Lifespan and healthspan benefits of exogenous H2S in C. elegans are independent from effects downstream of eat-2 mutation.
Caloric restriction (CR) is one of the most effective interventions to prolong lifespan and promote health. Recently, it has been suggested that hydrogen sulfide (H2S) may play a pivotal role in mediating some of these CR-associated benefits. While toxic at high concentrations, H2S at lower concentrations can be biologically advantageous. H2S levels can be artificially elevated via H2S-releasing donor drugs. In this study, we explored the function of a novel, slow-releasing H2S donor drug (FW1256) and used it as a tool to investigate H2S in the context of CR and as a potential CR mimetic. We show that exposure to FW1256 extends lifespan and promotes health in Caenorhabditis elegans (C. elegans) more robustly than some previous H2S-releasing compounds, including GYY4137. We looked at the extent to which FW1256 reproduces CR-associated physiological effects in normal-feeding C. elegans. We found that FW1256 promoted healthy longevity to a similar degree as CR but with fewer fitness costs. In contrast to CR, FW1256 actually enhanced overall reproductive capacity and did not reduce adult body length. FW1256 further extended the lifespan of already long-lived eat-2 mutants without further detriments in developmental timing or fertility, but these lifespan and healthspan benefits required H2S exposure to begin early in development. Taken together, these observations suggest that FW1256 delivers exogenous H2S efficiently and supports a role for H2S in mediating longevity benefits of CR. Delivery of H2S via FW1256, however, does not mimic CR perfectly, suggesting that the role of H2S in CR-associated longevity is likely more complex than previously described.


Target-enriched long-read sequencing (TELSeq) contextualizes antimicrobial resistance genes in metagenomes.
Metagenomic data can be used to profile high-importance genes within microbiomes. However, current metagenomic workflows produce data that suffer from low sensitivity and an inability to accurately reconstruct partial or full genomes, particularly those in low abundance. These limitations preclude colocalization analysis, i.e., characterizing the genomic context of genes and functions within a metagenomic sample. Genomic context is especially crucial for functions associated with horizontal gene transfer (HGT) via mobile genetic elements (MGEs), for example antimicrobial resistance (AMR). To overcome this current limitation of metagenomics, we present a method for comprehensive and accurate reconstruction of antimicrobial resistance genes (ARGs) and MGEs from metagenomic DNA, termed target-enriched long-read sequencing (TELSeq). Using technical replicates of diverse sample types, we compared TELSeq performance to that of non-enriched PacBio and short-read Illumina sequencing. TELSeq achieved much higher ARG recovery (>1,000-fold) and sensitivity than the other methods across diverse metagenomes, revealing an extensive resistome profile comprising many low-abundance ARGs, including some with public health importance. Using the long reads generated by TELSeq, we identified numerous MGEs and cargo genes flanking the low-abundance ARGs, indicating that these ARGs could be transferred across bacterial taxa via HGT. TELSeq can provide a nuanced view of the genomic context of microbial resistomes and thus has wide-ranging applications in public, animal, and human health, as well as environmental surveillance and monitoring of AMR. Thus, this technique represents a fundamental advancement for microbiome research and application. Video abstract.


Complicated Diverticulitis and Pelvic Radiation Leading to Colonic Stricture, Colorectal Fistula, and Anal Stenosis.
The purpose of this case is to highlight a rare case of sigmoid colon-to-rectum fistula. A 66-year-old man with a history of pelvic radiation and diverticulosis presented to the emergency department with a 3-week history of abdominal pain and watery diarrhea. Computed tomography (CT) imaging was significant for a sigmoid-to-rectum fistula with sigmoid stricture. The patient underwent a laparoscopic colectomy with end colostomy. Pathology revealed perforated diverticulitis. To date, there have been no cases reported in literature describing sigmoid-to-rectum fistula. In conclusion, it is important to consider the development of complex diverticular disease in patients with history of pelvic radiation.


Comparative and phylogenetic analysis of the complete chloroplast genomes of six Polygonatum species (Asparagaceae).
Polygonatum Miller belongs to the tribe Polygonateae of Asparagaceae. The horizontal creeping fleshy roots of several species in this genus serve as traditional Chinese medicine. Previous studies have mainly reported the size and gene contents of the plastomes, with little information on the comparative analysis of the plastid genomes of this genus. Additionally, there are still some species whose chloroplast genome information has not been reported. In this study, the complete plastomes of six Polygonatum were sequenced and assembled, among them, the chloroplast genome of P. campanulatum was reported for the first time. Comparative and phylogenetic analyses were then conducted with the published plastomes of three related species. Results indicated that the whole plastome length of the Polygonatum species ranged from 154,564 bp (P. multiflorum) to 156,028 bp (P. stenophyllum) having a quadripartite structure of LSC and SSC separated by two IR regions. A total of 113 unique genes were detected in each of the species. Comparative analysis revealed that gene content and total GC content in these species were highly identical. No significant contraction or expansion was observed in the IR boundaries among all the species except P. sibiricum1, in which the rps19 gene was pseudogenized owing to incomplete duplication. Abundant long dispersed repeats and SSRs were detected in each genome. There were five remarkably variable regions and 14 positively selected genes were identified among Polygonatum and Heteropolygonatum. Phylogenetic results based on chloroplast genome strongly supported the placement of P. campanulatum with alternate leaves in sect. Verticillata, a group characterized by whorled leaves. Moreover, P. verticillatum and P. cyrtonema were displayed as paraphyletic. This study revealed that the characters of plastomes in Polygonatum and Heteropolygonatum maintained a high degree of similarity. Five highly variable regions were found to be potential specific DNA barcodes in Polygonatum. Phylogenetic results suggested that leaf arrangement was not suitable as a basis for delimitation of subgeneric groups in Polygonatum and the definitions of P. cyrtonema and P. verticillatum require further study.


Gene expression profile changes in NB4 cells induced by realgar.
To compare the gene expression profiles of acute promyelocytic leukemia cell line NB(4) before and after 12 hours of realgar treatment using cDNA microarray. Two cDNA probes were prepared through reverse transcription from mRNA of both untreated and realgar treated NB(4) cells. The probes were labeled with Cy3 and Cy5 fluorescence dyes individually, hybridized with cDNA microarray representing 1003 different human genes, and scanned for fluorescent intensity. The genes were screened through the analysis of the difference in two gene expression profiles. The analysis of gene expression profiles indicates that 9 genes were up-regulated and 37 genes were down-regulated. Among the 9 up-regulated genes, 2 genes were involved in a proteasome degradation pathway. Some genes related to protein synthesis, signal transduction and cell receptors were down-regulated. PSMC2 and PSMD1 genes may play an important role in the apoptosis and partial differentiation of NB(4) cells.


Portal hypertension evolving from sickled hepatopathy: Could hepatic venous Doppler ultrasound be beneficial in its evaluation?
Sickle cell intrahepatic cholestasis involves sickling within hepatic sinusoids leading to vascular stasis and localized hypoxia resulting in ballooning of the hepatocytes causing a direct back pressure effect with resultant intracanalicular cholestasis. Vascular stasis may ultimately lead to portal hypertension. We proposed to document findings suggestive of portal hypertension evolving from hepatopathy in steady-state sickle cell disease (SCD) patients using hepatic venous Doppler ultrasound. This is a prospective case series of 6 SCD subjects in steady-state (median age, 30 years; range, 19-43), comprising of 3 males and 3 females, who underwent a routine Doppler ultrasound evaluation of their hepatic veins and were discovered to have an abnormal biphasic waveform pattern. Venous blood was obtained from all subjects to evaluate for P-selectin, homocysteine, foetal haemoglobin, haematocrit levels, white cell and platelet counts. Doppler ultrasound was also carried out on all subjects to evaluate for the hepatic waveform, right renal artery RI and PI along with the hepatic artery velocities. All the 6 subjects had reduced haematocrit (median value of 21.5%; range, 18-25%) and some degree of renal dysfunction (plasma cystatin-C ranged from 1.6 to 12.2 mg/L). Elevated white cell count, hyperhomocysteinemia, reduced SpO2(<94.0%) and reduced estimated GFR (eGFR < 90 ml/min) was also noted in 4 subjects (66.7%). Similarly, 4 subjects (66.7%) had elevated RI in the right kidneys while 3 subjects (50.0%) had elevated PI in the right kidney. Doppler ultrasound Hepatic vein waveform analysis may be a useful examination in the evaluation of patients with SCD as it may elicit feature of portal hypertension. Further studies are suggested to confirm this in a larger population of SCD patients using the gold standard.


Discrimination of normal and transformed cells in vitro by cytologic and morphologic analysis.
Malignant A-549 lung carcinoma and adenovirus-12 SV40 hybrid virus transformed non-tumorigenic human bronchial epithelial cells (BEAS-2B) were objectively discriminated from normal bronchial epithelial (BE) cells on the basis of Papanicolaou stained nuclear features (e.g. shape, chromatin texture, hyperchromasia) and nucleolar morphology (e.g. number per cell, irregular contours). Morphometric analysis indicated that significant differences in cellular morphology existed between BE, BEAS-2B, and A-549 cells. Similar analyses of transformed, tumorigenic cell lines demonstrated that nuclear features (i.e., chromatin texture, clearing of parachromatin, hyperchromasia, variation in thickness of the nuclear envelope, sharp indentations in the nuclear envelope), and nucleolar features (i.e., degree of roundness, presence of angular projections, number per cell) discriminated chemically and virally transformed cells from spontaneously transformed cells. Nuclear and nucleolar features were correlated with the growth rate of tumorigenic cell lines. These analytical approaches will be helpful in studies of the effects of various factors (e.g. vitamin A, phorbol ester, oncogene transfection) on cellular proliferation and/or differentiation.


