion in Children.
This retrospective case series was designed to evaluate the clinical experience of pediatric patients who had ingested water absorbent beads (WABs) and review the relevant literature. A retrospective chart analysis of 21 children who were admitted to the emergency service or outpatient clinic with complaints of swallowing WABs during a 10-year period was performed. The patients were divided into two groups: 6 patients who were hospitalized and 15 patients who were observed as outpatients for groups 1 and 2, respectively. All data were screened with respect to presentation, age, gender, clinical and radiological findings, modalities of treatment, and follow-up. All patients were asymptomatic with uneventful outcomes. Laboratory studies, X-ray, and ultrasound examinations revealed normal results. None of the patients underwent surgery or other medical procedures. Gastrointestinal system obstruction or any other pathological findings were not detected after WAB ingestion. No complications were identified during follow-up. This first clinical study showed that asymptomatic patients do not require inpatient observation and laboratory investigations, X-ray, and ultrasound. However, results of the present study cannot be generalized to children of all ages due to the few cases of bowel obstruction reported to date in small children.


Preoperative exercise training prevents functional decline after lung resection surgery: a randomized, single-blind controlled trial.
To investigate the effects of a preoperative pulmonary rehabilitation programme in patients with lung cancer undergoing video-assisted thoracic surgery. Randomized, single-blind controlled trial. Teaching hospital. Patients with suspected or confirmed lung cancer undergoing video-assisted thoracic surgery. Participants were randomized to either a prehabilitation group or a control group. Participants in the prehabilitation group underwent a combination of moderate endurance and resistance training plus breathing exercises three to five times per week. The primary outcome of the study was exercise capacity. Secondary outcomes were muscle strength (Senior Fitness Test), health-related quality of life (Short-Form 36) and the postoperative outcomes. Patients were evaluated at baseline (before randomization), presurgery (only the prehabilitation group), after surgery and three months post-operatively. A total of 40 patients were randomized and 22 finished the study (10 in the prehabilitation group and 12 in the control group). Three patients were lost to follow-up at three months. After the training, there was a statistically significant improvement in exercise tolerance (+397 seconds, p = 0.0001), the physical summary component of the SF-36 (+4.4 points, p = 0.008) and muscle strength ( p < 0.01). There were no significant differences between groups after surgery. However, three months postoperatively, significant differences were found in the mean change of exercise capacity ( p = 0.005), physical summary component ( p = 0.001) and upper and lower body strength ( p = 0.045 and p = 0.002). A pulmonary rehabilitation programme before video-assisted thoracic surgery seems to improve patients' preoperative condition and may prevent functional decline after surgery. Clinical Registration Number: NCT01963923 (Registration date 10/10/2013).


Using an Expert Panel to Develop Social Support Program Sequencing for Young Adults With Type 1 Diabetes.
Young adults living with type 1 diabetes (T1DM) often encounter poor health outcomes, such as uncontrolled blood glucose levels. Social support programs can be a helpful method to support T1DM self-management. Effectively planning and tailoring social support programs for young adults living with T1DM are crucial for improving these programs and associated outcomes. This study convened an expert panel primarily composed of young adults living with T1DM to generate ideas and key components for sequential inclusion in social support programs prioritizing them. Exploratory expert panel meetings consisting of four individuals living with T1DM were held where Nominal Group Technique and Ideawriting exercises were used to develop themes and discussion points. Six themes emerged from the meetings representing areas of difficulty for young adults living with T1DM. Topics such as following self-care recommendations, nutrition, handling stress, coping with social situations, and navigating the health care system were identified as important issues facing young adults. By incorporating this approach into new or existing support group improved program discussions for young adults can be achieved and pertinent issues addressed, thus leading to improved health care outcomes.


Effects of electroconvulsive shock on noradrenergic and serotonergic receptors in rat brain.
It is likely that both noradrenalin and serotonin neurotransmission are important in the pathophysiology of depression and to its treatment. In particular, certain receptors for these neurotransmitters are altered by repeated treatment with both antidepressant drugs and electroconvulsive shock. This paper reviews the effects of electroconvulsive shock on alpha-1-adrenergic, beta-adrenergic, and serotonin-2 receptors in rat brain, and compares these effects to those produced by anti-depressant drugs. The similarities and differences in the effects of antidepressant drugs and electroconvulsive shock in rat brain may provide clues for the development of more effective treatments for depression and could single out targets for future investigation of the pathophysiology of depression in patients when safe methods are developed.


