Age-related interference with Chlamydia pneumoniae microimmunofluorescence serology due to circulating rheumatoid factor.
Microimmunofluorescence (MIF) serology is commonly used in the diagnosis of chlamydial infections. In the MIF assay, Chlamydia pneumoniae elementary bodies were used to detect C. pneumoniae immunoglobulin G (IgG) and IgM antibodies in paired serum samples from 286 patients with respiratory illnesses. In 69 patients, MIF serology was compared with C. pneumoniae cultures. All C. pneumoniae cultures remained negative. However, 205 (71%) of 286 patients were C. pneumoniae antibody positive and 64 (22%) had MIF test results indicating recent infection; 11 showed a fourfold increase in IgG titer, 18 had IgG titers of greater than or equal to 1:512, and 41 had IgM titers of greater than or equal to 1:16. In 35 (55%) of 64 patients, a recent-infection diagnosis was based on C. pneumoniae IgM antibodies only. However, 78% of C. pneumoniae IgM-positive patients had circulating rheumatoid factor (RF) by rheumatoid arthritis latex assay. RF positivity increased with age. After absorption with anti-human IgG, all C. pneumoniae IgM-positive sera became C. pneumoniae IgM negative in the MIF assay. Twenty-five patients with active rheumatoid arthritis but without respiratory illness were also tested; 14 were C. pneumoniae IgG positive and C. pneumoniae IgM positive as well. Absorption of IgG from these RF-containing sera invariably resulted in disappearance of reactivity in the MIF IgM assay. We conclude that with age the serologic diagnosis of recent C. pneumoniae infection becomes increasingly prone to false-positive results unless sera are routinely absorbed prior to MIF IgM testing.


Epidermal Merkel cells are mechanosensory cells that tune mammalian touch receptors.
Touch submodalities, such as flutter and pressure, are mediated by somatosensory afferents whose terminal specializations extract tactile features and encode them as action potential trains with unique activity patterns. Whether non-neuronal cells tune touch receptors through active or passive mechanisms is debated. Terminal specializations are thought to function as passive mechanical filters analogous to the cochlea's basilar membrane, which deconstructs complex sounds into tones that are transduced by mechanosensory hair cells. The model that cutaneous specializations are merely passive has been recently challenged because epidermal cells express sensory ion channels and neurotransmitters; however, direct evidence that epidermal cells excite tactile afferents is lacking. Epidermal Merkel cells display features of sensory receptor cells and make 'synapse-like' contacts with slowly adapting type I (SAI) afferents. These complexes, which encode spatial features such as edges and texture, localize to skin regions with high tactile acuity, including whisker follicles, fingertips and touch domes. Here we show that Merkel cells actively participate in touch reception in mice. Merkel cells display fast, touch-evoked mechanotransduction currents. Optogenetic approaches in intact skin show that Merkel cells are both necessary and sufficient for sustained action-potential firing in tactile afferents. Recordings from touch-dome afferents lacking Merkel cells demonstrate that Merkel cells confer high-frequency responses to dynamic stimuli and enable sustained firing. These data are the first, to our knowledge, to directly demonstrate a functional, excitatory connection between epidermal cells and sensory neurons. Together, these findings indicate that Merkel cells actively tune mechanosensory responses to facilitate high spatio-temporal acuity. Moreover, our results indicate a division of labour in the Merkel cell-neurite complex: Merkel cells signal static stimuli, such as pressure, whereas sensory afferents transduce dynamic stimuli, such as moving gratings. Thus, the Merkel cell-neurite complex is an unique sensory structure composed of two different receptor cell types specialized for distinct elements of discriminative touch.


Alcohol consumption and utilization of health services in a health maintenance organization.
This study examined the relation between alcohol use and utilization of health services during a 3-year period in a sample of 4,264 adult respondents to a member health survey in a health maintenance organization. Respondents were categorized as abstainers (no drinks in the past year, n = 1,139), lighter drinkers (less than seven drinks/week, n = 2,330), moderate drinkers (seven to 13 drinks/week, n = 498), and heavier drinkers (> or =14 drinks/week, n = 297). Each drinker group was compared with abstainers on outpatient visits, hospital days, and number of hospitalizations controlling for age, race, and health plan membership. The mean number of outpatient visits was inversely related to the amount of alcohol consumed. Significant differences also were found for mean number of hospitalizations and mean days hospitalized per year. Compared with the three drinker groups, abstainers were significantly higher on both inpatient measures. These results might be explained by the inclusion in the abstainer group of exdrinkers who quit because of illness, inattention to health problems by heavier drinkers, or lower rates of illness among drinkers. The findings underscore the importance of replicating our study in other cohorts in which problem drinkers can be identified and compared with non-problem drinkers and in which lifelong abstainers can be separated from exdrinkers in the analysis.


Acute respiratory dysfunction after surgery for acute type A aortic dissection.
Acute respiratory dysfunction (ARD) can occur after acute type A aortic dissection, but relatively little is known about ARD in such patients. This study aims to analyse the clinical impact of ARD after surgery for acute type A aortic dissection and to assess possible treatment options. We reviewed our institutional database to identify patients who underwent surgery for acute type A dissection between October 1994 and January 2008 (n=276). Postoperative ARD was defined as oxygenation impairment (PaO(2)/FiO(2) <150) that occurred within 72 h of surgery and was not related to other documented causes of acute respiratory failure. A total of 37 patients (13%) (27 male, mean age 60.7+/-11 years) experienced ARD after surgery for acute type A dissection. Intensive care unit stay was significantly longer for patients with ARD than those without (18+/-11 days vs 7.5+/-6 days, respectively, p<0.0001). However, hospital mortality was not significantly different between groups (16% for ARD patients vs 19% for patients without ARD, p=0.6). Logistic regression analysis identified preoperative multiple malperfusion as the only risk factor for ARD (OR 3.2, 95% confidence interval (C.I.): 2.2-4.9). Peak C-reactive protein levels were significantly higher in ARD patients (17.7+/-6.7 vs 9.6+/-5.4 mg dl(-1), p=0.04). Prone positioning ventilation was performed in 15 patients (40%) with severely impaired oxygenation and resulted in an immediate increase in mean oxygenation index from 71.6+/-8.8 to 138+/-92.6 (p<0.001). There was a tendency towards a shorter total time of mechanical ventilation (355+/-188 h vs 433+/-318 h, p=0.2) and shorter ICU stay (405+/-198 h vs 505+/-265 h, p=0.2) in the prone positioning subgroup. ARD is a relatively common complication of surgery for acute type A dissection and is associated with increased morbidity and resource utilisation. Patients with preoperative malperfusion are at increased risk for development of ARD. Prone positioning is a viable treatment option that significantly improves pulmonary oxygenation.


Hydrolysis of synthetic pyrophosphoric esters by an isoenzyme of apyrase from Solanum tuberosum.
A highly purified isoenzyme of apyrase obtained from potatoes (Solanum tuberosum var. Pimpernel) exhibits a low specificity for the organic moiety of synthetic pyro- and triphosphates. Methyl di- and tri-phosphates were hydrolysed at higher rates than ADP and ATP, but their Km values were also higher. Steric hindrance at the carbon atom linked to the pyrophosphate chain decreases both binding and maximum rate, whereas length or polarity of the organic chain do not have systematic effects. t-Butyl diphosphate, inorganic pyrophosphate, adenosine 5'-[alpha,beta-methylene]triphosphate and adenosine 5'-[beta,gamma-methylene]triphosphate are competitive inhibitors of the hydrolysis of ATP and ADP.


[Experiences with the combination carboplatin/VP 16 in the treatment of recurrent tumors in children].
The efficiency and toxicity of Carboplatin, a derivate of cisplatin in clinical use since 1981, was investigated in combination with VP16 in 11 children with recurrent or refractory tumors. 10 patients received therapy as outpatients without hydration. 160 mg/m2/day of Carboplatin were administered days 1 to 3, or days 1 to 5 as one hour infusion, combined with VP16 100 mg/m2/day, days 1 to 3, or days 1 to 5 as one hour infusion. One patient received Carboplatin, 600 mg/m2, as 24 hrs. continuous infusion in combination with VM26, 150 mg/m2/day as one hour infusion, 8 hours after Carboplatin. 9 of the treated children had measurable disease. 3 of 9 patients achieved a tumor response of more than 50% (partial remission). 5 of 9 children showed tumor response of less than 50% (stable disease). 1 out of 9 patients had progressive disease. 8 of 9 patients reported improvement or disappearance of clinical symptoms after the first course. All patients had severe myelosuppression with different times to recover. The WBC nadir was 14 days after Carboplatin/VP16. The nadir platelet counts occurred 21 days after therapy with fast recovery. Non-hematological side effects were minimal.


Accounting for exhaust gas transport dynamics in instantaneous emission models via smooth transition regression.
Collecting and analyzing high frequency emission measurements has become very usual during the past decade as significantly more information with respect to formation conditions can be collected than from regulated bag measurements. A challenging issue for researchers is the accurate time-alignment between tailpipe measurements and engine operating variables. An alignment procedure should take into account both the reaction time of the analyzers and the dynamics of gas transport in the exhaust and measurement systems. This paper discusses a statistical modeling framework that compensates for variable exhaust transport delay while relating tailpipe measurements with engine operating covariates. Specifically it is shown that some variants of the smooth transition regression model allow for transport delays that vary smoothly as functions of the exhaust flow rate. These functions are characterized by a pair of coefficients that can be estimated via a least-squares procedure. The proposed models can be adapted to encompass inherent nonlinearities that were implicit in previous instantaneous emissions modeling efforts. This article describes the methodology and presents an illustrative application which uses data collected from a diesel bus under real-world driving conditions.


[Genetic variability and phylogenetic analysis of 39 short tandem repeat loci in Beijing Han population].
In this study, we studied the genetic polymorphisms of short tandem repeat (STR) loci from 13 CODIS and 26 non-CODIS system in Beijing Han population for the first time, and established a database of 39 STR loci whose forensic parameters were further evaluated. Our results demonstrated no significant deviation from the Hardy-Weinberg equilibrium of 39 STR loci and no pairwise linkage disequilibrium between them. The power of discriminations, expected heterozygosity, polymorphic information content, and power of exclusion of 39 STR loci ranged from 0.7740-0.9818, 0.6000-0.9350, 0.5317-0.9047 and 0.2909-0.8673. The cumulated discrimination power and cumulative probability of exclusion were 0.999999999999999999999999999999999999999964971 and 0.999999999973878, respectively. Moreover, the genetic distance was calculated based on allele frequency and phylogenetic tree was built using STR loci data from Beijing Han and other 11 Chinese ethnic groups.This study provides important basic data for Chinese forensic DNA database and population genetics database, and has important significance in carrying out forensic individual identification, paternity testing, and population genetic study.