Factors associated with prolonged duration of post-tympanoplasty local treatment in adult chronic otitis media patients: A retrospective observational study using a Japanese inpatient database.
The occurrence of persistent infection following tympanoplasty has been reported in many studies, and it is important to know the risks for site infection after tympanoplasty. In this study, we aimed to explore the factors affecting early wound complications after tympanoplasty for chronic otitis media. We conducted a retrospective cohort study using the Diagnosis Procedure Combination database. Data on a total of 13,094 adult patients from 420 acute-care hospitals who received tympanoplasty for chronic otitis media from 2010 to 2013 were extracted. The duration (days) of postsurgical local wound treatment was measured as an outcome, because this duration was assumed to be prolonged by the existence of wound infection. The associations between treatment duration and background characteristics (age, sex, body mass index, smoking status, diabetes mellitus, use of antithrombotic agents, with or without cholesteatoma, duration of anesthesia, academic hospital or not, and hospital volume) were assessed by multivariable linear regression analyses, fitted with a generalized estimating equation to adjust for within-hospital clustering. The median treatment duration in each hospital was 8 days (interquartile range: 7-11). Factors significantly associated with longer treatment duration were: older age (0.2 days for 10-year increase), use of antithrombotic agents during hospitalization (1.8 days), and prolonged duration of anesthesia (vs. <120min of anesthesia, additional 1, 2, 3, and 4 days for 120-179, 180-239, 240-299, and ≥300min of anesthesia, respectively). Body mass index and smoking status were not significantly associated with treatment duration. Older age, antithrombotic agents during hospitalization, and longer anesthesia time were independently associated with early local wound complications after tympanoplasty for chronic otitis media.


A preliminary assessment of microplastics in indoor dust of a developing country in South Asia.
Microplastics (MPs) pollution is an emerging global environmental concern. Considering the high fraction of time people spend indoors, the human population can be directly exposed to this contamination through indoor dust. This preliminary study evaluates MPs' abundance and human health risk assessment in the deposited indoor dust. A total of forty dust samples (n = 20) were collected from homes in two different cities (Pakistan) in steel mesh pouches using the vacuum cleaner. The identification and quantification of MPs were conducted with a stereo microscope, whereas the qualitative assessment was performed with Fourier transform infrared spectroscopy (FTIR). The US EPA parameters to calculate the human health risk assessment were used to determine MPs' risk per-day/month/year. Overall, microfibers were the dominant category, followed by microfilms, micro-fragments, and nurdles. The chemical categorization of MPs was revealed as polyester, polyethylene, copolymers of polypropylene, and polyurethane. In Lahore, an average abundance of 241.45 (items/m2) MPs were observed compared to Sahiwal, with 162.1 (items/m2). More than 90% of the identified MPs were microfibers, with higher detection frequency and abundance in Lahore than Sahiwal. The human health risk assessment revealed high exposure risk because of indoor MPs. Moreover, toddlers were more vulnerable as compared to adults at both low and high exposure risk scenarios. There is an imminent need to conduct in-depth risk assessment focusing on the respirable fraction of MPs.


Histological grade of invasive carcinoma of the breast assessed on needle core biopsy - modifications to mitotic count assessment to improve agreement with surgical specimens.
Histological grade assessed on core biopsy of mammary invasive carcinomas tends to underestimate the grade in the excision specimen. A major factor is underestimation of mitoses. The aim of this study was to improve the assessment of mitoses in core biopsies. In a test set (100 tumours), reducing the threshold for a mitotic score of 2 from 11 mitoses per 10 high-power fields (diameter 0.61 mm) to 6/10 resulted in improved agreement between core and excision from 69% to 73%. In tumours graded as T3 P3 M1 (few tubules, marked nuclear pleomorphism and few mitoses) on core (a group with poor agreement of grade between core and surgical specimens), this change in threshold also improved agreement in four tumours. Use of the mitotic count in the core of axillary nodal metastasis, if higher than the count in the breast core, improved agreement in two patients. Use of both approaches in a validation set (349 tumours) improved agreement from 65% to 70% in the whole group, and from 31% to 55% in tumours scored as T3 P3 M1 on the core. These strategies improve the accuracy of grading on core biopsy and are of particular value in T3 P3 M1 tumours.


Information technology for clinical, translational and comparative effectiveness research. Findings from the section clinical research informatics.
To summarize advances of excellent current research in the new emerging field of Clinical Research Informatics. Synopsis of four key articles selected for the IMIA Yearbook 2013. The selection was performed by querying PubMed and Web of Science with predefined keywords. From the original set of 590 papers, a first subset of 461 articles which was in the scope of Clinical Research Informatics was refined into a second subset of 79 relevant articles from which 15 articles were retained for peer-review. The four selected articles exemplify current research efforts conducted in the areas of data representation and management in clinical trials, secondary use of EHR data for clinical research, information technology platforms for translational and comparative effectiveness research and implementation of privacy control. The selected articles not only illustrate how innovative information technology supports classically organized randomized controlled trials but also demonstrate that the long promised benefits of electronic health care data for research are becoming a reality through concrete platforms and projects.


Predictors of retention in dual-focus self-help groups.
Attendance at 12-step groups has been found useful in maintaining abstinence from substance use; many members disengage early, missing out on potential benefits. New 12-step based groups have emerged to address the recovery needs of the many substance users with psychiatric comorbidity. Little is known about factors associated with retention in 12-step, especially in this population. This study sought to identify predictors of retention over a one-year period among members of a dual-focus 12-Step fellowship (N = 276). Using multivariate analysis, the following baseline characteristics were associated with greater retention one year later: older age, more lifetime arrests, abstinence in the pre-baseline year, more psychiatric symptoms in the pre-baseline year, not taking psychiatric medication, being more troubled by substance abuse than by mental health, and greater level of self-efficacy for recovery; residing in supported housing and being enrolled in outpatient treatment at follow-up were also significantly associated with better retention. Clinical implications to enhance retention in specialized 12-step groups are discussed.


Clinical and health services relationships between major depression, depressive symptoms, and general medical illness.
Patients with chronic medical illness have a high prevalence of major depressive illness. Major depression may decrease the ability to habituate to the aversive symptoms of chronic medical illness, such as pain. The progressive decrements in function associated with many chronic medical illnesses may cause depression, and depression is associated with additive functional impairment. Depression is also associated with an approximately 50% increase in medical costs of chronic medical illness, even after controlling for severity of physical illness. Increasing evidence suggests that both depressive symptoms and major depression may be associated with increased morbidity and mortality from such illnesses as diabetes and heart disease. The adverse effect of major depression on health habits, such as smoking, diet, over-eating, and sedentary lifestyle, its maladaptive effect on adherence to medical regimens, as well as direct adverse physiologic effects (i.e., decreased heart rate variability, increased adhesiveness of platelets) may explain this association with increased morbidity and mortality.


Cluster headache and "dynamite headache": blood flow velocities in the middle cerebral artery.
Nitroglycerin (NG) induces in cluster headache patients and controls an increase in systemic diastolic blood pressure and/or heart rate and a decrease in blood flow velocity in the middle cerebral artery (VMCA). Termination of NG induced cluster headache-like attack was correlated to an increase of VMCA compared to the VMCA before NG administration (p less than 0.01). This increase was not found in patients without attack or in controls. The NG induced "dynamite headache" in the controls subsided when blood pressure and heart rate were normalized, but the decrease of VMCA still prevailed. Orbital phlebograms have shown pathologic changes in cluster headache and in Tolosa-Hunt syndrome but not in controls. Ocular sympathetic nerves are involved in cluster headache but seldom in Tolosa-Hunt syndrome. It is suggested that the start of a cluster headache attack is due to an increase and the termination of the attack to a decrease of blood flow to the sympathoplegic phlebopathic cavernous sinus.


Next-Generation Sequencing in Tumor Diagnosis and Treatment.
Next-Generation Sequencing (NGS) allows for the sequencing of multiple genes at a very high depth of coverage [...].


Erectile dysfunction--the effect of sending a questionnaire to patients on consultations with their family doctor.
Erectile dysfunction (ED) is a common problem among male adults that generally has been ignored by family practitioners. Our aim was to assess the effect of a mailed questionnaire about ED on the readiness of patients to raise the subject with their family doctor. The study population included all men aged 40 years and over on the patient list of a family practitioner. A control group made up of males of similar ages was chosen in another family practice. The patient files were reviewed for ED. Anonymous questionnaires including questions about sexual dysfunction and satisfaction with sex life, as well as demographic and medical details, were sent to the study population. Patients who suffered from ED were invited to visit their family doctor. In the following 2 months, the study and control group populations' visits to the family practitioner were monitored for complaints of sexual dysfunction. In the 2 years prior to the study, 14/205 (6.8%) of the study population had complained to their family practitioner of ED in comparison with 6/205 (2.9%) in the control group (P = NS). In the 2 months following the sending of the questionnaire, 23 patients consulted their family practitioner with ED, 19 of whom had not discussed their problem with the family practitioner previously; only a further two patients went to discuss ED in the same period in the control group (P < 0.001). A total of 85/205 (41.5%) patients returned the questionnaire and 35/85 (42.5%) said they suffered from ED. Of 35 patients who reported ED, 15 had been for a consultation; only six of them consulted their family doctor. ED is reported infrequently to family doctors. Sending an anonymous questionnaire on the subject increases awareness of the problem and in turn increases the number of cases that can be treated.


Smart Responsive Luminescent Aptamer-Functionalized Covalent Organic Framework Hydrogel for High-Resolution Visualization and Security Protection of Latent Fingerprints.
Covalent organic frameworks (COFs) have been proposed as alternative candidates for "smart" materials due to their ordered π-columnar structures. However, it remains a challenge to develop external-stimuli-responsive luminescent COFs for confidential information protection. Here, we have designed and synthesized a water-dispersible and smart responsive luminescent carboxymethyl cellulose-COF hydrogel encapsulated 5-(dimethylamino)-N,N-bis (pyridin-2-ylmethyl) napthalene-1-sulfonamide, named CMC-COF-LZU1⊃DPYNS, for latent fingerprint imaging and encryption. We show that the fluorescence of CMC-COF-LZU1⊃DPYNS is reversibly switchable upon addition of Cu2+/H2O. This effect endows potential applications of tunable luminescent COFs based hydrogel as an invisible security probe for imaging, recording, storage, and security of latent fingerprint information. It is shown that the latent fingerprint information incubated by the aptamer-functionalized CMC-COF-LZU1⊃DPYNS hydrogel is invisible in the presence of Cu2+, but three levels of fingerprint features with high-resolution patterns could be readable upon addition of H2O under UV light. The design strategy provides a promising platform for the development of smart responsive luminescent COFs and their detection and protection of valuable latent fingerprint information.


