Expanding the substrate repertoire of a DNA polymerase by directed evolution.
Nucleic acid polymerases are the most important reagents in biotechnology. Unfortunately, their high substrate specificity severely limits their applications. Polymerases with tailored substrate repertoires would significantly expand their potential and allow enzymatic synthesis of unnatural polymers for in vivo and in vitro applications. For example, the ability to synthesize 2'-O-methyl-modified polymers would provide access to materials possessing properties that make them attractive for biotechnology and therapeutic applications, but unfortunately, no known polymerases are capable of efficiently accepting these modified substrates. To evolve such enzymes, we have developed an activity-based selection method which isolates polymerase mutants with the desired property from libraries of the enzyme displayed on phage. In this report, mutants that could efficiently synthesize an unnatural polymer from 2'-O-methyl ribonucleoside triphosphates were immobilized and isolated by means of their activity-dependent modification of a DNA oligonucleotide primer attached to the same phage particle. In each case, directed evolution resulted in relocating a critical side chain to a different position in the polypeptide, thus re-engineering the overall active site while preserving critical protein-DNA interactions. Remarkably, one evolved polymerase is shown to incorporate the modified substrates with an efficiency and fidelity equivalent to that of the wild-type enzyme with natural substrates.


The small molecule ISRIB reverses the effects of eIF2α phosphorylation on translation and stress granule assembly.
Previously, we identified ISRIB as a potent inhibitor of the integrated stress response (ISR) and showed that ISRIB makes cells resistant to the effects of eIF2α phosphorylation and enhances long-term memory in rodents (Sidrauski et al., 2013). Here, we show by genome-wide in vivo ribosome profiling that translation of a restricted subset of mRNAs is induced upon ISR activation. ISRIB substantially reversed the translational effects elicited by phosphorylation of eIF2α and induced no major changes in translation or mRNA levels in unstressed cells. eIF2α phosphorylation-induced stress granule (SG) formation was blocked by ISRIB. Strikingly, ISRIB addition to stressed cells with pre-formed SGs induced their rapid disassembly, liberating mRNAs into the actively translating pool. Restoration of mRNA translation and modulation of SG dynamics may be an effective treatment of neurodegenerative diseases characterized by eIF2α phosphorylation, SG formation, and cognitive loss.


A Retrospective Naturalistic Study Comparing the Efficacy of Ketamine and Repetitive Transcranial Magnetic Stimulation for Treatment-Resistant Depression.
Depression is a common mental disorder that affects many people worldwide, while a significant proportion of patients remain non-responsive to antidepressant medications. Alternative treatment options such as ketamine therapy and repetitive transcranial magnetic stimulation (rTMS) therapy are offered nowadays. This study aims to describe and compare the acute antidepressive efficacy of both, intramuscular ketamine and rTMS in depression patients seeking help in a naturalistic clinical mental health setting. The clinical records of 24 patients with treatment resistant depression were collected from the clinical base of a real life clinic. Twelve patients were treated with intramuscular ketamine, twice weekly for 8 sessions, and twelve patients were treated with 30 sessions of left dorsolateral prefrontal cortex - intermittent theta-burst stimulation (DLPFC-iTBS). Using three clinical assessments (HDRS, HAM-A, BDI-II), our data reveal that both therapies led to significant improvement in symptoms from pre- to post- treatment, as well as that the two experimental groups did not differ significantly with respect to pre- to post- depressive and anxiety symptoms, indicating that the effect of both experimental groups in our sample was equally effective. Furthermore, our results showed high remission and response rates in both groups, with no statistical differences between the patients of ketamine group and rTMS group in remission and response rates. We show a significant pre- to post- treatment reduction in depressive and anxiety symptoms, with no significant differences between the two experimental groups, indicating that the effect of both therapies was equally effective in our limited sample.


[Role of water-water circulation in excessive light energy dissipation of ginger leaves].
To investigate the photo-protection of water-water circulation in ginger leaves, the effects of different treatments such as natural light + water control (T1), shading 50% + water control (T2), natural light + 10 mmol x L(-1) IA (T3), shading 50% + 10 mmol x L(-1) IA (T4) on leaf chlorophyll fluorescence parameters, Mehler reaction and the activities of SOD, APX of potted ginger were studied. The results showed that the Pn and Fv/Fm of ginger leaves in T3 and T4 reduced constantly, but that of T1 and T2 had no significant change during treatment. For example, at the ninth day after treatment, the Pn of T3 and T4 decreased by 64% and 33.2% respectively, and the Fv/Fm decreased by 16.5% and 10.9% respectively, while Mehler reaction of T3 and T4 increased by 139.4% and 72.6% respectively, the activity of SOD and APX were significantly higher than those of the controls. At the sixth day after treatment, the leaf Pn and Fv/Fm in all treatments reduced significantly at noon, but Mehler reaction and the activities of SOD and APX increased markedly, and the largest amplitude was observed in T3, followed by T4, and then T1 and T2. So, it was clear that Mehler reaction and the activity of reactive-oxygen scavenging enzymes increased when leaf Pn was inhibited by exogenous IA. The results suggested that water-water circulation played an essential role in dissipating excessive light energy of ginger leaves.


[Ultrastructural analysis of reactive changes in the exo- and endocrine epithelium of embryonal pancreas in organ cultures].
The purpose of the work was to study reactive changes of endocrine and exocrine epithelii of the rat embryonic pancreas in organic culturation. The culturation was carried out at 34 degrees C and in the nutritional mixture of the following composition: medium 199--70%, cattle serum--20%, chicken embryo extract--10%, glucose--400 mg/100 ml, vitamin "C"--7 mg/100 ml, benzilpenicillin--50 U/ml. Three series of experiments were carried out with cultivation of the pancreatic gland of 18, 19 and 20-day-old embryos for 4--28 days. The electronograms obtained demonstrate a certain difference in reactivity, cytophysiology and differentiation of exocrine and endocrine epithelium under the conditions of organic cultivation. During the process, endocrine epithelium undergoes primary differentiation, secondary cytodifferentiation and, further, degenerative changes and necrosis develop. B-cells of the insular apparatus grow and function better than the exogenous epithelium. On the 28th day of cultivation, the embryonic pancreas of 19-day-old embryo demonstrates certain signs of synthesis and B-granule formation. In B-cells cytoplasm, granules of different stages of secrete formation are revealed.


MYC2 Orchestrates a Hierarchical Transcriptional Cascade That Regulates Jasmonate-Mediated Plant Immunity in Tomato.
The hormone jasmonate (JA), which functions in plant immunity, regulates resistance to pathogen infection and insect attack through triggering genome-wide transcriptional reprogramming in plants. We show that the basic helix-loop-helix transcription factor (TF) MYC2 in tomato (Solanum lycopersicum) acts downstream of the JA receptor to orchestrate JA-mediated activation of both the wounding and pathogen responses. Using chromatin immunoprecipitation sequencing (ChIP-seq) coupled with RNA sequencing (RNA-seq) assays, we identified 655 MYC2-targeted JA-responsive genes. These genes are highly enriched in Gene Ontology categories related to TFs and the early response to JA, indicating that MYC2 functions at a high hierarchical level to regulate JA-mediated gene transcription. We also identified a group of MYC2-targeted TFs (MTFs) that may directly regulate the JA-induced transcription of late defense genes. Our findings suggest that MYC2 and its downstream MTFs form a hierarchical transcriptional cascade during JA-mediated plant immunity that initiates and amplifies transcriptional output. As proof of concept, we showed that during plant resistance to the necrotrophic pathogen Botrytis cinerea, MYC2 and the MTF JA2-Like form a transcription module that preferentially regulates wounding-responsive genes, whereas MYC2 and the MTF ETHYLENE RESPONSE FACTOR.C3 form a transcription module that preferentially regulates pathogen-responsive genes.


Selenium and cancer: biomarkers of selenium status and molecular action of selenium supplements.
The relationship between selenium and cancer involves many different aspects. These include the forms of selenium present in the diet and in the body, their functions and mechanisms of action, and methods employed in assessing an individual's selenium nutritional status-both in general, and in epidemiological studies of the risk of cancer in relation to diet, as well as in connection with long-term trials for investigating the disease-preventive potential of selenium supplementation. To review different aspects on selenium metabolism, the occurrence of different selenoproteins and their use as biomarkers of selenium status, the results of intervention trials of the cancer-preventive effects of selenium supplementation, the mechanisms of action involved, together with epidemiological findings on relations between the selenium status in the body and risk of cancer. The rapid advance in the knowledge of different selenoproteins and their biological functions has opened up new possibilities for the understanding of the biological effects of selenium supplementation. A wide variety of effects of different forms and doses of selenium has been observed in a number of experimental systems, and it is at present difficult to pinpoint the mechanism that may explain the positive preventive effects of selenium supplementation observed in some human long-term trials. Moreover, additional such trials are needed to define the benefits and risks of different types and doses of selenium supplements which in the future may be implemented for public health reasons. Another necessary focus for future research is a better understanding of the mechanisms by which selenium interferes with the carcinogenesis process.


The effects of Tai Chi on the balance control of elderly persons with visual impairment: a randomised clinical trial.
balance control is a major problem for older individuals with poor vision. There are limitations, however, for visually impaired elderly persons wishing to participate in exercise programmes. The benefits of Tai Chi for balance control, muscle strength and preventing falls have been demonstrated with sighted elderly subjects. This study was designed to extend those findings to elderly persons with visual impairment. to investigate the effects of Tai Chi on the balance control of elderly persons with visual impairment. randomised clinical trial. residential care homes. forty visually impaired persons aged 70 or over. the participants were randomly divided into Tai Chi and control groups and assessed pre- and post-intervention using three tests: (i) passive knee joint repositioning to test knee proprioception; (ii) concentric isokinetic strength of the knee extensors and flexors and (iii) a sensory organisation test to quantify an individual's ability to maintain balance in a variety of complex sensory conditions. after intervention, the Tai Chi participants showed significant improvements in knee proprioception and in their visual and vestibular ratios compared with the control group. practicing Tai Chi can improve the balance control of visually impaired elderly persons.


Segment distraction to reduce a wide alveolar cleft before alveolar bone grafting.
To demonstrate a method for reduction of wide alveolar clefts prior to bone grafting. This method aims to facilitate bone grafting and achieve adequate soft tissue coverage of the graft with attached gingiva. Treatment of a patient with bilateral cleft lip and palate with a severe alveolar defect on the left side is illustrated. Distraction osteogenesis was used to mesialize the left segment so that the alveolar cleft was reduced to a minimum. After a 10-week retention period, bone grafting of the reduced alveolar defect was successfully performed. After consolidation of the bone graft, the alveolar cleft was found to have an osseous closure with adequate soft tissue coverage. In patients with a wide alveolar cleft, soft tissue coverage with keratinized mucosa can be more readily assured by reducing the alveolar cleft by callus distraction to mesialize the lateral segment prior to bone grafting.


