Alzheimer's disease: new horizons in diagnosis and treatment.
We are in the midst of an explosion in research in Alzheimer's disease. Advances in our scientific understanding of genetic factors and pathogenesis are leading to enhanced diagnostic capabilities and new presymptomatic and symptomatic therapies. Estrogen therapy in postmenopausal females and NSAIDs show early promise, while AChE inhibitors keep getting better. Armed with knowledge and new diagnostic and therapeutic tools, we can finally begin to fight this devastating disease.


A Density-Based Clustering Approach to Motor Unit Potential Characterizations to Support Diagnosis of Neuromuscular Disorders.
Electrophysiological muscle classification involves characterization of extracted motor unit potentials (MUPs) followed by the aggregation of these MUP characterizations. Existing techniques consider three classes (i.e., myopathic, neurogenic, and normal) for both MUP characterization and electrophysiological muscle classification. However, diseased-induced MUP changes are continuous in nature, which make it difficult to find distinct boundaries between normal, myopathic, and neurogenic MUPs. Hence, MUP characterization based on more than three classes is better able to represent the various effects of disease. Here, a novel, electrophysio- logical muscle classification system is proposed, which considers a dynamic number of classes for characterizing MUPs. To this end, a clustering algorithm called neighbor- hood distances entropy consistency is proposed to find clusters with arbitrary shapes and densities in an MUP feature space. These clusters represent several concepts of MUP normality and abnormality and are used for MUP characterization instead of the conventional three classes. An examined muscle is then classified by embedding its MUP characterizations in a feature vector fed to an ensemble of support vector machine and nearest neighbor classifiers. For 103 sets of MUPs recorded in tibialis anterior muscles, the proposed system had a 97% electro-physiological muscle classification accuracy, which is significantly higher than in previous works.


Minoxidil topical solution: an unsafe product for children.
Minoxidil hair formulation is commonly used for the treatment of male or female androgenic alopecia. This over-the-counter product is wrongly considered safe. The ingestion of a few milliliters by a child can lead to significant intoxication. We report a case of significant intoxication after the ingestion of topical minoxidil (Alopexy; Pierre Fabre Laboratoires, SA, Switzerland). A 7-year-old girl, who accidentally ingested a teaspoon of minoxidil hair solution, presented to the pediatric emergency department for emesis. At admission, she had a blood pressure of 86/56 mm Hg and a pulse of 149 beats per minute. Hypotension lasted 40 hours with the lowest value 24 hours after ingestion (79/33 mm Hg). She presented electrocardiogram changes (sinus tachycardia and flattening T-waves) but normal cardiac enzymes. Infusion of 20 mL/kg of normal saline fluid had no hemodynamic effect. Her blood pressure normalized on day 2. Minoxidil topical solution is an unsafe product for children. This formulation should be strictly kept out of reach of children and manufacturers should enhance child-resistance security of packaging. The over-the-counter availability must be questioned.


Endogenous DNA double-strand breaks: production, fidelity of repair, and induction of cancer.
This article extends our previous quantitative analysis of the relationship between the dynamics of the primary structure of DNA and mutagenesis associated with single-strand lesions to an analysis of the production and processing of endogenous double-strand breaks (EDSBs) and to their implications for oncogenesis. We estimate that in normal human cells approximately 1% of single-strand lesions are converted to approximately 50 EDSBs per cell per cell cycle. This number is similar to that for EDSBs produced by 1.5-2.0 Gy of sparsely ionizing radiation. Although EDSBs are usually repaired with high fidelity, errors in their repair contribute significantly to the rate of cancer in humans. The doubling dose for induced DSBs is similar to doubling doses for mutation and for the induction of carcinomas by ionizing radiation. We conclude that rates of production of EDSBs and of ensuing spontaneous mitotic recombination events can account for a substantial fraction of the earliest oncogenic events in human carcinomas.


Effects of drought stress on legume symbiotic nitrogen fixation: physiological mechanisms.
Drought stress is one of the major factors affecting nitrogen fixation by legume-rhizobium symbiosis. Several mechanisms have been previously reported to be involved in the physiological response of symbiotic nitrogen fixation to drought stress, i.e. carbon shortage and nodule carbon metabolism, oxygen limitation, and feedback regulation by the accumulation of N fixation products. The carbon shortage hypothesis was previously investigated by studying the combined effects of CO2 enrichment and water deficits on nodulation and N2 fixation in soybean. Under drought, in a genotype with drought tolerant N2 fixation, approximately four times the amount of 14C was allocated to nodules compared to a drought sensitive genotype. It was found that an important effect of CO2 enrichment of soybean under drought was an enhancement of photo assimilation, an increased partitioning of carbon to nodules, whose main effect was to sustain nodule growth, which helped sustain N2 rates under soil water deficits. The interaction of nodule permeability to O2 and drought stress with N2 fixation was examined in soybean nodules and led to the overall conclusion that O2 limitation seems to be involved only in the initial stages of water deficit stresses in decreasing nodule activity. The involvement of ureides in the drought response of N2 fixation was initially suspected by an increased ureide concentration in shoots and nodules under drought leading to a negative feedback response between ureides and nodule activity. Direct evidence for inhibition of nitrogenase activity by its products, ureides and amides, supported this hypothesis. The overall conclusion was that all three physiological mechanisms are important in understanding the regulation of N2 fixation and its response of to soil drying.


Smoking Modifies the Association Between Radon Exposure and Incident Ischemic Stroke: The REGARDS Study.
Exposure to radon has been linked to lung cancer and other lung diseases. Although biologically plausible, research of residential radon exposure in relation to stroke risk is scarce. Study participants were from the REGARDS (Reasons for Geographic and Racial Differences in Stroke) cohort (n=30 239), which consisted of male and female non-Hispanic Black and White adults aged 45 and older. After excluding participants with baseline stroke and transient ischemic attack, and missing information on exposure and outcome of interest, the final sample size was 26 950. The primary outcome was time to the first ischemic stroke through September 30, 2020. County-level radon measures from Lawrence Berkeley National Laboratory were linked to each participant based on their geocoded residential history. We used Cox proportional hazards regression models with a time-dependent exposure to estimate hazard ratios and 95% CIs for the association. After controlling for potential confounding factors including demographic, lifestyle, clinical variables, and PM2.5, radon exposure was significantly associated with incident ischemic stroke among never-smokers (hazard ratio, 1.39 [95% CI, 1.01-1.90]) but not ever-smokers. The results were generally consistent in the sensitivity analysis when using radon measures from state/Environmental Protection Agency residential radon survey. Findings from this study suggest that the association between residential radon exposure and incidence of ischemic stroke varies by smoking status and may be prominent in never-smokers. Further studies incorporating indoor-radon measures are needed to confirm these findings.


Identification of a Theileria mutans-specific antigen for use in an antibody and antigen detection ELISA.
Purified piroplasms of Theileria mutans were used to immunize BALB/c mice to generate monoclonal antibodies (MoAbs). The MoAbs recognized an antigen of a relative molecular mass of 32 kDa in Western blots. This antigen was also recognized by sera from cattle which had recovered naturally from experimental tick-transmission or infections induced by the blood stages of T. mutans. The MoAbs did not react, in indirect immunofluorescence or enzyme-linked immunosorbent assays (ELISA), with the common haemoparasites of cattle, namely, T. parva, T. annulata, Babesia bigemina, B. bovis, Anaplasma marginale, Trypanosoma congolense, T. vivax or T. brucei. An antigen capture ELISA was established with two of the MoAbs which recognized different epitopes on the 32 kDa molecule. Using this test it was possible to detect circulating antigens or immune complexes in sera collected from cattle during the acute or chronic phases of infection. When the purified 32 kDa protein was used as antigen in a micro-ELISA to detect circulating antibodies in both experimental and field cattle sera, it was found that the titres of antibodies ranged between 1:20 and 1:10,240. Results of this study indicate that the antigen and immune complex capture assays and the antibody detection ELISA can be complementary in the immunodiagnosis of acute and chronic T. mutans infections. Moreover, the tests are useful in the differential diagnosis of the disease and for epidemiological studies.


Multistate product traceforward investigation to link imported romaine lettuce to a US cyclosporiasis outbreak - Nebraska, Texas, and Florida, June-August 2013.
During June-August 2013, 25 US states reported 631 cyclosporiasis cases including Nebraska and Iowa where a regional investigation implicated common-source imported salad mix served in two chain restaurants. At least two common-origin growing fields were likely sources of contaminated romaine lettuce. Using producer- and distributor-provided data, we conducted a grower-specific traceforward investigation to reveal exposures of ill US residents elsewhere who reported symptom onset during 11 June-1 July 2013, the time period established in the Nebraska and Iowa investigation. Romaine lettuce shipped on 2-6 June from one of these Mexico-origin growing fields likely caused cyclosporiasis in 78 persons reporting illness onsets from 11 June to 1 July in Nebraska, Texas, and Florida. Nationwide, 97% (314/324) of persons confirmed with cyclosporiasis with symptom onset from 11 June to 1 July 2013 resided in 11 central and eastern US states receiving approximately two-thirds of romaine lettuce from this field. This grower's production practices should be investigated to determine potential sources of contamination and to develop recommendations to prevent future illnesses.


Immunohistochemical analysis of the ras oncogene protein product in human thyroid neoplasms.
The monoclonal antibody Y13 259 to the ras oncogene p21 product was used in an immunohistochemical study of ras expression in human thyroid neoplasms. Both papillary and follicular carcinomas showed consistently higher staining intensity than normal tissue adjacent to carcinomas. Some follicular, foetal, embryonal and oxyphil adenomas exhibited higher staining intensity than normal tissue. Our results suggest that elevated ras expression may be important in the development of some adenomas and in their conversion to carcinomas.


Predictive ability of the pooled cohort risk assessment for the incidence and progression of coronary artery calcification.
The aim of the present study was to evaluate the predictive ability of pooled cohort risk assessment (PRA) models for the incidence and progression of coronary artery calcification (CAC) over time comparing the Framingham risk score (FRS) in a healthy Korean population. The study cohort included 4194 individuals, aged 40 or older, without a history of cardiovascular disease or statin therapy, who underwent repeated cardiac computed tomography for CAC scoring from 2011 to 2014. Logistic regression models and Cox proportional hazards models were used. Of the 4194 individuals, 1244 had CAC progression and during over 5184 person-years of follow-up, 204 participants developed incident CAC. Compared with patients whose FRS was less than 10%, the odds ratios (ORs) [95% confidence intervals (CIs)] for CAC progression in patients whose FRS ranged from 10 to 20% and whose FRS was at least 20% were 2.54 (2.17-2.98) and 3.73 (2.69-5.15), respectively. Similarly, compared with the reference group, whose PRA was less than 5%, the ORs (95% CIs) for CAC progression in patients whose PRAs ranged from 5 to less than 7.5%, 7.5 to less than 10%, and at least 10% were 1.85 (95% CI 1.52-2.25), 2.63 (95% CI 2.01-3.46), and 3.58 (95% CI 2.73-4.70), respectively. Similarly, both PRA and FRS were associated positively with the development of CAC in a dose-response manner. In discriminating incident CAC and progression using receiver operating characteristic analysis, PRA was not significantly different from FRS. In a large sample of asymptomatic Korean adults without cardiovascular disease, both the PRA and FRS similarly predict the onset and progression of subclinical atherosclerosis over time.