Thermal conditions for successful breeding in Great Tits (Parus major L.) : I. Relation of growth and development of temperature regulation in nestling great tits.
The development of temperature regulation in relation to the growth and age of the nestlings is described in a way permitting use of the data in a model designed to predict the range of temperature tolerance of broods of Great Tits in the nestling stage. Such a model is described in a second paper. The physiological part of that model is made up mainly of six equations (nos. 6, 10, 11, 12, 14 and 15), which are all presented and discussed here. It is shown in this paper that the development of temperature regulation is a function of body weight rather than of age. The level of the basal metabolic rate of nestling Great Tits is lower than that of adult passerines of comparable size. The basal metabolic rate of a newly hatched Great Tit is only about one fourthe of the metabolic rate expected from Lasiewski and Dawson's equation for adult passerine birds. This discrepancy diminishes gradually during the nestling period and disapears shortly before fledging.Basal and maximum metabolic rates, as well as the body temperatures coinciding with these rates, are described in allometric equations as functions of nestling body weight. The evaporative heat loss of the nestlings is described as a function of body weight and body temperature, and an estimate of the maximum amount of water available to them for evaporative heat loss is given. A distinction is made between a long-term risk of hyperthermia, which results in mortality through dehydration of the nestling body, and an immediate risk of hyperthermia, which occurs when the maximum rate at which nestlings can evaporate water is insufficient to cope with the required heat loss by water evaporation. It is concluded that this immediate risk of hyperthermia is the most important of the factors affecting the upper limit of the range of temperature tolerance.


Effect of variations in temperature on antimuscarinic activity in guinea-pig atria.
The characteristics of the antimuscarinic activity of homatropine, gallamine and stercuronium in guinea-pig atria remained constant over the temperature range 22--37 degrees C in that a linear Arunlakshana-Schild plot was obtained with homatropine and nonlinear plots occurred with gallamine or stercuronium. A trend towards higher dose-ratios with reduction in temperature was only significant for gallamine.


TRPM2 mediates CaMKⅡ-Beclin-1 signaling in early cortical injury after induced subarachnoid hemorrhage in mice.
Though early brain injury (EBI) is the primary cause of poor outcomes among patients with subarachnoid hemorrhage (SAH), its exact molecular mechanisms remain unclear. Improved the understanding of how transient receptor potential melastatin-related 2 (TRPM2) is involved in SAH-induced EBI will help develop novel interventions. Wild type (WT) male C57BL/6J mice were subjected to SAH for 12 h, 24 h or 48 h, after which neurological scores and pathological changes in the hippocampus (CA3, DG, and CA1) and temporal base cortex were observed. Expressions of TRPM2, Ca2+/calmodulin (CaM)-dependent protein kinase Ⅱ (CaMKⅡ), and Beclin-1 in hippocampus (CA3, DG, and CA1) and temporal base cortex were compared across post-SAH timepoints. TRPM2-deficient (TRPM2-/-) male C57BL/6 J mice and a CaMKⅡ inhibitor (KN-93) were used to analyze the effects oTRPM2 on the CaMKⅡ-Beclin-1 signaling post SAH. Neurological and temporal base cortex deterioration were more severe with increased time post-SAH induction, whereas hippocampal damage was not observed. Post-SAH, TRPM2-CaMKⅡ-Beclin-1 cascade was activated in the temporal base cortex, but not the hippocampus. Using TRPM2-/- mice and KN-93 administration, SAH-induced EBI was improved, and CaMKⅡ and Beclin-1 expressions in the temporal base cortex were significantly decreased compared with WT mice. TRPM2-/- mice also showed better neurological improvement compared with KN-93 treated mice. TRPM2 mediates CaMKⅡ-Beclin-1 signaling that aggravates SAH-induced EBI in the temporal base cortex. TRPM2 may be an alternative therapy target in EBI after SAH. The datasets generated and/or analysed during the current study are available from the corresponding author.


Identification of the Yukb allo-antigen on platelet glycoprotein IIIa.
The glycoprotein (GP) localization of a new platelet-specific allo-antigen Yukb is described. The antibody was isolated from serum of a patient with neonatal allo-immune thrombocytopenia. In immunoblot procedure, it bound exclusively to platelet GP IIIa, like anti-PlA1, while the known anti-Baka and anti-Leka reacted with GP IIb. Analysis of GP from chymotrypsin-treated platelets with anti-Yukb revealed no binding in the 68-kilodalton position while anti-PlA1 did. Thus, unlike the PlA1 antigen, the Yukb determinant either resides on the 30-kilodalton fragment of GP IIIa or it is destroyed by chymotrypsin treatment.


Telmisartan attenuates aortic hypertrophy in hypertensive rats by the modulation of ACE2 and profilin-1 expression.
Profilin-1 has recently been linked to vascular hypertrophy and remodeling. Here, we assessed the hypothesis that angiotensin (Ang) II type I receptor antagonist telmisartan improves vascular hypertrophy by modulation of expression of profilin-1 and angiotensin-converting enzyme 2 (ACE2). Ten-week-old male spontaneously hypertensive rats (SHR) were received oral administration of telmisartan (5 or 10mg/kg; daily) or saline for 10 weeks. Compared with Wistar-Kyoto (WKY) rats, there were marked increases in systolic blood pressure and profilin-1 expression and reduced ACE2 and peroxisome proliferator activated receptor-γ (PPARγ) levels in aorta of SHR, associated with elevated extracellular-signal regulated kinase 1/2 (ERK1/2) and c-Jun N-terminal kinase (JNK) phosphorylation signaling and aortic hypertrophy characterized with increased media thickness, which were strikingly reversed by telmisartan. In cultured human umbilical artery smooth muscle cells (HUASMCs), Ang II induced a dose-dependent increase in profilin-1 expression, along with decreased ACE2 protein expression and elevated ERK1/2 and JNK phosphorylation. In addition, blockade of ERK1/2 or JNK by either specific inhibitor was able to abolish Ang II-induced ACE2 downregulation and profilin-1 upregulation in HUASMCs. Importantly, treatment with telmisartan (1 or 10 μM) or recombinant human ACE2 (2mg/ml) largely ameliorated Ang II-induced profilin-1 expression and ERK1/2 and JNK phosphorylation and augmented PPARγ expression in the cultured HUASMCs. In conclusion, telmisartan treatment attenuates vascular hypertrophy in SHR by the modulation of ACE2 and profilin-1 expression with a marked reversal of ERK1/2 and JNK phosphorylation signaling pathways.


Exercise training for claudication.
Exercise advice is a well established component of the conservative management of intermittent claudication. Supervised programmes of exercise remain relatively uncommon and are provided mainly in secondary care. This review outlines the evidence for the effectiveness of different exercise regimens and the relative benefits of exercise therapy, where comparisons have been made with medical therapy, angioplasty and surgery.


Histopathology of testis in acquired immune deficiency syndrome.
Histologic sections from the testes of 32 autopsied patients with acquired immune deficiency syndrome (AIDS) were examined. Almost invariably the testes displayed decreased spermatogenesis, and 20 of the 32 cases showed marked hypospermatogenesis with Sertoli cells predominantly lining the tubules. Although the seminiferous tubules were generally of normal size, the tunica propria at the periphery of the tubules was mildly to moderately thickened in 19 cases and markedly thickened in 10. The interstitial cells of Leydig were unaltered in most patients, with only 4 testes showing Leydig cell hyperplasia. The testicular blood vessels were slightly thickened in many patients, but 5 exhibited moderate to marked intimal proliferation with narrowing of the lumen. Mononuclear inflammatory infiltration of the testicular interstitium was slight in 11 cases, moderate in 6. Only 7 of the 28 AIDS patients with opportunistic infections had evidence of direct involvement of the testes by the infectious organisms. We concluded that the extragonadal endocrine balance of AIDS patients may be deranged due to the infectious process and so deserves clinical evaluation.


Masturbation: Scientific Evidence and Islam's View.
Masturbation is the stimulation of sexual organs usually to the point of orgasm with an essential autoerotic component. Due to the high prevalence of this sexual behavior, it was and still is a matter of debate if masturbation is a normal action without any side effects and even if it is advantageous or it is associated with side effects necessitating public education how to avoid it. In addition, it is a common question if masturbation is religiously lawful or not. In this study, I assess the results of scientific studies about this sexual behavior and also shed some light on the Islam's view about it.


Petiveria alleaceae L. (anamú). Study of the hypoglycemic effect.
The combined phytochemical and pharmaceutical study of Petiveria alleaceae L. (anamú) has shown the existence in the leaves and stems of the plant of a possible hypoglycemic active principle. Extracts from leaves and stem powder were found to produce a decrease of blood sugar concentration of more than 60% one hour after oral administration in male Balb/C mice weighing 20 g fasted for 48 hours.


Alteration of retinal rod outer segment membrane fluidity in a rat model of Smith-Lemli-Opitz syndrome.
Smith-Lemli-Opitz syndrome (SLOS) is caused by an inherited defect in the last step in cholesterol (Chol) biosynthesis, leading to abnormal accumulation of 7-dehydrocholesterol and decreased Chol levels. Progressive retinal degeneration occurs in an animal model of SLOS, induced by treating rats with AY9944, a selective inhibitor of the enzyme affected in SLOS. Here we evaluated alterations in the biochemical and physical properties of retinal rod outer segment (ROS) membranes in this animal model. At 1 month of AY9944 treatment, there were modest alterations in fatty acid composition, but no significant differences in cis-parinaric acid (cPA) spectroscopic parameters in ROS membranes from treated versus control rats. However, at 3 months, ROS docosahexaenoic acid (DHA) content was dramatically reduced, and cPA fluorescence anisotropy values were decreased, relative to controls. Also, 1,6-diphenyl-1,3,5-hexatriene exhibited decreased rotational motion and increased orientational order in ROS membranes from 3 month-old AY9944-treated rats, relative to controls. No significant changes in protein:lipid ratios were observed; however, rhodopsin regenerability was compromised by 3 months of treatment. These findings are consistent with reduced ROS membrane fluidity in the SLOS rat model, relative to controls, primarily due to the dramatic reduction in membrane DHA levels, rather than altered sterol composition.