COVID-19 and Thrombotic or Thromboembolic Disease: Implications for Prevention, Antithrombotic Therapy, and Follow-Up: JACC State-of-the-Art Review.
Coronavirus disease-2019 (COVID-19), a viral respiratory illness caused by the severe acute respiratory syndrome-coronavirus-2 (SARS-CoV-2), may predispose patients to thrombotic disease, both in the venous and arterial circulations, because of excessive inflammation, platelet activation, endothelial dysfunction, and stasis. In addition, many patients receiving antithrombotic therapy for thrombotic disease may develop COVID-19, which can have implications for choice, dosing, and laboratory monitoring of antithrombotic therapy. Moreover, during a time with much focus on COVID-19, it is critical to consider how to optimize the available technology to care for patients without COVID-19 who have thrombotic disease. Herein, the authors review the current understanding of the pathogenesis, epidemiology, management, and outcomes of patients with COVID-19 who develop venous or arterial thrombosis, of those with pre-existing thrombotic disease who develop COVID-19, or those who need prevention or care for their thrombotic disease during the COVID-19 pandemic.


Conservative management of hyperplastic and multicentric nephroblastomatosis.
The treatment of hyperplastic nephroblastomatosis remains controversial. We report the advantages of conservative management of hyperplastic and multicentric nephroblastomatosis associated with unilateral Wilms tumor (WT). During the last 10 years 48 children with unilateral WT were consecutively treated at our 2 institutions. Children with multiple solid renal masses on imaging were treated with 2-drug chemotherapy until disappearance of the lesions. Stabilization or progression of the lesions despite chemotherapy, as well as heterogeneity of the lesions on imaging, prompted nephron sparing surgery (NSS). Three female infants (12, 13 and 20 months old, respectively) presented with multiple solid renal tumors at imaging. Despite chemotherapy, small and unilateral WT developed in 2 cases of hyperplastic nephroblastomatosis, which was excised. One of these infants subsequently presented with a small contralateral metachronous WT, which was excised. Both infants are disease-free with 2 normal kidneys at followup of 6 and 2 years, respectively. The third infant, who presented with unilateral multicentric WT and unilateral hyperplastic nephroblastomatosis nodules, was successfully treated with preoperative chemotherapy and enucleation of 5 tumors. Subsequently, nephrectomy was performed at another institution because the abnormal kidney outline due to NSS was misinterpreted as a recurrence of WT. She was lost to followup. Hyperplastic and multicentric nephroblastomatosis is not a rare lesion and is most often associated, either initially or subsequently, with WT. In some infants with multiple solid renal masses on imaging chemotherapy and for developing WT NSS may safely allow maximum sparing of the parenchyma of both kidneys.


[Bacteriological quality of air in a ward for sterile pharmaceutical preparations].
An extremely clean area is required for preparation of sterile pharmaceutical compounds, in compliance with international standards, to minimize the probability of microbial contamination. To evaluate the bacteriological quality of the air in the Sterile Pharmaceutical Preparation Unit of the University of Chile's Clinical Hospital and to set up alerts and action levels of bacterial growth. We studied eight representative sites of our Unit on a daily basis from January to February 2005 and twice a week from June 2005 to February 2006. We collected 839 samples of air by impact in the Petri dish. 474 (56.5%) samples were positive; 17 (3.5%) of them had an inappropriate bacterial growth (2% of total samples). The samples from sites 1 and 2 (big and small biosafety cabinets) were negative. The countertop and transfer area occasionally exceeded the bacterial growth limits. The most frequently isolated bacteria were coagulase-negative staphylococci, Micrococcus spp and Corynebacterium spp, from skin microbiota, and Bacillus spp, an environmental bacteria. From a microbiological perspective, the air quality in our sterile preparation unit complied with international standards. Setting institutional alerts and action levels and appropriately identifying bacteria in sensitive areas permits quantification of the microbial load and application of preventive measures.


