Normal erythrocyte calpain I activity on membrane proteins under near-physiological conditions in patients with essential hypertension.
It has been reported that the equilibrium between the erythrocyte protease calpain I and its physiological inhibitor calpastatin is disrupted in patients with essential hypertension. To investigate the activity of non-purified calpain I in hemolysates against the erythrocytic membrane proteins, rather than against other substrates. Evaluation of calpain I red cell activity upon its own physiological substrates in hypertensive patients, in a near-physiological environment. LIM-23 and LIM-40 of Hospital das Clinicas of the Faculty of Medicine of USP. Patients with moderate primary hypertension over 21 years of age who were given amlodipine (n:10) and captopril (n:10) for 8 weeks, plus normal controls (n:10). Red cell membrane proteins were incubated with and without protease inhibitors and with and without calcium chloride and underwent polyacrylamide gel electrophoresis. Digestion of bands 2.1 and 4.1 was observed, indicating calpain I activity. No statistical differences regarding bands 2.1 and 4.1 were observed before treatment, between the controls and the hypertensive patients, either in ghosts prepared without calcium or with increasing concentrations of calcium. Nor were statistical differences observed after treatment, between the controls and the patients treated with amlodipine and captopril, or between the patients before and after treatment with both drugs. The final activity of non-purified calpain I upon its own physiological substrate, which was the approach utilized in this study, may more adequately reflect what happens in red cells. Under such conditions no imbalance favoring calpain I activity increase was observed. The protective factor provided by calpastatin against calpain I activity may diminish under hypertension.


Causal association between major depressive disorder and coronary heart disease: a two-sample bidirectional mendelian randomization study.
Major depressive disorder (MDD) is a highly heterogeneous mental illness and a major public health problem worldwide. A large number of observational studies have demonstrated a clear association between MDD and coronary heart disease (CHD), and some studies have even suggested that the relationship is bidirectional. However, it was unknown whether any causal relationship existed between them and whether causality was bidirectional in such an instance. Thus, we aimed to determine whether there is a bidirectional causal relationship between major depressive disorders and coronary heart disease. Our two-sample Bidirectional Mendelian Randomization Study consisted of two parts: forward MR analysis regarded MDD as exposure and CHD as the outcome, and reverse MR analysis considered CHD as exposure and MDD as the outcome. Summary data on MDD and CHD were obtained from the IEU Open GWAS database. After screening criteria(P < [Formula: see text]), 47 MDD-associated SNPs and 39 CHD-associated SNPs were identified. The inverse-variance weighted (IVW) method, ME-Egger regression, and weighted median method were used to estimate causality. In addition, sensitivity methods, including the heterogeneity test, horizontal pleiotropy test, and leave-one-out method, were applied to ensure the robustness of causal estimation. Based on the MR-Egger regression intercept test results, there did not appear to be any horizontal pleiotropy in this study (MDD: intercept = -0.0000376, P = 0.9996; CHD: intercept = -0.0002698, P = 0.920). Accordingly, IVW results suggested consistent estimates of causal effect values. The results showed that people with MDD increased the risk of CHD by 14.7% compared with those without MDD (OR = 1.147, 95%CI: 1.045-1.249, P = 0.009). But there was no direct evidence that CHD would increase the risk of MDD(OR = 1.008, 95%CI: 0.985-1.031, P = 0.490). The heterogeneity test and funnel plot showed no heterogeneity in 47 SNPs of MDD (Q = 42.28, [Formula: see text]=0, P = 0.629), but there was heterogeneity in 39 SNPs of CHD (Q = 62.48, [Formula: see text]=39.18%, P = 0.007). The leave-one-out method failed to identify instances where a single SNP was either biased toward or dependent on the causation. Our study supports a one-way causal relationship between MDD and CHD, but there is no bidirectional causal relationship. MDD increases the risk of CHD, but there is no evidence that CHD increases the risk of MDD. Therefore, the influence of psychological factors should also be considered in the prevention and treatment of CHD. For MDD patients, it is necessary to prevent cardiovascular diseases.


RNA-based technologies for insect control in plant production.
RNA interference (RNAi) is a biological process in which small RNA (sRNA) molecules sequence-specifically silence gene expression at the transcriptional or post-transcriptional level, either by directing inhibitory chromatin modifications or by decreasing the stability or translation potential of the targeted mRNA. The trigger for gene silencing is double-stranded RNA (dsRNA) generated from an endogenous genomic locus or a foreign source, such as a transgene or virus. The process of gene silencing can be exploited in agriculture to control plant diseases and pests. Of the pests that impact crop yield (including nematodes, arthropods, rodents, snails, slugs and birds), insects constitute the largest and most diverse group. Here, we review the "pros" and "cons" of using RNAi technology mediated by dsRNA-expressing transgenic plants (host-induced gene silencing, HIGS) or direct application of chemically synthesized dsRNA to control plant-damaging insects. Rapid progress in elucidating RNAi mechanisms has led to the first commercial products on the market. Given the high potential of RNAi strategies, their use in agriculture, horticulture, and forestry will likely be extensive in the future. However, further studies are needed to improve the efficacy of RNAi-based plant protection strategies and to assess their associated safety risks.


Safety and efficacy of trigonal BTX-A injections for children with neurological detrusor overactivity secondary to spinal cord injury.
To evaluate the efficacy and safety of trigonal botulinum toxin A (BTX-A) injections for children with neurological detrusor overactivity (NDO) secondary to spinal cord injury (SCI). From February 2012 to December 2018, children with NDO secondary to SCI were enrolled. All patients received 200U BTX-A intradetrusor injections including the trigone. Videourodynamic study was performed at baseline and 12 weeks after injection. The primary outcome measures were the presence of vesicoureteral reflux and standardized urodynamic measures. Secondary outcomes included incontinence quality of life questionnaire (I-QoL), voiding volume, urinary incontinence episodes, and complete dryness. A total of 33 pediatric inpatients (28 male and 5 female) completed the study. No one developed VUR at week 0 or week 12. At the first instance of NDO, maximum detrusor pressure (PdetmaxFNDO) and NDO duration were reduced by 29.8% and 31.8%, respectively, whereas NDO volume (VFNDO) increased by 50.5%,12 weeks after injection. Mean urinary incontinence episodes were reduced by 31.7%, whereas voiding volume and I-QOL were increased by 52.9% and 23.3%. 3 patients reported mild transient hematuria during the first week after injection. Our results suggest that the use of bladder-trigone-including intradetrusor BTX-A injection does not induce VUR, and is safe and effective in children with NDO secondary to SCI. A prospective self-controlled trial LEVEL OF EVIDENCE: Level II.


In vitro hatching of the tapeworm Moniezia expansa (Cestoda: Anoplocephalidae) and some properties of the egg membranes.
In vitro studies revealed that the hatching of oncospheres of Moniezia expansa requires the mechanical breakage of the eggshell and subshell membrane and enzymic digestion of the pyriform apparatus. Removal of the outer two egg membranes elicits the activation of most oncospheres. Between pH 5.0-7.8, there is no significant difference in numbers of oncospheres activated by eggshell removal and the addition of sodium bicarbonate has no effect. Solutions of more extreme pH values (2.0 and 10.0) are harmful and render oncospheres immobile. The subshell membrane forms a barrier to the passage of water in an osmotic gradient and to several molecular and ionic substances. Between the eggshell and subshell membrane is a layer of droplets which have a strong affinity for Sudan stains and which are partially removed by lipase. The eggshell is resistant to a variety of proteolytic enzymes, amylases and lipase. The pyriform apparatus is digested by chymotrypsin and pepsin, though not by trypsin. Both eggshell and pyriform apparatus are dissolved by solutions of sodium sulphide and sodium hypochlorite, indicating that their structures are stabilized by disulphide bonds and other covalent linkages.


Isolation and identification of a small molecular weight inhibitor of cyclic nucleotide phosphodiesterase from bovine brain.
In a search for endogenous regulators for cyclic nucleotide phosphodiesterase (3':5'-cyclic-AMP 5'-nucleotidohydrolase, EC 126.96.36.199), we found that the ultrafiltrate of bovine brain homogenate contained a cyclic nucleotide phosphodiesterase inhibitor. The inhibitor-containing fraction was further purified by ion-exchange column chromatography and gel filtration chromatography. The purified inhibitor was found to be a small molecular weight compound which had a maximum absorption at 248 nm. This compound was identified by thin-layer chromatography and high-pressure liquid chromatography as hypoxanthine. We suggest that hypoxanthine may serve as an endogenous regulator for the hydrolysis of cyclic nucleotide by cyclic nucleotide phosphodiesterase.


Nipple-sparing mastectomy with different approaches: surgical incisions, complications, and cosmetic results. Preliminary results of 100 consecutive patients at a single center.
Nipple- and areola-sparing mastectomy is a novel surgical approach that preserves the nipple-areolar complex. Patients with moderate and/or severe breast ptosis are usually not eligible for this surgical approach. In this study, we aimed to demonstrate the feasibility of nonconventional surgical approaches for nipple-sparing mastectomy. One hundred consecutive patients diagnosed with primary breast cancer (BC) were enrolled in this study. Clinical and pathological data such as body mass index, smoking status, breast ptosis, complications, and aesthetic satisfaction (Breast-Q test) were collected. According to different types of breast ptosis, surgical procedures were classified as (a) hemi-periareolar, (b) round block, (c) vertical pattern, and (d) wise pattern skin incisions. We performed statistical analysis to assess the correlation with complications, degree of ptosis, and breast-Q scores. Among the 117 surgical procedures performed in 100 patients with BC, no significant associations are verified considering clinical and pathological data, complications, pre- and postsurgery satisfactions, and other parameters. Different surgical approaches represent the evolution of "classic" nipple-sparing mastectomy, thus meeting the cosmetic and oncological results. These procedures are safe and also indicated in cases conventionally considered as not eligible for nipple-areola preservation.


Studies on the role of histamine and 5-hydroxytryptamine in immunity against the nematode Trichostrongylus colubriformis. V. Changes in amine levels in the intestine following infection of guinea pigs of different immune status.
A temporal relationship was established between the onset of expulsion of the parasitic nematode Trichostrongylus colubriformis from the intestine of both vaccinated and adoptively immunized guinea pigs with a sudden increase in small intestinal mucosal histamine to two or three times pre-infection levels. However, in guinea pigs whose capacity to expel T. colubriformis was inhibited by treatment with anti-lymphocyte serum, mucosal histamine remained at pre-infection levels. The results support previous findings which suggest an important role for histamine in the effector mechanism of the immune response of guinea pigs against T. colubriformis.