Motor timing and motor sequencing contribute differently to the preparation for voluntary movement.
Two crucial processes preceding voluntary action are determining the time for movement initiation and planning of the specific sequence of motor output. In this study we aimed to differentiate the neural activity related to motor timing and motor sequencing and to examine over what time periods they contribute to premovement activity during the readiness for voluntary action. Eighteen participants performed self-initiated voluntary finger movements in a readiness potential paradigm, both during EEG measurement and during fMRI. The finger movement task involved three conditions: (1) simple repetitive sequences; (2) increased demand on the sequencing of movement order; and (3) increased demand on the timing of movement initiation. Functional MRI and 64 channels EEG were conducted in two separate sessions. Motor timing and motor sequencing were found to involve different neural processes occurring at different times prior to movement initiation. Motor timing involved greater activation in lateral prefrontal regions over the earliest part of premovement activity, from 1200 ms before movement onset. Motor sequencing involved greater activation of dorsal premotor and parietal areas and was reflected in central and parietal scalp regions only over the later part of premovement activity, within 600 ms of movement onset. We suggest that different neural processes contribute to different aspects of the intended action over different time periods during the preparation for movement, and it is the coordinated activity of these multiple regions that is represented in premovement activity during the readiness for voluntary action.


The vascular basis of the radial forearm flap.
The radial forearm flap is one of several valuable fasciocutaneous flaps. Therefore, its arterial and venous supply has been studied by dissection of 56 cadavers using various techniques, such as ink, latex, and barium sulfate injections. The relevant arterial branches tend to be in three groups with three associated and anastomosing perfusion zones. The blood supply of the distal radius is shown by latex injections. The vessels to tendons and nerves are briefly discussed. The flap is drained by superficial veins or radial artery venae comitantes or both. Each arterial perforator has one or two associated veins through which blood can pass into the venae comitantes. It is confirmed that the veins have valves, through which reverse flow has to occur in the pedicle of distally based island flaps. An explanation of how this is possible is given. The relevance of the findings to fasciocutaneous flaps in general is discussed.


Mortality of workers potentially exposed to epichlorohydrin.
An epidemiological study was undertaken to determine whether the animal carcinogen, epichlorohydrin (ECH), produces cancer in man. A total of 863 workers with probable exposure to ECH at two chemical plants during 1948-65 were followed up for deaths up to 1983. Twenty years or more after first exposure the all cancer SMR was 112.2 (22 deaths) and the SMR for leukaemia was 500.0 (three deaths), which is statistically significant. All cancer, leukaemia, and most other causes of death were related to estimated levels of exposure to ECH, except violence. The most consistent (both plants) relation was between exposure level and heart disease. Overall, the heart disease SMR 20 years or more after first exposure was 39.2 (five deaths) for low exposure and 105.4 (17 deaths) for high exposure. Limited evidence of a cardiovascular disease relation to ECH production in one other epidemiological study is supported by this study. Allyl chloride used in the production of ECH may play a part. The relation of heart disease and exposure does not appear to be an artifact, although the fact that many other causes of death were also related to exposure argues against a causal relation.


Tissue Doppler imaging in Fabry disease.
The development of effective enzyme replacement/enhancement therapy makes of clinical relevance considering Fabry disease in the differential diagnosis of patients with hypertrophic cardiomyopathy. In particular the opportunity to significantly modify the clinical progression of the disease has reinforced the need for early diagnosis of Fabry cardiomyopathy. The study with tissue Doppler of Fabry patients with endomyocardial biopsy-proven cardiac involvement showed a reduction of both diastolic and systolic myocardial velocities recorded at septal and lateral corners of mitral annulus. Tissue Doppler abnormalities were present not only in patients with left ventricular hypertrophy but also in younger patients with normal cardiac wall thickness and represent the first sign of myocardial damage. Furthermore tissue Doppler studies have been shown useful in detecting cardiac involvement in female carriers with no systemic manifestations of Fabry disease. In patients already submitted to enzyme-replacement therapy tissue Doppler and strain rate imaging represent useful noninvasive tools in assessing treatment efficacy. Tissue Doppler imaging can provide early detection of cardiac involvement in Fabry disease and represents the most accurate and sensitive noninvasive tool for the diagnosis of myocardial dysfunction and for the assessment of cardiac improvement during enzyme replacement therapy. The detection of tissue Doppler abnormalities in female carriers may represent a hint for an invasive assessment of cardiac involvement.


[Drug interaction computer programs: which choice?].
Adverse drug reactions represent a partly preventable cause of morbidity. Computer-based tools may be useful for the prevention of those events resulting from drug interactions. While several such tools are currently available to practitioners, they have rarely been compared regarding their performances and limits. This article presents a benchmark evaluation of seven drug interaction databases which may be elected by physicians or pharmacists as an aid to prescription on a regular basis. None of the databases tested appears to be perfect, and the systems vary in their overall completeness and ease of use.


Rat brain inositol 1,4,5-trisphosphate 3-kinase. Ca2(+)-sensitivity, purification and antibody production.
Inositol 1,4,5-trisphosphate (InsP3) 3-kinase catalyses the ATP-dependent phosphorylation of InsP3 to inositol 1,3,4,5-tetrakisphosphate (InsP4). InsP3 3-kinase was purified from rat brain by Blue-Sepharose, phosphocellulose and calmodulin (CaM)-Sepharose affinity chromatography. The purified enzyme was stimulated by Ca2+/CaM by 3-6-fold as compared with the activity measured in the presence of EGTA. Rat brain InsP3 3-kinase activity was associated with two silver-stained bands of about equal activity which migrated with an apparent Mr of 50,000 on SDS/polyacrylamide gels. InsP3 3-kinase activity from rat brain could be immunoprecipitated by an antiserum against the SDS/PAGE-purified 50,000-Mr protein doublet. InsP3 kinase activity from bovine brain and the InsP3 5-phosphatase activity from rat brain were not immunoprecipitated. On Western blot, the human brain crude InsP3 3-kinase reacted specifically, but less strongly than the rat brain enzyme, with the antiserum.


Implementation of a system-wide policy for labor induction.
Standardization of key clinical protocols and policies in the inpatient obstetric setting has the potential to improve care processes, ultimately resulting in better patient outcomes and decreased professional liability. Increasingly hospitals and healthcare systems are realizing benefits with adoption of standardization as a strategy for quality care improvement. We describe successful system-wide development and implementation of a policy for labor induction including avoidance of elective births before 39 completed weeks of gestation and standardization of various aspects of the labor induction process, with the goal of providing safer care.


Design of amphiphilic oligopeptides as models for fine tuning peptide assembly with plasmid DNA.
We discuss the design of novel amphiphilic oligopeptides with hydrophobic and cationic amino acids to serve as models to understand peptide-DNA assembly. Biophysical and thermodynamic characterization of interaction of these amphiphilic peptides with plasmid DNA is presented. Peptides with at least +4 charges favor stable complex formation. Surface potential is dependent on the type of hydrophobic amino acid for a certain charge. Thermodynamically it is a spontaneous interaction between most of the peptides and plasmid DNA. Lys(7) and Tyr peptides with +4/+5 charges indicate cooperative binding with pDNA without saturation of interaction while Val(2)-Gly-Lys(4), Val-Gly-Lys(5), and Phe-Gly-Lys(5) lead to saturation of interaction indicating condensed pDNA within the range of N/Ps studied. We show that the biophysical properties of DNA-peptide complexes could be modulated by design and the peptides presented here could be used as building blocks for creating DNA-peptide complexes for various biomedical applications, mainly nucleic acid delivery.


Irritable Bowel Syndrome Symptoms in Nonalcoholic Fatty Liver Disease Patients Are an Indicator of Depression and Anxiety.
Determine factors associated with Irritable bowel syndrome (IBS) in nonalcoholic fatty liver disease (NAFLD) patients. IBS affects 10% to 15% of the adult population worldwide and is linked to anxiety and depression. The impact of IBS-type symptoms in NAFLD patients is not well described. A cross-sectional study of patients in the hepatology clinic at Houston Methodist Hospital was performed based on a respondent postal survey. IBS was defined by the Rome IV questionnaire, anxiety and depression were assessed with the Hospital Anxiety Depression scale. Patients with inflammatory bowel disease, colorectal carcinoma, or small bowel tumors were excluded. Patients were divided based on Rome IV diagnostic criteria for IBS and Hospital Anxiety Depression scale. 130 patients were included in the analysis, 38 satisfied Rome IV criteria for IBS (IBS group) versus 92 who did not (non-IBS group). Depression was more prevalent in the IBS group (18.4% vs 5.4%, P =0.01). Anxiety was also greater in the IBS group (31.6% vs 9.8%, P =0.002). Female sex, depression, and body mass index (BMI)>30 were independent predictors of IBS in NAFLD in 4 multiple logistic regression models. In newly diagnosed IBS patients, gamma-glutamyl transferase levels were lower (67.5 vs 28, P =0.04). Current abdominal pain was higher than 100% versus 81.3% ( P =0.045), as was pain associated with the change in stool frequency (96.3% vs 50%; P <0.001). Our study highlights the increased rate of IBS symptoms, depression, and anxiety in patients with NAFLD. Clinicians should be alert when IBS symptoms are reported by a NAFLD patient and be aware of the impact of these comorbidities on quality of life and response to therapy.


Comparison of the cloned H-2Kbm1 variant gene with the H-2Kb gene shows a cluster of seven nucleotide differences.
The gene for the H-2K class I antigen of the bm1 variant was cloned and analyzed at the DNA level and compared with the previously cloned parent B6/Kh gene. Sequence determination and comparative restriction endonuclease studies indicate that Kbm1 is derived from the Kb gene. Seven nucleotide changes within a 13-nucleotide stretch distinguish the mutant from the parent gene and result in amino acid differences at positions 152, 155, and 156 in the antigen. The data confirm previously reported changes at amino acid positions 155 and 156 (arginine to tyrosine and leucine to tyrosine, respectively) and extend the altered region to include two nucleotides encoding a glutamate to alanine substitution at amino acid 152, a change not detected by the protein studies because of limitations of the methods used. The DNA sequence encoding this region of the Kbm1 glycoprotein is identical to the DNA sequence of at least one other known class I gene in the mouse, a finding consistent with the hypothesis that the mutation was not a random event but may be the result of a block transfer of information by a copy mechanism analogous to gene conversion. As the sequence analysis of the coding region for the first 273 amino acid residues shows identity between parent and mutant except for the seven nucleotide changes, all variant-parent functional differences must depend only on the cluster of three amino acid differences in the second domain of the Kb glycoprotein.