Potentiometric glucose biosensor based on core-shell Fe3O4-enzyme-polypyrrole nanoparticles.
Core-shell Fe3O4-enzyme-polypyrrole (Ppy) nanoparticles with excellent magnetism and conductivity were successfully prepared via the surface modification and enzyme self-encapsulation within Ppy. A novel potentiometric glucose biosensor has been constructed by effectively attaching the proposed Fe3O4-enzyme-Ppy nanoparticles to the surface of the magnetic glassy carbon electrode (MGCE). The optimum biosensing conditions could be provided with polymerization time of pyrrole for 6h and 0.42 mg immobilization amount of Fe3O4-enzyme-Ppy nanoparticles on MGCE. The performance of the developed glucose biosensor was evaluated and the results indicated that a sensitive glucose biosensor could be fabricated. The obtained glucose biosensor presents shorter response time (6 s), wider linear range (0.5 μM to 34 mM), lower limit of detection (LOD, 0.3 μM), high-selectivity monitoring of glucose and good stability (with about 98.1% of the initial response signal retained after 20 days). The analytical application of the glucose biosensor confirms the feasibility of glucose detection in serum sample.


Reversing hypomyelination in BACE1-null mice with Akt-DD overexpression.
β-Site amyloid precursor protein convertase enzyme 1 (BACE1), a type I transmembrane aspartyl protease required to cleave amyloid precursor protein for releasing a toxic amyloid peptide, also cleaves type I and type III neuregulin-1 (Nrg-1). BACE1 deficiency in mice causes hypomyelination during development and impairs remyelination if injured. In BACE1-null mice, the abolished cleavage of neuregulin-1 by BACE1 is speculated to cause reduced myelin sheath thickness in both the central nervous system and peripheral nervous system because reduced cleavage of Nrg-1 correlates with reduced Akt phosphorylation, a downstream signaling molecule of the Nrg-1/ErbB pathway. Here we tested specifically whether increasing Akt activity alone in oligodendrocytes would be sufficient to reverse the hypomyelination phenotype in BACE1-null mice. BACE1-null mice were bred with transgenic mice expressing constitutively active Akt (Akt-DD; mutations with D(308)T and D(473)S) in oligodendrocytes. Relative to littermate BACE1-null controls, BACE1(-/-)/Akt-DD mice exhibited enhanced expression of myelin basic protein and promoter of proteolipid protein. The elevated expression of myelin proteins correlated with a thicker myelin sheath in optic nerves; comparison of quantified g ratios with statistic significance was used to confirm this reversion. However, it appeared that myelin sheath thickness in the sciatic nerves was not increased in BACE1(-/-)/Akt-DD mice, as the g ratio was not significantly different from the control. Hence, increased Akt activity in BACE1-null myelinating cells only compensates for the loss of BACE1 activity in the central nervous system, which is consistent with the observation that overexpression of Akt-DD in Schwann cells did not induce hypermyelination. Our results suggest that signaling activity other than Akt may also contribute to proper myelination in peripheral nerves.


A model-based study of passive joint properties on muscle effort during static stance.
This study examined the impact of lower extremity joint stiffnesses and simulated joint contractures on the muscle effort required to maintain static standing postures after a spinal cord injury (SCI). Static inverse computer simulations were performed with a three-dimensional 15 degree of freedom musculoskeletal model placed in 1600 different standing postures. The required lower extremity muscle forces were calculated through an optimization routine that minimized the sum of the muscle stresses squared, which was used as an index of the muscle effort required for each standing posture. Joint stiffnesses were increased and decreased by 100 percent of their nominal values, and contractures were simulated to determine their effects on the muscle effort for each posture. Nominal muscle and passive properties for an individual with a SCI determined the baseline muscle effort for comparisons. Stiffness changes for the ankle plantar flexion/dorsiflexion, hip flexion/extension, and hip abduction/adduction directions had the largest effect on reducing muscle effort by more than 5 percent, while changes in ankle inversion/eversion and knee flexion/extension had the least effect. For erect standing, muscle effort was reduced by more than 5 percent when stiffness was decreased at the ankle plantar flexion/dorsiflexion joint or hip flexion/extension joint. With simulated joint contractures, the postural workspace area decreased and muscle effort was not reduced by more than 5 percent for any posture. Using this knowledge, methods can be developed through the use of orthoses, physical therapy, surgery or other means to appropriately augment or diminish these passive moments during standing with a neuroprosthesis.


Association between cumulative cisplatin dose and reproductive and sexual functions in patients with malignant ovarian germ cell tumors treated with bleomycin, etoposide, and cisplatin therapy: a case series study.
The impact of cumulative dose of cisplatin on gonadal function has not been clarified. We evaluated whether the cumulative cisplatin dose affects the resumption of menses in patients treated with bleomycin, etoposide, and cisplatin (BEP). A case series study of women < 40 years with malignant ovarian germ cell tumors receiving BEP was conducted at Mie University Hospital. Using linear regression analysis, the correlation between the cumulative dose and resumption of menses was determined. Additionally, we compared the resumption of menses stratified by age (age < 20 years or ≥ 20 years). Ten women (median age: 20 [interquartile range: 15-26] years) have received a median of 4 cycles of BEP. The median period of resumption of menses was 5 months, which had no correlation with cumulative doses of bleomycin (143 mg/m2 [71-220], y = -0.0069 x + 6.15, r = 0.19, P = 0.60), etoposide (1,533 mg/m2 [900-2,000], y = 0.0004 x + 4.56, r = 0.08, P = 0.82), and cisplatin (363 mg/m2 [225-400], y = 0.01 x + 1.67, r = 0.35, P = 0.32). Although the resumption of menses was comparable across ages, the cumulative doses of cisplatin were higher in patients aged < 20 years than in those aged ≥ 20 years (400 mg/m2 [363-450] vs. 225 mg/m2 [225-350], P = 0.02). Similarly, patients aged < 20 years had a higher cumulative etoposide dose than those aged ≥ 20 years (2,000 mg/m2 [1,533-2,250] vs. 900 mg/m2[900-1,600], P = 0.03). Moreover, patients aged < 20 years received more cycles of BEP than those aged ≥ 20 years (4 cycles vs. 3 cycles, P = 0.03). All patients can recover menses after BEP, and the resumption of menses appeared at the median period of 5 months after BEP. The timing of menses resumption did not correlate with the cumulative doses of cisplatin.


External fixator followed by a plate for distraction, reduction, and fixation in neglected femoral diaphyseal fractures.
This study aims to investigate the outcome of a two-stage surgery in the treatment of neglected femoral diaphyseal fractures which are not uncommon in developing nations. Ten patients with neglected or late-presenting femoral diaphyseal fractures were considered in this study. All patients underwent a two-stage surgery, which consisted of distraction by an external fixator and open reduction by internal plate fixation. All patients received a supervised regimen of physiotherapy. Patients were followed up clinically and with radiographs at 2 months to assess union and at monthly intervals thereafter. All patients achieved bony union in an average of 3.7 months (2-6 months) with no one lost follow-up. Seven patients regained full range of motion, and the mean knee range of motion was 139.5°. No wound related or neurovascular complications were detected. One patient with stiff knee was re-admitted 1 year after surgery for metal removal, arthrolysis and quadricepsplasty for improving knee range of motion to 90° flexion. We conclude that the treating of neglected femoral diaphyseal fractures with a two-stage surgery is a satisfactory therapy showing reliable bony union, however continuing medical education is necessary for physicians in primary medical facilities as well as for patients with traditional views.


Role of hydration and water coordination in micellization of Pluronic block copolymers.
Raman, attenuated total reflectance FTIR, near-infrared spectroscopy, and DFT calculations have been used in a study of aqueous solutions of three tri-block copolymers poly(ethylene oxide)-poly(propylene oxide)-poly(ethylene oxide) or PEO-PPO-PEO with commercial names Pluronic PE6200, PE6400 and F68. It is shown that the process of micellization as a response to increased temperature is reflected in the hydroxyl stretching region of infrared and Raman spectra, which contains information both about restructuring of water and changes of polymer chains in polymer/water aggregates. Raman spectra exhibit differences between individual Pluronics even at temperatures below the critical micellization temperature (CMT). According to the attenuated total reflection (ATR) FTIR spectra, the same five water coordination types defined by the number of donated/accepted hydrogen bonds are present in interacting water as in bulk water. It indicates that models considering mixed states of water with different hydrogen bonding environments provide appropriate descriptions of bound water both below and above the CMT. Above the CMT, aggregate hydration increases in the order PE6400 < PE6200 < F68, although that does not fully correspond to the EO/PO ratio, and points to the differences in microstructure of aggregates formed by each copolymer. This study relates nanoscale phenomena (hydrophobic and hydrophilic hydration) with the mesoscale phenomenon of micellization.


Solving the Light's criteria misclassification rate of cardiac and hepatic transudates.
Pleural transudates are most commonly due to heart failure (HF) or hepatic hydrothorax (HH), but a number of these effusions are misclassified as exudates by standard (Light's) criteria. The aim of this study was to determine the prevalence of mislabelled transudates and to establish simple alternative parameters to correctly identify them. We retrospectively analysed the pleural fluid and serum protein, lactate dehydrogenase and albumin concentrations from 364 cardiac effusions and 102 HH. The serum-to-pleural fluid protein and albumin gradients (serum concentration minus pleural fluid concentration), as well as the pleural fluid-to-serum albumin ratio (pleural fluid concentration divided by the serum concentration) were calculated for the mislabelled transudates. Light's criteria had misclassified more HF-associated effusions than HH (29% vs 18%, P = 0.002). A serum-to-pleural fluid protein gradient >3.1 g/dL correctly identified 55% and 61% of the HF and HH false exudates, respectively. The figures for an albumin gradient >1.2 g/dL were 83% and 62%. Finally, a pleural fluid-to-serum albumin ratio <0.6 had identical accuracy for labelling miscategorized cardiac and liver-related effusions (78% and 77%, respectively). If the clinical picture is consistent with HF but the pleural fluid meets Light's exudative criteria, the measurement of the albumin rather than the protein gradient is recommended. In the context of cirrhosis, a potentially 'false' exudate is identified better by the pleural fluid-to-serum albumin ratio.