Finding related functional neuroimaging volumes.
We describe a content-based image retrieval technique for finding related functional neuroimaging experiments by voxelization of sets of stereotactic coordinates in Talairach space, comparing the volumes and reporting related volumes in a sorted list. Voxelization is accomplished by convolving each coordinate with a Gaussian kernel. The scheme allows us to compare experiments represented as either lists of coordinates or volumes, and we introduce alternative entrances to databases by image-based indices constructed via novelty measures and singular value decomposition.


Expression of importin-α isoforms in human nasal mucosa: implication for adaptation of avian influenza A viruses to human host.
Transportation into the host cell nucleus is crucial for replication and transcription of influenza virus. The classical nuclear import is regulated by specific cellular factor, importin-α. Seven isoforms of importin-α have been identified in human. The preference of importin-α3 of avian influenza virus and -α7 isoform of human strains during replication in human cells was previously identified. In addition, both avian and human influenza viruses were shown to use importin-α1 isoform for their replication. The mRNA levels of importin-α1, -α3, and -α7 isoforms in human respiratory tract was determined by real-time RT-PCR. The results indicate that mRNA level of importin-α7 was significantly higher than that of importin-α1 (p-value < 0.0001) and importin-α3 (p-value < 0.0001) isoforms in human nasal mucosa while importin-α1 was detected as the highest expression importin-α isoform in lung tissues. These results may explain the preference of importin-α7 isoforms in seasonal influenza viruses in human upper respiratory tract and may suggest a selective pressure toward importin-α7 in human respiratory tract infection of an avian virus.


Mannan-binding lectin deficiency - Good news, bad news, doesn't matter?
Mannan-binding lectin (MBL) deficiency has been classified as a commonly occurring immune disorder, affecting approximately 30% of the human population. MBL, being part of the innate immune system, supports the recognition of infectious pathogens by binding to carbohydrate moieties expressed on microorganisms and activates the lectin pathway of the complement system. MBL2 gene polymorphisms are associated with quantitative and qualitative MBL abnormalities in the serum. The clinical impact of MBL deficiency and its association to a wide variety of diseases has been extensively studied. The picture is puzzling as the studies suggest a detrimental or beneficial or no impact of low or high MBL serum levels on disease susceptibility. In this review we attempt to extract what is relevant from the literature and address controversial issues. We finally suggest that a comprehensive understanding of the role of MBL in human diseases requires considering its context-dependency.


Randomized Controlled Trial of Talactoferrin Oral Solution in Preterm Infants.
To evaluate the safety and explore the efficacy of recombinant human lactoferrin (talactoferrin [TLf]) to reduce infection. We conducted a randomized, double blind, placebo-controlled trial in infants with birth weight of 750-1500 g. Infants received enteral TLf (n = 60) or placebo (n = 60) on days 1 through 28 of life; the TLf dose was 150 mg/kg every 12 hours. Primary outcomes were bacteremia, pneumonia, urinary tract infection, meningitis, and necrotizing enterocolitis (NEC). Secondary outcomes were sepsis syndrome and suspected NEC. We recorded clinical, laboratory, and radiologic findings, along with diseases and adverse events, in a database used for statistical analyses. Demographic data were similar in the 2 groups of infants. We attributed no enteral or organ-specific adverse events to TLf. There were 2 deaths in the TLf group (1 each due to posterior fossa hemorrhage and postdischarge sudden infant death), and 1 death in the placebo group, due to NEC. The rate of hospital-acquired infections was 50% lower in the TLf group compared with the placebo group (P < .04), including fewer blood or line infections, urinary tract infections, and pneumonia. Fourteen infants in the TLf group weighing <1 kg at birth had no gram-negative infections, compared with only 3 of 14 such infants in the placebo group. Noninfectious outcomes were not statistically significantly different between the 2 groups, and there were no between-group differences in growth or neurodevelopment over a 1-year posthospitalization period. We found no clinical or laboratory toxicity and a trend toward less infectious morbidity in the infants treated with TLf. ClinicalTrials.gov: NCT00854633.


Smallpox vaccination and vaccine hesitancy in the Kingdom of the Two Sicilies (1801) and the great modernity of Ferdinand IV of Bourbon: a glimpse of the past in the era of the SARS-COV-2 (COVID-19) pandemic.
The current health emergency caused by COVID-19 disease shows several correspondences with well-known epidemics of the past. The knowledge of their management and overcoming could give us useful tools to face the present COVID-19 pandemic and future epidemics. On 1 March 1801, the first smallpox vaccinations were carried out in Palermo, and a few weeks later, the vaccine was also administered in Naples and the various provinces of the Kingdom. We aim to study the mass vaccination programme initiated by the Bourbon king Ferdinand IV that was the first large-scale campaign to be conducted in Italy and one of the first in Europe. The authors searched and examined historical testimony and different aspects linked to the public health issues on vaccination. It is a topical topic in the current period with the COVID pandemic. Albeit with the due differences determined by the passage of time and by the scientific and cultural advances of modern society, this testimony from the past can provide us with food for thought regarding how to face the present COVID-19 pandemic and to prepare for the future. Indeed, it shows us how the terrible smallpox epidemic was handled and finally overcome, thanks to vaccination.


Combining ventral buccal mucosal graft onlay and dorsal full thickness skin graft inlay decreases failure rates in long bulbar strictures (≥6 cm).
To evaluate the efficacy of augmenting ventral onlay buccal urethroplasty using a using a double-sided graft technique by adding a second, full-thickness skin graft (FTSG) dorsally in long strictures ≥4 cm, we hypothesized that a double-sided graft technique would improve surgical outcomes over buccal mucosal graft (BMG) ventral onlay urethroplasty alone. Retrospective chart review was performed comparing 15 patients who had undergone double-sided BMG + FTSG urethroplasty for long strictures ≥4 cm to a cohort group of 115 patients who had undergone BMG onlay urethroplasty for strictures of similar length. Comparisons of age, stricture lengths, time-to-failure, follow-up intervals, and failure rates were analyzed. Further analysis included age and stricture length matched control comparisons between BMG + FTSG patients to BMG only patients. Mean patient age, stricture length, and time to stricture recurrence were 44 years, 5.1 cm, and 10 months, respectively, in the BMG group. For the BMG + FTSG group, mean age was 52 years, stricture length 5.9 cm, and time to recurrence was 9 months. Overall, the BMG group had a decreased failure rate; 17% compared to 21%. Further analysis showed BMG ventral onlay had decreased failure rates for strictures <6 cm; 16% compared to 33%. However, for strictures ≥6 cm, the BMG + FTSG had a 0% failure rate compared to 24% in the BMG ventral onlay group (P = .005). Combined BMG and FTSG urethroplasty for very long bulbar urethral strictures ≥6 cm improve success rates compared to using BMG urethroplasty alone.


BacMam production of active recombinant lecithin-cholesterol acyltransferase: Expression, purification and characterization.
Lecithin-cholesterol acyltransferase (LCAT) is a key enzyme in the esterification of cholesterol and its subsequent incorporation into the core of high density lipoprotein (HDL) particles. It is also involved in reverse cholesterol transport (RCT), the mechanism by which cholesterol is removed from peripheral cells and transported to the liver for excretion. These processes are involved in the development of atherosclerosis and coronary heart disease (CHD) and may have therapeutic implications. This work describes the use of baculovirus as a transducing vector to express LCAT in mammalian cells, expression of the recombinant protein as a high-mannose glycoform suitable for deglycosylation by Endo H and its purification to homogeneity and characterization. The importance of producing underglycosylated forms of secreted glycoproteins to obtain high-resolution crystal structures is discussed.


An endogenous TNF-alpha antagonist induced by splice-switching oligonucleotides reduces inflammation in hepatitis and arthritis mouse models.
Tumor necrosis factor-alpha (TNF-alpha) is a key mediator of inflammatory diseases, including rheumatoid arthritis (RA), and anti-TNF-alpha drugs such as etanercept are effective treatments. Splice-switching oligonucleotides (SSOs) are a new class of drugs designed to induce therapeutically favorable splice variants of targeted genes. In this work, we used locked nucleic acid (LNA)-based SSOs to modulate splicing of TNF receptor 2 (TNFR2) pre-mRNA. The SSO induced skipping of TNFR2 exon 7, which codes the transmembrane domain (TM), switching endogenous expression from the membrane-bound, functional form to a soluble, secreted form (Delta7TNFR2). This decoy receptor protein accumulated in the circulation of treated mice, antagonized TNF-alpha, and altered disease in two mouse models: TNF-alpha-induced hepatitis and collagen-induced arthritis (CIA). This is the first report of upregulation of the endogenous, circulating TNF-alpha antagonist by oligonucleotide-induced splicing modulation.


Two developmentally regulated messenger RNAs differing in their coding region may exist for the myelin-associated glycoprotein.
Mouse and rat brain RNA were translated in vitro in a rabbit reticulocyte lysate translation system, and myelin-associated glycoprotein-related polypeptides were immunoprecipitated. Two products of Mr = 72,000 (p72MAG) and Mr = 67,000 (p67MAG) were specifically immunoprecipitated. These polypeptides were characterized as follows: 1) exogenously added purified myelin-associated glycoprotein (MAG) competitively eliminates their immunoprecipitation; 2) peptide maps obtained from them are very similar; 3) they are both destined to become glycoproteins, as assayed by insertion into dog pancreas microsomes and their subsequent sensitivity to endoglycosidase H. Translatable mRNA for the two polypeptides have different developmental expressions. p72MAG mRNA appears at an earlier age than p67MAG mRNA and remains the dominant MAG mRNA species throughout the period of rapid myelination. As the rate of myelination decreases, p67MAG mRNA becomes the dominant MAG mRNA species. Finally, two endoglycosidase H-sensitive polypeptides were specifically immunoprecipitated from mouse brain microsomes. Therefore, there exist two MAG proteins that differ slightly in their polypeptide structure and that are developmentally regulated. The possibility that the two polypeptides are synthesized de novo from two coordinately regulated mRNAs that differ in their coding region is discussed.


Integrating Plasmonic Supercrystals in Microfluidics for Ultrasensitive, Label-Free, and Selective Surface-Enhanced Raman Spectroscopy Detection.
Surface-enhanced Raman spectroscopy (SERS) microfluidic chips for label-free and ultrasensitive detection are fabricated by integrating a plasmonic supercrystal within microfluidic channels. This plasmonic platform allows the uniform infiltration of the analytes within the supercrystal, reaching the so-called hot spots. Moreover, state-of-the-art simulations performed using large-scale supercrystal models demonstrate that the excellent SERS response is due to the hierarchical nanoparticle organization, the interparticle separation (IPS), and the presence of supercrystal defects. Proof-of-concept experiments confirm the outstanding performance of the microfluidic chips for the ultradetection of (bio)molecules with no metal affinity. In fact, a limit of detection (LOD) as low as 10-19 M was reached for crystal violet. The SERS microfluidic chips show excellent sensitivity in the direct analysis of pyocyanin secreted by Pseudomonas aeruginosa grown in a liquid culture medium. Finally, the further integration of a silica-based column in the plasmonic microchip provides charge-selective SERS capabilities as demonstrated for a mixture of positively and negatively charged molecules.