Proteome Analysis of Pathogen-Responsive Proteins from Apple Leaves Induced by the Alternaria Blotch Alternaria alternata.
Understanding the defence mechanisms used by apple leaves against Alternaria alternate pathogen infection is important for breeding purposes. To investigate the ultrastructural differences between leaf tissues of susceptible and resistant seedlings, in vitro inoculation assays and transmission electron microscopy (TEM) analysis were conducted with two different inoculation assays. The results indicated that the resistant leaves may have certain antifungal activity against A. alternate that is lacking in susceptible leaves. To elucidate the two different host responses to A. alternate infection in apples, the proteomes of susceptible and resistant apple leaves that had or had not been infected with pathogen were characterised using two-dimensional electrophoresis (2-DE) and matrix-assisted laser desorption/ionisation time-of-flight tandem mass spectrometry (MALDI-TOF-TOF MS). MS identified 43 differentially expressed proteins in two different inoculation assays. The known proteins were categorised into 5 classes, among these proteins, some pathogenesis-related (PR) proteins, such as beta-1,3-glucanase, ascorbate peroxidase (APX), glutathione peroxidase (GPX) and mal d1, were identified in susceptible and resistant hosts and were associated with disease resistance of the apple host. In addition, the different levels of mal d1 in susceptible and resistant hosts may contribute to the outstanding anti-disease properties of resistant leaves against A. alternate. Taken together, the resistance mechanisms of the apple host against A. alternate may be a result of the PR proteins and other defence-related proteins. Given the complexity of the biology involved in the interaction between apple leaves and the A. alternate pathogen, further investigation will yield more valuable insights into the molecular mechanisms of suppression of the A. alternate pathogen. Overall, we outline several novel insights into the response of apple leaves to pathogen attacks. These findings increase our knowledge of pathogen resistance mechanisms, and the data will also promote further investigation into the regulation of the expression of these target proteins.


Multispectral image visualization through first-order fusion.
We present a new formalism for the treatment and understanding of multispectral images and multisensor imagery based on first-order contrast information. Although little attention has been paid to the utility of multispectral contrast, we develop a theory for multispectral contrast that enables us to produce an optimal grayscale visualization of the first-order contrast of an image with an arbitrary number of bands. We demonstrate how our technique can reveal significantly more interpretive information to an image analyst, who can use it in a number of image understanding algorithms. Existing grayscale visualization strategies are reviewed. A variety of experimental results are presented to support the performance of the new method.


Sensing in 15 s for aqueous fluoride anion by water-insoluble fluorescent probe incorporating hydrogel.
Anion recognition and sensing via artificial receptors have attracted a great deal of attention since they play a fundamental and important role in chemical, biological, medical, and environmental processes. Fluoride, as one of the smallest anions, is of particular interest because of its role in dental care and the analysis of drinking water. Herein, we invented a new method for F(-) detection by adopting the hydrogel as the supporter of reaction between a water insoluble fluorescent probe and F(-) in the water environment. This method is highly rapid, selective, and sensitive, which can determine F(-) levels in 15 s at the drinking water standard. A novel compound N-(3-(benzo[d]thiazol-2-yl)-4-(tert-butyldiphenylsilyloxy)phenyl) acetamide (BTBPA) was synthesized as the fluorescent probe because of the significant fluorescent color change from blue to green after the reaction with F(-). This method does not require the probe substances to be water-soluble, which greatly expands the range of the specific fluorescent molecules used in ion detection. Additionally, just a few microliter samples were required in the analysis procedures with this method.


Nano albumin bound-paclitaxel in pancreatic cancer: Current evidences and future directions.
Pancreatic cancer (PDAC) is an aggressive and chemoresistant disease, representing the fourth cause of cancer related deaths in western countries. Majority of patients have unresectable, locally advanced or metastatic disease at time of diagnosis and the 5-year survival rate in these conditions is extremely low. For more than a decade gemcitabine has been the cornerstone of metastatic PDAC treatment, although survival benefit was very poor. PDAC cells are surrounded by an intense desmoplastic reaction that may create a barrier to the drugs penetration within the tumor. Recently PDAC stroma has been addressed as a potential therapeutic target. Nano albumin bound (Nab)-paclitaxel is an innovative molecule depleting tumor stroma, through interaction between albumin and secreted protein acidic and rich in cysteine. Addition of nab-paclitaxel to gemcitabine has showed activity and efficacy in metastatic PDAC first-line treatment improving survival and overall response rate vs gemcitabine alone in the MPACT phase III study. This combination represents one of the standards of care in advanced PDAC therapy and is suitable to a broader spectrum of patients compared to other schedules. Nab-paclitaxel is under investigation as a backbone of chemotherapy in novel combinations with target agents or immunotherapy in locally advanced or metastatic PDAC. In this article, we provide an updated and critical overview about the role of nab-paclitaxel in PDAC treatment based on the latest advances in preclinical and clinical research. Furthermore, we focus on the use of nab-paclitaxel within the context of metastatic PDAC treatment landscape and we discuss about future implications in the light of current clinical ongoing trials.