Identification of putative sex chromosomes in the blue tilapia, Oreochromis aureus, through synaptonemal complex and FISH analysis.
Sex determination in the blue tilapia, Oreochromis aureus, is primarily a ZW female-ZZ male system. Here, by analysis of the pachytene meiotic chromosomes of O. aureus, we demonstrate the presence of two distinct regions of restricted pairing present only in heterogametic fish. The first, a subterminal region of the largest bivalent is located near to the region of unpairing found in the closely related species O. niloticus, while the second is in a small bivalent, most of which was unpaired. These results suggest that O. aureus has two separate pairs of sex chromosomes.


Between-groups study designs demand between-groups analyses: a response to hernandez-reif, shor-posner, baez, soto, mendoza, castillo, quintero, perez, and zhang.
The most recent massage therapy (MT) study by Hernandez-Reif et al. displays flaws persistent in this area of research that are attributable to MT researchers' frequent mistake of using within-group analyses of dependent variables in studies that are purported to be randomized control trials. This practise violates the logic of using randomization to create treatment and control groups, and thereby fails to control for the validity threats of spontaneous remission, placebo effects, and statistical regression. The result is that a clear understanding of what MT can and cannot do is seriously hampered.


Does the immobilizing effect of thiopental in brain exceed that of halothane?
Recent studies suggest that anesthetics such as isoflurane act in the spinal cord to suppress movement that occurs during noxious stimulation. The authors examined the effect of halothane and thiopental on suppression of noxious-evoked movement using a model of differential anesthetic delivery. They hypothesized that halothane and thiopental, similar to isoflurane, would suppress movement primarily via an action in spinal cord. Goats were anesthetized and prepared for differential anesthetic delivery. Anesthesia was maintained with halothane (n = 5) or thiopental (n = 5). Anesthetic requirements were determined (noxious clamp on a dewclaw for 1 min) during halothane or thiopental (via infusion) delivery to the whole body and delivery only to the head. Control (whole body) halothane requirement was 0.9 +/- 0.2%; halothane requirement in the head during differential delivery was 3.4 +/- 1% (P < 0.01). During selective halothane delivery, the electroencephalogram was greatly depressed or was isoelectric even though the animals moved during noxious stimulation. Control (whole body) plasma thiopental requirement was 20 +/- 10 microg/ml. When thiopental was selectively delivered to the head, the electroencephalogram was active in all five animals, and cranial thiopental requirement was 42 +/- 6 microg/ml (P < 0.01). These data suggest that halothane and thiopental, like isoflurane, act in spinal cord to suppress movement occurring with noxious stimulation. However, halothane appears to be less potent in the brain as evidenced by the electroencephalogram data, suggesting that action in spinal cord plays a more significant role for halothane than for thiopental.


Estimating the burden of influenza-associated hospitalizations and deaths in Central America.
Our objective was to estimate the incidence of influenza-associated hospitalizations and in-hospital deaths in Central American Region. We used hospital discharge records, influenza surveillance virology data, and population projections collected from Costa Rica, El Salvador, Guatemala, Honduras, and Nicaragua to estimate influenza-associated hospitalizations and in-hospital deaths. We performed a meta-analysis of influenza-associated hospitalizations and in-hospital deaths. The highest annual incidence was observed among children aged <5 years (136 influenza-associated hospitalizations per 100 000 persons). Annually, 7 625-11 289 influenza-associated hospitalizations and 352-594 deaths occurred in the subregion. Our results suggest that a substantive number of persons are annually hospitalized because of influenza. Health officials should estimate how many illnesses could be averted through increased influenza vaccination.


Loss of Immunization-Induced Epitope-Specific CD4 T-Cell Response following Anaplasma marginale Infection Requires Presence of the T-Cell Epitope on the Pathogen and Is Not Associated with an Increase in Lymphocytes Expressing Known Regulatory Cell Phenotypes.
We have shown that in cattle previously immunized with outer membrane proteins, infection with Anaplasma marginale induces a functionally exhausted CD4 T-cell response to the A. marginale immunogen. Furthermore, T-cell responses following infection in nonimmunized cattle had a delayed onset and were sporadic and transient during persistent infection. The induction of an exhausted T-cell response following infection presumably facilitates pathogen persistence. In the current study, we hypothesized that the loss of epitope-specific T-cell responses requires the presence of the immunizing epitope on the pathogen, and T-cell dysfunction correlates with the appearance of regulatory T cells. In limited studies in cattle, regulatory T cells have been shown to belong to γδ T-cell subsets rather than be CD4 T cells expressing forkhead box protein P3 (FoxP3). Cattle expressing the DRB3*1101 haplotype were immunized with a truncated A. marginale major surface protein (MSP) 1a that contains a DRB3*1101-restricted CD4 T-cell epitope, F2-5B. Cattle either remained unchallenged or were challenged with A. marginale bacteria that express the epitope or with A. marginale subsp. centrale that do not. Peripheral blood and spleen mononuclear cells were monitored for MSP1a epitope F2-5B-specfic T-cell proliferative responses and were stained for γδ T-cell subsets or CD4(+) CD25(+) FoxP3(+) T cells before and during infection. As hypothesized, the induction of T-cell exhaustion occurred only following infection with A. marginale, which did not correlate with an increase in either CD4(+) CD25(+) FoxP3(+) T cells or any γδ T-cell subset examined.


Desmopressin.
Desmopressin (dDAVP), a synthetic analog of the neurohypophyseal nonapeptide arginine vasopressin, has enhanced antidiuretic potency, markedly diminished pressor activity, and a prolonged half-life and duration of action compared to the natural hormone. Desmopressin is the treatment of choice for central diabetes insipidus and can be administered either intranasally or parenterally. A newly approved indication is treatment of mild classical hemophilia and von Willebrand's disease, in which deficient concentrations of factor VIII and von Willebrand's factor are transiently increased to levels that allow minor surgery.


Brain volume in Tanzanian children with sickle cell anaemia: A neuroimaging study.
Brain injury is a common complication of sickle cell anaemia (SCA). White matter (WM) and cortical and subcortical grey matter (GM), structures may have reduced volume in patients with SCA. This study focuses on whether silent cerebral infarction (SCI), vasculopathy or anaemia affects WM and regional GM volumes in children living in Africa. Children with SCA (n = 144; aged 5-20 years; 74 male) and sibling controls (n = 53; aged 5-17 years; 29 male) underwent magnetic resonance imaging. Effects of SCI (n = 37), vasculopathy (n = 15), and haemoglobin were assessed. Compared with controls, after adjusting for age, sex and intracranial volume, patients with SCA had smaller volumes for WM and cortical, subcortical and total GM, as well as bilateral cerebellar cortex, globus pallidus, amygdala and right thalamus. Left globus pallidus volume was further reduced in patients with vasculopathy. Putamen and hippocampus volumes were larger in patients with SCA without SCI or vasculopathy than in controls. Significant positive effects of haemoglobin on regional GM volumes were confined to the controls. Patients with SCA generally have reduced GM volumes compared with controls, although some subcortical regions may be spared. SCI and vasculopathy may affect the trajectory of change in subcortical GM and WM volume. Brain volume in non-SCA children may be vulnerable to contemporaneous anaemia.


Chromosome 5D instability in cell lines of Triticum tauschii and morphological variation in regenerated plants.
Immature embryos of Triticum tauschii cultured on L2 medium with 30 g∙L−1maltose gave rise to compact and highly structured callus that readily regenerated shoots when transferred to MS medium supplemented with zeatin and indole-3-acetic acid. Two cell suspensions were initiated from the callus induced on L2 medium. An analysis of chromosome number in these cell suspensions after 3 months of culture showed that 95 and 75% of the cells had a normal complement. After 5 months, in both lines the majority of cells had chromosome complements of 2n = 13 and many of these aneuploid cells possessed a dicentric chromosome. C-banding indicated that the dicentric was often formed from chromosomes 2D and 5D. Fifty-six plants regenerated from callus were grown to maturity. Variation was observed in tiller number, flowering time, and seed-set; in two cases, the variation in early flowering was shown to be heritable. All the regenerants analysed had normal chromosome counts (2n = 2x = 14) and meiotic analyses of 35 plants revealed no obvious structural rearrangements.


Hepatocellular carcinoma of diffuse type: MR imaging findings and clinical manifestations.
To assess MR imaging findings and clinical manifestations of diffuse-type hepatocellular carcinoma (HCC). We retrospectively reviewed our experience with diffuse HCC from November 1994 to October 2001. MR imaging findings and clinical features were assessed. Twenty-two consecutive patients with diffuse-type HCC (19 men and three women, age range 16-80 years [mean, 52 years]) were identified in a review of liver MR studies. This represented 13% of all patients with HCC imaged during this time period. Diffuse HCC showed a permeative, infiltrative pattern with ill-defined borders and no evidence of convex margination in all cases. At least 50% of the liver volume was involved with tumor. Diffuse-type HCC showed hypointensity in 15 patients, mixed intensity in three, and isointensity in four on T1-weighted images; heterogeneous hyperintensity in 16 patients; and homogeneous hyperintensity in six on T2-weighted MR images. Diffuse-type HCC showed patchy enhancement in 12 patients, miliary enhancement in nine, and minimal enhancement in one on postcontrast early-phase images, and showed heterogeneous wash-out in all patients on postcontrast late-phase images. Proximal portal venous tumor thrombosis was seen in all patients. Serum alpha-fetoprotein (AFP) value was elevated (>10 ng/mL) in 14 of 18 patients, and 13 showed a value greater than 500 ng/mL. The four patients who did not have elevated AFP had tumors which were indistinguishable from those in patients with elevated AFP; they also did not have a distinctive clinical history. Diffuse-type HCC was typically seen as an extensive, heterogeneous permeative hepatic tumor, with portal venous tumor thrombosis on MR images in all cases. Early enhancement, observed as patchy in 12 and miliary in nine of 22 patients, was a distinctive imaging feature. Elevated serum AFP value was a common finding; however, 22% had normal values.