Enhancing automatic closed-loop glucose control in type 1 diabetes with an adaptive meal bolus calculator - in silico evaluation under intra-day variability.
Current prototypes of closed-loop systems for glucose control in type 1 diabetes mellitus, also referred to as artificial pancreas systems, require a pre-meal insulin bolus to compensate for delays in subcutaneous insulin absorption in order to avoid initial post-prandial hyperglycemia. Computing such a meal bolus is a challenging task due to the high intra-subject variability of insulin requirements. Most closed-loop systems compute this pre-meal insulin dose by a standard bolus calculation, as is commonly found in insulin pumps. However, the performance of these calculators is limited due to a lack of adaptiveness in front of dynamic changes in insulin requirements. Despite some initial attempts to include adaptation within these calculators, challenges remain. In this paper we present a new technique to automatically adapt the meal-priming bolus within an artificial pancreas. The technique consists of using a novel adaptive bolus calculator based on Case-Based Reasoning and Run-To-Run control, within a closed-loop controller. Coordination between the adaptive bolus calculator and the controller was required to achieve the desired performance. For testing purposes, the clinically validated Imperial College Artificial Pancreas controller was employed. The proposed system was evaluated against itself but without bolus adaptation. The UVa-Padova T1DM v3.2 system was used to carry out a three-month in silico study on 11 adult and 11 adolescent virtual subjects taking into account inter-and intra-subject variability of insulin requirements and uncertainty on carbohydrate intake. Overall, the closed-loop controller enhanced by an adaptive bolus calculator improves glycemic control when compared to its non-adaptive counterpart. In particular, the following statistically significant improvements were found (non-adaptive vs. adaptive). Adults: mean glucose 142.2 ± 9.4vs. 131.8 ± 4.2mg/dl; percentage time in target [70, 180]mg/dl, 82.0 ± 7.0vs. 89.5 ± 4.2; percentage time above target 17.7 ± 7.0vs. 10.2 ± 4.1. Adolescents: mean glucose 158.2 ± 21.4vs. 140.5 ± 13.0mg/dl; percentage time in target, 65.9 ± 12.9vs. 77.5 ± 12.2; percentage time above target, 31.7 ± 13.1vs. 19.8 ± 10.2. Note that no increase in percentage time in hypoglycemia was observed. Using an adaptive meal bolus calculator within a closed-loop control system has the potential to improve glycemic control in type 1 diabetes when compared to its non-adaptive counterpart.


Sociodemographic and environmental health risk factor of COVID-19 in Jakarta, Indonesia: An ecological study.
Since December 2019, the COVID-19 pandemic has rapidly emerged on a global scale. Many factors have influenced the spread of COVID-19. This research studies the sociodemographic and environmental health risk factors associated with COVID-19. The study used an ecological study design with subdistricts as its unit of analysis. The total population was 44 subdistricts. Data analysis used correlation and linear regression tests. The study results showed that the average COVID-19 incident rate in Jakarta is 99.8 per 10,000 population. Risk factors for the spread of COVID-19 were associated with population's high level of education (B = 3.094, p value<0.001), population density (B = 0.275, p value = 0.029), and slum area (B = 0.404, p value<0.001). The main risk factor for the spread of COVID-19 in Jakarta is high level of education, which can reflect a higher economic status to the population and a tendency to be more mobile. The government needs to enforce a mobility restriction to lessen the spread of COVID-19.


New PET/CT Features for the Evaluation of Tumor Response.
With the emerging multi-modality imaging performed at multiple time points for each patient, it becomes more important to analyze the serial images quantitatively, select and combine both complementary and contradictory information from various sources, for accurate and personalized evaluation of tumor response to therapy.


Platelet-activating factor-induced polymorphonuclear neutrophil priming independent of CD11b adhesion.
Our previous studies have implicated phospholipase A2-dependent platelet-activating factor (PAF) production in the genesis of polymorphonuclear neutrophil (PMN)-mediated tissue injury after gut ischemia-reperfusion. Further, these studies have suggested a discordance of PMN sequestration and tissue injury. CD11B-dependent PMN-endothelial cell adhesion has been purported to play a dominant role in PMN-mediated tissue injury. We therefore undertook this study with the hypothesis that PAF-induced PMN superoxide production requires CD11B-mediated PMN-endothelial cell adherence. Human PMNs, isolated by Percoll gradient centrifugation, were exposed to PAF (10 ng/ml). At fixed times of exposure during 120 minutes, (1) superoxide production, (2) CD11B receptor expression, and (3) PMN adhesion to unstimulated human umbilical vein endothelial cell cultures were assayed. PAF induced prompt changes in PMN priming (increased superoxide production after N-formyl-methyl-leucyl-phenylalanine activation), adhesion to unstimulated endothelial cells, and CD11B receptor expression. Priming was temporally concordant with the rise and fall of CD11B expression but appeared to precede adhesion. CD11B blockade (F(Ab') 2 anti-CD11B [60.1] antibodies), before or at maximal PAF priming, reduced PMN adhesion but had no effect on superoxide production. In summary, PAF-induced PMN priming occurs in temporal concordance with the expression of CD11B and subsequent endothelial cell adherence, but CD11B-mediated adherence is not essential for this process.


(1Z,3Z)-1,4-Di(pyridin-2-yl)buta-1,3-diene-2,3-diol: the planar highly conjugated symmetrical enediol with multiple intramolecular hydrogen bonds.
1H, (13)C, and (15)N NMR spectral data show that in chloroform solution (1Z,3Z)-1,4-di(pyridin-2-yl)buta-1,3-diene-2,3-diol, OO, is in ca. 9:1 equilibrium with (3Z)-3-hydroxy-1,4-di(pyridin-2-yl)but-3-en-2-one, OK, while no 1,4-di(pyridin-2-yl)-2,3-butanedione, KK, was detected. The species present in the tautomeric mixture were identified by comparing their experimental chemical shifts with those known for similar compounds as well as with the theoretically calculated (GIAO-HF/DFT) values. Ab initio calculations show that OK and especially the highly conjugated OO forms are preferred in the tautomeric mixtures both in vacuo and in chloroform solution. Comparison of experimental (Arrhenius) and calculated (ab initio) energies of OK shows that the MP2/6-31G//RHF/6-31G method gives the most precise results. There are one and two strong O-H.N hydrogen bonds present in the molecules of the former and latter compound, respectively. Other tautomeric forms, e.g., dienaminedione [(1Z,4Z)-1,4-di[pyridin-2(1H)-ylidene]butane-2,3-dione], and their rotamers were found to have higher energies. The single-crystal X-ray diffraction studies show that dienediol OO is the only tautomeric form present in the crystal at 173 K. Its almost perfectly planar molecule is stabilized by two strong intramolecular O-H.N hydrogen bonds.


Study protocol for a randomized controlled trial for anterior inguinal hernia repair: transrectus sheath preperitoneal mesh repair compared to transinguinal preperitoneal procedure.
Anterior open treatment of the inguinal hernia with a tension-free mesh has reduced the incidence of hernia recurrence. The Lichtenstein procedure is the current reference technique for inguinal hernia treatment. Chronic pain has become the main postoperative complication after surgical inguinal hernia repair, especially following Lichtenstein. Preliminary experiences with a soft mesh positioned in the preperitoneal space (PPS) by transinguinal preperitoneal (TIPP) or total extraperitoneal (TEP) technique, showed promising results considering the reduction of postoperative chronic pain. Evolution of surgical innovations for inguinal hernia repair led to an open, direct approach with preperitoneal mesh position, such as TIPP. Based on the TIPP procedure, another preperitoneal repair has been recently developed, the transrectus sheath preperitoneal (TREPP) mesh repair. The ENTREPPMENT trial is a multicentre randomized clinical trial. Patients will be randomly allocated to anterior inguinal hernia repair according to the TREPP mesh repair or TIPP procedure. All patients with a primary unilateral inguinal hernia, eligible for operation, will be invited to participate in the trial. The primary outcome measure will be the number of patients with postoperative chronic pain. Secondary outcome measures will be serious adverse events (SAEs), including recurrence, hemorrhage, return to daily activities (for example work), operative time and hospital stay. Alongside the trial health status, an economic evaluation will be performed. To demonstrate that inguinal hernia repair according to the TREPP technique reduces the percentage of patients with postoperative chronic pain from 12% to <6%, a sample size of 800 patients is required (two-sided test, α = 0.05, 80% power).The ENTREPPMENT trial aims to evaluate the TREPP and TIPP procedures from patients' perspective. It is hypothesized that the TREPP technique may reduce the number of patients with any form of postoperative chronic pain by 50% compared to the TIPP procedure. Current Controlled Trials: ISRCTN18591339.


Base-Free Generation of Organic Electron Donors from Air-Stable Precursors.
Organic electron donors (OEDs) are powerful reducing agents recognized for their potential in the reduction of challenging substrates and in original applications. Nonetheless, their low stability in atmospheric oxygen or over time complicates their manipulation and storage. To overcome these constraints and enhance OED practicality, new air- and moisture-stable aminopyridinium carboxylate and carbonate precursors were synthesized and thermally activated to generate the potent electron donor in situ. Carboxylate adducts proved to be excellent latent OED systems, enabling the facile and efficient reduction of challenging substrates. Their reduction properties were correlated to their structural characteristics by thermogravimetric and spectroscopic analysis.