Assessing interventions available to internally displaced persons in Abia state, Nigeria.
Internally displaced persons are faced with several problems, such as sexual violence, and deserve appropriate intervention, especially in view of the increasing prevalence of HIV/AIDS and other infections in Nigeria. This study attempts to assess interventions offered by governmental authorities and organizations to internally displaced persons and to identify gaps in services as well as to identify what needs to be strengthened. The author reviewed relevant published and unpublished documents and collected data by interviews with semi-structured questions. Twenty-five organizations and government and police departments and 55 internally displaced persons were interviewed. None of the organizations, including governmental institutions, provided social services or assistance in prevention of HIV/AIDS to internally displaced persons. The main services provided by 17 (68%) organizations to 43 (78.2%) of internally displaced persons were provision of food, clothing and money, but these were provided on an ad hoc basis. Only 3 organizations (12%) included spiritual counseling and resolution of communal conflicts in their services. The fact that most organizations, including the government, do not have services for internally displaced persons indicates lack of support for internally displaced persons. The government should be urged to include these people in most prevention services, including HIV/AIDS prevention and treatment. This should help reduce the national prevalence of HIV/AIDS.


Low-molecular-weight heparin in patients with chronic renal insufficiency.
Low-molecular-weight heparin (LMWH) has largely replaced unfractionated heparin for the treatment of venous thromboembolism. The predictable anticoagulant effect of LMWH is seen across almost all patient populations, with few exceptions. However, because LMWH is primarily eliminated through the kidneys, patients with renal insufficiency are at risk of LMWH accumulation and bleeding complications. The risk of LMWH accumulation and bleeding is dependent on several factors including the degree of renal insufficiency, dose and type of LMWH. These risks are greatest when therapeutic doses of LMWH are used in patients with creatinine clearance less than 30 ml/min. Prophylactic dose LMWH does not appear to be associated with an increased bleeding risk, but has not been evaluated in large trials. LMWHs with a higher molecular weight may be less prone to accumulation and bleeding. LMWH must be used carefully in patients with renal insufficiency, particularly in those with severe renal impairment.


Predictors of high residual gradient after transcatheter aortic valve replacement in bicuspid aortic valve stenosis.
To define the incidence of high residual gradient (HRG) after transcatheter aortic valve replacement (TAVR) in BAVs and their impact on short term outcome and 1-year mortality. Transcatheter heart valves (THVs) offer good performance in tricuspid aortic valves with low rate of HRG. However, data regarding their performance in bicuspid aortic valves (BAV) are still lacking. The BEAT (Balloon vs Self-Expandable valve for the treatment of bicuspid Aortic valve sTenosis) registry included 353 consecutive patients who underwent TAVR (Evolut R/PRO or Sapien 3 valves) in BAV between June 2013 and October 2018. The primary endpoint was device unsuccess with post-procedural HRG (mean gradient ≥ 20 mmHg). The secondary endpoint was to identify the predictors of HRG following the procedure. Twenty patients (5.6%) showed HRG after TAVR. Patients with HRG presented higher body mass index (BMI) (30.7 ± 9.3 vs. 25.9 ± 4.8; p < 0.0001) and higher baseline aortic mean gradients (57.6 ± 13.4 mmHg vs. 47.7 ± 16.6, p = 0.013) and more often presented with BAV of Sievers type 0 than patients without HRG. At multivariate analysis, BMI [odds ratio (OR) 1.12; 95% confidence interval (CI) 1.05-1.20, p = 0.001] and BAV type 0 (OR 11.31, 95% CI 3.45-37.06, p < 0.0001) were confirmed as independent predictors of high gradient. HRG following TAVR in BAVs is not negligible and is higher among patients with high BMI and with BAV 0 anatomy.


Genetic variation in South Asia: assessing the influences of geography, language and ethnicity for understanding history and disease risk.
South Asia is home to more than 1.5 billion humans representing many diverse ethnicities, linguistic and religious groups and representing almost one-quarter of humanity. Modern humans arrived here soon after their departure from Africa approximately 50,000-70,000 years before present (YBP) and several subsequent human migrations and invasions, as well as the unique social structure of the region, have helped shape the pattern of genetic diversity currently observed in these populations. Over the last few decades population geneticists and molecular anthropologists have analyzed DNA variation in indigenous populations from this region in order to catalog their genetic relationships and histories. The emphasis is gradually shifting from the study of population origins to high resolution surveys of DNA variation to address issues of population stratification and genetic susceptibility or resistance to diseases in genome-wide association surveys. We present a historical overview of the genetic studies carried out on populations from this region in order to understand the influence of geographic, linguistic and religious factors on population diversity in this region, and discuss future prospects in light of developments in high throughput genotyping and next generation sequencing technologies.


Effects of biological type of beef steers on vitamin D, calcium, and phosphorus status.
Feedlot steers (n = 36) from three biological types (Bos indicus, Bos taurus-Continental, and Bos taurus-English) were used to determine the Ca, P, and vitamin D3 status of feedlot cattle. The USDA yield and quality grade traits were measured at slaughter, and the concentrations of vitamin D3 (VITD) and the metabolites 25-hydroxyvitamin D3 (25-OH D) and 1,25-dihydroxyvitamin D3 (1,25-(OH)2 D) were determined in LM, liver, kidney, and plasma. Plasma and muscle Ca and P concentrations also were determined. Biological type of cattle affected a number of carcass traits. Carcasses from Bos taurus-English cattle had more marbling, resulting in higher quality grades (P < 0.05). Carcasses from Bos taurus-Continental cattle had lower calculated yield grades (P < 0.05) than did carcasses from cattle in the other biological types. In general, differences in carcass traits resulting from biological type were consistent with other reports. Plasma and LM Ca and P concentrations were not affected (P = 0.06) by biological type of cattle, indicating that Ca and P homeostasis is a conserved trait across the different types of cattle. Plasma VITD and 25-OH D concentrations were not affected (P = 0.41) by biological type, whereas plasma 1,25-(OH)2 D concentration was lower (P < 0.05) in Bos taurus-English cattle than in Bos taurus-Continental and Bos indicus cattle. Liver VITD and 25-OH D were not affected by biological type (P = 0.76), but liver 1,25-(OH)2 D concentration was greater (P < 0.05) in Bos indicus cattle than in Bos taurus-Continental cattle. Kidney vitamin D metabolite concentrations were not affected by biological type of cattle (P = 0.21). Muscle VITD concentration was greater (P < 0.05) in Bos taurus-English cattle than in the other two biological types, and muscle 25-OH D concentrations were greater (P < 0.05) in Bos taurus-English cattle than in Bos indicus cattle. Muscle 1,25-(OH)2 D concentration was less (P < 0.05) in the Bos taurus-Continental cattle than in the other two biological types. Cooking eliminated vitamin D metabolite differences among the biological types. Our results suggest that Bos indicus cattle had greater 1,25-(OH)2 D (the biologically active form) in tissues, and greater 1,25-(OH)2 D plasma concentrations than Bos taurus cattle. Thus, the need for VITD supplementation and optimal levels of Ca and P in feedlot diets might differ between Bos indicus and Bos taurus cattle.


CRISPR/Cas9-targeted removal of unwanted sequences from small-RNA sequencing libraries.
In small RNA (smRNA) sequencing studies, highly abundant molecules such as adapter dimer products and tissue-specific microRNAs (miRNAs) inhibit accurate quantification of lowly expressed species. We previously developed a method to selectively deplete highly abundant miRNAs. However, this method does not deplete adapter dimer ligation products that, unless removed by gel-separation, comprise most of the library. Here, we have adapted and modified recently described methods for CRISPR/Cas9-based Depletion of Abundant Species by Hybridization ('DASH') to smRNA-seq, which we have termed miRNA and Adapter Dimer-DASH (MAD-DASH). In MAD-DASH, Cas9 is complexed with single guide RNAs (sgRNAs) targeting adapter dimer ligation products, alongside highly expressed tissue-specific smRNAs, for cleavage in vitro. This process dramatically reduces adapter dimer and targeted smRNA sequences, can be multiplexed, shows minimal off-target effects, improves the quantification of lowly expressed miRNAs from human plasma and tissue derived RNA, and obviates the need for gel-separation, greatly increasing sample throughput. Additionally, the method is fully customizable to other smRNA-seq preparation methods. Like depletion of ribosomal RNA for mRNA-seq and mitochondrial DNA for ATAC-seq, our method allows for greater proportional read-depth of non-targeted sequences.


Structure and Enzymatic Activity of an Intellectual Disability-Associated Ornithine Decarboxylase Variant, G84R.
Ornithine decarboxylase (ODC) is a rate-limiting enzyme for the synthesis of polyamines (PAs). PAs are required for proliferation, and increased ODC activity is associated with cancer and neural over-proliferation. ODC levels and activity are therefore tightly regulated, including through the ODC-specific inhibitor, antizyme AZ1. Recently, ODC G84R has been reported as a partial loss-of-function variant that is associated with intellectual disability and seizures. However, G84 is distant from both the catalytic center and the ODC homodimerization interface. To understand how G84R modulates ODC activity, we have determined the crystal structure of ODC G84R in both the presence and the absence of the cofactor pyridoxal 5-phosphate. The structures show that the replacement of G84 by arginine leads to hydrogen bond formation of R84 with F420, the last residue of the ODC C-terminal helix, a structural element that is involved in the AZ1-mediated proteasomal degradation of ODC. In contrast, the catalytic center is essentially indistinguishable from that of wildtype ODC. We therefore reanalyzed the catalytic activity of ODC G84R and found that it is rescued when the protein is purified in the presence of a reducing agent to mimic the reducing environment of the cytoplasm. This suggests that R84 may exert its neurological effects not through reducing ODC catalytic activity but through misregulation of its AZ1-mediated proteasomal degradation.