Electrochemical phase diagrams of Ni from ab initio simulations: role of exchange interactions on accuracy.
The stabilities of Ni metal and its derived compounds, including oxides, hydroxides, and oxyhydroxides under electrochemical conditions, can be readily predicted from the Ni Pourbaix diagram, where the formation free energies of the involved species are utilized to construct the phase stability map with respect to electrode potential and pH. We calculate and analyze the crystal structures, electronic structures, and thermodynamic energies of Ni metal and its compounds using different exchange-correlation functionals to density-functional-theory (DFT), including the semilocal LDA and GGA density functionals, the nonlocal metaGGA, and the hybrid density functionals. Next, we simulate the corresponding Ni Pourbaix diagrams to compare systematically the performance of the functional to each other and to experimental observations. We show that the structures and energies obtained from experimental databases may not be sufficiently accurate to describe direct electrochemical observations, and we explain how the electronic exchange within the density functionals plays a key role in determining the accuracy of the DFT calculated electronic, thermodynamic, and electrochemical properties. We find that only the hybrid density functional produces reliable results owing to the fractional contribution of exact Fock exchange included therein. Last, based on our accurate Ni Pourbaix diagram, we construct band-gap and magnetic electrochemical maps which can facilitate more experimental measurements and property assessments under variable potential and pH in the future.


5-Aminoisoquinolin-1(2H)-one, a water-soluble poly (ADP-ribose) polymerase (PARP) inhibitor reduces the evolution of experimental periodontitis in rats.
Poly (ADP-ribose) polymerase (PARP), a nuclear enzyme activated by strand breaks in DNA, plays an important role in the tissue injury associated with ischaemia-reperfusion and inflammation. Recent studies have demonstrated that PARP activation plays a crucial role in the pathogenesis of acute periodontal injury. We have investigated the effect of 5-aminoisoquinolin-1(2H)-one (5-AIQ), a water-soluble PARP inhibitor, in a rat model of periodontitis. Periodontitis was induced in rats by placing a 2/0 braided silk ligature around the lower left first molar. At day eight, the gingivomucosal tissue encircling the mandibular first molar was removed for biochemical and histological analysis. Ligation significantly induced an increased neutrophil infiltration and a positive staining for PARP activation. Ligation significantly increased Evans blue extravasation in gingivomucosal tissue and alveolar bone destruction. Intraperitonial injection of 5-aminoisoquinolin-1(2H)-one (5-AIQ) (5 mg/kg daily for eight days) significantly decreased all of the parameters of inflammation as described above. This suggests that inhibition of PARP may represent a novel approach for the treatment of periodontal disease.


Fulminant hepatic failure secondary to congestive heart failure.
An elderly female with an acute episode of congestive heart failure, unaccompanied by any periods of hypotension, developed fulminant hepatic failure with an accompanying coagulopathy. Attempts to establish an etiology for her acute hepatic insufficiency, other than cardiac failure, proved negative. Fulminant hepatic failure as a consequence of congestive heart failure, without prolonged periods of hypotension preceding alteration in hepatic function, has not heretofore been described. Liver function is adversely effected in congestive heart failure. Hepatic ammonia clearance is impaired in cardiac failure and may be diminished to the point of resulting in hepatic encephalopathy. Coagulopathy is a frequent concomitant of fulminant hepatic failure. Establishing a clear etiology for a coagulopathy in the face of concomitant liver disease is difficult, thus making any therapeutic intervention fraught with peril.


[Leptofibrils in rat ventricular cardiomyocytes in vitro].
The presence of leptofibrils in ventricular myocytes of the heart of the 2- and 14-day old rats in 8-day old cultures was shown and their structure was investigated. Leptofibrils are seen only in myocytes on a definite stage of differentiation. Leptofibrils are attached to Z-bands running parallel or perpendicular to myofibrils. The period of their cross striation varies from 135 to 215 nm. Possible functions of the leptofibrils are discussed.