Biological and tumor-promoting effects of dioxin-like and non-dioxin-like polychlorinated biphenyls in mouse liver after single or combined treatment.
To assess the impact of a mixture containing dioxin-like and non-dioxin-like polychlorinated biphenyls (PCBs), male mice were initiated with N-nitroso-diethylamine and subsequently treated with PCB126, an Ah-Receptor agonist, and PCB153, acting via activation of the constitutive androstane receptor. The two congeners were given at two dose levels: the low dose was adjusted to induce ~150-fold increases in cytochrome P450 (Cyp)1a1 (PCB126) and Cyp2b10 mRNAs (PCB153), and the high dose was chosen as twice the low dose. To keep the liver PCB levels constant, mice were given initial loading doses followed by weekly maintenance doses calculated on the basis of the PCBs' half-lives. Mice were treated with the individual congeners (low and high dose) or with a mixture consisting of the low doses of the 2 PCBs. The following results were obtained: (1) the 2 PCBs produced dose-dependent increases in Cyp1a1 and Cyp2b10 mRNA, protein, and activity when given individually; (2) combined treatment caused more than additive effects on Cyp1a1 mRNA expression, protein level, and ethoxyresurofin activity; (3) changes in the levels of several proteins were detected by proteome analysis in livers of PCB-treated mice; (4) besides these biological responses, the individual PCBs caused no significant increase in the number of glucose-6-phospatase (G6Pase)-deficient neoplastic lesions in liver, whereas a moderate significant effect occurred in the combination group. These results suggest weak but significant response-additive effects of the 2 PCBs when given in combination. They also suggest that the Cyp biomarkers tend to overestimate the carcinogenic response produced by the PCBs in mouse liver.


EGFR-induced cytoskeletal changes drive complex cell behaviors: The tip of the iceberg.
Cytoskeletal networks are dramatically reorganized upon EGF stimulation to enable complex cell behaviors such as cell-cell communication, migration and invasion, and cell division. In this issue of Science Signaling, Roth et al. and Pike et al. use proteomic methods to identify several effectors of EGF responses. The findings show the interdependent nature of growth factor signaling and the cytoskeleton and identify potential new therapeutic targets for treating cancer and other growth factor-driven diseases.


The impact of health worker absenteeism on patient health care seeking behavior, testing and treatment: A longitudinal analysis in Uganda.
Absenteeism of frontline health workers in public sector facilities is widespread in low-income countries. There is little quantitative evidence on how health worker absenteeism influences patient treatment seeking behavior, though low public sector utilization and heavy reliance on the informal sector are well documented in low-income settings. Using a unique panel dataset covering health facilities and households over a 10-month period in Uganda, we investigate the extent to which health worker absenteeism (defined as zero health workers present at a health facility) impacts patient care seeking behavior, testing, and treatment. We find high rates of health worker absenteeism at public sector health facilities, with most of the absenteeism occurring at lower level public health clinics. On average, no health worker was present in 42% of all days monitored in lowest level public health clinics, whereas this number was less than 5% in high level public hospitals and private facilities. In our preferred empirical model with household fixed effects, we find that health worker absenteeism reduces the odds that a patient seeks care in the public sector (OR = 0.65, 95% CI = 0.44-0.95) and receives malaria testing (OR = 0.73, 95% CI = 0.53-0.99) and increases the odds of paying out-of-pocket for treatment (OR = 1.41, 95% CI = 1.10-1.80). The estimated differences in care-seeking are larger for children under-five than for the overall study population. The impact of health worker absenteeism on the quality of care received as well as the financial burden faced by households in sub-Saharan Africa is substantial.


Injuries of the cervical spine in schoolboy rugby football.
Serious neck injury in rugby football is becoming more common, especially in schoolboys. We report five who presented with spinal cord damage between 1977 and 1983, two in the 1982-83 season. The mechanisms of injury, the treatment, and the outcome are discussed. We suggest measures to reduce the incidence of this most serious injury and draw attention to the appropriate management in the critical phase directly after injury.


Japan's challenges of translational regenerative medicine: Act on the safety of regenerative medicine.
The first issue of Nature Medicine published 20 years ago featured an article that reported Japan's critical situation regarding clinical trials, calling for major reform. Twenty years later, Japan has enacted three laws to promote the use of regenerative medicine as a national policy. The first law to be enacted was the Regenerative Medicine Promotion Act, which represents the country's determination to work toward the promotion of regenerative medicine. Subsequently, the Pharmaceuticals, Medical Devices, and Other Therapeutic Products Act (PMD Act) and the Act on the Safety of Regenerative Medicine (RM Act) came into effect. The PMD Act created a new category for regenerative medicine products, and established the process for obtaining approval for cell therapy and other regenerative therapies through the implementation of clinical trials. The RM Act specified the regulations that doctors, review committees, and cell culture/processing facilities must adhere to when providing regenerative medicine in medical care, not only in clinical research but also in private practice. Previously, researchers in regenerative medicine only had a set of guidelines to follow for conducting clinical research. Now, with the enactment of the RM Act, all areas for improvement that had been enumerated 20 years ago-such as the lack of appropriate review committees and governmental control-have been addressed by law, creating a system that gives the highest priority to patient safety. In this paper, we present the particularly noteworthy points of the RM Act, along with the actual current conditions of regenerative medicine in Japanese medical care.


TranScout: prediction of gene expression regulatory proteins from their sequences.
The advent of genomics yields thousands of reading frames in search of function. Identification of conserved functional motifs in protein sequences can be helpful for function prediction. A database and a classification of reported DNA-binding protein motifs has been designed. A program ('TranScout') has been developed for the detection and evaluation of conserved motifs in prokaryotic and eukaryotic sequences of proteins with a gene regulatory function. The efficiency of the program is shown in a benchmark against a database obtained from SWISS-PROT without the protein sequences used to train the program. All motifs were detected with a mean average sensitivity of 0.98 and a mean average specificity of 0.92. The program is freely available for use on the internet at http://luz.uab.es/transcout/. The user can find additional information at this site.


TEMPERATURE-SENSITIVE DEXTRANSUCRASE SYNTHESIS BY A LACTOBACILLUS.
Dunican, L. K. (Cornell University, Ithaca, New York), and H. W. Seeley, Jr. Temperature-sensitive dextransucrase synthesis by a lactobacillus. J. Bacteriol. 86:1079-1083. 1963.-Dextran synthesis was found to be temperature-dependent in Lactobacillus strain RWM-13. Dextran was not formed above 37 C, although growth of cells occurred up to 42 C. Logarithmically growing cells transferred from 30 C to 40 C ceased producing dextran while growth decreased nominally. An examination of the extracts of cells broken by sonic treatment showed that as the temperature of growth was increased above 37 C the production of dextransucrase decreased. By use of an inhibitor of invertase, 10(-4)m AgNO(3), it was shown that invertase replaced dextransucrase activity at temperatures above 37 C. In contrast to dextransucrase in Leuconostoc mesenteroides, the enzyme in Lactobacillus strain RWM-13 was constitutive and thus resembled that of Streptococcus bovis. Thermosensitivity of dextransucrase synthesis has not been observed in Leuconostoc or Streptococcus.


Lot-to-lot consistency, safety and immunogenicity of 3 lots of Haemophilus influenzae type b conjugate vaccine: results from a phase III randomized, multicenter study in infants.
Vaccination against Haemophilus influenzae type b (Hib) is included in routine pediatric immunization schedule in the United States. Previous vaccine shortages have created the need for additional options for Hib vaccination. This phase III, randomized, multi-centered study (NCT01000974) evaluated the safety and immunogenicity of a monovalent tetanus toxoid-conjugate Hib vaccine (Hib-TT) compared to a monovalent (Hib-TT control) and a combination Hib-TT vaccine. We hierarchically assessed lot-to-lot consistency of 3 Hib-TT lots and non-inferiority of Hib-TT to Hib-TT control. We co-administered routine pediatric vaccines with Hib-TT vaccines at 2, 4, 6months (primary vaccination) and 15-18months of age (booster vaccination). We recorded adverse events (AEs) for 4 (solicited) and 31days (unsolicited) post-vaccination and serious AEs (SAEs) throughout the study. Of 4009 enrolled children, 3086 completed booster phase. Lot-to-lot consistency was not demonstrated. The study met statistical criteria for non-inferiority of Hib-TT to Hib-TT control in terms of immune responses to Hib and co-administered vaccines' antigens, but not in terms of participants achieving post-primary vaccination anti-PRP levels ≥1µg/mL. Because of the hierarchical nature of the objectives, non-inferiority could not be established. In all groups, 92.5-96.7% and 99.6-100% of participants achieved anti-PRP levels ≥0.15µg/mL, while 78.3-89.8% and 97.9-99.1% had anti-PRP levels ≥1µg/mL, post-primary and post-booster vaccination, respectively. Immune responses to co-administered vaccines and reported incidence of AEs were comparable among groups. We recorded SAEs for 107/2963 (3.6%), 24/520 (4.6%), and 21/520 (4.0%) children post-primary vaccination, and 29/2337 (1.2%), 4/435 (0.9%), and 2/400 (0.5%) children post-booster vaccination with Hib-TT, Hib-TT control and combination Hib-TT vaccine, respectively; 6/5330 (0.1%) SAEs in the Hib-TT groups were considered vaccine-related. Hib-TT induced seroprotective antibody concentrations in the majority of participants and was well-tolerated when co-administered with routine pediatric vaccines according to a 3+1 schedule.


Serial determinations of Der p 1 and Der f 1 show predominance of one Dermatophagoides species.
One dust sample each month was collected for an average of 11 months throughout the year from the bedroom carpet of 39 mite allergic, asthmatic individuals living in the Tampa Bay area of Florida. Dust samples were collected from a square meter area from the bedroom carpet for 2 min using a portable vacuum cleaner and analyzed using two-site monoclonal antibody assays. The geometric mean of Dermatophagoides allergens Der p 1 and Der f 1 levels measured in each individual home was used in the statistical analysis of the results and reflects the average allergen level per year per home. Der p 1 levels were greater than Der f 1 in 21 homes; five homes had between 1.1 and 5 times more; one home had between 5 and 10; and 15 had between 10 and 175. Der f 1 levels were greater than Der p 1 in 18 homes; seven homes had between 1.1 and 5 times more; four had between 5 and 10; and seven homes had between 10 and 44. Spearman rank analysis of the results yielded a negative correlation coefficient (Rho) = -0.213 (p = 0.19) between Der p 1 and Der f 1 levels. These data suggest that when mite allergen levels reach a geometric mean value of approximately 2 micrograms/gram of dust throughout a year, one mite species predominates over the other.