Accuracy and reliability of cone beam computed tomography of the jaws for comparative forensic identification: a preliminary study.
Conventional computed tomography is an emerging modality in forensic identification but is not sufficiently accurate for use in dental identification primarily because of problems with metallic dental restoration-induced streak artifact. In this study, the accuracy and reliability of recording forensic information from cone beam computed tomography (CBCT) scans of the jaws when compared to conventional panoramic radiographs has been analyzed under experimental conditions. Information could be recorded with near-perfect repeatability and reproducibility. Information could also be recorded accurately, the sensitivity being 96.6% (95% CI, 95.1-98.1) and specificity being 98.4% (95% CI, 96.2-100). The metal dental restoration-induced streak artifact was at a level that permitted, in most cases, accurate observations. This is considered an important step in validating CBCT as a tool in comparative dental identification of bodies. It may have a role in mass fatalities and in chemical, biological, radiological, and nuclear incidents, but further studies are required to assess the feasibility of this.


Inactive membrane protein kinase Cs: a possible target for receptor signalling.
The activation of the multifunctional cell signalling enzymes, the protein kinase Cs (PKCs), is generally thought to result from the translocation of inactive cytosolic enzymes to activation sites in cell membranes. However, recent studies suggest that PKCs may also be stimulated in cells by processes independent of translocation. One possible mechanism is the modulation of the activity of PKCs already resident in membranes. A PKC assay that measures enzyme activity directly in isolated native membranes has revealed the presence of an activatable pool of PKCs resident in native membranes of various cells and tissues. In 3T3-L1 cells, some or all of this pool of membrane PKCs was stimulated within 10 min of exposing the cells to 10 ng/ml epidermal growth factor or 100 ng/ml fibroblast growth factor. Similar increases in PKC activity were observed in native membranes isolated from CTLL-2, WEHI-231 and S49 lymphoma cells that had been exposed to interleukin-2. These growth factors all stimulated membrane PKC activity without detectably translocating cytosolic enzymes to the membranes. In intact WEHI cells, low concentrations (5-10 microM) of a diacylglycerol, 1-oleoyl-2-acetyl-sn-glycerol (OAG), or low concentrations (2-10 nM) of phorbol 12-myristate 13-acetate sufficed to activate PKCs already resident in membranes, but much higher concentrations (50-100 microM and 50-100 nM respectively) were needed to detectably stimulate the translocation of cytosolic PKCs. A phosphatidylcholine-specific phospholipase C also selectively stimulated membrane PKCs in WEHI cells at concentrations that were much less than those needed to induce the translocation of cytosolic enzymes. Furthermore, interleukin-2 and low concentrations of OAG both stimulated the phosphorylation of the 85 kDa PKC-selective substrate protein in intact WEHI cells in which translocation of PKCs was not evident. These results suggest that the membranes of some cells maintain a pool of activatable PKCs that respond to lower levels of extracellular stimuli than cytosolic PKCs, and that can be stimulated by signals which produce diacylglycerols through the hydrolysis of phospholipids other than polyphosphoinositides.


Experiences of aftermath of COVID-19 in relation to social, financial and health related aspects among previously hospitalized patients: a qualitative exploration.
There is increasing evidence of long-term consequences of COVID-19. The world has seen multidimensional impact of the pandemic and Bangladesh is no exception to that. Policymakers in Bangladesh laid out strategies to curb the initial spread of COVID-19. However, long-term consequences of COVID-19 received little or no attention in the country. Evidence suggests that people presumed to be recovered face multidimensional post-covid consequences. This study aimed to describe the aftermath of COVID-19 in relation to social, financial and health related aspects among previously hospitalized patients. This descriptive qualitative study includes participants (n = 14) who were previously hospitalized for COVID-19 and returned home after recovery. The participants were part of a mixed method study from which they were purposively selected. Semi-structured in-depth interviews were conducted over telephone. Inductive content analysis was used to analyze the data. Twelve sub-categories emerged from the data analysis which converged into five main categories. The main categories included perspective on physical health, financial struggle, life adjustment, interplay between different domains, and spontaneous support. The lived experiences of COVID-19 recovered patients highlighted multidimensional impact on their daily lives. Physical and psychological wellbeing found to be related to the effort of restoring financial status. People's perception about life altered due to pandemic, for few the pandemic was an opportunity to grow while others found it difficult to accept the hardship. Such multidimensional post COVID-19 impact on people's lives and wellbeing holds considerable implication for response and mitigation plan for future related pandemics.


Identifying clinically meaningful severity categories for PROMIS pediatric measures of anxiety, mobility, fatigue, and depressive symptoms in juvenile idiopathic arthritis and childhood-onset systemic lupus erythematosus.
A key limitation to widespread adoption of patient-reported outcome (PRO) measures is the lack of interpretability of scores. We aim to identify clinical severity thresholds to distinguish categories of no problems, mild, moderate, and severe along the PROMIS® Pediatric T-score metric for measures of anxiety, mobility, fatigue, and depressive symptoms for use in populations with juvenile idiopathic arthritis (JIA) and childhood-onset systemic lupus erythematosus (cSLE). We used a modified standard setting methodology from educational testing to identify clinical severity thresholds (clinical cut scores). Using item response theory-based parameters from PROMIS item banks, we developed a series of clinical vignettes that represented different severity or ability levels along the PROMIS Pediatric T-score metric. In stakeholder workshops, participants worked individually and together to reach consensus on clinical cut scores. Median cut-score placements were taken when consensus was not reached. Focus groups were recorded and qualitative analysis was conducted to identify decision-making processes. Nine adolescents (age 13-17 years) with JIA (33% female) and their caregivers, five adolescents (age 14-16 years) with cSLE (100% female) and their caregivers, and 12 pediatric rheumatologists (75% female) participated in bookmarking workshops. Placement of thresholds for bookmarks was highly similar across stakeholder groups (differences from 0 to 5 points on the PROMIS t-score metric) for all but one bookmark placement. This study resulted in clinical thresholds for severity categories for PROMIS Pediatric measures of anxiety, mobility, fatigue, and depressive symptoms, providing greater interpretability of scores in JIA and cSLE populations.


[Pathogenesis of cachexia in cardiac insufficiency].
In 81 patients with circulatory insufficiency of the IIB-III stage some in dices of the lipids and protein metabolism along with the blood sugar level were studied. Changes in the lipids metabolism values are more pronounced in patients with mitral defects than in those with atherosclerotic cardiosclerosis. The authors attach definite importance to these derangements in the pathogenesis of cardiac cachexia secondary to the developing circulatory insufficiency.


Exercise training in pulmonary arterial hypertension associated with connective tissue diseases.
The objective of this prospective study was to assess short- and long-term efficacy of exercise training (ET) as add-on to medical therapy in patients with connective tissue disease-associated pulmonary arterial hypertension (CTD-APAH). Patients with invasively confirmed CTD-APAH received ET in-hospital for 3 weeks and continued at home for 12 weeks. Efficacy parameters have been evaluated at baseline and after 15 weeks by blinded-observers. Survival rate has been evaluated in a follow-up period of 2.9 ± 1.9 years. Twenty-one consecutive patients were included and assessed at baseline, and after 3 weeks, 14 after 15 weeks. Patients significantly improved the mean distance walked in 6 minutes compared to baseline by 67 ± 52 meters after 3 weeks (p < 0.001) and by 71 ± 35 meters after 15 weeks (p = 0.003), scores of quality of life (p < 0.05), heart rate at rest, peak oxygen consumption, oxygen saturation and maximal workload. Systolic pulmonary artery pressure and diastolic systemic blood pressure improved significantly after 3 weeks of ET. The 1- and 2-year overall-survival rates were 100%, the 3-year survival 73%. In one patient lung transplantation was performed 6 months after ET. ET as add-on to medical therapy is highly effective in patients with CTD-APAH to improve work capacity, quality of life and further prognostic relevant parameters and possibly improves the 1-, 2- and 3-year survival rate. Further randomized controlled studies are needed to confirm these results. ClinicalTrials.gov: NCT00491309.


Transformation of Bartonella bacilliformis by electroporation.
Bartonella bacilliformis is a member of the order Rickettsiales, family Bartonellaceae. The bacterium is an intracellular parasite of human erythrocytes. To date, members of the family Bartonellaceae have not been transformed by standard chemical methods. We report the first successful transformation of a member of the Bartonellaceae family, B. bacilliformis, by the method of electroporation. The optimal conditions for electroporation of B. bacilliformis include a field strength of 12.5 kV/cm and a time constant of 5 ms using 0.2-cm cuvettes. With these parameters and the cosmid pEST (RK2 replicon), a transformation efficiency of 7.8 x 10(5) was obtained. Transformants were readily cultured on medium containing kanamycin sulfate at concentrations ranging from 15 to 600 micrograms/mL. Bacterial survival was approximately 31% under optimal electroporation conditions, and the maximal number of transformants was obtained with 80 ng of pEST DNA. Bartonella bacilliformis was verified as the transformed organism by a comparison of transformant protein profiles with those of wild-type B. bacilliformis using sodium dodecyl sulfate polyacrylamide gel electrophoresis, and detection of the exogenous plasmid in DNA from the transformed bacteria by DNA hybridization. Transformations using the plasmids pMK20, pML31, and pUCK18 (containing the replicons ColE1, F, and pMB1, respectively) were not successful.


Prevalence and correlates of suicidal ideation and suicide attempts among veterans in primary care referred for a mental health evaluation.
The Veterans Health Administration has made concerted efforts to increase mental health services offered in primary care. However, few studies have evaluated correlates of suicidal ideation and suicide attempt in veterans in primary care-mental health integration (PCMHI). The purpose of the present study is to examine associations between suicidal ideation and suicide attempts as dependent variables and demographic and clinical factors as the independent variables. Veterans (n=3004) referred from primary care to PCMHI were contacted for further assessment, which included past-year severity of suicidal thoughts (none, low, high) and attempts using the Paykel Suicide Scale, mental health disorders, and illicit drug use. Multinomial logistic regression models were used to identify correlates of suicidal ideation and suicide attempts. Thoughts of taking one's life was endorsed by 24% of participants and suicide attempts were reported in 2%. In adjusted models, depression, psychosis, mania, PTSD and generalized anxiety disorder were associated with high severity suicidal ideation, but not suicide attempt. Illicit drug use was not associated with suicidal ideation, but was the only variable associated with suicide attempt. The study was cross-sectional, focused on one clinical setting, and the suicide attempt analyses had limited power. PCMHI is a critical setting to assess suicidal ideation and suicide attempt and researchers and clinicians should be aware that the differential correlates of these suicide-related factors. Future research is needed to identify prospective risk factors and assess the utility of follow-up care in preventing suicide.