Examining Hearts Containing Left Ventricular Assist Devices at Necropsy.
There are no publications describing hearts at necropsy containing left ventricular assist devices (LVADs). The purpose was to study the relation of the LVAD cannula to the left ventricular (LV) cavity and wall. We studied the hearts at necropsy of 15 adults who had an LVAD inserted from 4 to 1,423 days (median 60) earlier. In 13 patients, the cannula had been inserted at an angle to the major longitudinal axis of the LV chamber, and in 11 patients, the orifice margin of the cannulas contacted the LV mural endocardium. In 3 patients, the LVAD cannula was inserted into the posterior wall, and, in another into the anterior wall. In the remaining 11 patients, the cannula had been inserted into the LV apex. Despite the insertion of the cannulas into the LV apex, the direction of the insertion was not into the longitudinal axis of the LV cavity in 9 patients. These unusual insertions in some patients may have altered flow into the orifice of the cannula; in others, based on their long postoperative survival, physiologic consequences were almost certainly absent. The presence of considerable quantities of subepicardial adipose tissue and pericardial adhesions from previous cardiac procedures (mainly coronary bypass) potentially interfered with achieving proper alignment of the LVAD cannula during its insertion. Misalignment of the cannulas of the LVAD in the LV cavity appears to be rather frequent.


A strategic assessment of the reproductive health and responsible parenthood programme of Buenos Aires, Argentina.
Since 1991, Argentina has had provincial reproductive health laws, a far-reaching national programme and strong public consensus in support of reproductive health policies. Nevertheless, the challenges of strengthening public services, increasing the number of programme sites and resisting conservative attacks remain. This article describes an assessment of the reproductive health programme of the city of Buenos Aires, passed in 2000, whose objectives are to prevent unwanted pregnancies and sexually transmitted diseases/HIV and to train health personnel. The programme operates in every public hospital and primary health care centre in the city. The assessment was conducted jointly by the Ombudsperson's Office of Buenos Aires and the Centre for the Study of State and Society (CEDES). Hormonal contraceptives, IUDs and male condoms were mostly available, but emergency contraception, female condoms and other barrier methods were not Some health professionals and service users were knowledgeable about the new laws and the reproductive rights recognised under the law. Over 90% were satisfied with quality of care in service delivery but many professionals described excessive workloads, deficient infrastructure, and shortages of supplies and staff. Wanting help to obtain a tubal ligation was the most frequent reason for the claims lodged with the Ombudsperson's Office, followed by HIV, quality of care, and abortion. Information and training for both health care providers and women's and human rights NGOs was carried out.


Trends in multiplicity of Plasmodium falciparum infections among asymptomatic residents in the middle belt of Ghana.
Malaria is the most important cause of mortality and morbidity in children living in the Kintampo districts in the middle part of Ghana. This study has investigated the multiplicity of infection (MOI) within asymptomatic residents of the Kintampo districts, and the influence of age and seasonality on MOI, by studying the distribution of the polymorphic Plasmodium falciparum antigen merozoite surface protein 2 (MSP2). DNA was extracted from an asymptomatic cohort of children and adults infected with P. falciparum during the period November 2003 to October 2004. Polymerase chain reaction was carried out and multiplicity of infection (MOI) was determined. Children under 10 years of age had an average MOI of 2.3 while adults 18 years and above had an average MOI of 1.4. Children below five years had high and low average MOIs of 2.8 in the March/April survey and 0.9 in the May/June survey respectively. A similar trend in the monthly distribution of MOI was observed for the entire cohort. IC/3D7 strains outnumbered the FC27 strains throughout the year by a ratio of about 4:1 with the difference between the prevalence of the two strains being least marked in the March/April survey, at the beginning of the rainy season. MOI was not linked to the level of malaria transmission as measured by the entomological inoculation rate. The impact of interventions, introduced since this baseline study was carried out on the parasite diversity of asymptomatic residents will be the subject of further investigations.


Anatomy that must be taught to a medical undergraduate: an interview-based survey in an Indian medical school.
Confusion still exists about the anatomy to be taught to the medical undergraduate. We did an interview-based survey at the Melaka Manipal Medical College in Manipal, India, to try to evaluate the quantum of anatomy that should be taught to the medical undergraduate. The results suggest that excluding trivia and making anatomy more clinically oriented would be advantageous. A hybrid approach to anatomy including both problem-based learning and discipline-based curricula would be a better option than the regional or systemic approaches.


Expression of the multidrug resistance-associated protein (MRP) gene in non-small-cell lung cancer.
We examined the levels of expression of the multidrug resistance-associated protein (MRP) gene quantified by Northern blot analysis in comparison with those of the MDR1 gene determined by reverse transcription-polymerase chain reaction (RT-PCR) in 104 non-small-cell lung cancer (NSCLC) specimens [59 adenocarcinoma (Ad), 40 squamous cell carcinoma (Sq), four large cell carcinoma (La) and one adeno-squamous carcinoma (AdSq)]. Thirty-three (31.7%) of the 104 NSCLC expressed the MRP gene at various levels. The NSCLC showing high (++) levels of MRP gene expression (19 out of 33, 57.6%) were predominantly squamous cell carcinomas (Ad, 5; Sq, 13; La, 1) (P < 0.05). Six of the eight NSCLCs expressing high levels of MRP mRNA and no MDR1 (MRP ++, MDR1-) were squamous cell carcinomas. Sixty-one of the 104 NSCLC patients received chemotherapy with MRP-related anti-cancer drugs [vindesine (VDS) and etoposide (VP-16)]. Twenty-three patients (37.7%) with tumour expressing high or moderate levels of MRP showed significantly worse prognoses than those with non- or low-MRP-expressing tumours (P < 0.05). These results suggest that the level of MRP gene expression is related to the histopathology and prognosis of NSCLC.


Primary chemotherapy for intracranial nongerminomatous germ cell tumors: results of the second international CNS germ cell study group protocol.
The optimum therapy for intracranial nongerminomatous germ cell tumors (NGGCT) remains controversial. The primary objective of this study was to determine whether intensive cisplatin and cyclophosphamide-based combination chemotherapy was effective in patients with intracranial NGGCT. Twenty patients were enrolled, aged 5 to 41 years (median, 13 years). Initial therapy included two courses of Regimen A (cisplatin, etoposide, cyclophosphamide, and bleomycin). Patients achieving a complete remission (CR) then received two courses of Regimen B (carboplatin, etoposide, and bleomycin). Those in CR after four courses of treatment received one additional course of Regimen A and Regimen B, while those not in CR after four treatment courses underwent second-look surgery and/or irradiation. Sixteen of 17 patients assessable for response after two courses of treatment achieved a CR or partial response (CR + partial response, 0.94; 95% CI, 0.73 to 1.0). With a median follow-up of 6.3 years, 14 of 20 patients are alive without disease; eight patients were without relapse or progression, of whom three received local irradiation in first complete remission in violation of protocol, and six patients were in durable second or third complete remission after further chemotherapy and/or irradiation. The 5-year overall survival and event-free survival were 0.75 (95% CI, 0.56 to 0.94) and 0.36 (95% CI, 0.13 to 0.59), respectively. Intensive chemotherapy was effective in one-third of patients in this study. Salvage therapy, including irradiation, was feasible in patients with recurrent disease.


Meta-analysis: coeliac disease and the risk of all-cause mortality, any malignancy and lymphoid malignancy.
Coeliac disease has been associated with an increased risk of mortality and malignancy. However, the strength of this association is conflicting among different studies. To perform a systematic review and quantitative meta-analysis to determine the risk of all-cause mortality, any malignancy and lymphoid malignancy in coeliac disease patients. Four electronic databases (Medline, PubMed, Embase and Current Contents Connect) were searched to 4 January 2012, with no language restrictions. From 8698 citations identified, a total of 17 studies met our inclusion criteria. The all-cause mortality meta-analysis showed an increased risk for all-cause mortality in coeliac patients [odds ratio (OR) 1.24; 95% confidence interval (CI) 1.19-1.30]. A subgroup analysis showed that patients identified by positive serology alone were also at an increased risk of all-cause mortality (OR 1.16; 95% CI 1.02-1.31). The non-Hodgkin lymphoma (NHL) meta-analysis showed an increased risk for NHL in coeliac patients (OR 2.61; 95% CI 2.04-3.33). A subgroup analysis showed that patients identified by positive serology alone were also at an increased risk of NHL (OR 2.55; 95% CI 1.02-6.36). The T-cell non-Hodgkin lymphoma (TNHL) meta-analysis showed an increased risk of TNHL (OR 15.84; 95% CI 7.85-31.94). The any malignancy meta-analysis showed no increased risk (OR 1.07; 95% CI 0.89-1.29). Patients with coeliac disease are at an increased risk of mortality and non-Hodgkin lymphoma, particularly T-cell non-Hodgkin lymphoma; they do not have an increased risk of any malignancy overall. Serologically defined patients with coeliac disease have an elevated risk of mortality and non-Hodgkin lymphoma.


Apolipoprotein A-I synthesis and secretion by cultured human intestinal mucosa.
Cultured human duodenojejunal mucosa was used to study the synthesis and secretion of apoprotein A-I (apoA-I), the major protein constituent of plasma high density lipoproteins. ApoA-I, measured by radioimmunoassay, was secreted continuously into the culture medium over a period of 24 hr. The highest rate was found in the first two hours (34.6 +/- 3.0 ng/mg tissue X hr, mean +/- SE, n = 24). Secretion rate decreased with incubation time, while changes of culture medium increased the rate. ApoA-I secretion was enhanced three- to fourfold by micellar lipid solution and was inhibited by puromycin, ApoA-I synthesis was confirmed by incorporation of 14C-leucine into this apoprotein. The specific activity of apoA-I in medium ultracentrifugal fractions (d less than 1.019 and d = 1.063--1.21 g/ml) was calculated from quantitative data obtained by radioimmunoassay and radioassay of apoA-I separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and found to be about 12 mCi/mmol. These observations indicate that cultured human intestinal mucosa is capable of secreting newly synthesized apoA-I. Organ culture may thus serve as a tool for studying the regulating mechanisms for the synthesis and secretion of this important apoprotein.