Enhancing the functional maturity of induced pluripotent stem cell-derived human hepatocytes by controlled presentation of cell-cell interactions in vitro.
Induced pluripotent stem cell-derived human hepatocyte-like cells (iHeps) could provide a powerful tool for studying the mechanisms underlying human liver development and disease, testing the efficacy and safety of pharmaceuticals across different patients (i.e., personalized medicine), and enabling cell-based therapies in the clinic. However, current in vitro protocols that rely upon growth factors and extracellular matrices (ECMs) alone yield iHeps with low levels of liver functions relative to adult primary human hepatocytes (PHHs). Moreover, these low hepatic functions in iHeps are difficult to maintain for prolonged times (weeks to months) in culture. Here, we engineered a micropatterned coculture (iMPCC) platform in a multiwell format that, in contrast to conventional confluent cultures, significantly enhanced the functional maturation and longevity of iHeps in culture for at least 4 weeks in vitro when benchmarked against multiple donors of PHHs. In particular, iHeps were micropatterned onto collagen-coated domains of empirically optimized dimensions, surrounded by 3T3-J2 murine embryonic fibroblasts, and then sandwiched with a thin layer of ECM gel (Matrigel). We assessed iHep maturity by global gene expression profiles, hepatic polarity, secretion of albumin and urea, basal cytochrome P450 (CYP450) activities, phase II conjugation, drug-mediated CYP450 induction, and drug-induced hepatotoxicity. Controlling both homotypic interactions between iHeps and heterotypic interactions with stromal fibroblasts significantly matures iHep functions and maintains them for several weeks in culture. In the future, iMPCCs could prove useful for drug screening, studying molecular mechanisms underlying iHep differentiation, modeling liver diseases, and integration into human-on-a-chip systems being designed to assess multiorgan responses to compounds.


[Water quality model and its application in polluted river remediation by aeration].
Aeration is a main method to remediate polluted river water. Based on Shanghai Xingang River's aeration remediation, theoretical polluted water quality model was built. At the same time, parameters identification and solution were defined. Compared with observation values, the results from the model showed that: the dissolved oxygen had an average relative error of 11.38%, of which the relative error less than 20% accounted for 83.3%; BOD, had an average relative error of 9.16%, of which the relative error less than 20% accounted for 91.7%; ammonia had an average relative error of 11.15%, of which the relative error less than 20% accounted for 91.7%. With the exception of the individual points, most of the values from this model had relative error less than 20%. This model could describe well Xingang River's water quality under aeration conditions. Aeration intensity, velocity, sediment suspension and temperature simulation were analyzed by this model, which provides theoretical calculation methods and the basis for engineering design and operation of polluted river aeration remediation.


Role of diffusion-weighted imaging in the diagnosis of cerebral venous thrombosis.
To evaluate the hyperintense signal (HIS) performance on diffusion-weighted imaging (DWI) in diagnosing cerebral venous thrombosis (CVT). Seventy-eight patients with CVT hospitalized from January 2004 to January 2015 were retrospectively studied alongside 78 controls without intracranial organic diseases. Diagnostic accuracy indices of HIS on DWI or T2-weighted imaging (T2WI) to diagnose CVT at different sites and states were analyzed. The overall sensitivity of HIS on DWI for the diagnosis of CVT was significantly lower than that of HIS on T2WI (34.6% vs. 79.5%). HIS on T2WI was more sensitive than HIS on DWI in detecting thrombosis, especially in the superior sagittal sinus and transverse sinus. HIS on DWI was inversely related to the time between disease onset and imaging. Compared with HIS on T2WI, combining HIS on DWI and T2WI did not increase the sensitivity for detecting CVT. HIS on DWI was not detected in the control group, but HIS on T2WI was detected in 26.3% of control individuals. The specificity of HIS on DWI for CVT was higher than that of HIS on T2WI (97.4% vs. 76.9%). HIS on DWI has a lower sensitivity, but a higher specificity, than HIS on T2WI for diagnosing CVT.


Prevalence and determinants of obesity in an urban sample of Portuguese adults.
To evaluate the prevalence and the determinants of obesity, and the associated cardiovascular risk factors in a random sample of non-institutionalised adults. Cross-sectional study. A random sample of 1436 habitants of Porto (873 women and 563 men) aged 18-90 years. All participants answered a structured questionnaire comprising information on social, demographic, behavioural and clinical aspects. Anthropometric measures, blood pressure and fasting blood samples were obtained. Diet was assessed using a semi-quantitative food-frequency questionnaire, and physical activity was evaluated using a questionnaire exploring all professional, domestic and leisure-time activities. When the body mass index was > or =30 kg/m2, the subject was considered as 'obese'. Proportions were age adjusted for the European population. Odds ratios and 95% confidence intervals were computed using unconditional logistic regression. The prevalence of obesity was significantly higher in women (26.1%) than men (13.9%). Regardless of gender, obesity increased with age, decreased with education, and was more frequent in married blue-collar workers and unemployed subjects. Smoking was more prevalent in subjects of normal weight, and a higher proportion of those reporting no regular exercise were obese. In men, obesity prevalence increased with increasing quartiles of energy intake, but no such changes were found in women. Compared with subjects of normal weight, obese men showed a significantly higher prevalence of hypertension (53.3 vs 26.1%) and hypertriglyceridaemia (23.4 vs 9.0%). Also, hypertension (43.7 vs 30.7%), diabetes (7.6 vs 2.7%), hypertriglyceridaemia (27.1 vs 5.0%), and abnormal low-density lipoprotein (30.4 vs 21.4%) and high-density lipoprotein cholesterol concentration (15.0 vs 5.3%) were more frequent in obese women. Obesity is a major public health issue in urban Portuguese populations, and obese individuals have many features of metabolic syndrome. Education and relative deprivation are modifiable factors that are significantly associated with obesity. However, no clear-cut relationship was found between physical activity and energy intake.


Resveratrol Alleviates Cadmium-Induced Damage and Overexpression of Epidermal Growth Factor Receptor and its Downstream Signaling Proteins in the Reproductive System of Male Swiss Albino Mice.
Nowadays, exposure to heavy metals and their detrimental effects in humans are grave health concerns. In this study, we investigated the protective effect of resveratrol (RES) against CdCl2 (cadmium chloride)-induced impairment of spermatogenesis, histopathological alterations, and the up-regulation of epidermal growth factor receptor (EGFR) signaling cascade in Swiss albino mice. Two different doses of CdCl2 were injected intraperitoneally into two groups of mice, and in the third group RES was administered orally before injecting CdCl2 (3 times/wk) for 14 days. Sperm motility, count, vitality, and morphology were analyzed. Hematoxylin and eosin (H&E) staining, immunohistochemistry (IHC), and western blot analyses were performed on testis tissue. In CdCl2-administered animals, significant perturbations of spermatogenesis and histoarchitecture of seminiferous tubules were observed. p-EGFR, p-AKT, AKT1/2/3, NF-κβ (p50), and COX-2 of the EGFR cascade were up-regulated. Although there was significant negative correlation between percentage of motile cells and protein expression, we found positive correlation between morphologically abnormal cells and overexpression of proteins in CdCl2-only treated groups. Marked improvement of sperm parameters and histopathological damages as well as down-regulation of the EGFR signaling cascade were observed in the RES-pretreated mice. Hence, the present study elucidates that RES protects against CdCl2-induced perturbation of spermatogenesis and overexpression of EGFR and its downstream signaling proteins.


Between-day reliability of a cluster-based method for multisegment kinematic analysis of the foot-ankle complex.
Detailed description of foot pronation-supination requires multisegment evaluation of the kinematics of the foot-ankle complex. There are noninvasive methods with independent (single) tracking markers attached directly to the skin. However, these methods are inconsistent with the usual rigid segments assumption. In contrast, using clustered markers is compatible with this assumption and is necessary for analyses that need tracking markers to be distant from the foot (eg, shod walking). This study investigated the between-day reliability of a cluster-based method for multisegment analysis of foot-ankle angles related to pronation-supination. Ten healthy adults participated in the study. An anatomically based, three-dimensional model comprising the shank, calcaneus, and forefoot was created. Rigid clusters of tracking markers were used to determine the relative positions and motions of the segments. Mean positions were measured with the subtalar joint in neutral position during standing. Furthermore, mean angles, peaks, and timings of peaks were measured during the stance phase of walking. All of the variables were measured twice, with a 1-week interval. To evaluate reliability, intraclass correlation coefficients were calculated for discrete variables and coefficients of multiple correlation for entire gait curves. Intraclass correlation coefficients varied from 0.8 to 0.93 for the angles obtained when the subtalar joint was in neutral and from 0.76 to 0.9 for walking variables. Coefficients of multiple correlation varied from 0.93 to 0.97 for walking curves. The method described has good to high reliability and provides a systematic method for multisegment kinematic evaluation of foot-ankle pronation-supination.


Estimation of genetic trend in a selected population with and without the use of a control population.
Data from a selection experiment conducted with sheep at Massey University, New Zealand, were analyzed to obtain an evaluation of selection response. Selection was for heavy 14-mo greasy fleece weight. Approximately seven generations of selection were represented in the data. Three estimates of genetic superiority of the selected line to the control line were obtained. All three estimates were obtained from a mixed model evaluation using the individual animal model for predicting breeding values from own and relatives' records. The estimators were 1) deviation of selected line predicted yearly phenotypes from control line predicted yearly phenotypes, 2) deviation of the predicted yearly phenotype for the selected line from the year estimate in the control line and 3) the mean yearly breeding value from the analysis of the selected line only. The realized heritability using the first approach was .20. However, the control line was found to have a slight positive drift; hence, this estimate was biased downward. Using Approach 2, accounting for drift, the realized heritability was .23. The same realized heritability, .23, was obtained from an analysis of the selected line ignoring the control (Approach 3), when a prior heritability of .30 was assumed for the mixed model evaluation. The estimate of genetic trend from predicted breeding values in the latter approach is, however, quite dependent on the assumed heritability.


Individual and Combined Cytotoxic Effects of Co-Occurring Fumonisin Family Mycotoxins on Porcine Intestinal Epithelial Cell.
Human health is seriously threatened by mycotoxin contamination, yet health risk assessments are typically based on just one mycotoxin, potentially excluding the additive or competitive interactions between co-occurring mycotoxins. In this investigation, we evaluated the individual or combined toxicological effects of three fumonisin-family B mycotoxins: fumonisin B1 (FB1), fumonisin B2 (FB2), and fumonisin B3 (FB3), by using porcine intestinal epithelial cells (IPEC). IPEC cells were exposed to various concentrations (2.5-40 μM) for 48 h, and a cell counting kit (CCK8) was used to determine cell vitality. Firstly, we discovered that they might inhibit cell viability. Additionally, the cytotoxicity of FB1 was significantly greater than that of FB2 and FB3. The results also indicated that the combinations of FB1-FB2, FB2-FB3, and FB1-FB2-FB3 showed synergistically toxicological effects at the ID10-ID50 levels and antagonistic effects at the ID75-ID90 levels. In addition, the FB1-FB3 exposure was also synergistic at the ID10-ID25 level. We also found that myriocin and resveratrol alleviated the cytotoxicity induced by fumonisin in IPEC cells. In all, this study may contribute to the determination of legal limits, the optimization of risk assessment for fumonisins in food and feed, and the development of new methods to alleviate fumonisin toxicity.