Guizhi Fuling pills inhibit the proliferation, migration and invasion of human cutaneous malignant melanoma cells by regulating the molecular axis of LncRNA TPT1-AS1 / miR-671-5p.
The current research was aimed to explore the effects of Guizhi Fuling Pills on the proliferation, migration and invasion of human cutaneous malignant melanoma cells and its regulation on the molecular axis of LncRNA TPT1-AS1 / miR-671-5p. Human cutaneous malignant melanoma cells A375 were cultured in vitro and randomly divided into Con group, Guizhi Fuling pills-L group, Guizhi Fuling pills-M group, Guizhi Fuling pills-H group, Guizhi Fuling pills-H + pcDNA group, Guizhi Poria pills-H + pcDNA-TPT1-AS1 group. MTT was used to detect cell proliferation. The Transwell cell test was used to detect cell migration and invasion. qRT-PCR was used to detect the expression of TPT1-AS1 and miR-671-5p. The dual-luciferase report experiment verified the targeting relationship of TPT1-AS1, miR-671-5p. Western blot was used to detect the expression of Ki-67, PCNA, MMP-2 and MMP-9. Compared with Con group, Guizhi Fuling Pills could inhibit cell proliferation, migration and invasion (p<0.05), and also inhibit the expression of Ki-67, PCNA, MMP-2, MMP-9, TPT1-AS1 (p<0.05), promote the expression of miR-671-5p (p<0.05)., and the differences between the indexes of Guizhi Fuling Pill-L group, Guizhi Fuling Pill-M group and Guizhi Fuling Pill-H group were statistically significant (p<0.05). The dual-luciferase report experiment confirmed that TPT1-AS1 could target and bind to miR-671-5p and could regulate the expression and activity of miR-671-5p. Overexpression of TPT1-AS1 could reduce the inhibitory effect of Guizhi Fuling Pills on proliferation, migration and invasion of A375 cells. Guizhi Fuling Pill may reduce the proliferation, migration and invasion of human cutaneous malignant melanoma cells by down-regulating the expression of TPT1-AS1 and up-regulating the expression of miR-671-5p.


Pneumocystis carinii pneumonia: a major complication of immunosuppressive therapy in patients with Wegener's granulomatosis.
The risk factors and clinical and laboratory parameters in Pneumocystis carinii pneumonia in patients with Wegener's granulomatosis have not been well characterized. We undertook a retrospective chart review of all patients with a diagnosis of Wegener's granulomatosis and P. carinii pneumonia who were followed at the National Institute of Allergy and Infectious Diseases of the National Institutes of Health. The chart review focused on clinical, laboratory, and roentgenologic evidence of P. carinii pneumonia. Eleven cases of P. carinii pneumonia were diagnosed in some 180 patients with Wegener's granulomatosis, for an overall incidence of approximately 6%. All patients developed P. carinii pneumonia either during the initial course of treatment or during therapy for recurrent Wegener's granulomatosis. All patients were receiving daily glucocorticoids and a second immunosuppressive therapy. Lymphocytopenia was noted in all patients, with a mean +/- SEM total lymphocyte count of 303 +/- 69 cells/microL. All patients tested (10 of 11) were seronegative for human immunodeficiency virus (HIV) infection. Eight presented with worsening chest roentgenograms compared with baseline, whereas three presented with normal chest roentgenograms. We conclude that P. carinii is a common opportunistic pathogen in patients with Wegener's granulomatosis receiving immunosuppressive therapy. Therapeutic immunosuppression (daily glucocorticoids and immunosuppressive agents) and the resultant lymphocytopenia, as well as the lymphocyte and monocyte functional abnormalities caused by glucocorticoids, may be the most likely factors predisposing to P. carinii pneumonia in patients with Wegener's granulomatosis. Based on our data, all patients with Wegener's granulomatosis should be given chemoprophylaxis against P. carinii while they are receiving daily glucocorticoids.(ABSTRACT TRUNCATED AT 250 WORDS)


Desquamative interstitial pneumonitis in a patient with systemic lupus erythematosus.
Desquamative interstitial pneumonia (DIP) is a rare form of interstitial lung disease (ILD) commonly found among healthy smokers. ILD is a rare manifestation of systemic lupus erythematosus (SLE), and typically associated with a histopathological pattern of nonspecific interstitial pneumonia (NSIP). The present article describes an unusual case of DIP in a nonsmoking patient with SLE presenting as NSIP. DIP can occur in the context of SLE in patients with a negative smoking history, and clinicians should consider lung biopsy to correctly classify ILD with unusual presentation on computed tomography scan.


Phosphorylation and nucleotide-dependent dephosphorylation of hepatic polypeptides related to the plasma cell differentiation antigen PC-1.
A glycoprotein fraction was isolated from rat liver membranes by affinity chromatography on immobilized wheat-germ lectin. Incubation of this fraction with MgATP or MgGTP resulted in a sequential phosphorylation and dephosphorylation of a complex of three polypeptides (118, 128 and 197 kDa on SDS/PAGE) with N-linked sialyloligosaccharides. Each polypeptide was recognized by polyclonal antibodies against recombinant plasma cell differentiation antigen PC-1. The relationship of the 118 kDa and 128 kDa polypeptides with PC-1 was confirmed by observations that they are linked by disulphide bonds into a larger protein, and that they are exclusively phosphorylated on Thr residues. Phosphorylation of p118, p128 and p197 only occurred after a lag period (up to 90 min at 30 degrees C), which lasted until most of the ATP had been converted to adenosine and Pi, with ADP and AMP as intermediate products. The length of the latency period increased with the concentration of initially added ATP (5-1000 microM) and could be prolonged by a second addition of similar concentrations of ATP, ADP, AMP and various nucleotide analogues. Most potent were the alpha beta-methylene derivatives of ADP and ATP. Adenosine was poorly effective. AMP, ADP, and perhaps ATP, emerge as the direct determinants of the latency. After further purification of the lectin-purified membrane fraction on anion-exchange and molecular-sieve columns, the complex of p118, p128 and p197 was still capable of autophosphorylation and dephosphorylation. The dephosphorylation was not affected by classical inhibitors (NaF, okadaic acid, EDTA, EGTA, phenylalanine). It was stimulated about 20-fold by various adenine nucleotides and analogues, with the same order of efficiency as noted for the induction of the latency. A similar stimulation of dephosphorylation was caused by 0.5 mM Na3VO4, which also prevented the phosphorylation of the three polypeptides. The likely explanation for the latency that precedes the phosphorylation of the membrane proteins is that the action of a protein kinase is initially offset by nucleotide-stimulated dephosphorylation.


Post-thyroidectomy hypoparathyroidism.
The incidence of hypoparathyroidism following thyroid surgery in the series reported was 7.8 per cent. Temporary hypoparathyroidism was present in 15 patients (4.6%) and permanent hypoparathyroidism in 10 (3.2%). Persistently low levels of serum calcium and high levels of serum phosphorus at six to 10 weeks after thyroid operation in patients with temporary hypoparathyroidism, with no tendency toward normalization, suggests permanent hypoparathyroidism. Serum calcium below 5 mg per cent suggests permanent hypoparathyroidism.


Challenges and Scientific Prospects of the Newest Generation of mRNA-Based Vaccines against SARS-CoV-2.
In the context of the current COVID-19 pandemic, traditional, complex and lengthy methods of vaccine development and production would not have been able to ensure proper management of this global public health crisis. Hence, a number of technologies have been developed for obtaining a vaccine quickly and ensuring a large scale production, such as mRNA-based vaccine platforms. The use of mRNA is not a new concept in vaccine development but has leveraged on previous knowledge and technology. The great number of human resources and capital investements for mRNA vaccine development, along with the experience gained from previous studies on infectious diseases, allowed COVID-19 mRNA vaccines to be developed, conditionally approved and commercialy available in less than one year, thanks to decades of basic research. This review critically presents and discusses the COVID-19 mRNA vaccine-induced immunity, and it summarizes the most common anaphylactic and autoimmune adverse effects that have been identified until now after massive vaccination campaigns.


Generation of CFUC suppressor T cells in vitro: VII. T derived colony inhibitory activity (Td/CIA) has no suppressor effect on in vitro immunoglobulin production.
T derived colony inhibitory activity (Td/CIA) was obtained from unstimulated T cells from aplastic anemia patients (SAA), or from PWM primed normal T cells. Td/CIA suppressed CFUC growth of normal allogeneic marrow to less than 30% of expected growth. Td/CIA was then added to normal peripheral blood T and B cells, primed with PWM, to test whether it would interfere with in vitro immunoglobulin (Ig) production. When Td/CIA from normal T cells was added to cultures of T + B cells + PWM there was a 2-2.1-fold increase in Ig production. Similarly the addition of Td/CIA from SAA patients also resulted in a 1.4 up to 166-fold increase in Ig production. These results indicate that either (a) the targets for Td/CIA are expressed on hemopoietic but not on T and B cells, or (b) that Td/CIA inactivates an accessory cell which is essential for CFUC growth but not for the PWM driven in vitro B cell differentiation system.


[Hydration status and aerobic capacity: effects on plasmatic volume during strenuous physical exercise].
During exercise, water loss frequently occurs in the intracellular spaces and there is a decrease in the plasmatic volume, with a blood concentration as a secondary characteristic. Plasmatic volume losses provoke a decrease in the blood flow, which directly affects cardiac function. Physical performance decreases and aerobic capacity deteriorates. An observational, prospective, longitudinal, and comparative study was carried out to evaluate gender influence and the aerobic capacity level upon hydration and the plasmatic volume produced by maximal physical exercise. Twenty four individuals between 18 and 35 years old were included. All participated in aerobic physical exercise and changes in hemoglobin, hematocrit, plasmatic volume, and hydration state were evaluated. Gender showed a significant influence on plasmatic volume but not on the hydration state. The aerobic capacity presented a relationship with plasmatic volume, and the plasmatic volume with the hydration state. Women presented more plasmatic volume loss than men (p < 0.05). Individuals who have adequate aerobic capacity tend to lose less plasmatic volume during exercise. Maximal physical exercise during a short period also leads to important liquid loss; therefore, it is important that people who engage in exercise understand their liquid losses and re-hydrate themselves appropriately before, during, and after exercise.