Purification and characterization of naturally occurring benzodiazepine receptor ligands in rat and human brain.
Chemicals that are active at the benzodiazepine receptor (endozepines) are naturally present in the CNS. These substances are present in tissue from humans and animals and in plants and fungi. Using selective extraction protocols, HPLC purification, receptor binding displacement studies, and selective anti-benzodiazepine antibodies, we have identified six or seven peaks of endozepines in rat and human brain. All material could competitively displace [3H]flunitrazepam binding to cerebellar benzodiazepine binding sites. Two peaks also competitively displaced Ro 5-4864 binding to the mitochondrial benzodiazepine binding site. Total amounts of brain endozepines were estimated to be present in potentially physiological concentrations, based on their ability to displace [3H]flunitrazepam binding. Although endozepine peaks 1 and 2 had HPLC retention profiles similar to those of nordiazepam and diazepam, respectively, gas chromatography-mass spectrometry as well as high-performance TLC revealed biologically insignificant amounts of diazepam (less than 0.02 pg/g) and nordiazepam (less than 0.02 pg/g) in the purified material. Electrophysiologically, some purified endozepines positively modulated gamma-aminobutyric acid (GABA) action on Cl- conductance, monitored in patch-clamped cultured cortical neurons or in mammalian cells transfected with cDNA encoding various GABAA receptor subunits. These studies demonstrate that mammalian brains contain endozepines that could serve as potent endogenous positive allosteric modulators of GABAA receptors.


Acute ST-elevation myocardial infarction (STEMI) in a young woman with unknown mitral stenosis and atrial fibrillation: Case report.
Chronic rheumatic heart disease is the most common cause of mitral stenosis. It remains a major public health problem. In almost half of the cases, paroxysmal or chronic atrial fibrillation occurs during the evolution of mitral stenosis, thereby exposing to an increased risk of thrombo-embolic events.Whereas the most frequent site for embolism is the cerebral circulation, any organ may be involved, especially the coronary circulation, resulting in a myocardial infarction (MI). Here, we report a rare case of a 50-year-old patient, with no risk factors for cardiovascular disease, presenting an acute ST-elevation myocardial infarction (STEMI) as initial presentation of unknown mitral stenosis with atrial fibrillation and strongly suggesting an embolic origin. The diagnosis was made based on the national cerebral and cardiovascular center (NCVC) criteria for the clinical diagnosis of coronary artery embolism (CE). Coronary angiography showed a distal thrombus in the right coronary artery that has been medically treated. The outcome was favorable and the patient was referred after that for mitral valve replacement.


The Impact of Sexual Satisfaction, Functioning, and Perceived Contraceptive Effects on Sex Life on IUD and Implant Continuation at 1 Year.
Contraceptives improve women's lives and public health, but many women discontinue their contraceptive method owing to dissatisfaction. An underexamined aspect of contraceptive discontinuation is sexual acceptability, or how contraception affects sexual experiences. Investigators' aims were two-fold: 1) to document changes in multiple domains of women's sexual experiences with their intrauterine device (IUD) or contraceptive implant over time and 2) to examine whether these sexuality factors were associated with method continuation at 12 months. We enrolled 200 eligible family planning clients and collected data at baseline and at 1, 3, 6, and 12 months. Sexual acceptability measures included the Female Sexual Function Index-6, the New Sexual Satisfaction Scale, and participants' perceptions of whether their contraceptive method had had a neutral, positive, or negative effect on their sex life. Survival analysis and Cox regression with time-varying covariates related sexuality measures to method continuation over time while controlling for other relevant factors. Among 193 women who received an IUD or implant, 20% selected the copper IUD, 46% the levonorgestrel IUD, and 34% the etonogestrel implant. Ten percent discontinued their method during the year. Although changes in Female Sexual Function Index-6 and New Sexual Satisfaction Scale scores were not associated with discontinuation, individuals who perceived that their method detracted from their sexual experience had significantly higher removal rates than those who reported no sexual changes or positive sexual changes (adjusted hazard ratio, 8.04; 95% CI, 1.53-42.24), even when controlling for method type, bleeding changes, and a variety of covariates and controls. Although limited by the small sample of discontinuers, we found that women's perceptions of how their method affects their sex life were associated with contraceptive continuation over time. Sexual acceptability should receive more attention in both contraceptive research and counseling.


Impact of different magnification levels on visual caries detection with ICDAS.
The aim of this in vitro study was to examine the effect of different levels of magnification on the accuracy and reliability of visual caries detection using ICDAS criteria. Occlusal surfaces of 100 extracted molars were assessed by 14 examiners (3rd and the 4th year dental students and dentists) using no magnification aids, a 2.5× Galilean loupe, a 4.5× Keplerian loupe, or a surgical microscope with 10× magnification. The assessments were repeated on a different day. Sensitivity, specificity, AUC and reliabilities were calculated according to the gold standard of histology. We found that with increasing magnification, the number of surfaces rated as "sound" (ICDAS code 0) decreased, while the number of surfaces with a localized enamel breakdown (ICDAS code 3) increased. While the sensitivities increased, the values of the specificities decreased to an unacceptably low level irrespective of the clinical experience of the examiners. ICDAS seems to be optimized for natural vision up to 2.0× magnification and not for high magnifications. The use of powerful magnification in visual caries detection involves the risk of unnecessary and premature invasive treatment. This paper discusses when it does and does not make sense to use magnification devices for visual caries detection using ICDAS criteria. Strong magnifications should be refrained from for this purpose.


Thienothiopyransulfonamides as complexing agents for the preparation of dual carbonic anhydrase inhibitors.
Co(II); Zn(II) and Cu(II) complexes of two new sulfonamide carbonic anhydrase (CA) inhibitors, derivatives of thienothiopyran-2-sulfonamide, were prepared and characterized by analytic, spectroscopic, magnetic and conductimetric measurements. The new complexes are more potent CA inhibitors than the parent sulfonamides, with IC(50) values around 0.1 nM, against isozyme CA II.


1,2,4-Triazolyl octahydropyrrolo[2,3-b]pyrroles: A new series of potent and selective dopamine D3 receptor antagonists.
A novel series of 1,2,4-triazolyl octahydropyrrolo[2,3-b]pyrroles showing high affinity and selectivity at the DA D3 receptor is reported here. Compounds endowed with high selectivity over the hERG channel were identified and their pharmacokinetic properties thoroughly analyzed. A few derivatives with appropriate developability characteristics were selected for further studies and progression along the screening cascade. In particular, derivative 60a, (DA D3 pKi=8.4, DA D2 pKi=6.0 and hERG fpKi=5.2) showed a balanced profile and further refinements are in progress around this molecule.


Public Health Measures and Prevention of Alcohol-Associated Liver Disease.
Hazardous alcohol consumption causes approximately 4% of deaths globally, constituting one of the leading risk factors for the burden of the disease worldwide. Alcohol has several health consequences, such as alcohol-associated liver disease, hepatocellular carcinoma, nonliver neoplasms, physical injury, cardiac disease, and psychiatric disorders. Alcohol misuse significantly affects workforce productivity, with elevated direct and indirect economic costs. Due to the high impact of alcohol consumption on the population, public health has led to the development of a range of strategies to reduce its harmful effects. Regulatory public health policies (PHP) for alcohol can exist at the global, regional, international, national, or subnational levels. Effective strategies incorporate a multilevel, multicomponent approach, targeting multiple determinants of drinking and alcohol-related harms. The World Health Organization categorizes the PHP into eight categories: national plan to fight the harmful consequences of alcohol, national license and production and selling control, taxes control and pricing policies, limiting drinking age, restrictions on alcohol access, driving-related alcohol policies, control over advertising and promotion, and government monitoring systems. These policies are supported by evidence from different populations, demonstrating that determinants of alcohol use depend on several factors such as socioeconomic level, age, sex, ethnicity, production, availability, marketing, and others. Although most policies have a significant individual effect, a higher number of PHP are associated with a lower burden of disease due to alcohol. The excessive consequences of alcohol constitute a call for action, and clinicians should advocate for developing and implementing a new PHP on alcohol consumption.


Spermatogonia and the cycle of the seminiferous epithelium in the nine-banded armadillo.
The cycle of the seminiferous epithelium of the nine-banded armadillo can be divided into ten stages. As in most mammals, only one stage is observed per tubular cross-section. The process of spermiogenesis can be divided into thirteen steps according to the development of the acrosomal system and the flagellum. Four generations of spermatogonia are abserved in the germinal epithelium: 1) stem cells, 2)type "A", 3) intermediate, and 4) type "B" spermatogonia. The stem cell is characterized by a highly irregular nucleus and the presence of glycogen in its cytoplasm. The type "A" spermatogonium contains an oblong nucleus with one or two shallow infoldings of the nuclear membrane. The intermediate spermatogonium contins an ovoid nucleus characterized by one or two nuclei and heterochromatin scattered in the nuceoplasm. The nucleus of the type "B" spermatogonium is more spherically shaped with a centrally placed nucleolus and heterochromatin associated with the nuclear envelope.


Identification of multi-component metal ion mixtures in complex systems using fluorescence sensor arrays.
The growing co-contamination of multiple metal ions seriously influences human health due to their synergistic and additive toxicological effects, whereas the rapid discrimination of multiple heavy metal ions in complex aquatic systems remains a major challenge. Herein, a high- throughput fluorescence sensor array was fabricated based on three gold nanoclusters (GSH-Au NCs, OVA-Au NCs, and BSA-Au NCs) for the direct identification and quantification of seven heavy metal ions (Pb2+, Fe3+, Cu2+, Co2+, Ag+, Hg2+ and As3+) from environmental waters without sample pretreatment other than filtration. At the detection system, three gold nanoclusters with various ligands possessed distinct binding capacities against metal ions and induced aggregation-induced fluorescence enhancement and quenching, resulting in a unique pattern of fluorescence variations. Meanwhile, integrated the collected fluorescence fingerprints with linear discriminant analysis (LDA) and hierarchical cluster analysis (HCA), a discrete database was obtained for the accurate recognition and sensitive detection of metal ions. Under the optimized conditions, the limit of detection (LOD) of the proposed fluorescence sensor array for metal ions detection at nM concentration level along with a satisfactory accuracy. Importantly, our study indicated that the fluorescence sensor array could be widely used as a general platform in environmental monitoring against multiple targets at low concentrations.