Key role of phosphorylation sites in ATPase domain and Linker region of MLH1 for DNA binding and functionality of MutLα.
MutLα is essential for human DNA mismatch repair (MMR). It harbors a latent endonuclease, is responsible for recruitment of process associated proteins and is relevant for strand discrimination. Recently, we demonstrated that the MMR function of MutLα is regulated by phosphorylation of MLH1 at serine (S) 477. In the current study, we focused on S87 located in the ATPase domain of MLH1 and on S446, S456 and S477 located in its linker region. We analysed the phosphorylation-dependent impact of these amino acids on DNA binding, MMR ability and thermal stability of MutLα. We were able to demonstrate that phosphorylation at S87 of MLH1 inhibits DNA binding of MutLα. In addition, we detected that its MMR function seems to be regulated predominantly via phosphorylation of serines in the linker domain, which are also partially involved in the regulation of DNA binding. Furthermore, we found that the thermal stability of MutLα decreased in relation to its phosphorylation status implying that complete phosphorylation might lead to instability and degradation of MLH1. In summary, we showed here, for the first time, a phosphorylation-dependent regulation of DNA binding of MutLα and hypothesized that this might significantly impact its functional regulation during MMR in vivo.


Genetic diversity and population structure analysis for morphological traits in upland cotton (Gossypium hirsutum L.).
A total of 96 different genotypes of upland cotton (Gossypium hirsutum) were selected from the breeding material and germplasm available at CCS HAU, India, to find the novel marker-trait associations for morphological traits used for registration of variety in upland cotton. Twenty-three morphological traits of the selected genotypes were recorded in field trials conducted in two replication of randomized block design during Kharif 2018 and 2019. A total of 11 traits showed sufficient variations in the screened germplasm and the same were further used for association mapping. A total of 168 SSRs were used for genotyping, of which 97 SSRs showed polymorphism amplifying 293 different alleles with an average of 3.02 alleles per SSR. Clustering, principal component analysis, and population structure analysis advocated that the current germplasm panel has enough diversity to be considered for association mapping. A total of 20 significant marker-trait associations were identified by the mixed linear model (MLM) and compressed mixed linear model (CMLM), of which 15 were common to both models, hence considered as promising associations. To the best of our knowledge, it is a first attempt to identify the linked markers in relation to morphological traits for the cotton crop. Results of the present study will be highly useful in speeding up variety registration programmes of upland cotton complementing to Distinctiveness, Uniformity, and Stability (DUS) testing.


Towards understanding nuclear pore complex architecture and dynamics in the age of integrative structural analysis.
Determining the functional architecture of the nuclear pore complex, that remains only partially understood, requires bridging across different length scales. Recent technological advances in quantitative and cross-linking mass spectrometry, super-resolution fluorescence microscopy and electron microscopy have enormously accelerated the integration of different types of data into coherent structural models. Moreover, high-resolution structural analysis of nucleoporins and their in vitro reconstitution into complexes is now facilitated by the use of thermostable orthologs. In this review we highlight how the application of such technologies has led to novel insights into nuclear pore architecture and to a paradigm shift. Today nuclear pores are not anymore seen as static facilitators of nucleocytoplasmic transport but ensembles of multiple overlaying functional states that are involved in various cellular processes.


Neonatal resuscitation practices in Italy: a survey of the Italian Society of Neonatology (SIN) and the Union of European Neonatal and Perinatal Societies (UENPS).
Providing appropriate care at birth remains a crucial strategy for reducing neonatal mortality and morbidity. We aimed to evaluate the consistency of practice and the adherence to the international guidelines on neonatal resuscitation in level-I and level-II Italian birth hospitals. This was a cross-sectional electronic survey. A 91-item questionnaire focusing on current delivery room practices in neonatal resuscitation was sent to the directors of 418 Italian neonatal facilities. The response rate was 61.7% (258/418), comprising 95.6% (110/115) from level-II and 49.0% (148/303) from level-I centres. In 2018, approximately 300,000 births occurred at the participating hospitals, with a median of 1664 births/centre in level-II and 737 births/centre in level-I hospitals. Participating level-II hospitals provided nasal-CPAP and/or high-flow nasal cannulae (100%), mechanical ventilation (99.1%), HFOV (71.0%), inhaled nitric oxide (80.0%), therapeutic hypothermia (76.4%), and extracorporeal membrane oxygenation ECMO (8.2%). Nasal-CPAP and/or high-flow nasal cannulae and mechanical ventilation were available in 77.7 and 21.6% of the level-I centres, respectively. Multidisciplinary antenatal counselling was routinely offered to parents at 90.0% (90) of level-II hospitals, and 57.4% (85) of level-I hospitals (p < 0.001). Laryngeal masks were available in more than 90% of participating hospitals while an end-tidal CO2 detector was available in only 20%. Significant differences between level-II and level-I centres were found in the composition of resuscitation teams for high-risk deliveries, team briefings before resuscitation, providers qualified with full resuscitation skills, self-confidence, and use of sodium bicarbonate. This survey provides insight into neonatal resuscitation practices in a large sample of Italian hospitals. Overall, adherence to international guidelines on neonatal resuscitation was high, but differences in practice between the participating centres and the guidelines exist. Clinicians and stakeholders should consider this information when allocating resources and planning perinatal programs in Italy.


Chondroitin sulfate-mediated albumin corona nanoparticles for the treatment of breast cancer.
Chondroitin sulfate-mediated albumin corona nanoparticles were readily prepared without any chemical reaction, and their active tumor targeting and therapeutic effects were examined. Negatively charged chondroitin sulfate (CS) and positively charged doxorubicin (DOX) self-assembled into nanoparticles (CS-DOX-NPs) via electrostatic interactions. Bovine serum albumin (BSA) was then adsorbed on the surface of CS-DOX-NPs to form albumin corona nanoparticles (BC-DOX-NPs) protected from endogenous proteins. Due to the dual effect of BSA and CS, BC-DOX-NPs interacted with the gp60, SPARC and CD44 receptors on tumor cells, facilitating their rapid and efficient transcytosis and improving their accumulation and uptake within tumor tissues. The simultaneous presence of BSA and CS also allowed BC-DOX-NPs to target CD44 efficiently, leading to greater cellular uptake and cytotoxicity against 4T1 cells than CS-DOX-NPs or free DOX. Intravenous injection of BC-DOX-NPs into orthotopic 4T1 tumor-bearing mice led to greater drug accumulation at the tumor site than with CS-DOX-NPs or free DOX, resulting in significant inhibition of tumor growth and lower exposure of major organs to the drug.


[Diagnosis and treatment of osteoporosis in childhood].
The authors review osteoporosis definition, the techniques to quantify bone mass, the assessment of therapy efficacy and the drugs used to manage osteoporosis in paediatric ages.


Pulse oximetry fails to accurately detect low levels of arterial hemoglobin oxygen saturation in dogs.
The accuracy of two commercially available pulse oximeters (the Ohmeda Biox 3700, software version "J," and the Nellcor N-100) in detecting low levels of arterial hemoglobin oxygen saturation (SaO2) was evaluated in 10 dogs in which hypoxia was induced by stopping the fresh gas flow into the anesthesia machine circle system. Measurements made in vivo with the pulse oximeters, with detectors placed on the tongue, were compared with measurements made in vitro using an IL 282 CO-Oximeter as SaO2 decreased toward zero. Measurements from the two oximeters correlated poorly over the range from 0 to 100% SaO2 (r = 0.69). In this range, the correlation between Nellcor N-100 measurements and those of the CO-Oximeter had an r of 0.82, a regression line slope of 0.82, and a y intercept of 14.8; the correlation between the Ohmeda Biox 3700 and the CO-Oximeter had an r of 0.83, a regression line slope of 0.66, and a y intercept of 32.7. The correlation with the CO-Oximeter was similar for both the Ohmeda and the Nellcor pulse oximeters at an SaO2 of 80% or more. However, when SaO2 was less than 80%, measurements by pulse oximetry correlated less well with CO-Oximeter measurements (r = 0.62, slope = 0.64, and y intercept = 21.0 for Nellcor; r = 0.71, slope = 0.67, and y intercept = 32.4 for Ohmeda). When SaO2 was less than 60%, both oximeters inaccurately indicated the co-oximetry values (r = 0.36 and y intercept = 26.1 for the Nellcor; r = 0.48 and y intercept = 33.2 for the Ohmeda).(ABSTRACT TRUNCATED AT 250 WORDS)


Evolution of serum hyaluronan and syndecan levels in prognosis of sepsis patients.
Endothelial glycocalyx shedding has been recognized as a contributor in sepsis pathophysiology. Hence, we attempted to analyze hyaluronan and syndecan (glycocalyx components) as markers of morbidity and prognosis of sepsis by performing serial measurements in these patients. Subjects were community acquired sepsis, severe sepsis and septic shock patients (150) admitted to ICU of our tertiary care hospital and controls were 50 healthy volunteers. Serum concentrations of markers were measured on days 1, 3, 5, 7 of ICU admission. Survival was assessed after 90days. Statistical analysis was performed by SPSS version 17. Hyaluronan and syndecan levels were significantly elevated in all categories of sepsis patients as compared to healthy controls (p<0.001). Levels of both markers were increased in severe sepsis and septic shock patients as compared to sepsis patient group at all time-points. Hyaluronan and syndecan differentiated survivors from non-survivors (p<0.001). Unlike non-survivors, in the survivor group, median hyaluronan and syndecan levels decreased significantly (p<0.001) in subsequent measurements. ROC analysis for the prediction of mortality identified cut-offs of 441ng/ml and 898ng/ml for hyaluronan and syndecan respectively. The specificity and negative predictive values were 90% and 90% for hyaluronan and 86% and 91% for syndecan respectively. Kaplan Meier curves revealed similar results. Both markers correlated significantly with APACHE II and SOFA scores. These observations indicate that serial measurements of hyaluronan and syndecan are significant prognostic markers for morbidity and survival in sepsis. Future therapeutic interventional possibilities need to be explored in experimental interventional prospective multi-centric trials.