RNA binding protein IGF2BP1 synergizes with ETV6-RUNX1 to drive oncogenic signaling in B-cell Acute Lymphoblastic Leukemia.
Acute lymphoblastic leukemia (ALL) is the most common pediatric hematological malignancy, with ETV6::RUNX1 being the most prevalent translocation whose exact pathogenesis remains unclear. IGF2BP1 (Insulin-like Growth Factor 2 Binding Protein 1) is an oncofetal RNA binding protein seen to be specifically overexpressed in ETV6::RUNX1 positive B-ALL. In this study, we have studied the mechanistic role of IGF2BP1 in leukemogenesis and its synergism with the ETV6::RUNX1 fusion protein. Gene expression was analyzed from patient bone marrow RNA using Real Time RT-qPCR. Knockout cell lines were created using CRISPR-Cas9 based lentiviral vectors. RNA-Seq and RNA Immunoprecipitation sequencing (RIP-Seq) after IGF2BP1 pulldown were performed using the Illumina platform. Mouse experiments were done by retroviral overexpression of donor HSCs followed by lethal irradiation of recipients using a bone marrow transplant model. We observed specific overexpression of IGF2BP1 in ETV6::RUNX1 positive patients in an Indian cohort of pediatric ALL (n=167) with a positive correlation with prednisolone resistance. IGF2BP1 expression was essential for tumor cell survival in multiple ETV6::RUNX1 positive B-ALL cell lines. Integrated analysis of transcriptome sequencing after IGF2BP1 knockout and RIP-Seq after IGF2BP1 pulldown in Reh cell line revealed that IGF2BP1 targets encompass multiple pro-oncogenic signalling pathways including TNFα/NFκB and PI3K-Akt pathways. These pathways were also dysregulated in primary ETV6::RUNX1 positive B-ALL patient samples from our center as well as in public B-ALL patient datasets. IGF2BP1 showed binding and stabilization of the ETV6::RUNX1 fusion transcript itself. This positive feedback loop led to constitutive dysregulation of several oncogenic pathways. Enforced co-expression of ETV6::RUNX1 and IGF2BP1 in mouse bone marrow resulted in marrow hypercellularity which was characterized by multi-lineage progenitor expansion and strong Ki67 positivity. This pre-leukemic phenotype confirmed their synergism in-vivo. Clonal expansion of cells overexpressing both ETV6::RUNX1 and IGF2BP1 was clearly observed. These mice also developed splenomegaly indicating extramedullary hematopoiesis. Our data suggest a combined impact of the ETV6::RUNX1 fusion protein and RNA binding protein, IGF2BP1 in activating multiple oncogenic pathways in B-ALL which makes IGF2BP1 and these pathways as attractive therapeutic targets and biomarkers.


Characterization and Validation of In Vitro and In Vivo Models to Investigate TNF-α-Induced Inflammation in Retinal Diseases.
Inflammation is implicated in the etiology of diverse retinopathies including uveitis, age-related macular degeneration or diabetic retinopathy. Tumor necrosis factor alpha (TNF-α) is a well-known proinflammatory cytokine that is described as a biomarker for inflammation in diverse retinopathies and therefore emerged as an interesting target to treat inflammation in the eye by neutralizing anti-TNF-α antibodies. Recently, we have demonstrated that Adeno-associated virus (AAV)-mediated expression of human TNF-α in the murine eye induces retinal inflammation including vasculitis and fibrosis, thereby mimicking human disease-relevant pathologies. In a proof-of-mechanism study, we now tested whether AAV-TNF-α induced pathologies can be reversed by neutralizing TNF-α antibody treatment. Strikingly, a single intravitreal injection of the TNF-α antibody golimumab reduced AAV-TNF-α-induced retinal inflammation and retinal thickening. Furthermore, AAV-TNF-α-mediated impaired retinal function was partially rescued by golimumab as revealed by electroretinography recordings. Finally, to study TNF-α-induced vasculitis in human in vitro cell culture assays, we established a monocyte-to-endothelium adhesion co-culture system. Indeed, also in vitro TNF-α induced monocyte adhesion to human retinal endothelial cells, which was prevented by golimumab. Overall, our study describes valuable in vitro and in vivo approaches to study the function of TNF-α in retinal inflammation and demonstrated a preclinical proof-of-mechanism treatment with golimumab. The AAV-based model expressing human TNF-α allows us to investigate TNF-α-driven pathologies supporting research in mechanisms of retinal inflammation.


Multiple chaperonins in bacteria--why so many?
A significant proportion of bacteria express two or more chaperonin genes. Chaperonins are a group of molecular chaperones, defined by sequence similarity, required for the folding of some cellular proteins. Chaperonin monomers have a mass of c. 60 kDa, and are typically found as large protein complexes containing 14 subunits arranged in two rings. The mechanism of action of the Escherichia coli GroEL protein has been studied in great detail. It acts by binding to unfolded proteins and enabling them to fold in a protected environment where they do not interact with any other proteins. GroEL can assist the folding of many proteins of different sizes, sequences, and structures, and homologues from many different bacteria can functionally replace GroEL in E. coli. What then are the functions of multiple chaperonins? Do they provide a mechanism for cells to increase their general chaperoning ability, or have they become specialized to take on specific novel cellular roles? Here I will review the genetic, biochemical, and phylogenetic evidence that has a bearing on this question, and show that there is good evidence for at least some specificity of function in multiple chaperonin genes.


Murine models of Candida gastrointestinal colonization and dissemination.
Ninety-five percent of infectious agents enter through exposed mucosal surfaces, such as the respiratory and gastrointestinal (GI) tracts. The human GI tract is colonized with trillions of commensal microbes, including numerous Candida spp. Some commensal microbes in the GI tract can cause serious human infections under specific circumstances, typically involving changes in the gut environment and/or host immune conditions. Therefore, utilizing animal models of fungal GI colonization and dissemination can lead to significant insights into the complex pathophysiology of transformation from a commensal organism to a pathogen and host-pathogen interactions. This paper will review the methodologic approaches used for modeling GI colonization versus dissemination, the insights learned from these models, and finally, possible future directions using these animal modeling systems.


Unravelling the evolutionary history and future prospects of endemic species restricted to former glacial refugia.
The contemporary distribution and genetic composition of biodiversity bear a signature of species' evolutionary histories and the effects of past climatic oscillations. For many European species, the Mediterranean peninsulas of Iberia, Italy and the Balkans acted as glacial refugia and the source of range recolonization, and as a result, they contain disproportionately high levels of diversity. As these areas are particularly threatened by future climate change, it is important to understand how past climatic changes affected their biodiversity. We use an integrated approach, combining markers with different evolutionary rates and combining phylogenetic analysis with approximate Bayesian computation and species distribution modelling across temporal scales. We relate phylogeographic processes to patterns of genetic variation in Myotis escalerai, a bat species endemic to the Iberian Peninsula. We found a distinct population structure at the mitochondrial level with a strong geographic signature, indicating lineage divergence into separate glacial refugia within the Iberian refugium. However, microsatellite markers suggest higher levels of gene flow resulting in more limited structure at recent time frames. The evolutionary history of M. escalerai was shaped by the effects of climatic oscillations and changes in forest cover and composition, while its future is threatened by climatically induced range contractions and the role of ecological barriers due to competition interactions in restricting its distribution. This study warns that Mediterranean peninsulas, which provided refuge for European biodiversity during past glaciation events, may become a trap for limited dispersal and ecologically limited endemic species under future climate change, resulting in loss of entire lineages.


Pilots involved in multiple crashes: "accident proneness" revisited.
Analysis of crashes of air taxi and commuter flights explored the controversial issue of "accident proneness." There were 20 pilots who had 2 or more crashes during 1983-88. These pilots (repeaters) and their 42 crashes were compared with 534 pilots who were each involved in a single air taxi or commuter crash during the same period (nonrepeaters). Unexpectedly, repeaters were more experienced pilots with a mean total flight time of 7016 h vs. 5321 for nonrepeaters. Repeaters did not differ from nonrepeaters in the overall proportion of crashes in which pilot performance appeared to be a major factor. Repeaters differed significantly from nonrepeaters as to flight hours during the past 90 d (mean 215 vs. 183 h) and the proportion of their crashes that occurred in Alaska (48% vs. 24%). Alaska repeaters differed from non-Alaska repeaters with regard to the proportion of crashes on takeoff (40% vs 14%) and airport conditions as a factor (50% vs. 18%). The high proportion of repeaters involved in crashes in Alaska, where environmental conditions make flying more hazardous, and the substantially greater recent flight time suggest that the intensity and amount of occupational exposure are major determinants of pilot involvement in more than one crash.


Senescent CD4+CD28- T Lymphocytes as a Potential Driver of Th17/Treg Imbalance and Alveolar Bone Resorption during Periodontitis.
Senescent cells express a senescence-associated secretory phenotype (SASP) with a pro-inflammatory bias, which contributes to the chronicity of inflammation. During chronic inflammatory diseases, infiltrating CD4+ T lymphocytes can undergo cellular senescence and arrest the surface expression of CD28, have a response biased towards T-helper type-17 (Th17) of immunity, and show a remarkable ability to induce osteoclastogenesis. As a cellular counterpart, T regulatory lymphocytes (Tregs) can also undergo cellular senescence, and CD28- Tregs are able to express an SASP secretome, thus severely altering their immunosuppressive capacities. During periodontitis, the persistent microbial challenge and chronic inflammation favor the induction of cellular senescence. Therefore, senescence of Th17 and Treg lymphocytes could contribute to Th17/Treg imbalance and favor the tooth-supporting alveolar bone loss characteristic of the disease. In the present review, we describe the concept of cellular senescence; particularly, the one produced during chronic inflammation and persistent microbial antigen challenge. In addition, we detail the different markers used to identify senescent cells, proposing those specific to senescent T lymphocytes that can be used for periodontal research purposes. Finally, we discuss the existing literature that allows us to suggest the potential pathogenic role of senescent CD4+CD28- T lymphocytes in periodontitis.