Regulation of the on bipolar cell mGluR6 pathway by Ca2+.
Glutamate produces a hyperpolarizing synaptic potential in On bipolar cells by binding to the metabotropic glutamate receptor mGluR6, leading to closure of a cation channel. Here it is demonstrated that this cation channel is regulated by intracellular Ca(2+). Glutamate-evoked currents were recorded from On bipolar cells in light-adapted salamander retinal slices in the presence of 2 mm external Ca(2+). When glutamate was applied almost continuously, interrupted only briefly to measure the size of the response, the glutamate response remained robust. However, currents elicited by intermittent and brief applications of glutamate exhibited time-dependent run down. Run down of the glutamate response was also voltage dependent, because it was accelerated by membrane hyperpolarization. Run down was triggered, at least in part, by a rise in intracellular Ca(2+); measured as a function of time or voltage, it was attenuated by intracellular buffering of Ca(2+) with BAPTA or by omitting Ca(2+) from the bathing solution. Current-voltage measurements demonstrated that Ca(2+) induced run down of the glutamate response by downregulating cation channel function, rather than by preventing closure of the channel by glutamate and mGluR6. A major source of the Ca(2+) that mediated this inhibition is the cation channel itself, which was found to be permeable to Ca(2+), accounting for the use dependence of the run down. These results suggest that Ca(2+) influx through the cation channel during background illumination could provide a signal to close the cation channel and repolarize the membrane toward its dark potential, an adaptive mechanism for coping with changes in ambient light.


Effect of rehabilitation setting on dependence following stroke: an analysis of the French inpatient database.
In France in 2009, patients admitted to Multidisciplinary Inpatient Rehabilitation for stroke were sent to a neurological rehabilitation center (NRC) or a general or geriatric rehabilitation (GRC) service. To describe the functional outcome of stroke patients admitted for rehabilitation in France in 2009, both globally and as a function of the rehabilitation setting (GRC or NRC). Data from the French Hospital Discharge Diagnosis databases for 2009 were included. Two logistic regression models were used to analyze factors related to improvement in dependence score and discharge home. Odds ratios (ORs) were also calculated. Among the 83 505 survivors of acute stroke in 2009, 28 201 were admitted for rehabilitation (33.8%). Of these, 19 553 went to GRC (69%) and 8648 to NRC (31%). On average, patients admitted to GRC were older (78.6 years vs 66.4 years), P < .001). At the start of rehabilitation, 50% of NRC patients and 56% of GRC patients were heavily dependent, but level of dependence was similar within each age-group. Rehabilitation in NRC lead to a greater probability of functional improvement (OR = 1.75, P < .001) and home discharge (OR = 1.61, P < .001) after adjustment for gender, age, Charlson's comorbidity index, initial level of dependence, type of stroke, and total length of stay. This study confirms, on a national level, the functional benefit of specialized rehabilitation in NRC. These results should be useful in the improvement of care pathways, organization of rehabilitation, and discharge planning.


[3rd and 4th generation estroprogestatives French controversy: What effect on women's behavior?].
The primary objective of this study was to determine the reasons given by women who have changed or suspended their contraceptive method in 2013. What influence had the 3rd and 4th generation estroprogestatives (EP) French controversy and advices from doctors? Did they notice any consequences of the controversy on their lifestyle? Did they feel an impact on their behavior to health professionals? A study was conducted on women of childbearing age. Data collection took place between November 4 and December 16, 2013. The included subjects were between 18 and 55 years old females. Subjects were surveyed through an anonymous self-administered questionnaire distributed through pharmacies. The protocol called for the construction of clusters of subjects by having heard about, or not, of the controversy. Patients reporting having heard about the controversy have formed the "exposed" group while the others have formed the "unexposed" group. We compared two parameters between these two groups: the rate of subjects who reported having modified or suspended their contraceptive method in 2013 on one hand, and the rate of subjects who reported having made a change in their contraceptive method without or against advices from a doctor on the other hand. The sample included 988 subjects. The average age was 34 years. The level of exposure to media debates was 86.5%. Of the respondents, 19.8% were under 3rd or 4th generation EP in 2012, thus directly involved in the discussions and affected by health recommendations. Of the patients, 38.5% reported having changed their contraception in 2013. In these women, when they confirmed having heard about the controversy, 40.1% of them claimed to have conducted this process without or against advices from a doctor. This number dropped to 18.2% for those who had not heard of it. Media coverage of the 3rd and 4th generation EP vascular risk was not mentioned more by patients who suspended all medical contraceptive method than it was by patients who simply changed their method. In all, 52.1% of women who completely stopped their contraception indicated that this choice was at least partly due to a change in their sexuality or reproductive life. This reason was evoked only in 19.0% of women who have simply made a change of contraceptive method. Among the views of women, the EP controversy was one of the reasons for a change of contraception method in 41.8% of cases, while advices from a doctor did play a role in 19.9% of cases. One has seen a decline of 11.0% of oral contraception between 2012 and 2013, almost superimposed on that of oral 3rd and 4th generation EP (10.6%). However, the use of 1st and 2nd generation EP stagnated: their increase was 0.4%. Beyond any change of contraceptive method, 66.9% of women expressed the absence of any impact of the controversy on their behavior. Of the women, 6.1% showed a loss of confidence in health professionals. We have shown how patients who stopped or modified their contraception method during 2013 placed the controversy influence in the forefront of the reasons for their decision. Although a medical reason has been widely quoted, only one in five felt that the advice of her doctor had clearly contributed to her choice. The influence of the media seemed to encroach on the doctor-patient relationship, which is fundamental to the quality of healthcare in the long term.