High Magnitude of Social Anxiety Disorder in School Adolescents.
Introduction. Social phobia is the most prevalent and chronic type of anxiety disorder worldwide and it affects occupational, educational, and social affairs of the individual. Social phobia is also known for its association with depression and substance use disorder. Objective. The aim of this study was to assess the prevalence and associated factors of social phobia among high school students in Ethiopia. Methods. Cross-sectional study was conducted among 386 randomly selected students. Data were collected using pretested and self-administered questionnaire. Social phobia was assessed by using Social Phobia Inventory (SPIN). Logistic regression was used to analyze the data with 95% confidence interval and variables with p value less than 0.05 were considered as statistically significant. Results. From 386 study participants, 106 (27.5%) of them were positive for social phobia. Being female (AOR = 3.1; 95% CI: 1.82-5.27), current alcohol drinking (AOR = 1.75; 95% CI: 1.03-2.98), poor social support (AOR = 2.40; 95% CI: 1.17-4.92), and living with single parent (AOR = 5.72; 95% CI: 2.98-10.99) were significantly associated with social phobia. Conclusion. The proportion of social phobia was higher compared to previous evidences. School-based youth-friendly mental health services might be helpful to tackle this problem.


Haemoglobinuria caused by propylene glycol in sheep.
HAEMOGLOBINURIA OCCURRED IN SHEEP ANAESTHETIZED BY AN INTRAVENOUS INJECTION OF PENTOBARBITONE SODIUM CONTAINING PROPYLENE GLYCOL: an equivalent dose failed to cause haemoglobinuria in rabbits. Intravenous injection of an aqueous solution of 20% propylene glycol caused haemoglobinaemia and haemoglobinuria in sheep. Neither distilled water nor 20% glycerol in water administered under identical conditions produced these effects.Haemoglobinuria occurred on some occasions when an aqueous 20% solution of propylene glycol was administered to sheep after an injection of saline: it never occurred when a solution of 20% propylene glycol prepared with physiological saline was injected. It is suggested that saline may protect against the haemolytic action of propylene glycol in sheep and that propylene glycol should be avoided as a menstruum for pharmaceutical preparations to be used for injection into the blood stream of these animals.


[The problem of radiation exposure during radiography of the entire spine].
The stimulus for this investigation concerning radiation exposure during radiography of the spine, particularly in children with scoliosis, is the fact that there are no data in the literature. In 16 series of measurements we studied the effect of grids, kilovoltage, focus-film distance and film-screen systems using dosimeters. The use of grids led to a 5-7 times increase in radiation dose compared with exposures without grids. For this reason grids are not recommended for serial examinations in cases of scoliosis. It is easier to evaluate the spine on a single large film than on several small films and radiation dose is also halved. Therefore, the use of large single films is recommended, particularly for children and adolescents.


Multifaceted management of the postthrombotic syndrome.
The postthrombotic syndrome (PTS) is a frequent cause of chronic pain, swelling, ulceration, and disability in patients with lower extremity deep vein thrombosis (DVT). As interventional radiologists are consulted on more patients with chronic DVT and PTS, their management strategies must be informed by a balanced understanding of the different facets of chronic DVT care and the available treatment options. This article provides an overview of the important elements of a multifaceted approach to the management of patients with PTS that includes pharmacological, physiological, and endovascular aspects of care.


Miscarriage and ectopic pregnancy 1: an overview.
This two-part unit explores miscarriage and ectopic pregnancy. This article, part 1, discusses definitions, classifications and signs and symptoms of both.


DTI and VBM reveal white matter changes without associated gray matter changes in patients with idiopathic restless legs syndrome.
We evaluated cerebral white and gray matter changes in patients with iRLS in order to shed light on the pathophysiology of this disease. Twelve patients with iRLS were compared to 12 age- and sex-matched controls using whole-head diffusion tensor imaging (DTI) and voxel-based morphometry (VBM) techniques. Evaluation of the DTI scans included the voxelwise analysis of the fractional anisotropy (FA), radial diffusivity (RD), and axial diffusivity (AD). Diffusion tensor imaging revealed areas of altered FA in subcortical white matter bilaterally, mainly in temporal regions as well as in the right internal capsule, the pons, and the right cerebellum. These changes overlapped with changes in RD. Voxel-based morphometry did not reveal any gray matter alterations. We showed altered diffusion properties in several white matter regions in patients with iRLS. White matter changes could mainly be attributed to changes in RD, a parameter thought to reflect altered myelination. Areas with altered white matter microstructure included areas in the internal capsule which include the corticospinal tract to the lower limbs, thereby supporting studies that suggest changes in sensorimotor pathways associated with RLS.


Numerical analysis of tooth mobility: formulation of a non-linear constitutive law for the periodontal ligament.
The aim of the present work is the analysis of mobility of human dentition under the action of physiological short-term loading by using a numerical approach. The mobility of tooth is mainly attributed to the deformation of the periodontal ligament, hence particular attention is focused on the definition of a suitable constitutive model for this soft tissue. A numerical model of human upper incisor and periodontum is proposed by means of the finite element method. The mechanical response of the most deformable constituent, i.e. the periodontal ligament, is described by using a hyperelastic constitutive model. Numerical analyses show an effective correspondence with in vivo experimental tests, in literature, on tooth mobility caused by the application of intrusive loads. The model proves to offer valid results, leading to a general, reliable and efficient numerical approach. The hyperelastic approach allows the description of the mechanical response of the periodontum under short lasting loads, according to the characteristic non-linear behavior of the PDL. This approach overcomes the approximation caused by the adoption of simplified models, widely reported in literature, and leads to a valid definition of tooth movement by means of an effective numerical procedure.


QuickStats: Smoking Cessation* During Pregnancy - 46 States and the District of Columbia, 2014.
In 2014, 20.6% of pregnant women who smoked cigarettes during the first or second trimester, in a reporting area of 46 states and the District of Columbia, stopped smoking during pregnancy. Women in three states, South Dakota (31.3%), California (31.2%) and New Mexico (30.2%), as well as the District of Columbia (41.5%), reported the highest cessation rates during pregnancy. Kentucky (11.4%) and Maine (11.6%) reported the lowest cessation rates; cessation rates were generally lower for states in the Southeast. The reporting area included 3,819,113 births and represented 95% of all U.S. births in 2014.


Immigration as risk factor for non-suicidal self-injury and suicide attempts in adolescents in Germany.
Whereas non-suicidal self-injury (NSSI) and suicide attempts (SA) are rather common among adolescents, the description of risk factors has often failed to take migration into perspective. Our study aimed to describe immigration status in adolescents with regards to their lifetime history of NSSI and SA. We carried out a population based study in a school community of ninth-graders (N = 452, mean age 14.85, SD 0.58) in southern Germany. Data were collected via adolescent self report on sociodemographic variables and on NSSI and SA using the Self Harm Behavior Questionnaire. Adolescents born outside Germany showed an elevated rate of a lifetime history of NSSI and SA. When compared to German adolescents without a (family) history of migration (NSSI 19.16%, SA 3.24%), adolescents who were born in another country had an elevated risk for NSSI (42.86%, OR 3.36) and SA (17.86%, OR 6.78), which was higher than the risk of adolescents who had at least one parent who had emigrated from another country (NSSI 30.08%, OR 2.46 and SA 8.94%, OR 4.45). Our findings should inform intervention services and prevention programs for NSSI and suicidality in youth. Adopting such programs to include culturally sensible modules could improve the outcome in ethnically diverse adolescents.


Unusual presentation of a large leiomyoma of the esophagus.
A case is presented with a giant pedunculated leiomyoma of the esophagus. The difficulties of diagnosis are presented along with a review of the literature. Surgical removal is safe and curative.


Calpeptin is a potent cathepsin inhibitor and drug candidate for SARS-CoV-2 infections.
Several drug screening campaigns identified Calpeptin as a drug candidate against SARS-CoV-2. Initially reported to target the viral main protease (Mpro), its moderate activity in Mpro inhibition assays hints at a second target. Indeed, we show that Calpeptin is an extremely potent cysteine cathepsin inhibitor, a finding additionally supported by X-ray crystallography. Cell infection assays proved Calpeptin's efficacy against SARS-CoV-2. Treatment of SARS-CoV-2-infected Golden Syrian hamsters with sulfonated Calpeptin at a dose of 1 mg/kg body weight reduces the viral load in the trachea. Despite a higher risk of side effects, an intrinsic advantage in targeting host proteins is their mutational stability in contrast to highly mutable viral targets. Here we show that the inhibition of cathepsins, a protein family of the host organism, by calpeptin is a promising approach for the treatment of SARS-CoV-2 and potentially other viral infections.


Complete Neurologic Recovery of Cerebral Fat Embolism Syndrome in Sickle Cell Disease.
Sickle cell disease is one of the most common inherited hemoglobinopathies diagnosed in the United States. Patients often present with severe anemia, pain crises, infections, and vaso-occlusive phenomena. Complications of these disorders can lead to significant debilitating morbidity and mortality. Fat embolism syndrome (FES) is a rare and devastating complication of sickle cell disease. It usually presents with a rapidly deteriorating clinical course, and the prognosis is dismal. We report a case of FES in a 19-year-old African American male with a history of sickle cell disease who presented with tonic-clonic seizures and was found to have multi-organ failure. FES was diagnosed 20 days from a presentation based on blood cytopenias and magnetic resonance imaging findings that were obscured at the initial presentation. We describe in this report, the patient's course from presentation until diagnosis and resolution. Our case is peculiar as the patient had a very good outcome without the need for red blood cell (RBC) exchange; instead, supportive treatment and simple RBC transfusions were enough to change the clinical course of this almost fatal syndrome.