Recurrence after surgery for varices in the groin is not dependent on body mass index.
To investigate if the body mass index (BMI) differs between different groups of patients (incomplete ligation also defined as technical error, neo-revascularisation, uncertain and mixed) in recurrent same site inguinal varices after surgery (REVAS). During a six and half year time span, we retrospectively analysed 203 consecutive procedures in 153 patients undergoing recurrent same site vein surgery in the groin. Individual BMI was calculated and compared within the different REVAS nature of the source groups. The median BMI was 28 for patients undergoing recurrent vein surgery in the groin with no relevant difference in BMI between the different source groups (confidence interval for the difference of adjusted group means equals [-1.5, 2.6]). There is no relevant difference in BMI between the two commonest REVAS groups. This may be due to small sample size, but confidence limits for difference of.


Studies of the blood of Ascidia ceratodes. Total blood cell counts, differential blood cell counts, hematocrit values, seasonal variations, and fluorescent characteristics of blood cells.
Seasonal, and animal size and weight variations of the blood cells of the vanadium-containing ascidian. Ascidia ceratodes, were determined. The fluorescent properties of various cell types were ascertained, and discussed in terms of cell development, phylogenic position of the species, and chemicals in the cells.


Alcoholic liver disease presenting with marked elevation of serum alkaline phosphatase. A combined clinical and pathological study.
Twenty patients with longstanding alcoholism and biopsy-proven alcoholic liver disease presented with marked elevation of serum alkaline phosphatase (in excess of four times the upper limit of normal). None had a past or present history to suggest pancreatitis or biliary tract disease, nor had any of these patients recently taken medication which could be implicated in cholestatic jaundice. Thirteen (65%) of this group either had radiologic or post mortem confirmation of nonobstructed biliary systems. The histologic findings in this group of patients were compared with those of a group of patients with alcoholic liver disease and normal or only mild elevation of serum alkaline phosphatase. Significantly more hepatocellular necrosis (P less than 0.05), alcoholic hyaline (P less than 0.02), and cholestasis (P less than 0.002) were noted in the severely hyperphosphatasemic group. Minimal degrees of steatosis were found in both groups. These data indicate that intrahepatic cholestasis occurs in patients with alcoholic liver disease, and this may often be secondary to alcoholic hepatitis. Overemphasis has previously been given to alcoholic fatty liver as a cause of this syndrome.


Cytotoxic and apoptogenic effect of tea (Camellia sinensis var. assamica) root extract (TRE) and two of its steroidal saponins TS1 and TS2 on human leukemic cell lines K562 and U937 and on cells of CML and ALL patients.
The anticancer activity of di- and tri-terpenes and other polyphenolic compounds present in tea is already reported. We evaluated the cytotoxic and apoptogenic effect of tea root extract (TRE) and two of its steroidal saponins named as TS1 and TS2, on human cell lines and on cells from leukemic patients. It was found that TRE, TS1 and TS2 significantly decreased cell count and that TRE caused apoptosis, as confirmed morphologically by confocal microscopy and by flow-cytometric analysis using Annexin-V FITC and propidium iodide (PI). Cell count and MTT assay in normal white blood cells (WBC) of healthy volunteers revealed that TRE produced insignificant reduction in cell count and cytotoxicity.


On the Use of Covariate Supersets for Identification Conditions.
The union of distinct covariate sets, or the superset, is often used in proofs for the identification or the statistical consistency of an estimator when multiple sources of bias are present. However, the use of a superset can obscure important nuances. Here, we provide two illustrative examples: one in the context of missing data on outcomes, and one in which the average causal effect is transported to another target population. As these examples demonstrate, the use of supersets may indicate a parameter is not identifiable when the parameter is indeed identified. Furthermore, a series of exchangeability conditions may lead to successively weaker conditions. Future work on approaches to address multiple biases can avoid these pitfalls by considering the more general case of nonoverlapping covariate sets.


Outpatient Thyroidectomy: Is it Safe?
Outpatient thyroid surgery is controversial because of concerns over life-threatening cervical hematoma. Despite this concern, outpatient thyroidectomy is becoming increasingly common, especially among high-volume endocrine surgeons. Multiple studies have now demonstrated that careful patient selection combined with surgeon experience can result in successful and safe surgery without a full inpatient admission. This article reviews the data on safety and outcomes for outpatient thyroidectomy and discusses several techniques used to minimize risk to patients.


Surgical treatment of locally advanced (T3) prostatic carcinoma: early results.
The fate of 48 patients with clinical stage T3 prostatic carcinoma after attempted curative surgical management was studied. In 23 of these patients positive frozen sections of the lymph nodes were found at pelvic lymphadenectomy and orchiectomy was performed. The median interval to progression was 61 months. Radical prostatectomy was performed in the remaining 25 patients. In 4 of these patients positive lymph nodes were found on paraffin sections but no additional treatment was given. Over-all, total tumor removal as defined by negative lymph nodes and negative margins of resection could be achieved in 14 of the 48 patients (29 per cent). During the same period 34 patients with clinical state T less than 3 prostatic carcinoma were treated in a similar manner. Orchiectomy was done in 4 patients because of positive frozen sections of the lymph nodes and radical prostatectomy was done in 30, including 1 in whom positive paraffin sections of the lymph nodes were found but no additional treatment was given. An attempt was made to study the impact of several prognostic factors by comparing the probability of progression between patients with stage pT3 disease with (T3pT3N0) or without (T less than 3pT3N0) extracapsular tumor growth as determined by preoperative rectal examination (36 versus 27 per cent progression at 3 years), with or without positive margins of resection (45 versus 20 per cent progression at 3 years) and with or without involvement of the seminal vesicles (47 versus 18 per cent progression at 3 years). Our results suggest that a certain proportion of patients with clinical stage T3 disease will benefit from radical prostatectomy. This is to be expected especially in patients with stage T3pT3N0 cancer and negative margins.


Comparison of the electronic properties, and thermodynamic and kinetic parameters of the aquation of selected platinum(II) derivatives with their anticancer IC50 indexes.
Three potential anticancer agents {trans-[PtCl(2)(NH(3))(thiazole)], cis-[PtCl(2)(NH(3))(piperidine)], and PtCl(2)(NH(3))(cyclohexylamine) (JM118)} were explored and compared with cisplatin and the inactive [PtCl(dien)](+) complex. Basic electronic properties, bonding and stabilization energies were determined, and thermodynamic and kinetic parameters for the aquation reaction were estimated at the B3LYP/6-311++G(2df,2pd) level of theory. Since the aquation process represents activation of these agents, the obtained rate constants were compared with the experimental IC(50) values for several tumor cells. Despite the fact that the processes in which these drugs are involved and the way in which they affect cells are very complex, some correlations can be deduced.


Architecture and active motif engineering of N-CoS2@C yolk-shell nanoreactor for boosted tetracycline removal via peroxymonosulfate activation: Performance, mechanism and destruction pathways.
Rational construction of yolk-shell architecture with regulated binding configuration is crucially important but challengeable for antibiotic degradation via peroxymonosulfate (PMS) activation. In this study, we report the utilization of yolk-shell hollow architecture consisted of nitrogen-doped cobalt pyrite integrated carbon spheres (N-CoS2@C) as PMS activator to boost tetracycline hydrochloride (TCH) degradation. The creation of yolk-shell hollow structure and nitrogen-regulated active site engineering of CoS2 endow the resulted N-CoS2@C nanoreactor with high activity for PMS activating toward TCH degradation. Intriguingly, the N-CoS2@C nanoreactor exhibits an optimal degradation performance with a rate constant of 0.194 min-1 toward TCH via PMS activation. The 1O2 and SO4•- species are demonstrated as the dominant active substances for TCH degradation through quenching experiments and electron spin resonance characterization. The possible degradation mechanism, intermediates and degradation pathways for TCH removal over the N-CoS2@C/PMS nanoreactor are unveiled. Graphitic N, sp2-hybrid carbon, oxygenated group (C-OH) and Co species are verified as the possible catalytic sites of N-CoS2@C for PMS activation toward TCH removal. This study offers a unique strategy to engineer sulfides as highly efficient and promising PMS activators for antibiotic degradation.


Effects of two inhaled corticosteroid/long-acting beta-agonist combinations on small-airway dysfunction in mild asthmatics measured by impulse oscillometry.
We previously showed that the long-acting beta agonist (LABA) salmeterol as inhalation powder or metered-dose inhaler improves lung-function parameters assessed by impulse oscillometry (IOS) in 2- to 5-year-old children with reversible-airway disease within 15 minutes. We studied 12- to 45-year-olds with mild persistent asthma in order to compare the onset and extent of peripheral airway effects following the first dose and after 4 weeks dosing with two inhaled corticosteroid (ICS)/LABA combinations: fluticasone propionate/salmeterol 115/21 and budesonide/formoterol 160/4.5. Thirty subjects with mild persistent asthma using only an as-needed short-acting beta-agonist (albuterol) who had at least a 40% change in integrated low-frequency reactance postalbuterol were selected and randomized to receive either fluticasone propionate/salmeterol or budesonide/formoterol (15 subjects each). We collected three to six IOS replicates at baseline, at 5, 20, 40, 60, 120, and 240 minutes postdose at randomization, and after 4 weeks of twice-daily dosing. Blinded investigators calculated IOS frequency-dependent resistance and reactance (R5-R20 and AX), indicative of small-airway dysfunction, and also estimated the peripheral airway resistance (Rp ) and peripheral airway compliance (Cp ), using a respiratory-impedance model. At randomization visits, onset of action was detected as early as 5 minutes (t-test, P < 0.05) after fluticasone propionate/salmeterol by Cp , and within 5 minutes after budesonide/formoterol by R5-R20, AX, Rp , and Cp . However, after 4 weeks of dosing, only Rp was significantly different (from 60 to 120 minutes) after fluticasone propionate/salmeterol, while R5-R20, AX, Rp , and Cp were not significantly different within 240 minutes after budesonide/formoterol. These two ICS/LABA combinations initially improved the peripheral airway function of 12- to 45-year-old asthmatics significantly in about 5 minutes or less, as measured by R5-R20, AX, Rp , and/or Cp . After regular dosing for 4 weeks, pre- to postdose differences in these parameters had diminished significantly due to improved predose status of peripheral airways. Single dosing with ICS/LABA combinations in mild persistent asthma improves small-airway function, and the effect is maintained over a 12-hour interval by regular use for 4 weeks.