Electroacupuncture at the Zusanli (ST-36) Acupoint Induces a Hypoglycemic Effect by Stimulating the Cholinergic Nerve in a Rat Model of Streptozotocine-Induced Insulin-Dependent Diabetes Mellitus.
Animal studies have shown that electroacupuncture (EA) at Zusanli (ST-36) and Zhongwan (CV-12) acupoints reduces plasma glucose concentrations in rats with type II diabetes. However, whether EA reduces plasma glucose levels in type I diabetes is still unknown. In this study, we explore the various non-insulin-dependent pathways involved in EA-induced lowering of plasma glucose. Streptozotocin (STZ) (60 mg kg(-1), i.v.) was administered via the femoral vein to induce insulin-dependent diabetes in non-adrenalectomized and in adrenalectomomized rats. EA (15 Hz) was applied for 30 min to bilateral ST-36 acupoints after administration of Atropine (0.1 mg kg(-1) i.p.), Eserine (0.01 mg kg(-1) i.p.), or Hemicholinium-3 (5 μg kg(-1) i.p.) in non-adrenalectomized rats. Rats administered acetylcholine (0.01 mg kg(-1) i.v.) did not undergo EA. Adrenalectomized rats underwent EA at bilateral ST-36 acupoints without further treatment. Blood samples were drawn from all rats before and after EA to measure changes in plasma glucose levels. Expression of insulin signaling proteins (IRS1, AKT2) in atropine-exposed rats before and after EA was measured by western blot. Atropine and hemicholinium-3 completely blocked the plasma glucose lowering effects of EA, whereas eserine led to a significant hypoglycemic response. In addition, plasma glucose levels after administration of acetylcholine were significantly lower than the fasting glucose levels. In STZ-adrenalectomized rats, EA did not induce a hypoglycemic response. EA stimulated the expression of IRS1 and AKT2 and atropine treatment blocked the EA-induced expression of those insulin signaling proteins. Taken together, EA at the ST-36 acupoint reduces plasma glucose concentrations by stimulating the cholinergic nerves.


Bone mineral density in haemophilia patients. A meta-analysis.
Osteoporosis is caused by bone mineral density (BMD) reduction. Haemophilia patients are at increased risk of osteoporosis because of decreased physical activity and blood-borne virus infections. This systematic review of the literature aims at evaluating BMD reduction in severe haemophilia patients and its correlation with patients' characteristics. Seven case-control studies evaluating lumbar BMD values [g/cm2] (all studies), BMI (5/7 studies), and hepatitis C virus (HCV) seropositivity (6/7 studies) in severe haemophilia patients and controls were meta-analysed. Standardised mean difference (SMD) of BMD was used to compare cases and controls. The effect of body mass index (BMI) and HCV infection was investigated by meta-regression. One hundred one adult cases (age 33 +/- 8.9) with 101 controls and 111 paediatric cases (age 8 +/- 3.6) with 307 controls were available for analysis. Lumbar BMD was significantly lower in severe haemophilia patients than in controls, both in adult (pooled SMD -1.379, 95% confidence interval [CI] -2.355 to -0.403, p=0.006) and children (pooled SMD -0.438, 95% CI -0.686 to -0.189, p=0.001). The reduction in BMD in patients versus controls was not significantly correlated with the reduction in BMI or with the percentage of HCV-infected patients. This meta-analysis confirms the association between severe haemophilia and low BMD. Future studies should investigate fracture rates and interventions to prevent bone loss in persons with haemophilia.


Small-molecule inhibitor binding to an N-acyl-homoserine lactone synthase.
Quorum sensing (QS) controls certain behaviors of bacteria in response to population density. In gram-negative bacteria, QS is often mediated by N-acyl-L-homoserine lactones (acyl-HSLs). Because QS influences the virulence of many pathogenic bacteria, synthetic inhibitors of acyl-HSL synthases might be useful therapeutically for controlling pathogens. However, rational design of a potent QS antagonist has been thwarted by the lack of information concerning the binding interactions between acyl-HSL synthases and their ligands. In the gram-negative bacterium Burkholderia glumae, QS controls virulence, motility, and protein secretion and is mediated by the binding of N-octanoyl-L-HSL (C8-HSL) to its cognate receptor, TofR. C8-HSL is synthesized by the acyl-HSL synthase TofI. In this study, we characterized two previously unknown QS inhibitors identified in a focused library of acyl-HSL analogs. Our functional and X-ray crystal structure analyses show that the first inhibitor, J8-C8, binds to TofI, occupying the binding site for the acyl chain of the TofI cognate substrate, acylated acyl-carrier protein. Moreover, the reaction byproduct, 5'-methylthioadenosine, independently binds to the binding site for a second substrate, S-adenosyl-L-methionine. Closer inspection of the mode of J8-C8 binding to TofI provides a likely molecular basis for the various substrate specificities of acyl-HSL synthases. The second inhibitor, E9C-3oxoC6, competitively inhibits C8-HSL binding to TofR. Our analysis of the binding of an inhibitor and a reaction byproduct to an acyl-HSL synthase may facilitate the design of a new class of QS-inhibiting therapeutic agents.


Synthesis and properties of isomerically pure anthrabisbenzothiophenes.
The synthesis of three heptacyclic heteroacenes is described, namely anthra[2,3-b:7,6-b']bis[1]benzothiophenes (ABBTs). A stepwise sequence of aldol reactions provides regiochemical control, affording only the syn-isomer. The ABBTs are characterized by X-ray crystallography, UV-vis absorption, and emission spectroscopy, as well as cyclic voltammetry. Field effect transistors based on solution-cast thin films of ABBT derivatives exhibit charge-carrier mobilities of as high as 0.013 cm(2)/(V s).


Label-free detection of telomerase activity using guanine electrochemical oxidation signal.
Telomerase is an important biomarker for cancer cells and its activation in 85% of all cancer types confers a clinical diagnostic value. A label-free electrochemical assay based on guanine oxidation signal to measure telomerase activity is described. This developed technology combined with a disposable sensor, carbon graphite electrode (CGE), and differential pulse voltammetry (DPV) was performed by using PCR amplicons with/without telomeric repeats as the guanine oxidation signal observed at +1.0 V measured after the immobilization of PCR products. Guanine oxidation signal was chosen as a measure of telomerase activity because a substantial increase in the number of guanines was introduced by the action of telomerase which adds hexameric repeats (TTAGGG)n that contain 50% guanine. The developed assay was shown to specifically measure telomerase activity from cell extracts, and elongation rates increased linearly in a concentration dependent manner. Telomerase activity could be detected in cell extracts containing as low as 100 ng/microL of protein. All of the electrochemical measurements were also confirmed with the conventional TRAP-silver staining assay. Rapidity, simplicity, and the label-free nature of the developed assay make it suitable for practical use in quantitative determination of telomerase activity from clinical samples for diagnosis of cancer.


Prognosis of colo-rectal carcinoma in Sudan.
Forty-two histologically confirmed cases of colo-rectal carcinoma were analysed. Thirty-three per cent were below the age of 30 years. Patients presented at an advanced stage of the disease evidenced by lympth node and/or hepatic involvement, ascites and intestinal obstruction. Nearly half of the patients had low grade tumours. Emergency and multi-staged operations were often necessary. The hospital mortality rate was 38.2 per cent. The problems of diagnosis and management under the conditions prevailing in the Sudan were discussed. It is suggested that a better health education of the people and more awareness of the medical profession are necessary to improve the present results.


Gene therapy and stem cell transplantation: strategies for hearing restoration.
Strategies to restore sensorineural hearing loss focus on the replacement of lost hair cells, the specialized mechanoreceptors in the organ of Corti that convert the mechanical energy of sound into electrical energy. Hair cells in mammalian systems do not have the capacity to regenerate, but two exciting lines of research hold promise in restoring inner ear function. Here we review basic principles of gene therapy and discuss its application in the inner ear. We survey the various viral vectors and routes of delivery into the inner ear. Applications of gene therapy in the inner include hair cell protection in the face of chemical or noise-induced ototoxicity, spiral ganglion cell survival following hair cell death or injury, and hair cell regeneration. More recently, the viability of gene therapy in human inner ear tissue has been reported. Transplantation of progenitor cells that can differentiate into functioning hair cells with the appropriate connections to their corresponding spiral ganglion cells is yet another strategy to restore sensorineural hearing loss. Neonatal or embryonic stem cells, adult mouse inner ear stem cells, and stem cells from the central nervous system have been shown to differentiate into cells containing hair cell markers and proteins. Prospects for stem cell therapy in the inner ear, and its limitations, will also be examined.


Representation of cloned genomic sequences in two sequencing vectors: correlation of DNA sequence and subclone distribution.
Representation of subcloned Caenorhabditis elegans and human DNA sequences in both M13 and pUC sequencing vectors was determined in the context of large scale genomic sequencing. In many cases, regions of subclone under-representation correlated with the occurrence of repeat sequences, and in some cases the under-representation was orientation specific. Factors which affected subclone representation included the nature and complexity of the repeat sequence, as well as the length of the repeat region. In some but not all cases, notable differences between the M13 and pUC subclone distributions existed. However, in all regions lacking one type of subclone (either M13 or pUC), an alternate subclone was identified in at least one orientation. This suggests that complementary use of M13 and pUC subclones would provide the most comprehensive subclone coverage of a given genomic sequence.


Influence of subclinical hypothyroidism on metabolic parameters in obese children and adolescents.
Subclinical hypothyroidism (SH) is a common condition in obese children. However, its effect on glucose and lipid metabolism in obese children remains controversial. The present study aimed to investigate the association between SH and metabolic parameters. A total of 215 obese children and adolescents aged 6-18 years were included in this retrospective cross-sectional study. The patients' anthropometric measurements such as thyrotropin (TSH), free thyroxine (fT4), fasting plasma glucose, and insulin levels, as well as homeostasis model assessment for insulin resistance (HOMA-IR) index, and lipid profiles were evaluated. The patients were allocated to the SH group (fT4 normal, TSH 5-10 mIU/L) (n=77) or the control group (fT4 normal, TSH<5 mIU/L) (n=138). The glucose and lipid metabolisms of the 2 groups were compared. SH was identified in 77 of 215 patients (36%). Mean body mass index was similar in both groups. The mean serum insulin, HOMA-IR, and triglyceride (TG) levels were higher and the mean high-density lipoprotein cholesterol level was lower in the SH group than in the control group (P=0.007, P=0.004, P=0.01, and P=0.02, respectively). A positive correlation was observed between TSH level and insulin level, HOMA-IR, and TG level. SH was identified in some of the obese children and adolescents. A clear association was observed between SH, insulin resistance, and dyslipidemia in obese children.