Targeting ß2 adrenergic receptors regulate human T cell function directly and indirectly.
It is well-established that central nervous system activation affects peripheral blood mononuclear cell (PBMCs) function through the release of the catecholamines (Epi) and norepinephrine (NE), which act on ß2-adrenergic receptors (ß2AR). However, most studies have used non-specific stimulation of cells rather than antigen-specific responses. Likewise, few studies have parsed out the direct effects of ß2AR stimulation on T cells versus indirect effects via adrenergic stimulation of antigen presenting cells (APC). Here we report the effect of salmeterol (Sal), a selective ß2AR agonist, on IFN-γ(+) CD4 and IFN-γ(+) CD8 T cells following stimulation with Cytomegalovirus lysate (CMVL-strain AD169) or individual peptides spanning the entire region of the HCMV pp65 protein (pp65). Cells were also stimulated with Staphylococcal enterotoxin B. Additionally, we investigated the effect of Epi and Sal on cytotoxic cell killing of transfected target cells at the single cell level using the CD107a assay. The results show that Sal reduced the percentage of IFN-γ(+) CD4 and IFN-γ(+) CD8 T cells both when applied directly to isolated T cells, and indirectly via treatment of APC. These inhibitory effects were mediated via a ß2 adrenergic-dependent pathway and were stronger for CD8 as compared to CD4 T cells. Similarly, the results show that Sal suppressed cytotoxicity of both CD8 T and NK cells in vitro following stimulation with Chinese hamster ovary cell line transfected with MICA(*009) (T-CHO) and the human erythromyeloblastoid leukemic (K562) cell line. The inhibitory effect on cytotoxicity following stimulation with T-CHO was stronger in NK cells compared with CD8 T cells. Thus, targeting the ß2AR on lymphocytes and on APC leads to inhibition of inflammatory cytokine production and target cell killing. Moreover, there is a hierarchy of responses, with CD8 T cells and NK cells inhibited more effectively than CD4 T cells.


Precut sphincterotomy: a reliable salvage for difficult biliary cannulation.
Even experienced endoscopists have 90% success in achieving deep biliary cannulation with standard methods. Biliary cannulation may become difficult in 10%-15% of patients with biliary obstruction and precut (access) sphincterotomy is frequently chosen as a rescue treatment in these cases. Generally, precut sphincterotomy ensures a rate of 90%-100% successful deep biliary cannulation. The precut technique has been performed as either a fistulotomy with a needle knife sphincterotome or as a transpapillary septotomy with a standard sphincterotome. Both methods have similar efficacy and complication rates when administered to the proper patient. Although precut sphincterotomy ensures over 90% success of biliary cannulation, it has been characterized as an independent risk factor for pancreatitis. The complications of the precut technique are not limited to pancreatitis. Two more important ones, bleeding and perforation, are also reported in some publications as being observed more commonly than during standard sphincterotomy. It is also reported that precut sphincterotomy increases morbidity when performed in patients without dilatation of their biliary tract. Nevertheless, precut sphincterotomy is a good alternative as a rescue method in the setting of a failed standard cannulation method. This paper discusses the technical details, timing, efficacy and potential complications of precut sphincterotomy.


Safety and efficacy of SHR4640 combined with febuxostat for primary hyperuricemia: a multicenter, randomized, double-blind, phase II study.
To evaluate the safety, tolerability, and efficacy of SHR4640, a highly selective urate transporter-1 inhibitor, in combination with febuxostat, in patients with primary hyperuricemia. In this randomized, double-blind, parallel-controlled phase II study, patients whose fasting serum uric acid (sUA) levels were ⩾ 480 μmol/L at screening with gout or sUA levels were ⩾ 420 μmol/L lasting for at least 3 months without gout, either with sUA levels ⩾ 540 μmol/L at screening or sUA levels ⩾ 480 μmol/L with comorbidities at screening, were enrolled. Patients were randomized (1:1:1) to receive SHR4640 10 mg plus febuxostat 80 mg, SHR4640 10 mg plus febuxostat 40 mg, and SHR4640 5 mg plus febuxostat 20 mg orally once daily. The primary end point was the incidence of treatment-emergent adverse events (TEAEs). A total of 93 patients were randomized and received treatment. TEAEs occurred in 55.9% of patients. The incidence of TEAEs was comparable among all the groups. Serious TEAEs occurred in one patient (1.1%), with no deaths observed. The proportion of patients who achieved the target sUA levels by week 4 was 79.3%, 96.6%, and 75.0% in the SHR4640 10 mg plus febuxostat 80 mg, SHR4640 10 mg plus febuxostat 40 mg, and SHR4640 5 mg plus febuxostat 20 mg groups, respectively. The mean percent reduction of sUA was 59.7%, 63.7%, and 41.8%, respectively. SHR4640 plus febuxostat exhibited a tolerable safety profile and substantial sUA lowering activity in patients with primary hyperuricemia. www.chinadrugtrials.org.cn; CTR 20192429.


Synergic activity of combined prostacyclin: dimethyl sulfoxide in experimental brain ischemia.
We examined whether drugs that block calcium, prostaglandins, free radicals, and endorphin release could modify cerebral blood flow or nerve tissue pathology following a focal cerebrovascular lesion. Cats were randomly divided into six groups and were subjected to standard middle cerebral artery occlusion (MCAO) performed using a transorbital approach. One hour after MCAO, cats received the following compounds intravenously: (i) saline (CS), 1.5 mL/kg or polyethylene glycol, 300 micrograms (CP); (ii) naloxone (NX), 2 mg/kg; (iii) nimodipine (NM), 1 microgram.kg-1.min-1 x 60 min; (iv) dimethyl sulfoxide (DS), 0.9 g/kg in a 40% solution; (v) prostacyclin (PGI2), 200 ng.kg-1.min-1 for 60 min; or (vi) DS-PGI2 combined. At 1-h intervals, local CBF was recorded from the cortical tissue proximal and distal to the MCAO site using the hydrogen clearance method. Five hours after MCAO, cortical tissue was removed for catecholamine histofluorescence or perfused for tyrosine hydroxylase immunoreactive axon examination. Treatment with NX, NM, CP, or CS had no effect on either CBF or cortical tissue neurotransmitter morphology. PGI2 showed a transiently modest but significant increase of CBF, while DS provided moderate protection of catecholaminergic fibers and increased CBF by 27% after MCAO. The combination of DS-PGI2 resulted in significant cytoprotection of cortical catecholaminergic fibers and generated a sustained CBF increase of 68% of control values. These findings suggest that combining DS with PGI2 can yield a synergic effect with respect to cortical neurotransmitter and CBF protection after MCAO.


Climacteric Symptoms and Sexual Dysfunction: Association between the Blatt-Kupperman Index and the Female Sexual Function Index.
Objective To investigate the association between the intensity of climacteric symptoms and sexual dysfunction in women aged 40 to 65 years. Methods Observational, analytic, cross-sectional study conducted with 63 women aged 40 to 65 treated at the gynecology outpatient clinic of a public hospital in northeastern Brazil. A questionnaire was used to collect identification data, clinical information, gynecological-obstetric data, lifestyle traits and information on chronic diseases. Climacteric symptoms and sexual function were evaluated by means of the Blatt-Kupperman index and the Female Sexual Function Index (FSFI) respectively. The association between the two indices was investigated using the chi-squared test; the difference in mean scores on the FSFI as a function of menopausal status was evaluated by Student's t-test. The significance level was set to p < 0.05. Results The mean value of the Blatt-Kupperman index was 26.42 (standard deviation [SD]: 4.52); 36.51% of the women exhibited severe symptoms. The mean score on the FSFI was 21.84 (SD: 4.11). More than half of the analyzed women (58.73%) exhibited sexual dysfunction (FSFI ≤ 26.5). Regarding the association between the Blatt-Kupperman index and the FSFI, the greater the intensity of the climacteric symptoms (Blatt-Kupperman), the higher the frequency of sexual dysfunction (FSFI). Sexual dysfunction was exhibited by 100% of the participants with severe climacteric symptoms, 70.59% of those with moderate symptoms, and only 9.09% with mild symptoms (p < 0.001). Conclusions The application of the Blatt-Kupperman index and of the FSFI allowed the detection of an association between the severity of climacteric symptoms and the prevalence of sexual dysfunction.


Multi-camera imaging to characterize jet and liner uniformity on the Plasma Liner Experiment (PLX).
At Plasma Liner Experiment, a set of 36 coaxial plasma guns are deployed quasi-uniformly over a 9 ft diameter spherical chamber and are used to form a high-Z spherically compressive plasma liner. Simulations indicate that for the concept to ultimately achieve optimal target density and temperature, a high degree of timing uniformity is required between all guns. To aid in quantifying and correcting gun-to-gun nonuniformities, a key diagnostic will consist of up to six fisheye-view CCD cameras positioned inside the main chamber such that each has all plasma guns within its view. The individual cameras can be triggered at different times to determine each plasma jet's muzzle velocity and structure for different operating conditions. This camera array is currently under development, and the implementation needs and challenges for this camera array are discussed here. Additionally, we detail the analysis methodology for determining jet-to-jet uniformity deviations and how we can correct them, thereby improving overall liner uniformity.


The threshold force required for femoral impaction grafting in revision hip surgery.
Femoral impaction grafting requires vigorous impaction to obtain adequate stability without risk of fracture, but the force of impaction has not been determined. We determined this threshold force in a preliminary study using animal femurs. Adult sow femurs were used because of their morphological similarity to human femurs in revision hip arthroplasty. 35 sow femurs were impacted with morselized bone chips and an increasing force was applied until the femur fractured. This allowed a threshold force to be established. 5 other femurs were impacted to this force and an Exeter stem was cemented into the neomedullary canal. A 28-mm Exeter head was attached and loaded by direct contact with a hydraulic testing machine. Axial cyclic loading was performed and the position sensor of the hydraulic testing machine measured the prosthetic head subsidence. 29 tests were completed successfully. The threshold force was found to be 4 kN. There was no statistically significant correlation between the load at fracture and the cortex-to-canal ratio or the bone mineral density. Following impaction with a maximum force of 4 kN, the average axial subsidence was 0.28 mm. We achieved a stable construct without fracture. Further studies using human cadaveric femurs should be done to determine the threshold force required for femoral impaction grafting in revision hip surgery.


Rate constant for diffusion-influenced ligand binding to receptors of arbitrary shape on a cell surface.
The theory of ligand binding to receptors on a cell surface suggested by Berg and Purcell and generalized by Zwanzig and Szabo uses the assumption that receptors are circular absorbing disks on an otherwise reflecting sphere. One of the key ingredients of this theory is a solution for the rate constant for ligand binding to a single circular receptor on a reflecting plane. We give an exact solution for the rate constant for binding to a single elliptic receptor and an approximate solution for binding to a single receptor of more general shape. The latter was tested by Brownian dynamics simulations. We found that the approximate formula predicted the rate constant with better than 10% accuracy for all studied receptor shapes. Using our solutions one can find the rate constant for ligand binding to a cell covered by N noncircular receptors by means of the Zwanzig-Szabo formula.