Identification of a novel vga(E) gene variant that confers resistance to pleuromutilins, lincosamides and streptogramin A antibiotics in staphylococci of porcine origin.
To investigate the genetic basis of pleuromutilin resistance in coagulase-negative staphylococci of porcine origin that do not carry known pleuromutilin resistance genes and to determine the localization and genetic environment of the identified resistance gene. Plasmid DNA of two pleuromutilin-resistant Staphylococcus cohnii and Staphylococcus simulans isolates was transformed into Staphylococcus aureus RN4220. The identified resistance plasmids were sequenced completely. The candidate gene for pleuromutilin resistance was cloned into shuttle vector pAM401. S. aureus RN4220 transformants carrying this recombinant shuttle vector were tested for their MICs. S. cohnii isolate SA-7 and S. simulans isolate SSI1 carried the same plasmid of 5584 bp, designated pSA-7. A variant of the vga(E) gene was detected, which encodes a 524 amino acid ATP-binding cassette protein. The variant gene shared 85.7% nucleotide sequence identity and the variant protein 85.3% amino acid sequence identity with the original vga(E) gene and Vga(E) protein, respectively. The Vga(E) variant conferred cross-resistance to pleuromutilins, lincosamides and streptogramin A antibiotics. Plasmid pSA-7 showed an organization similar to that of the apmA-carrying plasmid pKKS49 from methicillin-resistant S. aureus and the dfrK-carrying plasmid pKKS966 from Staphylococcus hyicus. Sequence comparisons suggested that recombination events may have played a role in the acquisition of this vga(E) variant. A novel vga(E) gene variant was identified, which was located on a small plasmid and was not associated with the transposon Tn6133 [in contrast to the original vga(E) gene]. The plasmid location may enable its further dissemination to other staphylococci and possibly also to other bacteria.


Kaempferol, a dietary flavonoid, ameliorates acute inflammatory and nociceptive symptoms in gastritis, pancreatitis, and abdominal pain.
Kaempferol (KF) is the most abundant polyphenol in tea, fruits, vegetables, and beans. However, little is known about its in vivo anti-inflammatory efficacy and mechanisms of action. To study these, several acute mouse inflammatory and nociceptive models, including gastritis, pancreatitis, and abdominal pain were employed. Kaempferol was shown to attenuate the expansion of inflammatory lesions seen in ethanol (EtOH)/HCl- and aspirin-induced gastritis, LPS/caerulein (CA) triggered pancreatitis, and acetic acid-induced writhing.


Impact of the location of the fenestration on Fontan circulation haemodynamics: a three-dimensional, computational model study.
There is no consensus or theoretical explanation regarding the optimal location for the fenestration during the Fontan operation. We investigated the impact of the location of the fenestration on Fontan haemodynamics using a three-dimensional Fontan model in various physiological conditions. A three-dimensional Fontan model was constructed on the basis of CT images, and a 4-mm-diameter fenestration was located between the extracardiac Fontan conduit and the right atrium at three positions: superior, middle, and inferior part of the conduit. Haemodynamics in the Fontan route were analysed using a three-dimensional computational fluid dynamic model in realistic physiological conditions, which were predicted using a lumped parameter model of the cardiovascular system. The respiratory effect of the caval flow was taken into account. The flow rate through the fenestration, the effect of lowering the central venous pressure, and wall shear stress in the Fontan circuit were evaluated under central venous pressures of 10, 15, and 20 mmHg. The pulse power index and pulsatile energy loss index were calculated as energy loss indices. Under all central venous pressures, the middle-part fenestration demonstrated the most significant effect on enhancing the flow rate through the fenestration while lowering the central venous pressure. The middle-part fenestration produced the highest time-averaged wall shear stress, pressure pulse index, and pulsatile energy loss index. Despite slightly elevated energy loss, the middle-part fenestration most significantly increased cardiac output and lowered central venous pressure under respiration in the Fontan circulation.


Role of acetylcholinesterase on the structure and function of cholinergic synapses: insights gained from studies on knockout mice.
Electrophysiological and ultrastructural studies were performed on phrenic nerve-hemidiaphragm preparations isolated from wild-type and acetylcholinesterase (AChE) knockout (KO) mice to determine the compensatory mechanisms manifested by the neuromuscular junction to excess acetylcholine (ACh). The diaphragm was selected since it is the primary muscle of respiration, and it must adapt to allow for survival of the organism in the absence of AChE. Nerve-elicited muscle contractions, miniature endplate potentials (MEPPs) and evoked endplate potentials (EPPs) were recorded by conventional electrophysiological techniques from phrenic nerve-hemidiaphragm preparations isolated from 1.5- to 2-month-old wild-type (AChE(+/+)) or AChE KO (AChE(-/-)) mice. These recordings were chosen to provide a comprehensive assessment of functional alterations of the diaphragm muscle resulting from the absence of AChE. Tension measurements from AChE(-/-) mice revealed that the amplitude of twitch tensions was potentiated, but tetanic tensions underwent a use-dependent decline at frequencies below 70 Hz and above 100 Hz. MEPPs recorded from hemidiaphragms of AChE(-/-) mice showed a reduction in frequency and a prolongation in decay (37%) but no change in amplitude compared to values observed in age-matched wild-type littermates. In contrast, MEPPs recorded from hemidiaphragms of wild-type mice that were exposed for 30 min to the selective AChE inhibitor 5-bis(4-allyldimethyl-ammoniumphenyl)pentane-3-one (BW284C51) exhibited a pronounced increase in amplitude (42%) and a more marked prolongation in decay (76%). The difference between MEPP amplitudes and decays in AChE(-/-) hemidiaphragms and in wild-type hemidiaphragms treated with BW284C51 represents effective adaptation by the former to a high ACh environment. Electron microscopic examination revealed that diaphragm muscles of AChE(-/-) mice had smaller nerve terminals and diminished pre- and post-synaptic surface contacts relative to neuromuscular junctions of AChE(+/+) mice. The morphological changes are suggested to account, in part, for the ability of muscle from AChE(-/-) mice to function in the complete absence of AChE.


Clinical evaluation of a portable near-infrared device for detection of traumatic intracranial hematomas.
The purpose of this multicenter observational clinical study was to evaluate the performance of a near-infrared (NIR)-based, non-invasive, portable device to screen for traumatic intracranial hematomas. Five trauma centers collected data using the portable NIR device at the time a computed tomography (CT) scan was performed to evaluate a suspected traumatic brain injury (TBI). The CT scans were read by an independent neuroradiologist who was blinded to the NIR measurements. Of 431 patients enrolled, 365 patients were included in the per-protocol population analyzed. Of the 365 patients, 96 were determined by CT scan to have intracranial hemorrhages of various sizes, depths, and anatomical locations. The NIR device demonstrated sensitivity of 88% (95% confidence interval [CI] 74.9,95.0%), and specificity of 90.7% (95% CI 86.4,93.7%), in detecting the 50 intracranial hematomas that were large enough to be clinically important (larger than 3.5 mL in volume), and that were less than 2.5 cm from the surface of the brain. For all 96 cases with intracranial hemorrhage, regardless of size and type of hemorrhage, the sensitivity was 68.7% (CI 58.3,77.6%), and specificity was 90.7% (CI 86.4,93.7%). These results confirm the results of previous studies that indicate that a NIR-based portable device can reliably screen for intracranial hematomas that are superficial and of a size likely to be of clinical importance. The NIR device cannot replace CT scanning in the diagnosis of TBI, but the device might be useful to supplement clinical information used to triage TBI patients, and in situations in which CT scanning is not readily available.


Developmental Training Model for the Sport Specialized Youth Athlete: A Dynamic Strategy for Individualizing Load-Response During Maturation.
Most available data on athletic development training models focus on adult or professional athletes, where increasing workload capacity and performance is a primary goal. Development pathways in youth athletes generally emphasize multisport participation rather than sport specialization to optimize motor skill acquisition and to minimize injury risk. Other models emphasize the need for accumulation of sport- and skill-specific hours to develop elite-level status. Despite recommendations against sport specialization, many youth athletes still specialize and need guidance on training and competition. Medical and sport professionals also recommend progressive, gradual increases in workloads to enhance resilience to the demands of high-level competition. There is no accepted model of risk stratification and return to play for training a specialized youth athlete through periods of injury and maturation. In this review, we present individualized training models for specialized youth athletes that (1) prioritize performance for healthy, resilient youth athletes and (2) are adaptable through vulnerable maturational periods and injury. Nonsystematic review with critical appraisal of existing literature. Clinical review. Level 4. A number of factors must be considered when developing training programs for young athletes: (1) the effect of sport specialization on athlete development and injury, (2) biological maturation, (3) motor and coordination deficits in specialized youth athletes, and (4) workload progressions and response to load. Load-sensitive athletes with multiple risk factors may need medical evaluation, frequent monitoring, and a program designed to restore local tissue and sport-specific capacity. Load-naive athletes, who are often skeletally immature, will likely benefit from serial monitoring and should train and compete with caution, while load-tolerant athletes may only need occasional monitoring and progress to optimum loads. B.


Sleep Treatment Education Program for Young Adult Cancer Survivors (STEP-YA): Protocol for an Efficacy Trial.
Young adult cancer survivors (YACS) are at elevated risk for chronic insomnia, even years after completing treatment. In addition to potential health consequences, insomnia can interrupt social, educational, and vocational development just as they are trying to "make up" for time lost to cancer. Cognitive behavioral therapy for insomnia (CBTI) is recommended as first-line treatment for insomnia but remains largely unavailable to YACS due to several barriers (ie, shortage of trained providers, geographic limitations, financial limitations). Traditional CBTI has not been adapted to meet YACS' unique developmental and circadian challenges. To improve availability of effective behavioral insomnia treatment for this population, we developed the Sleep Treatment Education Program for Young Adult Cancer Survivors (STEP-YA), a low-intensity educational intervention delivered virtually online. In this phase 2 "proof of concept" trial, primary aims are to test the efficacy of STEP-YA to improve insomnia symptoms and mood in YACS and assess the utility of individualized coaching to improve treatment effects. A secondary aim will explore participant variables associated with clinically significant response to STEP-YA. This 2-arm randomized prospective trial will enroll 74 off-treatment YACS aged 20 years to 39 years with clinically significant insomnia. Each participant completes the STEP-YA intervention in a 1-on-1 synchronous online session led by a trained interventionist following a structured outline. The 90-minute intervention presents educational information on the development of insomnia after cancer and offers specific suggestions for improving insomnia symptoms. During the session, participants review the suggestions and develop a personalized sleep action plan for implementing them. After the session, participants are randomized to either the coaching condition, in which they receive 2 telephone coaching sessions, or the no-coaching condition, which offers no subsequent coaching. The Insomnia Severity Index (ISI) and the Profile of Mood States: Short Form (POMS-SF) are assessed at baseline and 4 and 8 weeks postintervention. Enrollment began in November 2022, with 28 participants currently enrolled. We anticipate recruitment will be completed in 2024. The primary endpoint is a change in ISI score from baseline to 8 weeks postintervention. The secondary endpoint is change in mood symptoms (POMS-SF) from baseline to 8 weeks postintervention. Change scores will be treated as continuous variables. Primary analyses will use ANOVA methods. A within-subjects analysis will examine if the STEP-YA intervention is associated with significant changes in insomnia and mood over time. A 2-way ANOVA will be used to evaluate the utility of coaching sessions to improve treatment effects. Chronic insomnia has significant negative effects on YACS' medical, educational, and psychological functioning. STEP-YA aims to address their needs; study results will determine if the intervention warrants future effectiveness and dissemination studies and if individualized coaching is necessary for adequate treatment response. ClinicalTrials.gov NCT05358951: https://clinicaltrials.gov/study/NCT05358951. DERR1-10.2196/52315.