Mitotic arrest-associated enhancement of O-linked glycosylation and phosphorylation of human keratins 8 and 18.
Arrest of the human colonic cell line HT29 at the G2/M phase of the cell cycle resulted in changes in keratin assembly that were coupled with a significant increase in the O-linked glycosylation and serine phosphorylation of keratin polypeptides 8 and 18 (K8/18). With mitotic arrest, enhanced keratin phosphorylation occurred preferentially on K8, whereas K18 showed a higher glycosylation level than K8. Removal of the arresting agent allowed cells to proceed through the cell cycle with a concomitant decrease in K8/18 glycosylation. In contrast, keratins isolated from S phase-enriched cells, obtained after synchronization with aphidicolin, did not show enhanced glycosylation. Tryptic peptide analysis of keratins in G2/M-arrested cells showed changes in the glycopeptide pattern of K8 and in the phosphopeptide patterns of K8 and K18. Labeling of K8/18 immunoprecipitates, isolated from G2/M-arrested cells, with [3H]galactose followed by beta-elimination showed that K8/18 glycosylation consisted of single N-acetylglucosamine residues. Threonine was identified as the site of glycosylation after comparing acid hydrolysis products of beta-eliminated and non-beta-eliminated K8 and K18. Specific cleavage at tryptophan residues indicated that K18 glycosylation and phosphorylation were restricted to the head and proximal rod domains, whereas K8 did not show the same restriction. Our results show a unique association of the single O-linked N-acetylglucosamine type of modification of keratins with mitotic arrest in HT29 cells. There was no reciprocal relationship between K8/18 glycosylation and phosphorylation, and each keratin showed a preferential G2/M cell cycle-associated increase in either serine phosphorylation or threonine glycosylation.


Effects of cardiac aglycones on the sodium efflux in barnacle muscle fibres.
1. External application of 10(-4) M digitoxigenin caused an 80% fall in the Na efflux in barnacle muscle fibres. The observed inhibition was irreversible and could be produced by as little as 10(-7) M digitoxigenin. 2. Internal application of 10(-2) M digitoxigenin lowered the Na efflux by 46% but was without effect following addition of external ouabain or K removal. 3. External application of digitoxigenin (or ouabain) following internal application of the aglycone was without effect on the Na efflux. 4. The efflux of [3H]digitoxigenin was fairly rapid, whereas that of [3H]ouabain was slow. 5. Inhibition of the Na efflux by internal application of digitoxigenin is probably the result of an action of the aglycone on the external side of the membrane following its leakage from the fibre interior.


Cordycepin inhibits LPS-induced inflammatory responses by modulating NOD-Like Receptor Protein 3 inflammasome activation.
The aim of this study is to examine the effects of cordycepin (CRD) on LPS-induced inflammatory response in macrophages and further investigate the underlying molecular mechanisms. Cultured mouse RAW264.7 macrophages and human THP-1-derived macrophages were used in this study. The mRNA and protein expression levels of cytokine IL-1β, IL-6, TNF-α, and MCP-1 were detected by real time RT-PCR, ELISA and Western blot, respectively. The activation of NOD-Like Receptor Protein 3 (NLRP3) inflammasome was analyzed with Western blot analysis and immunofluorescence staining. The ERK1/2 activation was assessed by Western blot analysis. The results demonstrated that pretreatment with CRD (6.25, 12.5, 25, 50μmol/L) dose-dependently inhibited LPS-induced expression of proinflammatory cytokines (IL-1β, IL-6, TNF-α, MCP-1) and cyclooxygenase 2 (COX-2) in mouse RAW264.7 cells. Similar results were obtained in human THP-1-derived macrophages with CRD. Furthermore, CRD remarkably inhibited LPS-induced activation of the NLRP3 inflammasome and ERK1/2 signaling pathway in RAW264.7 cells. CRD exerts anti-inflammatory effects in LPS-induced murine and human macrophages at least in part by inhibiting the activation of NLRP3 inflammasome and ERK1/2 signaling pathway as well as inhibition of COX2-mediated inflammatory response.


Relationship between laboratory-measured variables and heart rate during an ultra-endurance triathlon.
The aim of the present study was to examine the relationship between the performance heart rate during an ultra-endurance triathlon and the heart rate corresponding to several demarcation points measured during laboratory-based progressive cycle ergometry and treadmill running. Less than one month before an ultra-endurance triathlon, 21 well-trained ultra-endurance triathletes (mean +/- s: age 35 +/- 6 years, height 1.77 +/- 0.05 m, mass 74.0 +/- 6.9 kg, = 4.75 +/- 0.42 l x min(-1)) performed progressive exercise tests of cycle ergometry and treadmill running for the determination of peak oxygen uptake (VO2peak), heart rate corresponding to the first and second ventilatory thresholds, as well as the heart rate deflection point. Portable telemetry units recorded heart rate at 60 s increments throughout the ultra-endurance triathlon. Heart rate during the cycle and run phases of the ultra-endurance triathlon (148 +/- 9 and 143 +/- 13 beats x min(-1) respectively) were significantly (P < 0.05) less than the second ventilatory thresholds (160 +/- 13 and 165 +/- 14 beats x min(-1) respectively) and heart rate deflection points (170 +/- 13 and 179 +/- 9 beats x min(-1) respectively). However, mean heart rate during the cycle and run phases of the ultra-endurance triathlon were significantly related to (r = 0.76 and 0.66; P < 0.01), and not significantly different from, the first ventilatory thresholds (146 +/- 12 and 148 +/- 15 beats x min(-1) respectively). Furthermore, the difference between heart rate during the cycle phase of the ultra-endurance triathlon and heart rate at the first ventilatory threshold was related to marathon run time (r = 0.61; P < 0.01) and overall ultra-endurance triathlon time (r = 0.45; P < 0.05). The results suggest that triathletes perform the cycle and run phases of the ultra-endurance triathlon at an exercise intensity near their first ventilatory threshold.


Occurrence and evolution of homogeneously staining regions may be due to breakage-fusion-bridge cycles following telomere loss.
Chromosomes with homogeneously staining regions (HSR) were analysed in a subclone of the H4 rat hepatoma cell line, where they represent amplification of the ribosomal RNA (rRNA) genes. Detailed G-band analysis of the subclone revealed that an HSR on the short arm of chromosome 3 became unstable and changed its position within the chromosome. The evolution of this marker chromosome was associated with the terminal deletion of the normal long arm of the HSR-bearing chromosome 3 and may have involved ring formation as a result of fusion between the HSR on the short arm and the broken end of the long arm. Evidence was obtained for breakage at different sites within the ring, producing chromosomes with HSRs located terminally on either the long arms or both arms. The terminally located HSR underwent elongation in some cells presumably as a result of a breakage-fusion-bridge cycle characteristic of instability due to telomeric loss. It is suggested that terminally located HSRs may generally occur this way.


Refinement of an automatic method for indexing medical literature--a preliminary study.
to rank according to their significance MeSH terms automatically extracted from Internet sites in the framework of a French project, VUMeF, a contribution to the NLM' UMLS project. scores are affected to key-words of a given document on the basis of the Semantic Network of the UMLS and frequencies of co-occurring major terms in the Medline literature. If N is the number of major terms of a document, and n is the number of major terms retrieved in the N first terms ranked in descending order according to their scores, the measure of the achievement of the method is n/N. a set of 1444 randomized documents have been extracted from Medline. For each document we computed the retrieved major terms among the first N terms with two methods: a statistical method using only frequencies given by co-occurrences, and our method that uses furthermore the UMLS semantic network. In 34% of cases corresponding to documents indexed by about 16 key-words, about 3 major terms among them, our method produces a better precision (7%) than the statistical method. the rough calculation of the proportion of retrieved major terms should be enhanced by the use of a probability law allowing to enlarge the list of terms to select taking into account both the number of major terms and the total number of key-words used to index each document.


Atrial fibrillation and QT corrected. What is the best formula to use?
QT interval varies with the heart rate (HR), so a correction in QT calculation is needed (QTc). Atrial fibrillation (AF) is associated with elevated HR and beat-to-beat variation. To find best correlation between QTc in atrial fibrillation (AF) versus restored sinus rhytm (SR) after electrical cardioversion (ECV) (primary end point) and to determine which correction formula and method are the best to determine QTc in AF (secondary end point). During a 3-month period, we considered patients who underwent 12-lead ECG recording and received an AF diagnosis with indication for ECV. Exclusion criteria were as follows: QRS duration >120 ms, therapy with QT-prolonging drugs, a rate control strategy and a nonelectrical cardioversion. The QT interval was corrected using Bazzett's, Framingham, Fridericia and Hodges formulas during the last ECG during AF and the first one immediately after ECV. QTc mean was calculated as mQTc (average of 10 QTc calculated beat per beat) and as QTcM (QTc calculated from the average of 10 raw QT and RR for each beat). Fifty consecutive patients were enrolled in the study. Bazett's formula showed a significant change in mean QTc value between the two rhythms (421.5 ± 33.9 vs. 446.1 ± 31.9; p < 0.001 for mQTc and 420.9 ± 34.1 vs. 441.8 ± 30.9; p = 0.003 for QTcM). On the contrary, in patients with SR, QTc assessed by the Framingham, Fridericia, and Hodges formulas was similar to that in AF. Furthermore, good correlations between mQTc and QTcM are present for each formula, even in AF or SR. During AF, Bazzett's formula, seems to be the most imprecise in QTc estimation.


[Analysis of the causes of fatal outcome in fractures of the upper third of the femur].
Analysis of 5-year mortality of 1348 patients with fractures of the proximal end of the femur is presented. The effectiveness of measures aimed at the prevention of developing complications has been assessed. The data obtained demonstrate the effectiveness of these measures.


Casimir switch: steering optical transparency with vacuum forces.
The Casimir force, originating from vacuum zero-point energy, is one of the most intriguing purely quantum effects. It has attracted renewed interests in current field of nanomechanics, due to the rapid size decrease of on-chip devices. Here we study the optomechanically-induced transparency (OMIT) with a tunable Casimir force. We find that the optical output rate can be significantly altered by the vacuum force, even terminated and then restored, indicating a highly-controlled optical switch. Our result addresses the possibility of designing exotic optical nano-devices by harnessing the power of vacuum.


Arginine-supplemented diets improve survival in gutderived sepsis and peritonitis by modulating bacterial clearance: the role of nitric oxide Gianotti L, Alexander J W, Pyle T, and Fukushima R. Ann Surg 1993; 217: 644-654.
The effects of arginine on survival rates and host defence mechanisms were studied using two models of infection that included allogeneic transfusion-induced immunosuppression. Mice were fed for 10 days with chow, or a semi-purified commercial diet (AIN-76A), which was either unsupplemented or supplemented with arginine (2%; test diet) or glycine. After allogeneic blood transfusions and feeding for a further 5 days, the mice were subjected to caecal ligation and puncture (CLP), or 20% burn injury plus gavage with 10(10), (14)C-radiolabelled E. coli. Survival rate after CLP (AIN-76A diets) was 56% in the arginine supplemented group (AIN-76A) versus 28% in the AIN-76A unsupplemented group and 20% in the animals fed chow (P 0.02). After burn injury and gavage survival was 100% in the arginine-supplemented group versus 50% in both the chow and AIN-76A glycine supplemented group, and 35% in the AIN-76A unsupplemented group (P 0.01). Quantitative colony counts and % viable bacteria remaining in mesenteric lymph nodes, liver, and spleen suggested that the ability to kill translocated bacteria was significantly enhanced in animals receiving arginine. Treatment with the nitric oxide inhibitor, Nw-Nitro-L-arginine, reversed the beneficial effects or arginine. It is concluded that the benefit of arginine appears to be mediated by improved bactericidal mechanisms via the arginine nitric-oxide pathway.