Effect of weekly physical activity frequency on weight loss in healthy overweight and obese women attending a weight loss program: a randomized controlled trial.
The effect of intensity and duration of physical activity (PA) on weight loss has been well described. However, the effect of the frequency of weekly PA on weight loss is still unknown. The purpose of this study was to evaluate the effect of the frequency of weekly PA sessions while maintaining the same total activity time on weight loss during a 24-wk weight loss program. Overweight and obese women [n = 75; body mass index (BMI; in kg/m2): 27-37; age: 18-40 y] who had a normally sedentary lifestyle were randomly allocated to 1 of 2 intervention groups: a high-frequency physical activity (HF) or a low-frequency physical activity (LF) group. The HF group included 50 min/d PA, 6 d/wk (300 min/wk). The LF group included 100 min/d PA, 3 d/wk (300 min/wk). Both groups were advised to follow the same dietary weight loss program. Both groups showed a significant decrease in anthropometric measurements and significant improvements in cardiometabolic disease risk characteristics over the 24 wk of the study. Compared with the HF group, the LF group had a greater decrease in weight (mean ± SD; LF: 9.58 ± 3.77 kg; HF: 7.78 ± 2.68 kg; P = 0.028), BMI (LF: 3.62 ± 1.56; HF: 2.97 ± 1.02; P = 0.029) and waist circumference (LF: 9.36 ± 4.02 cm; HF: 7.86 ± 2.41 cm; P = 0.031). However, there were no significant differences in carbohydrate metabolism characteristics or lipid profile after the 24 wk of intervention. Weekly PA undertaken over fewer sessions of longer duration during the week could be more effective for weight loss than when undertaken as more frequent shorter sessions in overweight and obese women on a weight loss program. This may be helpful for those who are neither willing nor able to schedule time for PA almost every day to achieve weight loss. This trial was registered at www.irct.ir as IRCT201402157754N4.


Multi-Channel 3D Deep Feature Learning for Survival Time Prediction of Brain Tumor Patients Using Multi-Modal Neuroimages.
High-grade gliomas are the most aggressive malignant brain tumors. Accurate pre-operative prognosis for this cohort can lead to better treatment planning. Conventional survival prediction based on clinical information is subjective and could be inaccurate. Recent radiomics studies have shown better prognosis by using carefully-engineered image features from magnetic resonance images (MRI). However, feature engineering is usually time consuming, laborious and subjective. Most importantly, the engineered features cannot effectively encode other predictive but implicit information provided by multi-modal neuroimages. We propose a two-stage learning-based method to predict the overall survival (OS) time of high-grade gliomas patient. At the first stage, we adopt deep learning, a recently dominant technique of artificial intelligence, to automatically extract implicit and high-level features from multi-modal, multi-channel preoperative MRI such that the features are competent of predicting survival time. Specifically, we utilize not only contrast-enhanced T1 MRI, but also diffusion tensor imaging (DTI) and resting-state functional MRI (rs-fMRI), for computing multiple metric maps (including various diffusivity metric maps derived from DTI, and also the frequency-specific brain fluctuation amplitude maps and local functional connectivity anisotropy-related metric maps derived from rs-fMRI) from 68 high-grade glioma patients with different survival time. We propose a multi-channel architecture of 3D convolutional neural networks (CNNs) for deep learning upon those metric maps, from which high-level predictive features are extracted for each individual patch of these maps. At the second stage, those deeply learned features along with the pivotal limited demographic and tumor-related features (such as age, tumor size and histological type) are fed into a support vector machine (SVM) to generate the final prediction result (i.e., long or short overall survival time). The experimental results demonstrate that this multi-model, multi-channel deep survival prediction framework achieves an accuracy of 90.66%, outperforming all the competing methods. This study indicates highly demanded effectiveness on prognosis of deep learning technique in neuro-oncological applications for better individualized treatment planning towards precision medicine.


Oral vaccination of mice with attenuated Salmonella encoding Trichinella spiralis calreticulin and serine protease 1.1 confers protective immunity in BALB/c mice.
Trichinella spiralis is a foodborne parasitic nematode which is a serious risk to meat safety. Development of anti-Trichinella vaccine is needed to control Trichinella infection in food animals. In this study, two novel T. spiralis genes (calreticulin and serine protease 1.1) in combination were used to construct oral DNA vaccines, and their induced protective immunity was evaluated in a murine model. TsCRT+TsSP1.1, TsCRT and TsSP1.1 DNA were transformed into attenuated Salmonella typhimurium ΔcyaSL1344. Oral vaccination of mice with TsCRT+TsSP1.1, TsCRT and TsSP1.1 DNA vaccines elicited a gut local mucosal sIgA response and systemic Th1/Th2 mixed response. Oral vaccination with TsCRT+TsSP1.1 induced obviously higher level of serum specific antibodies, mucosal sIgA and cellular immune response than either of single TsCRT or TsSP1.1 DNA vaccination. Oral vaccination of mice with TsCRT+TsSP1.1 exhibited a 53.4% reduction of enteral adult worms and a 46.05% reduction of muscle larvae, conferred a higher immune protection than either of individual TsCRT (44.28 and 42.46%) or TsSP1.1 DNA vaccine (35.43 and 29.29%) alone. Oral vaccination with TsCRT+TsSP1.1, TsCRT and TsSP1.1 also obviously ameliorated inflammation of intestinal mucosa and skeletal muscles of vaccinated mice after challenge. TsCRT and TsSP1.1 might be regarded the novel potential targets for anti-Trichinella vaccines. Attenuated Salmonella-delivered DNA vaccine provided a prospective approach to control T. spiralis infection in food animals.


Clinical Presentation, Quality of Care, Risk Factors and Outcomes in Women with Acute ST-Elevation Myocardial Infarction (STEMI): An Observational Report from Six Middle Eastern Countries.
Most of the available literature on ST-Elevated myocardial infarction (STEMI) in women was conducted in the developed world and data from Middle-East countries was limited. To examine the clinical presentation, patient management, quality of care, risk factors and inhospital outcomes of women with acute STEMI compared with men using data from a large STEMI registry from the Middle East. Data were derived from the third Gulf Registry of Acute Coronary Events (Gulf RACE-3Ps), a prospective, multinational study of adults with acute STEMI from 36 hospitals in 6 Middle-Eastern countries. The study included 2928 patients; 296 women (10.1%) and 2632 men (89.9%). Clinical presentations, management and in-hospital outcomes were compared between the 2 groups. Women were 10 years older and more likely to have diabetes mellitus, hypertension, and hyperlipidemia compared with men who were more likely to be smokers (all p<0.001). Women had longer median symptom-onset to emergency department (ED) arrival times (230 vs. 170 min, p<0.001) and ED to diagnostic ECG (8 vs. 6 min., p<0.001). When primary percutaneous coronary intervention (PPCI) was performed, women had longer door-to-balloon time (DBT) (86 vs. 73 min., p=0.009). When thrombolytic therapy was not administered, women were less likely to receive PPCI (69.7 vs. 76.7%, p=0.036). The mean duration of hospital stay was longer in women (6.03 ± 22.51 vs. 3.41 ± 19.45 days, p=0.032) and the crude in-hospital mortality rate was higher in women (10.4 vs. 5.2%, p<0.001). However, after adjustments, multivariate analysis revealed a statistically non-significant trend of higher inhospital mortality among women than men (6.4 vs. 4.6%), (p=0.145). Our study demonstrates that women in our region have almost double the mortality from STEMI compared with men. Although this can partially be explained by older age and higher risk profiles in women, however, correction of identified gaps in quality of care should be attempted to reduce the high morbidity and mortality of STEMI in our women.


Key stakeholder perspectives on obstacles to an autism-specific Medicaid waiver service delivery model.
Children and youth with autism spectrum disorder have significant, multi-system needs that require supports, such as the autism waiver (AW) service delivery model. This study aimed to identify and describe characteristics of the AW, define obstacles and strengths in the provision of adequate services and provide recommendations for improving overall effectiveness. This mixed-methods exploratory study used an electronic survey to gain information and perceptions of AW provider agency directors (n = 27) and service coordinators (n = 30). The key barriers reported were the shortage of qualified staff, inadequate staff training, complexity of cases or symptom severity of clients and lack of communication at multiple levels throughout the agency as well as with parents. Recommendations include reinforcing the workforce through higher salaries, greater training and communication interventions. These strategies may reduce staff turnover and shortage, lighten the caseload, reduce the waitlist period and improve the effectiveness and responsiveness of AW services.


Polymerase chain reaction amplification and functional characterization of sockeye salmon histone H3, metallothionein-B, and protamine promoters.
The histone H3 (H3), metallothionein-B (MT-B), protamine (PT) promoters of sockeye salmon were cloned by PCR using primers based on the DNA sequences of the rainbow trout promoters. Comparison of the DNA sequences of the sockeye salmon and the rainbow trout histone H3, MT-B, and PT promoters revealed that their DNA sequences and putative transcriptional cis-elements are remarkably conserved. Promoter activity of the sockeye salmon H3, MT-B, and PT promoters were examined by transfection studies using cell lines from fish and human. The H3 and MT-B promoters were shown to be active in all fish cell lines but were only weakly active in HeLa and GM637 cells. The MT-B promoter was also inducible by zinc and cadmium in RTH-149 cells. In contrast, the PT promoter was inactive in all fish cell lines. Transfection experiments also established that the HCMV IE promoter has very strong activity in the various fish cell lines. Our results suggest that the HCMV IE promoter and the sockeye salmon H3 and MT-B promoters are useful for the construction of expression vectors for transgenic and gene transfer studies in fishes.


Preference judgements involve a network of structures within frontal, cingulate and insula cortices.
Environmental stimuli constantly compete for human attention and in many cases decisions are made based on the affective meaning they convey. Although the network of structures involved in processing affective value has been well described, the specific contribution of these structures to the process by which affective value guides decision making is less well understood and is the focus of the present study. Thus, subjects read descriptions of individually tailored holidays, varying in incentive value and then made preference judgements, cognitive judgements or no decision. Choices made from an affective perspective, compared with those made from a cognitive perspective, activated a region of the anterior insula/operculum and also the anterior cingulate cortex. Furthermore, activity in perigenual, anterior cingulate cortex was correlated with subjective ratings of incentive value. In contrast, medial orbitofrontal cortex (OFC) and a region of posterior ventrolateral prefrontal cortex (PFC), bordering on the insula, were found to be more active when affective stimuli guided response selection than when no selection was made. However, only the activity in the ventrolateral PFC was specific to response selection based on affective compared with cognitive judgements. It is proposed that the necessary introspection required to make subjective preference judgements is provided by the insula and cingulate cortices, while the medial OFC and posterior ventrolateral PFC/insula cortices contribute to stimulus evaluation and motivational aspects of response selection, respectively.