Population Genetic Structure of the Cayo Santiago Colony of Rhesus Macaques (Macaca mulatta).
The rhesus macaque population at Cayo Santiago increases annually and is in urgent need of control. In-depth assessments of the colony's population genetic and pedigree structures provide a starting point for improving the colony's long-term management program. We evaluated the degree of genetic variation and coefficients of inbreeding and kinship of the Cayo Santiago colony by using pedigree and short tandem repeat (STR) data from 4738 rhesus macaques, which represent 7 extant social groups and a group of migrant males. Information on each animal's parentage, sex, birth date, and date of death or removal from the island were used to generate estimates of mean kinship, kinship value, gene value, genome uniqueness (GU), founder equivalents (fe), and founder genome equivalents (fg). Pedigree and STR analyses revealed that the social groups have not differentiated genetically from each other due to male-mediated gene flow (that is, FST estimates were in the negative range) and exhibit sufficient genetic variation, with mean estimates of allele numbers and observed and expected heterozygosity of 6.57, 0.72, and 0.70, respectively. Estimates of GU, fe, and fg show that a high effective number of founders has affected the colony's current genetic structure in a positive manner. As demographic changes occur, genetic and pedigree matrices need to be monitored consistently to ensure the health and wellbeing of the Cayo Santiago colony.


Sequence-ready contig for the 1.4-cM ductal carcinoma in situ loss of heterozygosity region on chromosome 8p22-p23.
We report the construction of an approximately 1.7-Mb sequence-ready YAC/BAC clone contig of 8p22-p23. This chromosomal region has been associated with frequent loss of heterozygosity (LOH) in breast, ovarian, prostate, head and neck, and liver cancer. We first constructed a meiotic linkage map for 8p to resolve previously reported conflicting map orders from the literature. The target region containing a putative tumor suppressor gene was defined by allelotyping 65 cases of sporadic ductal carcinoma in situ with 18 polymorphic markers from 8p. The minimal region of loss encompassed the interval between D8S520 and D8S261, and one tumor had loss of D8S550 only. We chose to begin physical mapping of this minimal LOH region by concentrating on the distal end, which includes D8S550. A fine-structure radiation hybrid map for the region that extends from D8S520 (distal) to D8S1759 (proximal) was prepared, followed by construction of a single, integrated YAC/BAC contig for the interval. The approximately 1730-kb contig consists of 13 YACs and 27 BACs. Fifty-four sequence-tagged sites (STSs) developed from BAC insert end-sequences and 11 expressed sequence tags were localized within the contig by STS content mapping. In addition, four unique cDNA clones from the region were isolated and fully sequenced. This integrated YAC/BAC resource provides the starting point for transcription mapping, genomic sequencing, and positional cloning of this region.