GENETIC SUBDIVISIONS AMONG SMALL CANIDS: MITOCHONDRIAL DNA DIFFERENTIATION OF SWIFT, KIT, AND ARCTIC FOXES.
Gene flow can effectively suppress genetic divergence among widely separated populations in highly mobile species. However, the same may not be true of species that typically disperse over shorter distances. Using mtDNA restriction-site and sequence analyses, we evaluate the extent of divergence among populations of two small relatively sedentary North American canids, the kit and swift foxes (genus Vulpes). We determine the significance of genetic differentiation among populations separated by distance and those separated by discrete topographic barriers. Our results show the among-population component of genetic variation in kit and swift foxes is large and similar to that of small rodents with limited dispersal ability. In addition, we found two distinct groupings of genotypes, separated by the Rocky Mountains, corresponding to the traditional division between kit and swift fox populations. Previous workers have characterized these morphologically similar populations either as separate species or subspecies. Our mtDNA data also suggest that kit and swift fox populations hybridize over a limited geographic area. However, the sequence divergence between kit and swift foxes is similar to that between these taxa and the arctic fox (Alopex lagopus), a morphologically distinct species commonly placed in a separate genus. This result presents a dilemma for species concepts, and we conclude that kit and swift foxes should be recognized as separate species.


Probability-based current dipole localization from biomagnetic fields.
Focal biomagnetic sources are described as pointlike current dipoles. The dipole parameters, position, and moment coordinates are commonly determined from biomagnetic data using iterative nonlinear optimization algorithms such as the Levenberg-Marquardt algorithm. However, even for single-dipole sources, mislocalizations can occur due to side minima of the cost function or due to a wrong choice of the start vector. This can be shown by introducing a cost function where the independent variables are only the position coordinates instead of position and moment coordinates. This dimensional reduction--which is also possible for multiple dipole sources--is achieved by calculating the cost function at each position with the position and data-dependent, optimum dipole moments. We call these dipoles with--in a least squares sense--optimum moments, locally optimal dipoles. The visualization of such a single-dipole cost function and of the iteration steps of the Levenberg-Marquardt algorithm show why mislocalizations cannot be avoided. Therefore, we propose an alternative noniterative localization algorithm for single-dipole sources without this drawback. It uses localization probabilities calculated by means of the locally optimal dipoles. Besides the determination of the dipole parameters, the proposed algorithm furnishes a reliable error for each localization. Its effectiveness is shown with simulated and real patient data.


The management of dog bites and dog bite infections to the hand.
Dog bites account for approximately one of 200 emergency room visits. The majority of bites are from dogs known to the victim. Due to the numerous small compartments and the thin covering of soft tissue over the bones and joints, bite wounds to the hand are more likely to become infected that are bites to the arm, leg, or face. Improper management can result in significant morbidity and prolonged treatment. This article discusses the microbiology of dog bite wounds, antibiotic selection, and proper wound management.


Prenatal ethanol exposure enhances glutamate release stimulated by quisqualate in rat cerebellar granule cell cultures.
Effects of prenatal ethanol exposure on extracellular glutamate accumulation stimulated by glutamate receptor agonists were studied in rat cerebellar granule cell cultures. The prenatal exposure to ethanol was achieved via maternal consumption of a Sustacal liquid diet containing either 5% ethanol or isocaloric sucrose (pair-fed) substituted for ethanol from gestation d 11 until the day of parturition. Neither the basal level of extracellular glutamate nor the increased accumulation of glutamate stimulated by KCl (40 mM) or by ionotropic glutamate receptor agonists, N-methyl-D-aspartate (NMDA) or kainate (KA) (100 microM each), in cells prepared from the ethanol-fed group was significantly different from that in cells prepared from the pair-fed group. Glutamate accumulation stimulated by quisqualate (QA, 100 microM) or by trans-(+/-)-1-amino-1,3-cyclopentanedicarboxylic acid (t-ACPD, 250 microM) in the ethanol-fed group was higher than that in the pair-fed group by 116 and 36%, respectively. In the presence of 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX, 100 microM), an ionotropic QA receptor antagonist, the QA-induced accumulation of glutamate in the ethanol-fed group was still higher than that in the pair-fed group. In the presence of MK-801 (5 microM), an antagonist of the NMDA receptor, the enhanced accumulation of glutamate stimulated by either QA or t-ACPD was still observable in the ethanol-fed group as compared to the pair-fed group. Addition of (RS)-alpha-methyl-4-carboxyphenylglycine (MCPG, 500 microM), a selective antagonist of the metabotropic glutamate receptor, abolished the enhanced accumulation of glutamate stimulated by either QA or t-ACPD in the ethanol-fed group. Although immunoblotting of mGluR1 and mGluR2/3 did not show apparent differences between the pair-fed and the ethanol-fed groups, the overall results suggest that the effect of prenatal ethanol exposure was selectively through a pathway mediated by the metabotropic glutamate receptor.


Epstein-Barr virus associated hepatic smooth muscle tumor in a patient with acquired immunodeficiency syndrome: A case report.
Epstein-Barr virus (EBV) associated smooth muscle tumors (SMTs) usually present under the condition of immunosuppression, including congenital immunodeficiency syndrome-SMT, post-transplantation-SMT and HIV-SMT. HIV-SMTs are most likely to invade the central nervous system, followed by the liver, lungs, and other locations. Many laboratory techniques, including serological techniques, polymerase chain reaction and immunohistochemistry (IHC), are employed to determine the aetiologies of these tumours. With respect to therapy, surgical resection is the main treatment. In patients with immunodeficiency, improving immune status is significant for defending against other viruses. We describe a case of the primary focus of SMT in the liver of HIV-positive patient without any metastasis. A young male HIV-positive patient complained of fever and abdominal pain for 2 months. IHC of liver tissue confirmed the finding: EBV-related smooth muscle tumor. Given the patient's general condition, he was not a suitable candidate for surgical resection. He was given antibiotics, antifungal agents and EBV-directed agents to control infection as well as highly active antiretroviral therapy to enhance the immunity. The patient's symptoms improved. He was discharged. In conclusion, EBV-related HIV-SMTs is a rare neoplasm found in the liver among immunodeficient patients. This case highlights that a variety of examinations such as IHC for smooth muscle markers (smooth muscle actin and desmin) and EBER, as well as polymerase chain reaction for EBV DNA should be done when diagnoses are ambiguous.


Hemisensory loss in myasthenia gravis.
A 73-year-old white man presents with left-sided ptosis and diplopia in the absence of ophthalmoplegia, with left hemibody paresthesia. He reports intermittent dysphagia and dizziness for 1 month and diarrhoea for 2 months. Serum and electrodiagnostic studies confirmed the diagnosis of myasthenia gravis. This case highlights the non-classic presentation of myasthenia gravis in the absence of ophthalmoplegia with a unique unexplained hemisensory deficit.


Adenylosuccinase deficiency with neonatal onset severe epileptic seizures and sudden death.
We report a male infant with adenylosuccinase deficiency who developed epileptic seizures on the second day of life. Growth was normal and seizures were well controlled with anti-epileptic drugs. Despite axial hypotonia associated with peripheral hypertonicity he presented some development until seven months of age, when he developed high fever and died within a few hours. Although clinical heterogeneity in this disorder of purine synthesis and interconversion is well-known, in 14 out of 17 cases who experienced epilepsy seizures started after the first year of life. The early presentation in our index patient followed by his sudden death at the age of 7 months has not been described before. A search for disorders of purine metabolism should be included in the screening programme for every child with severe neonatal convulsions.


Underutilization of Implantable Cardioverter Defibrillator in Primary Prevention of Sudden Cardiac Arrest.
The aim of this study was to evaluate the overall use of implantable cardioverter defibrillators (ICD) for primary prevention of sudden cardiac arrest (SCA), among eligible patients from an outpatient cardiology clinic and to determine what factors might contribute to underutilization of ICDs. This report was a retrospective chart review of patients with ischemic or non-ischemic cardiomyopathy and left ventricular ejection fraction ≤ 35% from an outpatient cardiology practice from January 2005 to May 2008. These patients met the eligibility criteria for ICD implantation for primary prevention of SCA. A detailed review of medical records captured distribution of ICD implantation including future plans for ICD implant, patient preference against ICD use, presence of severe co-morbidities, and any other documented reasons/contraindications regarding ICD implantation. Of the 275 patients who were eligible for ICD for primary prevention of SCA, 119 (43%) had an ICD implantation. ICDs were used in 84 (48%) eligible men and 35 (35%) eligible women (P 0.02). Among 156 (57%) patients who did not receive ICD, 79 (28%) had severe co-morbidities precluding them from having ICD. Twenty-six patients (10%) refused to have ICD implanted. The remaining 51 (19%) patient charts did not include any documentation regarding ICD use (future plan or contraindication). ICDs are underutilized for primary prevention of SCA, with rates of use being lowest among eligible women. This underutilization exists even after accounting for patient preferences and presence of severe co-morbid conditions that might make an otherwise eligible patient not a suitable candidate for ICD implantation.


[Quantitative studies of urine sediment morphology during the course of pregnancy].
Bacterial infections of the urinary system during pregnancy occur relatively frequently and exert unfavourable effect on its course while their constant and sensitive indicators are leucocyturia and bacteriuria. The studies were carried out in 30 healthy women in the second phase of menstrual cycle and in 86 pregnant women in various stages of pregnancy (I, II and III trimester). The subject of own analysis was leucocyte count in 1 ml of morning urine portion in correlation with the result of quantitative and qualitative urine culture. As significant bacteriuria 10(4) bacteria in 1 ml of urine was accepted. Urine sediment examination was carried out by the Mc Geachie-Kennedy method. The own studies demonstrated an increase of morphotic element count in 1 ml of urine with the progression of pregnancy. This increase was statistically significant in the group of pregnant women with significant bacteriuria. The results of own studies were compared with those of other authors.


Submergence-induced leaf acclimation in terrestrial species varying in flooding tolerance.
Earlier work on the submergence-tolerant species Rumex palustris revealed that leaf anatomical and morphological changes induced by submergence enhance underwater gas exchange considerably. Here, the hypothesis is tested that these plastic responses are typical properties of submergence-tolerant species. Submergence-induced plasticity in leaf mass area (LMA) and leaf, cell wall and cuticle thickness was investigated in nine plant species differing considerably in tolerance to complete submergence. The functionality of the responses for underwater gas exchange was evaluated by recording oxygen partial pressures inside the petioles when plants were submerged. Acclimation to submergence resulted in a decrease in all leaf parameters, including cuticle thickness, in all species irrespective of flooding tolerance. Consequently, internal oxygen partial pressures (pO(2)) increased significantly in all species until values were close to air saturation. Only in nonacclimated leaves in darkness did intolerant species have a significantly lower pO(2) than tolerant species. These results suggest that submergence-induced leaf plasticity, albeit a prerequisite for underwater survival, does not discriminate tolerant from intolerant species. It is hypothesized that these plastic leaf responses may be induced in all species by several signals present during submergence; for example, low LMA may be a response to low photosynthate concentrations and a thin cuticle may be a response to high relative humidity.