Crimes of passion: When romantic obsession leads to abusive relationships.
This research examines the relationship between passion for a romantic partner and obsessive relational intrusion (ORI) which is defined as repeated and unwanted behaviors toward a partner that invade his/her personal privacy and cause fear for his/her safety. Based on the dualistic model of passion, we predicted that obsessive passion (OP) would be associated with fear of abandonment, which in turn would be associated with ORI. In contrast, harmonious passion (HP) should not be associated with ORI. Four studies (N = 1257) supported these predictions. Study 1 found cross-sectional support for these hypotheses from the victim's perspective. Study 2 replicated Study 1 and extended it to the perpetrator's perspective by demonstrating that the relationship between passion and ORI is mediated by fear of abandonment. Study 3 replicated these findings using an experimental manipulation of passion. Study 4 found similar results by experimentally manipulating fear of abandonment, the mediator, to demonstrate its causal influence on ORI. Taken together, the present results demonstrate that passion and fear of abandonment play an important role in the type of behavior people are willing to pursue to maintain their romantic relationship.


Updates in the Assessment and Management of Agitation.
Agitated patients can be dangerous to themselves and others. In fact, severe medical complications and death can occur with severe agitation. Because of this, agitation is considered a medical and psychiatric emergency. Regardless of the treatment setting, identifying the agitated patient early is a necessary skill. The authors review relevant literature regarding the identification and management of agitation and summarize current recommendations for adults, as well as children and adolescents.


Challenges Caused by Imported Cases Abroad for the Prevention and Control of COVID-19 in China.
Background: Overseas imported cases of COVID-19 continue to increase in China, so we conducted this study to review the epidemiological characteristics of these patients. Methods: From February 26 to April 4, 2020, the imported cases from abroad were enrolled in this study. The effect of prevention countermeasures in curbing the spread of COVID-19 was assessed in this study. Moreover, we defined incubation period and confirmed time as from the date of leaving the epicenter to date of symptom onset and date of final diagnosed, respectively, and the interval of symptom onset to final diagnosed time was defined as diagnostic time. Categorical variables were summarized as numbers and percentages, and the difference among the variables were analyzed. Results: For 670 cases imported from abroad, 555 were Chinese and 115 were foreigners. Apparently, confirmed cases had significantly decreased after China was compelled to temporarily suspend the entry of foreign passport holders with valid visas or residence permits; 6 days after implement of controlled measures, the daily new confirmed cases were reduced to 13 cases. Moreover, about 84.3% of patients (166/197) presented symptoms 1 week after leaving the epicenter, and notably seven patients (3.6%) had symptoms 2 weeks after leaving the epicenter. The median incubation period was 3.0 days (inter quartile range, 1.0 to 6.0), the 95th percentile was 11.6 days. Additionally, most of cases (92.9%) were detected positively of nucleic acid after symptom onset with 4 days, the median diagnostic time was 2.0 days (interquartile range, 1.0 to 3.0), and the 95th percentile of the distribution was 5.0 days. Finally, about 5.8% of patients were healthy carriers, and the median confirmed time of asymptomatic patients was 4.0 days (interquartile range, 2.0 to 9.0). The following variables might be associated with confirmed time: symptom type (P = 0.005), exported regions (P < 0.001), and symptom onset time (P < 0.001). Conclusions: The prevention countermeasures for imported cases implemented by the Chinese government played an indispensable role in curbing the spread of COVID-19; the time of departure from epicenter could provide an estimate of the incubation period; and a confirmed time, 2-week quarantine period might need to be prolonged, while asymptomatic patients should be closely monitored.


Exclusion probabilities of 22 bovine microsatellite markers in fluorescent multiplexes for semiautomated parentage testing.
Six multiplexes developed for semiautomated fluorescence genotyping were evaluated for parentage testing. These multiplexes contained primer pairs for the amplification of 22 microsatellites on 17 bovine autosomes. Exclusion probabilities were determined from genotypes of 1022 Holstein cattle and 311 beef cattle belonging to five breeds. Two cases were considered: case 1, genotypes are known for an alleged parent and an offspring but genotypes of a confirmed parent are unknown; and case 2, genotypes are known for an alleged parent, a confirmed parent and an offspring. If the alleged parent is not the true parent, then the 22 markers will exclude the alleged parent with a probability of > 0.9986 for case 1 and with a probability of > 0.99999 for case 2. On the basis of these exclusion probabilities, the probability that an alleged parent will be falsely included as a parent is in the range of 1/716 to 1/2845 for case 1 and 1/1.2 million to 1/159753 for case 2. In addition to these results, a rapid and efficient non-organic method for extraction of DNA from semen is described.


Multiple B-vitamin inadequacy amplifies alterations induced by folate depletion in p53 expression and its downstream effector MDM2.
Folate is required for biological methylation and nucleotide synthesis, aberrations of which are thought to be the mechanisms that enhance colorectal carcinogenesis produced by folate inadequacy. These functions of folate also depend on the availability of other B-vitamins that participate in "one-carbon metabolism," including B2, B6 and B12. Our study therefore investigated whether combined dietary restriction of these vitamins amplifies aberrations in the epigenetic and genetic integrity of the p53 gene that is induced by folate depletion alone. Ninety-six mice were group pair-fed diets with different combinations of B-vitamin depletion over 10 weeks. DNA and RNA were extracted from epithelial cells isolated from the colon. Within the hypermutable region of p53 (exons 5-8), DNA strand breaks were induced within exons 6 and 8 by folate combined with B2, B6 and B12 restriction (p < 0.05); such effects were not significantly induced by mild folate depletion alone. Similarly, a minor degree of hypomethylation of exon 6 produced by isolated folate depletion was significantly amplified (p < or = 0.05) by simultaneous depletion of all 4 B-vitamins. Furthermore, the expression of p53 and MDM2 were significantly decreased (p < or = 0.05) by the combined depletion state but not by folate depletion alone. These data indicate that inadequacies of other 1-carbon vitamins may amplify aberrations of the p53 gene induced by folate depletion alone, implying that concurrent inadequacies in several of these vitamins may have added tumorigenic potential beyond that observed with isolated folate depletion.


Preservation of Smooth Muscle Cell Integrity and Function: A Target for Limiting Abdominal Aortic Aneurysm Expansion?
(1) Abdominal aortic aneurysm (AAA) is a silent, progressive disease with significant mortality from rupture. Whilst screening programmes are now able to detect this pathology early in its development, no therapeutic intervention has yet been identified to halt or retard aortic expansion. The inability to obtain aortic tissue from humans at early stages has created a necessity for laboratory models, yet it is essential to create a timeline of events from EARLY to END stage AAA progression. (2) We used a previously validated ex vivo porcine bioreactor model pre-treated with protease enzyme to create "aneurysm" tissue. Mechanical properties, histological changes in the intact vessel wall, and phenotype/function of vascular smooth muscle cells (SMC) cultured from the same vessels were investigated. (3) The principal finding was significant hyperproliferation of SMC from EARLY stage vessels, but without obvious histological or SMC aberrancies. END stage tissue exhibited histological loss of α-smooth muscle actin and elastin; mechanical impairment; and, in SMC, multiple indications of senescence. (4) Aortic SMC may offer a therapeutic target for intervention, although detailed studies incorporating intervening time points between EARLY and END stage are required. Such investigations may reveal mechanisms of SMC dysfunction in AAA development and hence a therapeutic window during which SMC differentiation could be preserved or reinstated.


Interactions between hemoglobin and cod muscle constituents following treatment at extreme pH values.
Acid and/or alkaline solubilization is a recent method developed to separate proteins from muscle foods with good functional properties. However, exposure of the muscle and its components to low pH values has been shown to promote lipid oxidation, limiting therefore the applications of this novel method. This research aimed primarily to study the physicochemical changes of the fish membranes brought about during acid or alkali solubilization processes. The effect on lipid oxidation and the possible role of the water soluble fraction of the muscle (press juice) as a potent antioxidant were also investigated. Model systems comprising minced cod muscle or cod microsomal suspensions were used. Results showed that acid or alkaline treatment (pH < 3.5 or pH > 10.5) of cod membranes significantly delayed lipid oxidation. Added triacylglycerols to washed cod system treated at low pH did not enhance hemoglobin-mediated lipid oxidation. Decreased precipitation of hemoglobin was observed with the alkali-treated membranes at all protein concentrations compared to the acid-treated and the untreated membranes. Finally, the addition of press juice to washed cod muscle tissue or to the membrane model system, significantly delayed hemoglobin lipid oxidation. The results of this study can be used to improve pH-shifting technologies to avoid or decrease lipid oxidation problems. Also, the use of press-juice from cod muscle as means of protecting the muscle against lipid oxidation is suggested.


Emission properties of oxyluciferin and its derivatives in water: revealing the nature of the emissive species in firefly bioluminescence.
The first systematic steady-state and time-resolved emission study of firefly oxyluciferin (emitter in firefly bioluminescence) and its analogues in aqueous buffers provided the individual emission spectra of all chemical forms of the emitter and the excited-state equilibrium constants in strongly polar environment with strong hydrogen bonding potential. The results confirmed the earlier hypothesis that excited-state proton transfer from the enol group is favored over proton transfer from the phenol group. In water, the phenol-keto form is the strongest photoacid among the isomers and its conjugate base (phenolate-keto) has the lowest emission energy (634 nm). Furthermore, for the first time we observed green emission (525 nm) from a neutral phenol-keto isomer constrained to the keto form by cyclopropyl substitution. The order of emission energies indicates that in aqueous solution a second deprotonation at the phenol group after the enol group had dissociated (that is, deprotonation of the phenol-enolate) does not occur in the first excited state. The pH-dependent emission spectra and the time-resolved fluorescence parameters revealed that the keto-enol tautomerism reaction, which can occur in a nonpolar environment (toluene) in the presence of a base, is not favored in water.