Cerebral dysfunction in chronic alcoholism: role of alcoholic liver disease.
Evidence suggests that liver disease per se may contribute to the cognitive and motor impairments encountered in chronic alcoholics. Neuropathologic studies reveal astrocytic changes (Alzheimer type II astrocytosis) in the brains of alcoholic cirrhotic patients who died in hepatic coma. Pathophysiologic mechanisms responsible for hepatic (portal-systemic) encephalopathy in alcoholics include the loss of neuron-astrocytic metabolic trafficking as well as selective alterations of serotoninergic and dopaminergic function. In addition, there is evidence to suggest that endogenous ligands for both central-type (GABA-related) and "peripheral-type" (astrocytic) benzodiazepine receptors are implicated in the pathogenesis of hepatic encephalopathy in these patients. Chronic liver disease may also interfere with brain thiamine homeostasis and thus contribute to the pathogenesis of the Wernicke-Korsakoff syndrome in chronic alcoholism.


Flow of mitochondrial DNA across a species boundary.
Restriction analysis shows that wild Scandinavian mice belonging to the species Mus musculus contain the mitochondrial DNA of a neighboring species, M. domesticus. This demonstration results from comparisons of Scandinavian mice with authentic M. domesticus and M. musculus from other parts of Europe. Electrophoretic and immunological analysis of eight diagnostic proteins confirms that mice from north of the hybrid zone in Denmark are M. musculus in regard to their nuclear genes. In contrast, the mice tested from this region and a nearby part of Sweden have exclusively M. domesticus types of mitochondrial DNA. Phylogenetic analysis of the restriction maps suggests that the mitochondrial DNAs found in Scandinavian M. musculus could stem from a single M. domesticus female.


A degenerate primer MOB typing (DPMT) method to classify gamma-proteobacterial plasmids in clinical and environmental settings.
Transmissible plasmids are responsible for the spread of genetic determinants, such as antibiotic resistance or virulence traits, causing a large ecological and epidemiological impact. Transmissible plasmids, either conjugative or mobilizable, have in common the presence of a relaxase gene. Relaxases were previously classified in six protein families according to their phylogeny. Degenerate primers hybridizing to coding sequences of conserved amino acid motifs were designed to amplify related relaxase genes from γ-Proteobacterial plasmids. Specificity and sensitivity of a selected set of 19 primer pairs were first tested using a collection of 33 reference relaxases, representing the diversity of γ-Proteobacterial plasmids. The validated set was then applied to the analysis of two plasmid collections obtained from clinical isolates. The relaxase screening method, which we call "Degenerate Primer MOB Typing" or DPMT, detected not only most known Inc/Rep groups, but also a plethora of plasmids not previously assigned to any Inc group or Rep-type.


Heme oxygenase-1 and heat shock protein 70 induction in glia and neurons throughout rat brain after experimental intracerebral hemorrhage.
Current experimental evidence demonstrates the development of ischemic regions adjacent to and spatially remote from an intracerebral hematoma. The cause of this ischemia is uncertain. Because ischemia is a known inducer of stress genes, we investigated the induction of two stress proteins, heme oxygenase (HO)-1 and heat shock protein (Hsp) 70, after intracerebral hemorrhage in the rat. Immunocytochemistry for HO-1, Hsp70, and HO-2, the constitutive isoform of the HO enzyme, was performed 1, 2, and 4 days after striatal injection of saline, whole blood, or lysed blood. Immunocytochemistry for HO-1, HO-2, and Hsp70 was also performed 1 day after cortical injection of saline, whole blood, or lysed blood. After striatal injection of lysed and whole blood, the HO-1 protein was induced in glia throughout the hemisphere ipsilateral to the hematoma, and HO-1 immunoreactivity persisted for at least 4 days. After cortical injection of lysed and whole blood, HO-1 was induced in glia throughout the neocortex. Neuronal induction of HO-1 was also observed after cortical injection of lysed blood but not whole blood or saline. After striatal injection of lysed blood, Hsp70 was induced in glia surrounding the hematoma and in neurons from the neocortex overlying the hematoma and the striatum adjacent to the hematoma. After cortical injection of lysed blood, Hsp70 was induced in neurons throughout the neocortex and hippocampus bilaterally. In contrast, after whole blood and saline injection into cortex, Hsp70 induction was observed only in scattered neurons surrounding the hematoma cavity. Our results demonstrate that blood in the brain parenchyma induces the HO-1 stress protein but does not significantly alter HO-2 immunostaining. Our results also demonstrate that lysed blood induces Hsp70 in multiple regions of the brain and that the stress response of the brain differs depending on whether lysed blood is injected into the cortex or striatum. These results suggest that blood lysis may play an unforeseen role in the stress response of the brain to intracerebral hemorrhage.