Monocyte Chemotactic Protein-1-Interleukin-6-Osteopontin Pathway of Intra-Aneurysmal Tissue Healing.
We have previously demonstrated that the local delivery of monocyte chemotactic protein-1 (MCP-1) via an MCP-1-releasing poly(lactic-co-glycolic acid)-coated coil promotes intra-aneurysmal tissue healing. In this study, we demonstrate that interleukin-6 (IL-6) and osteopontin are downstream mediators in the MCP-1-mediated aneurysm-healing pathway. Murine carotid aneurysms were created in C57BL/6 mice. Drug-releasing coils (MCP-1, IL-6, and osteopontin) and control poly(lactic-co-glycolic acid) coils were created and then implanted into the aneurysms to evaluate their intra-aneurismal-healing capacity. To investigate the downstream mediators for aneurysm healing, blocking antibodies for IL-6 receptor and osteopontin were given to the mice implanted with the MCP-1-releasing coils. A histological analysis of both murine and human aneurysms was utilized to cross-validate the data. We observed increased expression of IL-6 in MCP-1-coil-treated aneurysms and not in control-poly(lactic-co-glycolic acid)-only-treated aneurysms. MCP-1-mediated intra-aneurysmal healing is inhibited in mice given blocking antibody to IL-6 receptor. MCP-1-mediated intra-aneurysmal healing is also inhibited by blocking antibody to osteopontin. The role of IL-6 in intra-aneurysmal healing is in recruiting of endothelial cells and fibroblasts. Local delivery of osteopontin to murine carotid aneurysms via osteopontin-releasing coil significantly promotes intra-aneurysmal healing, but IL-6-releasing coil does not, suggesting that IL-6 cannot promote aneurysm healing independent of MCP-1. In the MCP-1-mediated aneurysm healing, osteopontin expression is dependent on IL-6; inhibition of IL-6 receptor significantly inhibits osteopontin expression in MCP-1-mediated aneurysm healing. Our findings suggest that IL-6 and osteopontin are key downstream mediators of MCP-1-mediated intra-aneurysmal healing.


Failure of the sterile air-flow component of a protected environment detected by demonstration of Chaetomium species colonization of four consecutive immunosuppressed occupants.
Four bone marrow transplant recipients consecutively occupying the same room on our Oncology-Hematology Special Care Unit (OHSCU) became colonized with Chaetomium species between January and April, 1987. These patients, aged 27 to 43 years, were immunocompromised as a result of intensive chemotherapy, and were consequently at increased risk for development of invasive fungal infection. At the time of Chaetomium colonization, all patients were febrile, two had transient new infiltrates on chest x-ray, and three were receiving amphotericin B therapy. Subsequent environmental cultures revealed Chaetomium contamination of the OHSCU air-handling system, including the HEPA (high-efficiency particulate air) filters in seven of the nine rooms comprising the unit. Because fungal colonization of HEPA filters used to create a "protective environment" for immunocompromised patients can occur and can serve as a source for patient infections, guidelines concerning proper surveillance of these HEPA filters should be established. We suggest that before a new patient enters a "protected" room, the clean side of the HEPA filter should be cultured. If fungi are recovered from that culture, we would recommend changing the filter.


[Ventricular pre-excitation in Wolff Parkinson White syndrome in the aged].
To evaluate any differences in ventricular pre-excitation secondary to Wolff Parkinson White syndrome in the aged compared to young and adult patients. a clinical study was performed using a comparative prospective criterion with retrospective analysis. The duration of follow-up ranged between one and ten years. the series was collected from the Cardiology Clinic of the Health District and the Cardiology Division of Gorizia, both forming part of no. 2 Isontina Health Service. the series included 17 patients suffering from Wolff Parkinson White syndrome who were divided into two study groups: 9 elderly patients and 8 young patients. The latter were subdivided into a first subgroup of 4 cases with Wolff Parkinson White syndrome with ECG positive for the presence of delta waves, and a second subgroup also with Wolff Parkinson White syndrome secondary to bundle. some young patients with Wolff Parkinson White syndrome who were symptomatic for tachycardia underwent ablative surgery with radiofrequency of the bundle. all patients underwent cardiological screening focused in particular on surface electrocardiogram. Those cases with Wolff-Parkinson White syndrome with occult bundle underwent transesophageal electrostimulation to find the conduction threshold of the anomalous bundle. Adult-elderly patients: six subjects were diagnosed with antero-septal and left ventricular Kent's bundle (type B common) and 3 cases with Mahaim-Wiston bundle (type A rare). Surface ECG revealed the presence of left ventricular hypertrophy in 6 cases, left anterior hemiblock and total block of the left branch in 3 cases, as well as myocardial pseudonecrosis correlated to Wolff Parkinson White syndrome. Young patients: four out of this group were affected by Kent's bundle with type B Wolff Parkinson White syndrome and the same number suffered from the same syndrome caused by occult bundle. Patients in the first subgroup showed an antero-septal, transitional and left ventricular orientation of Kent's bundle, with the onset of 2 cases of orthodromic and antidromic reciprocal rhythm respectively and 1 case of atrial fibrillation. The refractory nature of the anomalous pathway was not very high in 2 cases, equal to 60 milliseconds and 240 milliseconds with the proposed ablation of the anomalous bundle.


Discovery and Biochemical Characterization of a Thermostable Glucose-1-phosphate Nucleotidyltransferase from Thermodesulfatator indicus.
The biosynthesis of NDP-glucoses is based on the nucleotide transfer from NTP donor substrates to glucose-1-phosphates catalyzed by glucose-1-phosphate nucleotidyltransferases. The cloning and biochemical characterization of a glucose-1-phosphate nucleotidyltransferase (TiGPNT) from the deep sea bacterium Thermodesulfatator indicus. The biochemical parameters of recombinant TiGPNT were determined using a plate reader-based coupled enzymatic assay, in which the reaction product UDP-glucose is oxidized in the presence of NAD+ forming UDP-Glucuronic acid and NADH. The substrate promiscuity of the enzyme was determined using thin-layer chromatography and MALDI-ToF mass spectrometry. TiGPNT was recombinantly expressed under the control of the T7 promoter in Escherichia coli and could be successfully enriched by heat treatment at 80°C for 30 min. The obtained enzyme worked best at pH 7.5 and the optimum reaction temperature was determined to be 50°C. Interestingly, TiGPNT could fully retain its activity even after extended incubation periods at temperatures of up to 80°C. The enzyme was strongly inhibited in the presence of Cu2+ and Fe2+ ions and EDTA. Among the tested glycosyl donor substrates, TiGPNT showed strict specificity towards glucose-1-phosphate. At the same time, TiGPNT was highly promiscuous towards all tested nucleotide donor substrates. TiGPNT shows comparable biochemical features in regards to pH optima, temperature optima and the substrate specificity to characterized glucose-1-phosphate nucleotidyltransferase from other species. The enzyme was capable of utilizing glucose-1-phosphate and all tested nucleoside triphosphate donors as substrates. The high activity of the enzyme and the simple purification protocol make TiGPNT an interesting new biocatalyst for the synthesis of glucose-diphospho nucleosides.


Evaluation of Antimicrobial and Antifungal efficacy of Chitosan as endodontic irrigant against Enterococcus Faecalis and Candida Albicans Biofilm formed on tooth substrate.
Bacterial biofilms formed on the root canal wall are often difficult to remove. This study aimed to evaluate the cytotoxic effect and antibacterial efficacy of chitosan when used as root canal irrigant against E. Faecalis and Candida albicans biofilm formed on tooth substrate. The present study evaluated antibacterial effect of 0.25% Chitosan, 0.5% Chitosan, 2% chlorhexidine and 3% sodium hypochlorite against Enterococcus faecalis and Candida Albicans. Agar-well diffusion methods, minimal inhibitory concentration tests and biofilm susceptibility assays were used to determine antibacterial activity. Teeth specimens were sectioned to obtain a standardized tooth length of 12mm. Specimens were inoculated with 10 mL of the freshly prepared E. Faecalis suspension and Candida albicans for 4 weeks. The specimens were then instrumented with ProTaper rotary files F3 size. After irrigation with test solution, three sterile paper points were placed into one canal, left for 60 s and transferred to a test tube containing 1 mL of reduced transport fluid. The number of CFU in 1 mL was determined. 3-week biofilm qualitative assay showed complete inhibition of bacterial growth with 3% Sodium hypochlorite, 2% Chlorhexidine and Chitosan except saline, which showed presence of bacterial growth. Significant reduction of colony forming units (CFU)/mL was observed for the chitosan groups and the antibacterial activity of the chitosan groups was at par with 3% NaOCl and 2% Chlorhexidine. It was observed that the chitosan showed no cytotoxicity at 3mg/ml and 10% cytotoxicity at 6mg/ml. The use of chitosan as a root canal irrigant might be an alternative considering the various undesirable properties of NaOCl and chlorhexidine. Key words:Biofilm, Candida albicans, Chitosan, Cytotoxicity, Enterococcus faecalis.


Cornea refractive surgery and scanning laser polarimetry.
The use of modern GDx technology (GDx-VCC and GDx-ECC) provides sufficient technical background for polarimetric diagnosis of glaucoma in eyes previously undergone corneal refractive surgery. This is of clinical importance, since the number of young myopic individuals treated with LASIK for ametropia correction is large. Most of them have not yet reached the age in which the prevalence of ahead of the ophthalmologists. Myopic persons have higher risk for glaucoma, and for those who have undergone LASIK, intraocular pressure with the usual applanation tonometric methods is underestimated. Thus the clinical importance of routine use of imaging techniques in this population is especially high.


Global 3' UTR shortening has a limited effect on protein abundance in proliferating T cells.
Alternative polyadenylation is a cellular mechanism that generates mRNA isoforms differing in their 3' untranslated regions (3' UTRs). Changes in polyadenylation site usage have been described upon induction of proliferation in resting cells, but the underlying mechanism and functional significance of this phenomenon remain largely unknown. To understand the functional consequences of shortened 3' UTR isoforms in a physiological setting, we used 3' end sequencing and quantitative mass spectrometry to determine polyadenylation site usage, mRNA and protein levels in murine and human naive and activated T cells. Although 3' UTR shortening in proliferating cells is conserved between human and mouse, orthologous genes do not exhibit similar expression of alternative 3' UTR isoforms. We generally find that 3' UTR shortening is not accompanied by a corresponding change in mRNA and protein levels. This suggests that although 3' UTR shortening may lead to changes in the RNA-binding protein interactome, it has limited effects on protein output.