MiRNA 26a expression in a novel panel of African American prostate cancer cell lines.
African American men have disproportionately high incidence and mortality rates of prostate cancer when compared to other ethnic groups in the United States. The identification of molecular factors that contribute to this disparity could improve diagnosis and therapeutic intervention. Therefore, the purpose of this study was to determine the miRNA 26a expression profile in novel African American and Caucasian prostate cell lines at each clinical stage of prostate cancer progression. The miR-26a expression profile was investigated using novel African American and Caucasian prostate cell lines representing each pathological stage: non-malignant, malignant, and metastatic tumors. Relative miRNA expression was determined by qRT-PCR. Our data showed a 2.25 fold increase for miR-26a in the non-malignant, a 13.3 fold increase in malignant and 2.38 fold increase in metastatic tumors, when comparing African American and Caucasian prostate cell lines of similar clinical stage and pathological grade. African American malignant prostate cancer cell lines showed the most significant fold difference in expression among all cell lines tested. Furthermore, there was a general increase in miR-26a expression toward the more aggressive cell lines in both African American and Caucasian prostate cell lines. To date, we are unaware of any studies that compare the miRNA profile at different stages of prostate cancer among two racial groups. Although a gene target for miR-26a has not been identified, our data show a possible role for miRNA regulation of gene expression in prostate cancer progression. Furthermore, this study suggests that miRNAs could possibly contribute to the aggressiveness associated in African American patients with prostate cancer.


67 th annual meeting of the American Diabetes Association.
The 67 th meeting of the American Diabetes Association was held in Chicago on 22 - 26 June. This annual meeting continues to grow in size and scope and is a unique combination of basic science and medical science but also incorporates all aspects of healthcare and pharmaceutical business relating to the treatment of diabetes. The meeting was composed of general sessions, symposia summarizing the status of various fields of study and medical practice, together with both oral and poster presentations of new, previously unpublished research. The abstracts are published in Diabetes and a collection of the information can be found online with very useful summaries from the final day. These contain personalized summaries of key findings of the meetings as seen by key researches in the field. In this Meeting Highlights article, the key take-away messages are summarized from the author's point of view.


Injectable bioactive glass/sodium alginate hydrogel with immunomodulatory and angiogenic properties for enhanced tendon healing.
Tendon healing is a complex process involving inflammation, proliferation, and remodeling, eventually achieving a state of hypocellularity and hypovascularity. Currently, few treatments can satisfactorily restore the structure and function of native tendon. Bioactive glass (BG) has been shown to possess immunomodulatory and angiogenic properties. In this study, we investigated whether an injectable hydrogel fabricated of BG and sodium alginate (SA) could be applied to enhance tenogenesis following suture repair of injured tendon. We demonstrated that BG/SA hydrogel significantly accelerated tenogenesis without inducing heterotopic ossification based on histological analysis. The therapeutic effect could attribute to increased angiogenesis and M1 to M2 phenotypic switch of macrophages within 7 days post-surgery. Morphological characterization demonstrated that BG/SA hydrogel partially reverted the pathological changes of Achilles tendon, including increased length and cross-sectional area (CSA). Finally, biomechanical test showed that BG/SA hydrogel significantly improved ultimate load, failure stress, and tensile modulus of the repaired tendon. In conclusion, administration of an injectable BG/SA hydrogel can be a novel and promising therapeutic approach to augment Achilles tendon healing in conjunction with surgical intervention.


Knee pain amongst the poor and affluent in Pakistan.
The frequency of joint symptoms was determined amongst 2022 affluent and 2210 poor adults in Karachi, Pakistan. Joint pain was significantly (P = 0.025) more common amongst the affluent (6.6%) compared with the poor (5%) and this was due to a significantly greater frequency of knee pain in the richer community (3% vs 1.8%; P = 0.008). The prevalence increased with age and was more common in females. Almost half were associated with varus deformity, suggesting the presence of associated OA in a high proportion. The overall frequency of knee pain seemed no greater than in series reported from the West. Compared with age- and sex-matched controls, body weight was significantly greater amongst those with knee pain, both amongst the affluent (P = 0.005) and the poor (P = 0.02). Control subjects were heavier in the affluent population, suggesting that the greater frequency of knee symptoms in this community was due to their relative obesity. Knee bending at prayer was most common amongst the affluent controls and may indicate that religious observance also contributed to the problem in the richer population. Squatting was a characteristic of the poor who had less knee pain than the affluent. Knee flexing could not therefore be confidently implicated. No relationship could be demonstrated between knee pain and joint laxity.


GABAergic projection from the ventral tegmental area and substantia nigra to the periaqueductal gray region and the dorsal raphe nucleus.
Previous studies have shown that neurons in the ventral tegmental area (VTA) and substantia nigra (SN) project to the ventrolateral periaqueductal gray (PAGvl) and dorsal raphe nucleus (DR). Research has also shown that stimulation of neurons in the VTA/SN elicits cardiovascular depressor responses that are mediated by a projection to the PAGvl/DR. Anatomic and physiological experiments were done in the present study to determine the neurochemical identity of the VTA/SN projection to the PAGvl/DR. Experiments were done to characterize the origin and chemical nature of this projection by combining cholera toxin B tracing with immunofluorescence for the 67K isoform of glutamic acid decarboxylase (GAD) and tyrosine hydroxylase. The PAGvl/DR region was found to receive a substantial input from neurons in the VTA, SN, and deep mesencephalic nucleus. The DR was preferentially innervated by neurons in the VTA, whereas the PAGvl was preferentially innervated by neurons in the SN. A proportion of neurons in the VTA and the reticular portion of the SN found to project to the PAGvl/DR were GAD positive. In addition, experiments were done in urethane-anesthetized rats to determine whether injections of a gamma-aminobutyric acid (GABA) antagonist in the region of the PAGvl/DR attenuated the cardiovascular depressor responses produced by glutamate stimulation of the VTA/SN. Injections of the GABA-blocking agent picrotoxin (2.5 nmol, 500 nl) into the PAGvl/DR eliminated the cardiovascular responses from stimulation of the VTA/SN (0.01 M, 50 nl). The results of the present investigation provide evidence for a GABAergic projection from the VTA/SN to the PAGvl/DR. This projection may be an important regulator of the PAGvl/DR, an area of the midbrain involved in the production of behavioral and physiological responses to pain and stress.


Use of an ophthalmic knife for middle-ear microsurgery.
Traditionally, micro-scissors, hooks, sickle knives and needles are used in the ear for the dissection and release of fibrous adhesions. The major limitation of these instruments is that they are often bent or blunt, as they are reusable. We describe an ophthalmic knife that offers precision in middle-ear microsurgery.


Antibiotic resistance of pathogens causing community-acquired pneumonia.
Community-acquired pneumonia remains an important cause of disease and death in both developed and developing countries and therefore continues to have a major medical impact. The mortality remains high despite the ready availability of potent antimicrobial agents to which the organisms are susceptible. However, management of these infections is potentially complicated by the emerging resistance of many of the common pathogens to the different classes of antibiotics that are usually prescribed. Furthermore, it is also being recognized that antibiotic resistance or treatment failures may occur not only through traditional microbial antibiotic resistance mechanisms but also through less well defined mechanisms, particularly those developed by the microbes in relation to their quorum sensing/biofilm machinery. Much recent research in this field has been focused on evaluating the clinical impact of antibiotic resistance on optimal antibiotic treatment and antimicrobial choices, as well as alternative strategies to deal with antibiotic resistance and treatment failures.


Genome Microscale Heterogeneity among Wild Potatoes Revealed by Diversity Arrays Technology Marker Sequences.
Tuber-bearing potato species possess several genes that can be exploited to improve the genetic background of the cultivated potato Solanum tuberosum. Among them, S. bulbocastanum and S. commersonii are well known for their strong resistance to environmental stresses. However, scant information is available for these species in terms of genome organization, gene function, and regulatory networks. Consequently, genomic tools to assist breeding are meager, and efficient exploitation of these species has been limited so far. In this paper, we employed the reference genome sequences from cultivated potato and tomato and a collection of sequences of 1,423 potato Diversity Arrays Technology (DArT) markers that show polymorphic representation across the genomes of S. bulbocastanum and/or S. commersonii genotypes. Our results highlighted microscale genome sequence heterogeneity that may play a significant role in functional and structural divergence between related species. Our analytical approach provides knowledge of genome structural and sequence variability that could not be detected by transcriptome and proteome approaches.


[Current theories on the mechanism of the effect of glucagon on carbohydrate metabolism].
Glucagon is considered for mechanisms of its action on the carbon metabolism, its significance in the complex polyhormonal regulation of the glucose synthesis and metabolism being studied. Glucagon exerts its effect on cells through specific receptors arranged on the plasma membrane, while its effect on the carbohydrate metabolism is mediated, mainly, by cAMP-dependent phosphorylation os some proteins participating in regulation of carbohydrate synthesis and metabolism including the proteins controlling expression of many genes. The glucagon effect on the level and efficiency of the action of insulin and some other hormones is an important link in the glucagon action on the carbohydrate metabolism.


Race, Poverty, and Foster Care Placement in the United States: Longitudinal and Cross-Sectional Perspectives.
Although the connections between race, poverty, and foster care placement seem obvious, the link has not in fact been studied extensively. To address this gap, we view poverty and placement through longitudinal and cross-sectional lenses to more accurately capture how changes in poverty rates relate to changes in placement frequency. The longitudinal study examines the relationship between poverty rate changes and changes in the placement of Black and White children between 2000 and 2015. The cross-sectional study extends the longitudinal analysis by using a richer measure of socio-ecological diversity and more recent foster care data. Using Poisson regression models, we assess the extent to which changes in race-differentiated child poverty rates are correlated with Black and White child placement frequencies and placement disparities. Regardless of whether one looks longitudinally or cross-sectionally, we find that Black children are placed in foster care more often than White children. Higher White child poverty rates are associated with substantially reduced placement differences; however, higher Black child poverty rates are associated with relatively small changes in placement disparity. Black and White child placement rates are more similar in counties with the fewest socio-ecological assets.


Light-mediated C-C sigma-bond driven crystallization of a phenalenyl radical dimer.
Polymorphism-the phenomenon that a given compound forms more than one crystalline arrangement of the molecules in the solid state- plays a crucial role in understanding organic conductors, superconductors, and magnets. We have found that solutions of a new phenalenyl radical can give rise to two (nonpolymorphic) crystalline forms depending on whether the crystallization is allowed to proceed in the presence or absence of light. In both cases the crystals take the form of black shining blades and are indistinguishable by optical microscopy. We have fully characterized these crystalline forms, and we show that they differ by the presence or absence of a C-C sigma-bond between the unpaired electrons of the parent radical. These molecular forms crystallize from the same solvent to give rise to a sigma-dimerized insulator and a monomeric radical semiconductor as dictated by the presence or absence of light.