A novel Xenopus laevis SRY-related gene, xSox33.
The sex-determining region Y (SRY) gene and its related Sox genes encode transcriptional regulatory factors. In this study, we isolated and sequenced a novel Sox cDNA from African clawed frog (Xenopus laevis). The Sox gene was named xSox33. xSox33 was revealed to encode 244 amino acids. Reverse transcription-polymerase chain reaction (RT-PCR) showed that xSox33 was expressed at very low levels in some frog tissues including lung, ovary, skeletal muscle, testis, brain and heart. Its embryonic expression was also studied by RT-PCR. After the mid-blastula transition, the zygotic expression was initiated during gastrulation and the level was elevated as the embryogenesis proceeded. Electrophoretic mobility shift assay (EMSA) indicated that a recombinant xSox33 polypeptide was capable of binding to the nucleotide sequence AACAAT.


Synthesis of monooxime-monocarbamoyl bispyridinium compounds bearing (E)-but-2-ene linker and evaluation of their reactivation activity against tabun- and paraoxon-inhibited acetylcholinesterase.
Six AChE monooxime-monocarbamoyl reactivators with an (E)-but-2-ene linker were synthesized using modification of currently known synthetic pathways. Their potency to reactivate AChE inhibited by the nerve agent tabun and insecticide paraoxon was tested in vitro. The reactivation efficacies of pralidoxime, HI-6, obidoxime, K048, K075 and the newly prepared reactivators were compared. According to the results obtained, one reactivator seems to be promising against tabun-inhibited AChE and two reactivators against paraoxon-inhibited AChE. The best results were obtained for bisquaternary substances with at least one oxime group in position four.


Chimeric foot-and-mouth disease viruses: evaluation of their efficacy as potential marker vaccines in cattle.
Previous work in pigs, has demonstrated that full protection against foot-and-mouth disease (FMD) can be achieved following vaccination with chimeric foot-and-mouth disease virus (FMDV) vaccines, in which the VP1 G-H loop had been substituted with that from another serotype. If proven to be effective in other economically important species such as cattle, such vaccine constructs could be trialed as potential marker vaccines. Here, we determine if G-H loop chimera FMDV vaccines can: (i) protect cattle from virus challenge and (ii) induce an antibody response that would enable the identification of infection, regardless of vaccination status. Inactivated, oil adjuvanated, chimeric vaccine constructs, based on the backbone sequence of the A(12)119 serotype virus, fully protected cattle from challenge 21 days post-vaccination. Differentiation assays developed for use in this study were able to identify sub-clinical infection, which in one vaccinated animal, persisted beyond day 32 post-challenge. This paper emphasises the importance of epitopes outside of the VP1 G-H loop for protective immunity in cattle, and demonstrates that chimeric FMDV vaccines could prove to be useful marker vaccines for the future.


Influence of in utero fetal death on perineal tears in vaginal deliveries.
The aim of this work was to evaluate and compare the incidence of perineal tears and Obstetrical anal sphincter injuries (OASIS) after vaginal delivery following a in utero fetal death (IUFD) compared with those with a live-birth. We conducted a single-center, retrospective cohort study using a database of all women who underwent a spontaneous vaginal delivery in the level III maternity ward. Exclusion criteria were breech presentation, cesarean section birth, instrumental delivery, multiple pregnancy, delivery before 24 + 6 weeks of gestation (WG) and termination of pregnancy for medical reasons. Women from the database were divided into two groups: an "in utero fetal death" (IUFD) group and a control group. Women were included in the IUFD group if they had a spontaneous vaginal delivery following a fetal demise after 24 + 6 WG in cephalic presentation between January 2006 and June 2020. Women in the "control" group were selected from the same database and were included if they underwent a spontaneous vaginal delivery of a live fetus in cephalic presentation, after 24 + 6 WG, during the same period. Each woman in the "IUFD" group was matched to two women (ratio 1:2) in the control group for parity, maternal age, body mass index, gestation and birth weight. The primary outcome was the presence of a sutured or unsutured perineal tear. During the study period, 31,208 patients delivered at a level III maternity hospital. Among them, 215 and 430 women were included in the IUFD group and the control group respectively. The two groups were comparable for all demographic and clinical characteristics except for an epidural analgesia (92% versus 70% in the control group, p < 0.01) and labor induction (86% versus 17% in the control group, p < 0.01). The incidence of any perineal tears was 13% (28/15) in the IUFD group versus 16% (70/430) in the control group. Relative risk of any perineal tears was non significative (RR = 0.8 IC95% [0.5-1.2]). The incidence of first-degree perineal tears was 10% in the IUFD group and 11% in the control group. The incidence of second-degree perineal tears was 18% in the IUFD group and 28% in the control group. Relative risk of first-degree perineal tears (RR = 0.88 95% CI [0.5-1.4]) and second-degree tears (RR = 0.51 95% CI [0.2-1.4]) were non significative. No obstetrical anal sphincter injury was found in either group. Vaginal delivery following a fetal demise did not appear to be either a risk factor or a protective factor for perineal tears. But there as a trend toward a lower incidence of second degree perineal tears in this context.


Proceedings of conference on Mechanism-Based Development of Natural Products for Human Health.
Mechanism-Based Development of Natural Products in Human Health.


Social participation in older adults with joint pain and comorbidity; testing the measurement properties of the Dutch Keele Assessment of Participation.
The Keele Assessment of Participation (KAP) questionnaire measures person-perceived participation in 11 aspects of life. Participation allows fulfilment of valued life activities and social roles, which are important to older adults. Since we aimed to use the KAP in a larger Dutch cohort, we examined the measurement properties of KAP in a Dutch sample of older adults with joint pain and comorbidity. Cohort study. A community-based sample in Amsterdam, the Netherlands and North Staffordshire, UK. Participants were aged 65 years and over, had at least two chronic diseases (identified through general practice consultation) and reported joint pain on most days (questionnaire). The Dutch cohort provided baseline data (n=407), follow-up data at 6 months (n=364) and test-retest data 2 weeks after 6 months (n=122). The UK cohort provided comparable data (n=404). The primary outcome was person-perceived participation, as measured with the KAP. The measurement properties examined were the following: structural validity (factor analysis), internal consistency (Cronbach's α), reliability (intraclass correlation coefficients; ICC), construct validity (hypothesis testing), responsiveness (hypothesis testing and area under the curve) and cross-cultural validity (differential item functioning; DIF). Factor analysis revealed two domains: KAPd1: 'participation in basic activities' and KAPd2: 'participation in complex activities', with Cronbach's α of 0.74 and 0.57 and moderate test-retest reliability: ICC of 0.63 and 0.57, respectively. Further analyses of KAPd1 showed poor construct validity and responsiveness. Despite the uniform DIF in item 'interpersonal relations', the total KAPd1 score seemed comparable between the Dutch and UK sample. Only KAP domain 'participation in basic activities' showed good internal consistency and sufficient reliability. KAPd2 lacks sufficient measurement properties for application in studies, although items may be used as single items. Further development of the concept 'participation' may help the development and validation of instruments to measure participation.


Microorganism-regulated mechanisms of temperature effects on the performance of anaerobic digestion.
Temperature is an important factor determining the performance and stability of the anaerobic digestion process. However, the microorganism-regulated mechanisms of temperature effects on the performance of anaerobic digestion systems remain further elusive. To address this issue, we investigated the changes in composition, diversity and activities of microbial communities under temperature gradient from 25 to 55 °C using 16S rRNA gene amplicon sequencing approach based on genomic DNA (refer to as "16S rDNA") and total RNA (refer to as "16S rRNA"). Microbial community structure and activities changed dramatically along the temperature gradient, which corresponded to the variations in digestion performance (e.g., daily CH4 production, total biogas production and volatile fatty acids concentration). The ratios of 16S rRNA to 16S rDNA of microbial taxa, as an indicator of the potentially relative activities in situ, and whole activities of microbial community assessed by the similarity between microbial community based on 16S rDNA and rRNA, varied strongly along the temperature gradient, reflecting different metabolic activities. The daily CH4 production increased with temperature from 25 to 50 °C and declined at 55 °C. Among all the examined microbial properties, the whole activities of microbial community and alpha-diversity indices of both microbial communities and potentially relative activities showed highest correlations to the performance. The whole activities of microbial community and alpha-diversity indices of both microbial communities and potentially relative activities were sensitive indicators for the performance of anaerobic digestion systems under temperature gradient, while beta-diversity could predict functional differences. Microorganism-regulated mechanisms of temperature effects on anaerobic digestion performance were likely realized through increasing alpha-diversity of both microbial communities and potentially relative activities to supply more functional pathways and activities for metabolic network, and increasing the whole activities of microbial community, especially methanogenesis, to improve the strength and efficiency in anaerobic digestion process.


Cardiac Surgery and the Blood-Brain Barrier.
Neurologic abnormality after cardiac surgery is common, and neurologic complications after cardiac surgery are among the most devastating problems that can occur in the postoperative period. Disruption of the blood-brain barrier (BBB) plays an important role in these complications. Assessment of the BBB integrity relies on cognitive testing, MRI, and measurement of brain biomarkers. In applying these methods, up to 50% of cardiac patients show some degree of BBB disruption and most of these abnormalities are short lived. To date there is no single test or measure that can predict BBB disruption in cardiac surgery.


Anti-M9 antibodies in sera from patients with primary biliary cirrhosis recognize an epitope of glycogen phosphorylase.
Anti-M9 antibodies in sera from patients with primary biliary cirrhosis (PBC) were previously found to recognize two antigenic determinants at 98 and 59 kD, using a purified antigen fraction derived from rat liver mitochondria in the Western blot. Here we show that these antibodies are directed against an epitope of the enzyme glycogen phosphorylase. By Western blotting, a determinant at 98 kD was obtained testing anti-M9 positive sera against phosphorylase from skeletal muscle, and after plasmin treatment a degradation product appeared at 59 kD. Both determinants were identical to the M9-specific determinants 98 and 59 kD as shown by absorption studies. When these antibodies were eluted from the 98 and 59 kD determinants of the M9 antigen after immunoblotting, they again recognized the same epitopes on plasmin-treated phosphorylase. Furthermore, phosphorylase enzyme activity could be also demonstrated in the purified M9 fraction, and anti-M9-positive/anti-M2-negative but not anti-M9-negative/anti-M2-positive sera could be shown to stimulate phosphorylase activity. Testing sera from 1189 patients with different hepatic and non-hepatic disorders against M9 and phosphorylase from skeletal muscle by ELISA, 20% were positive with phosphorylase and only 2% with the M9 fraction. These data indicate that the commercially available phosphorylase from skeletal muscle cannot be recommended as M9 source. It may still contain non-PBC-specific epitopes which are probably recognized by naturally occurring antibodies directed against this highly conserved protein.