[Use of preparations of microbial origin for stimulating antitumor resistance].
Results of application of the preparations made of mycobacteria, clostridia, staphylococci and streptococci, lactobacteria, Salmonella, Streptomyces and other are analyzed. Researchers in different countries pay great attention to the antitumour activity of capsular and somatic antigens of a microbic cell, its separate anatomical structures, enzymes, lipids, polypeptides and especially polysaccharides. The action of bacterial preparations and factors restraining their application in clinical oncology are considered.


Ictal vocalizations are relatively common in myoclonic-atonic seizures associated with Doose syndrome: an audio-video-polygraphic analysis.
The aim of this study was to investigate ictal vocalizations associated with myoclonic (MS) and myoclonic-atonic seizures (MAS) in patients with myoclonic epilepsy in infants (MEI) and epilepsy with myoclonic-atonic seizures (EMAS, Doose syndrome), respectively. Subjects were retrospectively recruited among patients with MEI and EMAS for whom ictal video-polygraphs were recorded between 1990 and 2019. We reviewed all MS and MAS in order to estimate how often they were associated with vocalizations, and analyze the temporal relationship between vocalizations and spike-wave complexes (SWCs) and myoclonic EMG potentials based on simultaneous examination of the polygraphs and sound signals. Ictal video-polygraphs from 15 patients with MEI (2-34 MS per patient) and 26 with EMAS (2-26 MAS per patient) were examined. Ictal vocalizations were audible in two patients with MEI (11%; 3-18 MS per patient) and nine with EMAS (35%; 2-11 MAS per patient). Sounds were always non-speech and were immediately followed by head or body dropping in the case of MAS. Detailed analysis based on simultaneous and synchronous examination of video-polygraphs and sound signals in one patient with MEI and five patients with EMAS demonstrated that the onset of the ictal vocalizations corresponded to that of the myoclonic EMG potentials and negative spike components of SWC. Comparison of the length of myoclonic EMG potentials as well as the strength of drop seizures between MAS with and without vocalizations revealed that MAS with vocalizations were associated with longer myoclonic EMG potentials and stronger drop seizures than MAS without vocalizations (p<0.05), suggesting that the vocalizations result from strong contraction of axial muscles. Ictal vocalizations due to massive motor seizure activity are a relatively common finding in MAS in Doose syndrome, which may help in the differential diagnosis of epileptic drop attacks.


COVID-19, Diabetes, and Associated Health Outcomes in China: Results from a Nationwide Survey of 10 545 Adults.
This study examined the associations between diabetes and self-reported/familial COVID-19 infection and investigated health-related outcomes among those with diabetes during China's nationwide quarantine. The 2020 China COVID-19 Survey was administered anonymously via social media (WeChat). It was completed by 10 545 adults in all of mainland China's 31 provinces. The survey consisted of 74 items covering sociodemographic characteristics, preventive measures for COVID-19, lifestyle behaviors, and health-related outcomes during the period of quarantine. Regression models examined associations among study variables. Diabetes was associated with a six-fold increased risk of reporting COVID-19 infection among respondents or their family members. Among people with diabetes, individuals who rarely wore masks had double the risk of suspected COVID-19 infection compared with those who always wore masks, with an inverse J-shaped relationship between face mask wearing and suspected COVID-19 infection. People with diabetes tended to have both poor knowledge of COVID-19 and poor compliance with preventive measures, despite perceiving a high risk of personal infection (40.0% among respondents reporting diabetes and 8.0% without diabetes). Only 54-55% of these respondents claimed to consistently practice preventive measures, including wearing face masks. Almost 60% of those with diabetes experienced food or medication shortages during the quarantine period, which was much higher than those without diabetes. Importantly, respondents who experienced medication shortages reported a 63% higher COVID-19 infection rate. Diabetes was associated with an increased risk of self-reported personal and family member COVID-19 infection, which is mitigated by consistent use of face masks.


[The organization of work in the pediatric department of an urban ophthalmologic hospital].
The paper analyses the work of a children ophthalmological department and of a centre of microsurgery in children created on its basis, that are attributed to the structure of the town ophthalmological clinical hospital in Kharkov. The paper presents data on the structure of eye pathology, surgical activity (it makes up 60%), suggestions for optimizing the work of the department. A positive role in optimization of work in children department is its belonging to a big ophthalmological hospital. This allows to widely use for children the whole store of diagnostic means, different surgical equipment, other technical means. Of importance is a permanent contact with the chair of children ophthalmology, enrollment of physicians into research work, constant improvement of their skill.


Uneven distribution of ventilation in acute respiratory distress syndrome.
The aim of this study was to assess the volume of gas being poorly ventilated or non-ventilated within the lungs of patients treated with mechanical ventilation and suffering from acute respiratory distress syndrome (ARDS). A prospective, descriptive study was performed of 25 sedated and paralysed ARDS patients, mechanically ventilated with a positive end-expiratory pressure (PEEP) of 5 cmH2O in a multidisciplinary intensive care unit of a tertiary university hospital. The volume of poorly ventilated or non-ventilated gas was assumed to correspond to a difference between the ventilated gas volume, determined as the end-expiratory lung volume by rebreathing of sulphur hexafluoride (EELVSF6), and the total gas volume, calculated from computed tomography images in the end-expiratory position (EELVCT). The methods used were validated by similar measurements in 20 healthy subjects in whom no poorly ventilated or non-ventilated gas is expected to be found. EELVSF6 was 66% of EELVCT, corresponding to a mean difference of 0.71 litre. EELVSF6 and EELVCT were significantly correlated (r2 = 0.72; P < 0.001). In the healthy subjects, the two methods yielded almost identical results. About one-third of the total pulmonary gas volume seems poorly ventilated or non-ventilated in sedated and paralysed ARDS patients when mechanically ventilated with a PEEP of 5 cmH2O. Uneven distribution of ventilation due to airway closure and/or obstruction is likely to be involved.


Attenuation of Aversive Properties of Peripheral Shock by Hypothalamic Stimulation.
Clinical reports of alleviation of pain with positive brain stimulation were investigated experimnentally. Rats in a two-compartment testing chamber sought out hypothalamic stimulation and escaped from aversive foot shock delivered through a grid-scrambling device. Animals also sought out paired hypothalamic stimulation and foot shock. Control experiments demonstrating that animals did not discriminate between hypothalamic stimulcation and paired hypothalamic stimulation and foot shock supported the view that hypothalamic stimulation attenuates the aversive properties of foot shock.


Chromosome rearrangements in Dictyostelium discoideum.
A tandem duplication (D350(III,III] of the whiB to radB interval of linkage group III has been characterized. The gene order on the duplication-bearing chromosome is: centromere, whiB500, radB+, whiB+, radB24, bsgA5, acrC4. Slow-growing, duplication-bearing strains (yellow-spored, radiation-resistant) produced four classes of faster growing sectors involving the whiB and radB loci: white-spored, radiation-sensitive (whiB500, radB24); white-spored, radiation-resistant (whiB500, radB+); yellow-spored, radiation-sensitive (whiB+, radB24); and yellow-spored, radiation-resistant. The first three classes can be explained as the products of single recombination events in which one copy of the whiB to radB interval was lost. The yellow-spored, radiation-resistant sectors probably arose by mutation elsewhere in the genome, but alternatively may represent multiple recombination events or deletion of part of one copy of the duplicated region. Loss of the duplicated segment was enhanced by irradiation with ultraviolet light (254 nm). Heterozygosity for a DNA repair mutation at the radB locus may have been involved in the formation of the duplication. It is proposed that translocations are a major cause of nonrandom segregation patterns such as the cosegregation of unlinked markers in Dictyostelium discoideum. Translocations involving all known linkage groups are tabulated and DNA damage by N-methyl-N'-nitro-N-nitrosoguanidine is implicated in the formation of translocations in D. discoideum.


The relative contributions of CYP3A4 and CYP3A5 to the metabolism of vinorelbine.
Vinorelbine is a semisynthetic vinca alkaloid used in the treatment of advanced breast and non-small cell lung cancers. Vincristine, a related vinca alkaloid, is 9-fold more efficiently metabolized by CYP3A5 than by CYP3A4 in vitro. This study quantified the relative contribution of CYP3A4 and CYP3A5 to the metabolism of vinorelbine in vitro using cDNA-expressed human cytochrome P450s (P450s) and human liver microsomes (HLMs). CYP3A4 and CYP3A5 were identified as the P450s capable of oxidizing vinorelbine using a panel of human enzymes and selective P450 inhibitors in HLMs. For CYP3A4 coexpressed with cytochrome b5 (CYP3A4+b5) and CYP3A5+b5, the Michaelis-Menten constants for vinorelbine were 2.6 and 3.6 μM, respectively, but the Vmax of 1.4 pmol/min/pmol was common to both enzymes. In HLMs, the intrinsic clearance of vinorelbine metabolism was highly correlated with CYP3A4 activity, and there was no significant difference in intrinsic clearance between CYP3A5 high and low expressers. When radiolabeled vinorelbine substrate was used, there were clear qualitative differences in metabolite formation fingerprints between CYP3A4+b5 and CYP3A5+b5 as determined by NMR and mass spectrometry analysis. One major metabolite (M2), a didehydro-vinorelbine, was present in both recombinant and microsomal systems but was more abundant in CYP3A4+b5 incubations. We conclude that despite the equivalent efficiency of recombinant CYP3A4 and CYP3A5 in vinorelbine metabolism the polymorphic expression of CYP3A5, as shown by the kinetics with HLMs, may have a minimal effect on systemic clearance of vinorelbine.