Infrared irradiation alters the expression of matrix metalloproteinases and glycosaminoglycans in the cornea and crystalline lens.
Prolonged exposure to infrared (IR) radiation is associated with different types of damage to cornea and lens. The aim of our study was to investigate the effect of acute and chronic exposure to IR radiation on the activity of matrix metalloproteinase-2 (MMP-2) and MMP-9 and on the expression of glycosaminoglycans (GAG) in the rabbit cornea and crystalline lens. New Zealand rabbits were subjected to IR radiation for 4 months (chronic exposure to IR) or to normal light (control group). In experiments regarding acute exposure, animals were subjected to IR radiation or normal light for 12 h, in the presence of 0.1% diclofenac sodium (eye drops instilled in the right eye of animals) or saline (instilled in the left eye of animals). The cornea and lens were dissected away and homogenized. The activity of MMP-2 and MMP-9 was assayed by gelatine zymography. Total GAG were isolated from tissue specimens after lipid extraction and extensive digestion with pronase and DNase and characterized by treatment with GAG-degrading enzymes, followed by electrophoresis on cellulose acetate membranes. Acute or chronic exposure to IR radiation induced the activity of MMP-2 in cornea and lens, whereas only acute IR radiation increased the content of heparan sulphate in crystalline lens. Local administration of diclofenac sodium did not prevent the above effects of acute IR radiation. The detrimental effects of excessive or prolonged exposure of the eyes to IR radiation are associated with induced activity of MMP-2 in cornea and lens and alterations in the content of heparan sulphate in lens. Thus, MMP and GAG may offer alternative targets for pharmacological intervention to confront ocular damages associated with IR radiation.


SPARKS 2 and SP3 servers in CASP6.
Two single-method servers, SPARKS 2 and SP3, participated in automatic-server predictions in CASP6. The overall results for all as well as detailed performance in comparative modeling targets are presented. It is shown that both SPARKS 2 and SP3 are able to recognize their corresponding best templates for all easy comparative modeling targets. The alignment accuracy, however, is not always the best among all the servers. Possible factors are discussed. SPARKS 2 and SP3 fold recognition servers, as well as their executables, are freely available for all academic users on http://theory.med.buffalo.edu.


Endograft infection leading to rupture of aortic aneurysm.
Aortic endografting has quickly been accepted as a less morbid method of aneurysm repair. However, preservation of the aortic sac after endografting remains a liability of this procedure. Late rupture has occurred, albeit rarely. Graft infections are another rare complication of endografting. We present the first reported case, to our knowledge, of aortic rupture secondary to infection of an aortic endograft.


Relationship between morphological and physiological responses to waterlogging and salinity in Sporobolus virginicus (L.) Kunth.
The effects of waterlogging and salinity on morphological and physiological responses in the marsh grass Sporobolus virginicus (L.) Kunth were investigated in a 4×2 factorial experiment. Plants were subjected to four salinity levels (0, 100, 200 and 400 mol m-3 NaCl) and two soil inundation conditions (drained and flooded) for 42 days. Flooding at 0 mol m-3 NaCl caused initiation of adventitious surface roots, increased internal acration and plant height, induced alcohol dehydrogenase activity (ADH), and decreased belowground biomass and the number of culms per plant. Salinity increase from 0 to 400 mol m-3 NaCl under drained conditions increased leaf and root proline concentrations and decreased photosynthesis, aboveground biomass, number of culms per plant and number of internodes per culm. Concurrent waterlogging and salinity induced ADH activity and adventitious surface roots but decreased plant height and aboveground biomass. Internal air space increased with waterlogging from 0 to 100 mol m-3 NaCl but further increases in salinity to 400 mol m-3 reduced air space. Combined waterlogging and salinity stresses, however, had no effect on photosynthesis or on the concentrations of proline in leaves or roots. These results are discussed in relation to the widespread colonization by S. virginicus of a wide range of coastal environments varying in soil salinity and in the frequency and intensity of waterlogging.


Consumption of Yeast-Fermented Wheat and Rye Breads Increases Colitis and Mortality in a Mouse Model of Colitis.
Cereals are known to trigger for wheat allergy, celiac disease and non-celiac wheat sensitivity (NCWS). Inflammatory processes and intestinal barrier impairment are suspected to be involved in NCWS, although the molecular triggers are unclear. We were interested if different bread types influence inflammatory processes and intestinal barrier function in a mouse model of inflammatory bowel disease. Epithelial caspase-8 gene knockout (Casp8ΔIEC) and control (Casp8fl) mice were randomized to eight groups, respectively. The groups received different diets for 28 days (gluten-free diet, gluten-rich diet 5 g%, or different types of bread at 50 g%). Breads varied regarding grain, milling and fermentation. All diets were isocaloric. Regardless of the diet, Casp8ΔIEC mice showed pronounced inflammation in colon compared to ileum, whereas Casp8fl mice were hardly inflamed. Casp8fl mice could tolerate all bread types. Especially yeast fermented rye and wheat bread from superfine flour but not pure gluten challenge increased colitis and mortality in Casp8ΔIEC mice. Hepatic expression of lipopolysaccharide-binding protein and colonic expression of tumor necrosis factor-α genes were inversely related to survival. The bread diets, but not the gluten-rich diet, also decreased colonic tight junction expression to variable degrees, without clear association to survival and inflammation. Bread components, especially those from yeast-fermented breads from wheat and rye, increase colitis and mortality in Casp8ΔIEC mice highly susceptible to intestinal inflammation, whereas control mice can tolerate all types of bread without inflammation. Yet unidentified bread components other than gluten seem to play the major role.


Antitumour activity on extrinsic apoptotic targets of the triterpenoid maslinic acid in p53-deficient Caco-2 adenocarcinoma cells.
We report that a novel triterpenoid, (2a,3b)-2,3-dihydroxyolean-12-en-28-oic acid (maslinic acid), isolated from olive pomace from Olea europaea, triggers primarily the extrinsic and later the intrinsic apoptotic pathways in Caco-2 human colon-cancer cells. Apoptosis induced by maslinic acid was confirmed by FACS analysis using annexine-V FICT staining. This induction of apoptosis was correlated with the early activation of caspase-8 and caspase-3, the activation of caspase-8 was also correlated with higher levels of Bid cleavage and decreased Bcl-2, but with no change in Bax expression. Maslinic acid also induced a sustained activation of c-Jun N-terminal kinase (JNK). Incubation with maslinic acid also resulted in the later activation of caspase-9, which, together with the lack of any Bax activation, suggests that the mitochondrial pathway is not required for apoptosis induced by maslinic acid in this cell line. In this study we found that the mechanism of apoptotic activation in p53-deficient Caco-2 cells differs significantly from that found in HT-29 cells. Natural agents able to activate both the extrinsic and intrinsic apoptotic pathways by avoiding the mitochondrial resistance mechanisms may be useful for treatment against colon cancer regardless of its aetiology.


Rituximab in autoimmune pemphigoid diseases: Indications, optimized regimens, and practice gaps.
Rituximab is a monoclonal antibody targeting CD20 on B cells with proven efficacy for pemphigus vulgaris, now an FDA-approved indication. Other autoimmune bullous diseases can be challenging to treat and have significant associated morbidity and mortality, but data supporting the use of rituximab in pemphigoid group diseases remain limited. Although rituximab demonstrates efficacy for clinical improvement and remission in pemphigoid, concern for adverse events may also limit the use of this medication. We review the current evidence fo rthe use of rituximab in pemphigoid diseases, pertinent dosing schedules and laboratory monitoring, and the associated common and rare adverse events. Review of the literature to date not only supports consideration of rituximab for treatment of refractory pemphigoid group diseases but also reflects tolerability and an acceptable safety profile.


MCM family in gastrointestinal cancer and other malignancies: From functional characterization to clinical implication.
Despite the recent advances in cancer research and treatment, gastrointestinal (GI) cancers remain the most common deadly disease worldwide. The aberrant DNA replication serves as a major source of genomic instability and enhances cell proliferation that contributes to tumor initiation and progression. Minichromosome maintenance family (MCMs) is a well-recognized group of proteins responsible for DNA synthesis. Recent studies suggested that dysregulated MCMs lead to tumor initiation, progression, and chemoresistance via modulating cell cycle and DNA replication stress. Their underlying mechanisms in various cancer types have been gradually identified. Furthermore, multiple studies have investigated the association between MCMs expression and clinicopathological features of cancer patients, implying that MCMs might serve as prominent prognostic biomarkers for GI cancers. This review summarizes the current knowledge on the oncogenic role of MCM proteins and highlights their clinical implications in various malignancies, especially in GI cancers. Targeting MCMs might shed light on the potential for identifying novel therapeutic strategies.


The deubiquitinating enzyme USP37 promotes keloid fibroblasts proliferation and collagen production by regulating the c-Myc expression.
Previous research testifies that c-Myc may promote keloid fibroblast proliferation and collagen accumulation. Ubiquitin-specific peptidase 37 (USP37)-mediated deubiquitination and stabilisation of c-Myc are vital for lung cancer proliferation, while the potential role of USP37 in keloid fibroblasts is not investigated. Elevated USP37, c-Myc, and Collagen I content were detected in keloid tissue with RT-PCR or ELISA assay. USP37 over-expression plasmids or USP37 short hairpin RNAs (shRNAs) were transfected into keloid fibroblasts with Lipofectamine 3000 to decipher the role of USP37 in keloid fibroblasts. USP37 overexpression could promote the proliferation of keloid fibroblasts with increased c-Myc and Collagen I expression. On the other hand, USP37 shRNAs inhibited the proliferation of keloid fibroblasts with diminished c-Myc and Collagen I expression. It was worth noting that C-Myc overexpression promoted the proliferation of keloid fibroblasts inhibited by USP37 shRNAs with increasing Collagen I expression. All of these results demonstrate that USP37 could regulate c-Myc to promote the proliferation and collagen deposit of keloid fibroblasts, and USP37 could be targeted in future keloid therapy.


Complete Remission of Post-transplantation Recurrence of Focal Segmental Glomerulosclerosis With the Use of Adrenocorticotrophic Hormone Gel: Case Report.
Post-transplantation recurrence of primary focal and segmental glomerulosclerosis (FSGS) is estimated to occur in 30%-50% of cases and doubles the risk of allograft failure. Treatment of recurrent FSGS is challenging because specific pathogenic targets are unknown and available therapeutic options have limited efficacy. We report a case of recurrent FSGS with nephrotic-range proteinuria (urine protein creatinine ratio [UPCR], >50) and debilitating edema that was resistant to rituximab and plasmapheresis. The patient had a remarkable response to adrenocorticotropic hormone (ACTH) gel and achieved complete remission (UPCR, 0.5; serum albumin, 4.1 g/dL; serum creatinine, 1.0 mg/dL) which was maintained over 10 months on this treatment. We conclude that ACTH gel is a potential therapeutic option for post-transplantation recurrence of FSGS and warrants further evaluation.