DNA methylation in polycystic ovary syndrome: Emerging evidence and challenges.
Polycystic ovary syndrome (PCOS) is a disease related to reproductive endocrine abnormalities in women of reproductive age, often accompanied by metabolic diseases such as hyperandrogenemia, insulin resistance and dyslipidemia. However, the etiology and mechanism of PCOS are still unclear. In recent years, more and more studies have found that epigenetic factors play an important role in PCOS. DNA methylation is the most widely studied epigenetic modification. At present, changes of DNA methylation have been found in serum, ovarian, hypothalamus, skeletal muscle, adipose tissue of PCOS patients, and these changes are closely related to insulin resistance, lipid metabolism and follicular development of PCOS. Although the current research on DNA methylation in PCOS is not in-depth, it indicated up a good direction for future research on the etiology and mechanism of PCOS. This review discussed the relationship between DNA methylation and PCOS. It is expected to help accelerate the application of DNA methylation in the diagnosis and treatment of PCOS.


Ethnic differences in cardiovascular risks and mortality in atherothrombotic disease: insights from the Reduction of Atherothrombosis for Continued Health (REACH) registry.
To determine whether ethnic-specific differences in the prevalence of cardiovascular risk factors and outcomes exist worldwide among individuals with stable arterial disease. From December 1, 2003, to June 30, 2004, the prospective, observational REduction of Atherothrombosis for Continued Health (REACH) Registry enrolled 49,602 out-patients with coronary artery disease, cerebrovascular disease, and/or peripheral arterial disease from 7 predefined ethnic/racial groups: white, Hispanic, East Asian, South Asian, Other Asian, black, and Other (comprising any race distinct from those specified). The baseline demographic and risk factor profiles, medication use, and 2-year cardiovascular outcomes were assessed among these groups. The prevalence of traditional atherothrombotic risk factors varied significantly among the ethnic/racial groups. The use of medical therapies to reduce risk was comparable among all groups. At 2-year follow-up, the rate of cardiovascular death was significantly higher in blacks (6.1%) compared with all other ethnic/racial groups (3.9%; P=.01). Cardiovascular death rates were significantly lower in all 3 Asian ethnic/racial groups (overall, 2.1%) compared with the other groups (4.5%; P<.001). The REACH Registry, a large international study of individuals with atherothrombotic disease, documents the important ethnic-specific differences in cardiovascular risk factors and variations in cardiovascular mortality that currently exist worldwide.


High production of heterologous proteins in Escherichia coli using the thermo-regulated T7 expression system.
The exclusive use of isopropyl beta-D-thiogalactopyranoside to activate the T7 promoter for protein production has limited the general use of the expression system. We have sought an alternative by constructing a recombinant Escherichia coli strain, BL21 (G2), to carry a chromosomal copy of T7 gene 1 fused to the lambdaPL and lambdaP(R) tandem promoter. As a result, the recombinant strain harboring the carbamoylase gene from Agrobacterium radiobacter NRRL B11291 was shown to display various levels of.protein production in response to different degrees of heat shock. In particular, the system remained inactive at 30 degrees C and exhibited high sensitivity to heat such that a detectable carbamoylase activity could be measured after exposure to 33 degrees C. Moreover, heating in two steps - elevating the temperature from 30 degrees C to 39 degrees C and holding for a brief period, followed by reducing to 37 degrees C--was found to be the most potent method for protein production in this case. Using this approach, the recombinant protein accounted for 20% of total protein content of the cell. These results reveal the advantages of this expression system: responsiveness to thermal modulation and high-level production capability. In an attempt to enhance the total protein yield, a fed-batch fermentation process was carried out to control the cell growth rate by adjusting the substrate inflow. By applying the two-step temperature change. a carbamoylase yield with enzyme activity corresponding to 14,256 units was obtained. This production yield is a 10-fold increase in comparison with that at the batch-fermentation scale and 2,000-fold higher than that achieved at the shake-flask scale. Overall, it illustrates the promise of the newly constructed T7 system based on heat inducibility for industrial scale production of recombinant proteins.


Usefulness of FTA® cards as a Pneumocystis-DNA extraction method in bronchoalveolar lavage samples.
FTA® cards (Fast Technology for Analysis of Nucleic Acids) are an alternative DNA extraction method in bronchoalveolar lavage (BAL) samples for Pneumocystis jirovecii molecular analyses. The goal was to evaluate the usefulness of FTA® cards to detect P. jirovecii-DNA by PCR in BAL samples compared to silica adsorption chromatography (SAC). This study used 134 BAL samples from immunocompromised patients previously studied to establish microbiological aetiology of pneumonia, among them 15 cases of Pneumocystis pneumonia (PCP) documented by staining and 119 with other alternative diagnoses. The FTA® system and SAC were used for DNA extraction and then amplified by nested PCR to detect P. jirovecii. Performance and concordance of the two DNA extraction methods compared to P. jirovecii microscopy were calculated. The influence of the macroscopic characteristics, transportation of samples and the duration of the FTA® card storage (1, 7, 10 or 12 months) were also evaluated. Among 134 BAL samples, 56% were positive for P. jirovecii-DNA by SAC and 27% by FTA®. All 15 diagnosed by microscopy were detected by FTA® and SAC. Specificity of the FTA® system and SAC were 82.4% and 49.6%, respectively. Compared to SAC, positivity by FTA® decreased with the presence of blood in BAL (62% vs 13.5%). The agreement between samples at 7, 10 and 12 months was 92.5% for FTA®. Positive cases by FTA® remained the same after shipment by mail. Results suggest that FTA® is a practical, safe and economical method to preserve P. jirovecii-DNA in BAL samples for molecular studies.


Electrochemiluinescence monitoring the interaction between human serum albumin and amyloid-β peptide.
Human serum albumin (HSA) is a major interaction-partner of amyloid-β (Aβ) peptide in the plasma and can modulate the aggregation of Aβ. In this work, an efficient smart electrochemiluminescence (ECL) sensing platform was applied to determine the ability of HSA to bind different Aβ40 aggregate species. Specifically, HSA was firstly adopted as a linker and self-assembled on the gold electrode, Aβ peptide were subsequently anchored onto the HSA-modified electrode through the recognition by HSA. CdS quantum dots (CdS QDs) modified with Aβ40 aptamer as the ECL substrate were then captured on the electrode via the specific coordination between Aβ40 and the aptamer. The present ECL sensor showed different ECL response in the presence of Aβ40 with different aggregation states. The ECL platform exhibited a signal-on response with the introduction of Aβ40 monomer in a concentration-dependent manner from 10 fM to 0.1 nM. Furthermore, the inhibition effect of several phenols on the aggregation of Aβ40 was also investigated. This work potentially provided a feasible strategy of sensing platforms for Aβ detection and further understanding of the mechanism by which HSA modulates the aggregation of Aβ.


Trends in incidence of recorded diagnosis of osteoporosis, osteopenia, and fragility fractures in people aged 50 years and above: retrospective cohort study using UK primary care data.
This study used primary care data to estimate the incidence of recorded diagnosis of osteoporosis, osteopenia, and fragility fracture in the UK during 2000-2018 accounting for age, sex, calendar year and social deprivation. More than 3 million people aged 50-99 years were included. We found that men living in the most deprived areas had a 45% higher risk of being diagnosed with osteoporosis and 50% higher risk of fragility fracture compared to men living in the least deprived areas. a) To estimate the incidence trends of a recorded diagnosis of osteoporosis, osteopenia, and fragility fracture in the UK over time; b) to describe differences according to age, sex, and social deprivation. This is a longitudinal population-based cohort study using routinely collected primary care data obtained via IQVIA Medical Research Database (IMRD). All patients aged 50-99 years registered with a practice participating in THIN (The Health Improvement Network) between 2000-2018 were included. The first recorded diagnosis of osteoporosis, osteopenia, or fragility fracture was used to estimate incidence rates (IR) per 10,000 person-years at risk. Poisson regression was used to provide Incidence Rate Ratios (IRR) adjusted by age, sex, social deprivation, calendar year, and practice effect. The year-specific adjusted IRR of recorded osteoporosis was highest in 2009 in women [IRR 1.44(95%CI 1.38-1.50)], whereas in men it was highest in 2013-2014 [IRR 1.94(95%CI 1.72-2.18)] compared to 2000. The year-specific adjusted IRR of fragility fracture was highest in 2012 in women [IRR 1.77(95%CI 1.69-1.85)], whereas in men it was highest in 2013 [IRR 1.64(95%CI 1.51-1.78)] compared to 2000. Men in the most deprived areas had a higher risk of being diagnosed with osteoporosis [IRR 1.45(95%CI 1.38-1.53)], osteopenia [IRR 1.17(95%CI 1.09-1.26)], and fragility fracture [IRR 1.50(95%CI 1.44-1.56)] compared to those living in the least deprived areas, but smaller differences were seen in women. Use of fracture risk assessment tools may enhance the detection of osteoporosis cases in primary care. Further research is needed on the effect of social deprivation on diagnosis of osteoporosis and fractures.


Activation of 'immediate-early' genes by estrogen is not sufficient to achieve stimulation of DNA synthesis in rat uterus.
17 beta-estradiol, a long acting estrogen that is mitogenic for rat uterus in vivo, or the short acting estrogens estriol and 16 alpha-estradiol, not mitogenic on their own, were injected into adult, castrated rats and their effect on uterine gene expression and rate of DNA synthesis were compared. All three compounds increased steady-state mRNA concentration of c-fos, c-jun and c-myc proto-oncogenes to comparable levels (2 hrs after treatment), whereas only 17 beta-estradiol was found to stimulate significantly DNA synthesis (20-22 hrs later). Based on the different retention time of the tested estrogens in rat tissues, it is concluded that a short exposure to the hormone is sufficient to render uterine cells competent to progress through the cell cycle, via activation of 'immediate-early' genes expression, but that stimulation of DNA synthesis requires further changes, achieved via a prolonged exposure of the cells to the estrogenic stimulus.