A needs and resource assessment of continuing medical education in Haiti.
Haiti has a chronic physician shortage, and the country has been facing an increased disease burden since the 2010 earthquake and the subsequent introduction of cholera. In such resource-challenged settings, access to postgraduate medical education often is limited due to inadequate financial, structural, and academic resources. A crucial component to improved health in Haiti is the expansion of continuing medical education (CME). To our knowledge there have been no previous studies investigating the continuing professional development needs of Haitian physicians working in this context. The objectives of this study are to describe the educational resources available to Haitian physicians and to understand their continuing professional development needs. We performed a needs and resource assessment of CME available to Haitian physicians using surveys and focus groups. We surveyed 62 physicians and led 3 focus groups. Questions gathered data on physicians' access to educational resources. Descriptive statistics were calculated from surveys, and focus group transcripts were manually reviewed for themes. In all, 82 conference attendees were invited to participate. Of these, 62 physicians completed the needs and resource assessment survey. Of the participants, 16% had a medical library at work and 31% had access to a computer at work. Educational conferences were available at work for 27% of participants, and 50% attended conferences outside of work. Less than half (45%) identified a clinical mentor. Focus group participants described inadequate tangible and reference resources, lack of colleague support, and lack of avenues for specialty training and employment. In this needs assessment, Haitian physicians identified lack of support for clinical decision making, poor access to CME activities, limited professional development, and absence of employment opportunities as key areas of need in support of their clinical and professional work.


Molarization of the lower second premolars.
This paper presents a case of extreme tooth variation. The patient was first observed during the mixed dentition period, when she presented a mild Class II malocclusion with increased overjet and acceptable overbite. In a panoramic radiograph, the presence of lower second premolars of disproportionate dimensions was discovered. When these oversized premolars erupted, the Class I malocclusion tended toward Class III, with an edge-to-edge bite. This created an unstable occlusion and the possible need for extractions.


Cardiac contractile dysfunction in insulin-resistant rats fed a high-fat diet is associated with elevated CD36-mediated fatty acid uptake and esterification.
Changes in cardiac substrate utilisation leading to altered energy metabolism may underlie the development of diabetic cardiomyopathy. We studied cardiomyocyte substrate uptake and utilisation and the role of the fatty acid translocase CD36 in relation to in vivo cardiac function in rats fed a high-fat diet (HFD). Rats were exposed to an HFD or a low-fat diet (LFD). In vivo cardiac function was monitored by echocardiography. Substrate uptake and utilisation were determined in isolated cardiomyocytes. Feeding an HFD for 8 weeks induced left ventricular dilation in the systolic phase and decreased fractional shortening and the ejection fraction. Insulin-stimulated glucose uptake and proline-rich Akt substrate 40 phosphorylation were 41% (p < 0.001) and 45% (p < 0.05) lower, respectively, in cardiomyocytes from rats on the HFD. However, long-chain fatty acid (LCFA) uptake was 1.4-fold increased (p < 0.001) and LCFA esterification into triacylglycerols and phospholipids was increased 1.4- and 1.5-fold, respectively (both p < 0.05), in cardiomyocytes from HFD compared with LFD hearts. In the presence of the CD36 inhibitor sulfo-N-succinimidyloleate, LCFA uptake and esterification were similar in LFD and HFD cardiomyocytes. In HFD hearts CD36 was relocated to the sarcolemma, and basal phosphorylation of a mediator of CD36-trafficking, i.e. protein kinase B (PKB/Akt), was increased. Feeding rats an HFD induced cardiac contractile dysfunction, which was accompanied by the relocation of CD36 to the sarcolemma, and elevated basal levels of phosphorylated PKB/Akt. The permanent presence of CD36 at the sarcolemma resulted in enhanced rates of LCFA uptake and myocardial triacylglycerol accumulation, and may contribute to the development of insulin resistance and diabetic cardiomyopathy.


[Licensure in nursing: a near future prospect].
The Spanish Association of Education in Nursing convoked an important meeting in Seville, this past March, to clarify ideas and generate constructive debate regarding the future of a licentiate degree in Nursing in our country. Maria Durán, Rosemaría Alberdi, José A. Martos and Jesús Sanz Villorejo attended this meeting. Over the course of the ensuing debate, the whole evolutionary process up to the present time was explained and the conferees expressed the need to form a consensus among all the institutions involved in the Spanish nursing profession.


Phenotyping of ABCA4 Retinopathy by Machine Learning Analysis of Full-Field Electroretinography.
Biallelic pathogenic variants in ABCA4 are the commonest cause of monogenic retinal disease. The full-field electroretinogram (ERG) quantifies severity of retinal dysfunction. We explored application of machine learning in ERG interpretation and in genotype-phenotype correlations. International standard ERGs in 597 cases of ABCA4 retinopathy were classified into three functional phenotypes by human experts: macular dysfunction alone (group 1), or with additional generalized cone dysfunction (group 2), or both cone and rod dysfunction (group 3). Algorithms were developed for automatic selection and measurement of ERG components and for classification of ERG phenotype. Elastic-net regression was used to quantify severity of specific ABCA4 variants based on effect on retinal function. Of the cohort, 57.6%, 7.4%, and 35.0% fell into groups 1, 2, and 3 respectively. Compared with human experts, automated classification showed overall accuracy of 91.8% (SE, 0.169), and 96.7%, 39.3%, and 93.8% for groups 1, 2, and 3. When groups 2 and 3 were combined, the average holdout group accuracy was 93.6% (SE, 0.142). A regression model yielded phenotypic severity scores for the 47 commonest ABCA4 variants. This study quantifies prevalence of phenotypic groups based on retinal function in a uniquely large single-center cohort of patients with electrophysiologically characterized ABCA4 retinopathy and shows applicability of machine learning. Novel regression-based analyses of ABCA4 variant severity could identify individuals predisposed to severe disease. Machine learning can yield meaningful classifications of ERG data, and data-driven scoring of genetic variants can identify patients likely to benefit most from future therapies.


Glassy behaviour of mechanically amorphised ZIF-62 isomorphs.
Zeolitic imidazolate frameworks (ZIFs) can be melt-quenched to form glasses. Here, we present an alternative route to glassy ZIFs via mechanically induced amorphisation. This approach allows various glassy ZIFs to be produced in under 30 minutes at room temperature, without the need for melt-quenching.


Efficient materiel handling and distribution: a design perspective.
This article offers a condensed examination of facility design and the interface between owners, hospital managers, and architects in planning for hospital materiel handling and storage needs. A brief history is given of the evolution of materiel management and progress today in efficient ordering, storing, distributing, and inventory and billing. The article closes with a discussion of work redesign and personnel vs. equipment concerns. Two case studies exemplify similar goals from different approaches.


Haemopoietic progenitor cells measured by limiting dilution in microwell culture.
Microwell culture assays, termed the "negative" well technique and the "positive" well technique have been used to measure the concentration of murine haemopoietic progenitor cells (HPC) by limiting dilution. Using morphologic examination and 3H-TdR uptake, the number of HPC of Balb/c mice per 10(5) bone marrow cells was 33 and 31 respectively. the linear relation between the number of harvested cells and input cells seen on the 4th and 7th days of culture reflected the growth of HPC in microwells. Besides measuring the concentration of HPC, this culture system may facilitate the analysis of effects on proliferation and differentiation of HPC of culture conditions, haemopoietic factors as well as cytotoxic agents.


Patient experience with non-invasive vagus nerve stimulator: gammaCore patient registry.
gammaCore is cleared by the FDA for acute and preventive treatment of cluster headache and the acute treatment of migraine in adults. Previously, only 2 treatments were approved for acute treatment of cluster headache while none were approved for preventive treatment. Following the initial FDA clearance, based on the ACT-1 and ACT-2 studies, a gammaCore Patient Registry (GPR) was designed to provide insights on the use of gammaCore and prescription patterns in the real-world setting and to characterize respective benefits and challenges during the acute treatment of episodic cluster headache. GPR was a prospective observational registry in which patients with episodic cluster headache (3rd edition of the International Classification of Headache Disorders criteria) who were prescribed gammaCore were invited to voluntarily enroll and provide information on their experiences between July 2017 and June 2018. Participants provided baseline information and were trained to self-administer treatment with gammaCore for cluster pain. Participants were also requested to record information for each cluster attack. Of the 182 patients who provided baseline demographic and cluster headache characteristics, 152 provided health index baseline data using EuroQol Health Index tool, 5-level format (EQ5D-5L) and 17 patients provided attack data on a total of 192 cluster headache attacks. The mean age was 49 years; 65% were male and 82% were white; the mean number of months of known diagnosis of cluster headache was 57; the mean number of attacks per cluster headache 4-week period was 14; and the mean pain score was 3.7 (0-4 scale) with a mean attack duration of 74 minutes. Sixty-seven percent of patients had used preventive treatments and 83% had used abortive treatments for cluster headaches; 25% of participants reported at least 1 comorbidity. The mean EQ5D-5L score (scale 0-1) was 0.83. Of the 192 cluster headache attacks reported, gammaCore was used in 116 (60%) attacks. Within this group, the mean pain score at the start of the attacks was 2.7, the mean number of stimulations used was 3.6, and the pain score after 30 minutes was 1.3. At 30 minutes, the pain of 81 (70%) attacks was reduced to none (27%) or mild (43%) (a pain score of 0 or 1) and in 94 (81%) attacks, patients experienced a reduction of at least 1 point in the pain score. This real-world observational evidence suggests that gammaCore adds clinically meaningful value to patients with episodic cluster headache by providing rapid pain relief and confirms that there is significant interest among prescribers in providing this new treatment and technology. This evidence further supports the need to redefine gammaCore as no longer investigational or experimental during considerations for reimbursement.


Emergency physicians' knowledge of cannabinoid designer drugs.
The use of synthetic drugs of abuse in the United States has grown in the last few years, with little information available on how much physicians know about these drugs and how they are treating patients using them. The objective of this study was to assess emergency physician (EP) knowledge of synthetic cannabinoids (SC). A self-administered internet-based survey of resident and attending EPs at a large urban emergency department (ED) was administered to assess familiarity with the terms Spice or K2 and basic knowledge of SC, and to describe some practice patterns when managing SC intoxication in the ED. Of the 83 physicians invited to participate, 73 (88%) completed surveys. The terms "Spice" and "K2" for SC were known to 25/73 (34%) and 36/73 (49%) of respondents. Knowledge of SC came most commonly (72%) from non-medical sources, with lay publications and the internet providing most respondents with information. Among those with previous knowledge of synthetic cannabinoids, 25% were not aware that SC are synthetic drugs, and 17% did not know they are chemically most similar to marijuana. Among all participants, 80% felt unprepared caring for a patient in the ED who had used synthetic cannabinoids. Clinically active EPs are unfamiliar with synthetic cannabinoids. Even those who stated they had heard of synthetic cannabinoids answered poorly on basic knowledge questions. More education is needed among EPs of all ages and levels of training on synthetic cannabinoids.


Decreased survival of human breast cancer cells expressing HER2/neu on in vitro incubation with an anti-HER2/neu antibody fused to C5a or C5a desArg.
Treatment of human epidermal growth factor receptor 2 (HER2/neu)-expressing breast cancer patients with a monoclonal antibody (mAb) directed against HER2/neu improves the outcome of chemotherapy. In cases in which remission is observed, antibody-dependent cell-mediated cytotoxicity (ADCC) seems to be one of the main mechanisms of anti-HER2/neu mAb action, implicating Fc gamma receptors (Fc gamma Rs) in this tumoricidal activity. In vitro and in vivo studies have revealed that anti-HER2/neu-mediated ADCC is mainly accomplished by polymorphonuclear granulocytes (PMN). C5a, a cleavage product of the complement component C5, modulates Fc gamma R expression via upregulation of activating and downregulation of inhibitory Fc gamma Rs. C5a also recruits PMNs to sites of inflammation and increases PMN survival. To enhance the recruitment and activation of C5a receptor-bearing cells into the tumor microenvironment, we developed antibody fusion proteins composed of a human IgG3 anti-HER2/neu antibody genetically fused to C5a [anti-HER2/neu IgG3-(C5a)] or to its derivative, C5a(desArg) [anti-HER2/neu IgG3-(C5a(desArg))]. Both fusion proteins were expressed, properly assembled, and secreted by murine myeloma cells, and displayed chemotactic activity on human PMN. Under comparable conditions, anti-HER2/neu IgG3-(C5a(desArg)) increased the survival of PMN more efficiently than anti-HER2/neu IgG3-(C5a) or C5a(desArg). Surprisingly, incubation of the fusion proteins with breast cancer cells that overexpress HER2/neu (SK-BR-3) induced cell death at a dose at which the anti-HER2/neu IgG3 antibody was innocuous. In the presence of human peripheral blood leukocytes as effector cells, both fusion proteins induced tumor cell death more efficiently than anti-HER2/neu IgG3. These data suggest that anti-HER2/neu IgG3-(C5a) and anti-HER2/neu IgG3-(C5a(desArg)) fusion proteins possess novel properties that could be useful in cancer immunotherapy.


Toward Accurate Post-Born-Oppenheimer Molecular Simulations on Quantum Computers: An Adaptive Variational Eigensolver with Nuclear-Electronic Frozen Natural Orbitals.
Nuclear quantum effects such as zero-point energy and hydrogen tunneling play a central role in many biological and chemical processes. The nuclear-electronic orbital (NEO) approach captures these effects by treating selected nuclei quantum mechanically on the same footing as electrons. On classical computers, the resources required for an exact solution of NEO-based models grow exponentially with system size. By contrast, quantum computers offer a means of solving this problem with polynomial scaling. However, due to the limitations of current quantum devices, NEO simulations are confined to the smallest systems described by minimal basis sets, whereas realistic simulations beyond the Born-Oppenheimer approximation require more sophisticated basis sets. For this purpose, we herein extend a hardware-efficient ADAPT-VQE method to the NEO framework in the frozen natural orbital (FNO) basis. We demonstrate on H2 and D2 molecules that the NEO-FNO-ADAPT-VQE method reduces the CNOT count by several orders of magnitude relative to the NEO unitary coupled cluster method with singles and doubles while maintaining the desired accuracy. This extreme reduction in the CNOT gate count is sufficient to permit practical computations employing the NEO method─an important step toward accurate simulations involving nonclassical nuclei and non-Born-Oppenheimer effects on near-term quantum devices. We further show that the method can capture isotope effects, and we demonstrate that inclusion of correlation energy systematically improves the prediction of difference in the zero-point energy (ΔZPE) between isotopes.


Understanding indocyanine green angiography in polypoidal choroidal vasculopathy: the group experience with digital fundus photography and confocal scanning laser ophthalmoscopy.
To evaluate the angiographic features in using fundus camera-based versus confocal scanning laser ophthalmoscope (cSLO)-based indocyanine green angiography in differentiating polypoidal choroidal vasculopathy (PCV) from typical age-related macular degeneration. Sixty-five eyes of 44 patients with exudative maculopathy due to PCV or typical age-related macular degeneration were prospectively imaged with indocyanine green angiography using fundus camera and cSLO. Images were graded independently by retinal specialists. The main outcome measure was agreement between cSLO and fundus camera for the diagnosis of PCV. The rate of detection and area under the receiver operating characteristic curve of 7 preselected individual features were also compared. The diagnosis of PCV was made with the cSLO system in 36 eyes (55.4%) and typical age-related macular degeneration in 29 eyes (44.6%), whereas the fundus camera diagnosed PCV in 39 eyes (60.0%) and typical age-related macular degeneration in 26 eyes (40.0%). There was moderate agreement between the two indocyanine green angiography systems (Kappa = 0.53). Using cSLO as the gold standard, fundus camera has a sensitivity and specificity of 83.3% and 69.0%, respectively. Typical nodular appearance was the most commonly detected feature (median, 88.9% for cSLO, 80.6% for fundus camera, P = 0.63) and had the highest area under the curve for the diagnosis of PCV in both systems (median, 80.2% for cSLO, 73.2% for fundus camera, P = 0.13). Confocal scanning laser ophthalmoscope was more sensitive in detecting branching vascular network and late hyperfluorescent plaque. Both systems detected >80% of PCV based on typical nodular appearance of polyps. However, the cSLO is superior in detecting additional features, particularly branching vascular network.


Clustered Regularly Interspaced Short Palindromic Repeats Are emm Type-Specific in Highly Prevalent Group A Streptococci.
Clustered regularly interspaced short palindromic repeats (CRISPR) are the bacterial adaptive immune system against foreign nucleic acids. Given the variable nature of CRISPR, it could be a good marker for molecular epidemiology. Group A streptococcus is one of the major human pathogens. It has two CRISPR loci, including CRISPR01 and CRISPR02. The aim of this study was to analyze the distribution of CRISPR-associated gene cassettes (cas) and CRISPR arrays in highly prevalent emm types. The cas cassette and CRISPR array in two CRISPR loci were analyzed in a total of 332 strains, including emm1, emm3, emm4, emm12, and emm28 strains. The CRISPR type was defined by the spacer content of each CRISPR array. All strains had at least one cas cassette or CRISPR array. More than 90% of the spacers were found in one emm type, specifically. Comparing the consistency between emm and CRISPR types by Simpson's index of diversity and the adjusted Wallace coefficient, CRISPR01 type was concordant to emm type, and CRISPR02 showed unidirectional congruence to emm type, suggesting that at least for the majority of isolates causing infection in high income countries, the emm type can be inferred from CRISPR analysis, which can further discriminate isolates sharing the same emm type.


A pilot study on the corneal curvatures and ocular dimensions of horses less than one year of age.
Intraocular lenses (IOLs) have been implanted in adult equine eyes after cataract surgery. Foals and weanlings comprise a large proportion of those horses undergoing cataract surgery. Due to potential differences in the size and corneal curvature of the juvenile eye, it is not currently known whether implantation of adult IOLs is appropriate in foals and weanlings. The objective of the study was to measure the anterior chamber depth (ACD), central lens thickness (CLT), vitreous chamber depth (VCD), axial globe length (AGL) and corneal curvature of horses less than one year of age. The axial dimensions from one eye of 10 foals were measured using simultaneous A and B scan ultrasonography. The corneal curvature from one eye of 7 weanlings was determined using a modified photokeratometer. Ultrasonography revealed a mean ACD of 4.94 mm, mean CLT of 9.38 mm, mean VCD of 18.96 mm and mean AGL of 33.32 mm for the foals. The mean corneal curvature was 15.4 diopters (D). The mean ACD, CLT, VCD and AGL of the foals were less than the measurements reported in the literature for adult horses. The mean corneal curvature was similar to the values reported by some authors for adult horses. Due to the differences in axial dimensions between adult and juvenile eyes, an IOL that corrects vision in an adult horse might not adequately correct vision in a horse less than one year of age.


[Analysis of designer drugs detected in the products purchased in fiscal year 2006].
Many psychotropic substances are easily available in Japan via the Internet, thus the spread of drug abuse is becoming more serious problem. To avoid drug abuse, 32 substances have been controlled in Japan since April in 2007 by the Pharmaceutical Affairs Law as designated substances (Shitei-Yakubutsu, classified as 11 tryptamines, 11 phenethylamines, 2 piperazines, 6 alkyl nitrites, 1 diterpene and 1 plant). Although the distributions of these drugs have been decreased through this regulation, new designer drugs are still being found. In this study, we detected 7 designer drugs in 15 products, which purchased just before the amendment of the law, by NMR, GC-MS and LC-MS analyses. Three methylone derivertives (1-(3,4-methylenedioxyphenyl-2-(pyrrolidin-1-yl)-1-pentanone: MDPV, 2-methylamino-1-(3,4-methylenedioxyphenyl)butan-1-one: bk-MBDB, 2-ethylamino-1-(3,4-methylenedioxyphenyl)propan-1-one): bk-MDEA, a MDMA derivative (N-hydroxy-1-(3,4-methylenedioxyphenyl)-2-aminopropane: N-OH MDMA), a methamphetamine derivative (N-methyl-1-(4-fluorophenyl)propan-2-amine: N-Me-4-FMP), a tryptamine derivative (5-methoxy-N-ethyl-N-isopropyltryptamine: 5-MeO-EIPT) and indan-2-amine were detected. 5-MeO-EIPT was newly identified in this study.


Analysis of the crown fractures and crown-root fractures due to dental trauma assisted by the Integrated Clinic from 1992 to 2002.
The purpose of this study was to analyze crown fractures and crown-root fractures due to dentoalveolar trauma, treated in the Integrated Clinic comprehensive dental care at Aracatuba School of Dentistry (UNESP), from January 1992 to July 2002. The data were obtained from files of trauma cases. On the analysis period, 293 patients had crown fractures or crown-root fractures, in 605 teeth. Sixty-nine percent were males and 31% were females. Adolescents between 11 and 18 years old were the most prevalent group (41.6%) and the maxillary arch was the most commonly traumatized (83%). The most commonly affected tooth was the maxillary central incisor (58.3%). The most frequent causes were falls from bicycles (30.8%). It was concluded that the reality of the local service is similar to the published data.


Rheumatoid arthritis: relationship with HLA-D.
Understanding of the genetic component of rheumatoid arthritis has been greatly enhanced by the discovery of the association of risk for development of this disease with certain histocompatibility antigens. The HLA-D region, at the centromeric end of the HLA gene complex, on the short arm of chromosome number 6, is concerned with important functions in the regulation of the immune response. The products of the HLA-D region (DR, SB, MT, and the like) function as "restriction elements" for antigen presentation, mediators of immune response and immune suppressor genes, stimulators of graft rejection, and of other allogenic effects. Rheumatoid arthritis is associated with Dw4 and DR4 in Caucasians. The association with DR4 has been observed also in Blacks and Orientals. HLA-DR4 is associated with more severe, rheumatoid factor-positive disease. Clinical and genetic heterogeneity has been revealed by studies of patients with juvenile arthritis. Conditions often associated with rheumatoid arthritis and also thought to have an immunologic basis, such as Sjögren's syndrome, Felty's syndrome, and rheumatoid vasculitis, and reactions to medications, such as gold and penicillamine, reveal an interesting spectrum of immunogenetic relationships. Thus, HLA studies have shed light on the classification of patients with rheumatoid diseases. In view of the role of HLA-D in the immune response, these studies are expected to further our understanding of the mechanism underlying predisposition for development of rheumatoid arthritis and related conditions.


Genetic manipulation of Neospora caninum to express the bradyzoite-specific protein NcSAG4 in tachyzoites.
Neospora caninum is an apicomplexan parasite and the aetiological agent of bovine neosporosis, one of the main causes of reproductive failure worldwide. We have generated 2 independent transgenic knock-in clones, Nc-1SAG4c1.1 and Nc-1SAG4c2.1, that express the bradyzoite stage-specific protein NcSAG4 in the tachyzoite stage. These clones have similar growth rates in vitro as the wild-type (WT) strain Nc-1. Studies in a cerebral mouse model of infection revealed a slightly lower rate of detection of the transgenic strains in brains during the chronic phase of infection. However, a pregnant mouse model of infection revealed a reduction in the virulence of the Nc-1SAG4c1.1 strain despite the same tachyzoite expression of NcSAG4 and a similar anti-NcSAG4 response displayed by mice inoculated with Nc-1 SAG4c1.1 or Nc-1 SAG4c2.1 parasites. This behaviour may be related to the reduced ability of the Nc-1SAG4c1.1 parasites to invade host cells, which was observed in in vitro assays. The apparent reduction in persistence and the high growth rate of the transgenic strains, together with their constitutive expression of the protein NcSAG4, may be useful features for future immunoprophylaxis trials based on a safe live attenuated vaccine.


Dependability and interrater reliability of two types of rating scales.
The reliability of a behaviorally anchored rating scale and the reliability of a bare trait rating scale measuring the same categories of expected behavior of medical laboratory students were studied. Moderate to high interrater reliability was shown for both scales, with little difference between the two. Internal consistency measures and dependability coefficients were high. The implications of these results for student behavior rating scales are discussed.


Quick method (FT-NIR) for the determination of oil and major fatty acids content in whole achenes of milk thistle (Silybum marianum (L.) Gaertn.).
Calibration models for the Fourier transform-near infrared (FT-NIR) instrument were developed for quick and non-destructive determination of oil and fatty acids in whole achenes of milk thistle. Samples with a range of oil and fatty acid levels were collected and their transmittance spectra were obtained by the FT-NIR instrument. Based on these spectra and data gained by the means of the reference method - Soxhlet extraction and gas chromatography (GC) - calibration models were created by means of partial least square (PLS) regression analysis. Precision and accuracy of the calibration models was verified via the cross-validation of validation samples whose spectra were not part of the calibration model and also according to the root mean square error of prediction (RMSEP), root mean square error of calibration (RMSEC), root mean square error of cross-validation (RMSECV) and the validation coefficient of determination (R(2) ). R(2) for whole seeds were 0.96, 0.96, 0.83 and 0.67 and the RMSEP values were 0.76, 1.68, 1.24, 0.54 for oil, linoleic (C18:2), oleic (C18:1) and palmitic (C16:0) acids, respectively. The calibration models are appropriate for the non-destructive determination of oil and fatty acids levels in whole seeds of milk thistle.


Structure-based drug design and AutoDock study of potential protein tyrosine kinase inhibitors.
Different classes of compounds were investigated for their binding affinities into different protein tyrosine kinases (PTKs) employing a novel flexible ligand docking approach by using AutoDock 3.05 and 4. These compounds include many flavin analogs, which were developed in our group with varying degrees of cytotoxic activity (comparable or moderately superior to cisplatin and ara-c), and database selected analogs. They were docked onto twelve different families of PTKs retrieved from the Protein Data Bank. These proteins are representatives of plausible models of interactions with chemotherapeutic agents. A comparative study of the intact co-crystallized ligands of various types of PTKs was carried out. Results revealed that the new class of 5-deazapteridine and steroid hybrid compounds VIa,b, and d, and the vertical-type bispyridodipyrimidine with n-hexyl chain junction between its N-10 and N-10 atoms Xa, exhibited non-selective PTK binding capacities, with the lowest (Gb). On the other hand, 2-amino benzoic acid analog IIa, phenoxypyrido [3, 4-d]pyrimidine derivative IVc, tyrosine containing tripeptide Vd, and the one from Sumisho data base 831 are proposed to have selective PTK binding affinities to certain classes of tyrosine kinases, namely, HGFR (c-met), ZAP-70, insulin receptor kinase, EGFR, respectively. All These compounds of highest affinities were docked within the binding sites of PTKs with reasonable RMSD and 1-5 hydrogen bonds.


Airway response of asthmatic subjects to inhaled allergen after exposure to pollutants.
Recent studies have suggested that air pollutants resulting from vehicle exhaust emissions and burning of fossil fuels, either in combination or individually, may enhance the airway response of asthmatic subjects to inhaled allergen. It was hypothesised that the airway response to inhaled allergen after exposure to a combination of 400 ppb nitrogen dioxide (NO2) and 200 ppb sulphur dioxide (SO2) is increased 24-48 hours after exposure. Thirteen mild atopic asthmatic volunteers were exposed for six hours to a single exposure of air and three exposures of the combination of 400 ppb NO2 + 200 ppb SO2 in randomised order, and then challenged with increasing concentrations of Dermatophagoides pteronyssinus allergen either immediately after exposure to air, or immediately, 24 hours or 48 hours after exposure to the combination of the two pollutants, until a 20% fall in forced expiratory volume in one second (FEV1) was recorded. Exposure to 400 ppb NO2 + 200 ppb SO2 significantly decreased the dose of D pteronyssinus allergen required to produce a 20% fall in FEV1 (PD20FEV1) at all times after exposure when compared with air. The mean percentage changes in allergen PD20FEV1 immediately, 24 hours, and 48 hours after exposure to 400 ppb NO2 + 200 ppb SO2 were -37% (95% confidence intervals (CI) -50 to -23), -63% (CI -75 to -51), and -49% (CI -75 to -28.8), respectively, when compared with the PD20FEV1 after air exposure and were significant at all time points studied. The allergen PD20FEV1 at 24 hours after exposure to the combination of the two pollutants was also found to be significantly lower when compared with that immediately after exposure to the two pollutants. These results demonstrate that exposure to a combination of NO2 and SO2, at concentrations which can be encountered during episodes of increased outdoor and indoor air pollution, enhances the airway response to inhaled allergen in asthmatic subjects. This effect persists over a period of 24-48 hours and is maximal 24 hours after exposure to these air pollutants.


Effects of lipid peroxidation-related protein modifications on RPE lysosomal functions and POS phagocytosis.
Lipofuscin accumulation in the RPE is a common downstream pathogenic pathway in various monogenic and complex retinal diseases including age-related macular degeneration (AMD). Lipid peroxidation-induced modification of proteins is thought to play a role in lipofuscinogenesis and may contribute to RPE dysfunction. A prior study demonstrated that a variety of lipofuscin-associated proteins are damaged by aberrant covalent modifications of malondialdehyde (MDA) and 4-hydroxynonenal (HNE). The present study was conducted to test the hypothesis that these damaged proteins are more resistant to proteolytic attack and act as protease inhibitors. Isolated photoreceptor outer segments (POS) were radioactively labeled and in vitro modified with MDA and HNE. Pure lysosomal fractions isolated from human RPE were tested for their proteolytic activities toward modified and unmodified POS proteins. In parallel, modified and radiolabeled POS were fed to RPE cell cultures for phagocytosis and their lysosomal degradation as well as intracellular accumulation was compared with unmodified POS. Both experimental approaches revealed that MDA or HNE modifications strikingly increase the resistance of POS proteins to the attack by lysosomal proteases. When cultured RPE cells were fed with modified or unmodified POS the amount of degraded POS proteins was reduced by approximately 60% to 70% for the modified POS compared with those in normal control subjects. Some of the modified proteins remained undegraded in the lysosomal compartment of cultured RPE cells and were still detectable 3 weeks after feeding, whereas unmodified POS were completely degraded within 1 week after feeding. Moreover, modified proteins had the potential to impair degradation of unmodified proteins, indicating their efficacy as proteolytic antagonists. The results indicate that lipid peroxidation-derived protein modifications are involved in lipofuscinogenesis and may contribute to cell damaging effects of lipofuscin in retinal diseases such as AMD.


A patient with oeosinophilic gastroenteritis and severe malnutrition improved with home parenteral nutrition.
Oeosinophilic gastroenteritis is a chronic and rare disorder characterised by massive oeosinophilic tissue infiltration involving one or more segments of the digestive tract. The management of patients with oeosinophilic gastroenteritis is complex and the therapeutic response often poor. Here we discuss the clinical case and management of a 23-year-old man with oeosinophilic gastroenteritis since the first year of life and the decision to prescribe lifelong total parenteral nutrition.


Acute effects of aerobic exercise and Hatha yoga on craving to smoke.
Recent studies have examined the effects of physical activity on craving to smoke and smoking withdrawal. The current study was designed to compare and contrast the effects of 2 different forms of physical activity on general and cue-elicited craving to smoke. Following 1-hr nicotine abstinence, 76 daily smokers were randomly assigned to engage in a 30-min bout of cardiovascular exercise (CE; brisk walk on a treadmill), Hatha yoga (HY), or a nonactivity control condition. Participants completed measures of craving and mood, and a smoking cue reactivity assessment, before, immediately following, and approximately 20 min after the physical activity or control conditions. Compared with the control condition, participants in each of the physical activity groups reported a decrease in craving to smoke, an increase in positive affect, and a decrease in negative affect. In addition, craving in response to smoking cues was specifically reduced among those who engaged in CE, whereas those who engaged in HY reported a general decrease in cravings. This study provides further support for the use of exercise bouts for attenuating cigarette cravings during temporary nicotine abstinence. Results also suggest that CE can attenuate cravings in response to smoking cues. There are several areas for further research that may improve integration of exercise within smoking cessation treatment.