Shoulder load during synchronous handcycling and handrim wheelchair propulsion in persons with paraplegia.
To compare the shoulder load during handcycling and wheelchair propulsion under similar conditions of external power in persons with spinal cord injury. Cross-sectional. Eight men with spinal cord injury. Kinetics and kinematics were measured during handbike and wheelchair propulsion at 25, 35, 45 and 55 W on a treadmill. Shoulder load (glenohumeral contact forces, relative muscle forces) was calculated with the Delft Shoulder and Elbow Model. At all power output levels, glenohumeral contact forces were significantly lower during handcycling compared with wheelchair propulsion (p < 0.001). At 55 W, the mean glenohumeral contact force was 345 N for hand-cycling, whereas it was 585 N for wheelchair propulsion. Also, relative muscle forces were lower during handcycling. The largest differences between handbike and wheelchair propulsion were found in the supraspinatus (4.5% vs. 20.7%), infraspinatus (3.7% vs. 16.5%) and biceps (5.0% vs. 17.7%). Due to continuous force application in hand-cycling, shoulder load was lower compared with wheelchair propulsion. Furthermore, muscles that are prone to overuse injuries were less stressed during handcycling. Therefore, handcycling may be a good alternative for outdoor mobility and may help prevent overuse injuries of the shoulder complex.


Toxoplasma-safe meat: close to reality?
In 2008, the centennial of the discovery of Toxoplasma gondii was celebrated. However, toxoplasmosis is still seen as a neglected and underreported disease, despite having a disease burden similar to that of salmonellosis and campylobacteriosis. Human vaccines are not available and current antiparasitic treatment is disappointing. This has led to an urge to focus more on prevention. Food, soil or water contaminated with oocysts from cat faeces and undercooked meat from infected intermediate hosts are important routes of infection. Oocyst contamination is difficult to control, whereas in Western countries, the control of T. gondii in meat should be feasible. Here, we discuss strategies aimed at developing a Toxoplasma-safe meat chain.


Influence of race and ethnicity on in vitro fertilization outcomes: systematic review.
We conducted a systematic review to evaluate the influence of race and ethnicity on clinical pregnancy and live birth outcomes after in vitro fertilization (IVF). We searched PubMed, EMBASE, Web of Science, CINAHL, POPLINE, and Cochrane Central, and hand-searched relevant articles published through July 22, 2015. Two reviewers independently evaluated abstracts to identify studies that compared clinical pregnancy rates and live birth rates for ≥2 racial and/or ethnic groups after nondonor IVF cycles. Twenty-four studies were included. All 5 US registry-based studies showed that black, Hispanic, and Asian women had lower clinical pregnancy rates and/or live birth rates after IVF, compared with white women. Similarly, most clinic-specific studies reported significant disparities in these primary outcomes, potentially attributable to differences in infertility diagnosis, spontaneous abortion, and obesity. Studies varied with respect to definitions of race/ethnicity, inclusion of first cycles vs multiple cycles for individual women, and collected covariates. Most studies were limited by sample size, inadequate adjustment for confounding, selection bias, and extensive missing data. Although current evidence points to race and ethnicity, especially black race, as strong predictors of poorer outcomes after IVF, the utility of results is constrained by the limitations described.


Chitosan nanocomposite films based on halloysite nanotubes modification for potential biomedical applications.
Chitosan is attracting increasing attention for biomedical applications because of its biocompatibility. In the present study, raw halloysite nanotubes (RHNTs) were functionalised with (3-aminopropyl) triethoxysilane (APTS) and then a sequence of novel chitosan biofilms were prepared by adding amino-modified halloysite nanotubes (HNTs-NH2) as a reinforcing material and ethylene glycol diglycidyl ether (EGDE) as a cross-linking agent. The reaction between the APTS and the RHNTs was demonstrated through characterisation of the HNTs-NH2. Fourier transform infra-red spectroscopy (FTIR) and X-ray diffraction (XRD) results confirmed the interaction of HNTs-NH2 with chitosan and EGDE. Scanning electron microscopy (SEM) showed a transformation of the surface morphology of the chitosan films. Measurement of the mechanical and thermal properties showed that the nanocomposite films exhibited substantial improvements in tensile strength, elongation at break and thermal stability compared with those of the pure chitosan films. However, the swelling rate of the nanocomposite films decreased upon incorporation of the HNTs-NH2 and EGDE. In addition, the water vapour transmission rate (WVTR) of the nanocomposite films was also improved. Given the aforementioned results, chitosan nanocomposites are promising biomedical materials.


[Basophilles degranulation test. Its value in an overseas allergologic dispensary (author's transl)].
Specific Ig.E fixed on basophilic leucocytes and mastocytes give way to a release of cytoplasmic granulations when antigen is present. This is the base of a clinical laboratory test easy to perform and consequently of great value in tropical areas. Report of first results in Republic of Djibouti.


Evaluation of the antimalarial properties and standardization of tablets of Azadirachta indica (Meliaceae) in mice.
The antimalarial activities of the tablet suspension of the bark and leaf of Azadirachta indica were evaluated on Plasmodium yoelli nigeriensis infected mice. The tablet suspensions exhibited high prophylactic, mode-rate suppressive and a very minimal curative schizonticidal effect. No animal was cured of the infection in the curative test and there was not much increase in the survival time of the animals compared with the control. The tablet suspensions from the leaf and bark at a concentration of 800 mg/kg and chloroquine at a concentration of 62.5 mg/kg body weight produced average percentage (%) parasitaemia of 79.6%, 68.2% and 99.5% for leaf, bark and chloroquine, respectively, in chemosuppression. Also in the prophylactic treatment, the tablet suspensions at 800 mg/kg and pyrimethamine at a concentration of 0.35 mg/kg gave an average parasitaemia reduction of 75.3%, 65.6% and 98.3% for the leaf, bark and pyrimethamine, respectively. There was a clear indication that both tablet suspensions from the leaf and bark possess antimalarial activity and a suspension from the former is relatively more effective than the bark. Extrapolation of the results from the antimalarial activity of the tablet suspension of the crude plant parts showed that an adult human would need to ingest a minimum of 48 g of the powdered plant material per day, an amount that is impracticable. A survival index value of 0.33 was obtained with the 800 mg/kg dose level, indicating that the tablet suspension has some moderate beneficial effect.


Ionic interactions. Subnanoscale hydrophobic modulation of salt bridges in aqueous media.
Polar interactions such as electrostatic forces and hydrogen bonds play an essential role in biological molecular recognition. On a protein surface, polar interactions occur mostly in a hydrophobic environment because nonpolar amino acid residues cover ~75% of the protein surface. We report that ionic interactions on a hydrophobic surface are modulated by their subnanoscale distance to the surface. We developed a series of ionic head groups-appended self-assembled monolayers with C2, C6, C8, and C12 space-filling alkyl chains, which capture a dendritic guest via the formation of multiple salt bridges. The guest release upon protonolysis is progressively suppressed when its distance from the background hydrophobe changes from 1.2 (C2) to 0.2 (C12) nanometers, with an increase in salt bridge strength of ~3.9 kilocalories per mole.


Detection of single-nucleotide polymorphisms with novel leaky surface acoustic wave biosensors, DNA ligation and enzymatic signal amplification.
This manuscript describes a new technique for detecting single-nucleotide polymorphisms (SNPs) by integrating a leaky surface acoustic wave (LSAW) biosensor, enzymatic DNA ligation and enzymatic signal amplification. In this technique, the DNA target is hybridized with a capture probe immobilized on the surface of a LSAW biosensor. Then, the hybridized sequence is ligated to biotinylated allele-specific detection probe using Taq DNA ligase. The ligation does not take place if there is a single-nucleotide mismatch between the target and the capture probe. The ligated detection probe is transformed into a streptavidin-horseradish peroxidase (SA-HRP) terminal group via a biotin-streptavidin complex. Then, the SA-HRP group catalyzes the polymerization of 3,3-diaminobenzidine (DAB) to form a surface precipitate, thus effectively increasing the sensitivity of detecting surface mass changes and allowing detection of SNPs. Optimal detection conditions were found to be: 0.3 mol/L sodium ion concentration in PBS, pH 7.6, capture probe concentration 0.5 μmol/L and target sequence concentration 1.0 μmol/L. The detection limit was found to be 1 × 10(-12)mol/L. Using this technique, we were able to detect a single-point mutation at nucleotide A2293G in Japanese encephalitis virus.


Histocultures of patient head and neck tumors for pharmacodynamics studies.
This investigation was to establish a clinically relevant experimental model to evaluate the pharmacodynamics of drugs used for head and neck cancers. A total of 83 surgical samples of primary and lymph nodal metastatic tumors was obtained from 66 patients. Fragments of these tumors were cultured on a collagen gel matrix. The tumor cell labeling index (LI) was determined by [3H]thymidine incorporation and autoradiography. Seventeen tumors (20%) were contaminated. About 80% of the remaining 65 tumors were successfully cultured for at least 2 weeks. The cultured tumor fragments retained the morphology and architecture of the freshly removed specimens; both tumor and stromal cells were present. The tumor cell LI after 2-3 weeks in culture, determined from the most proliferative area of the tissue, averaged 77 +/- 12% for primary tumors and 78 +/- 12% for nodal metastases. The activity of three clinically active agents, 5-fluorouracil (FU), cisplatin (DDP), and mitomycin C (MMC), was evaluated in 47 tumors. All three drugs inhibited the tumor LI. The concentrations needed to produce a 50% inhibition of the tumor LI (IC50) were within the clinically achievable concentration range. The intertumor variation in the IC50 for FU (60-fold) was considerably greater than that for DDP and MMC (7- to 8-fold). The nodal metastatic tumors appeared to be less sensitive to FU than the primary tumors, while there were no apparent differences for DDP or MMC.(ABSTRACT TRUNCATED AT 250 WORDS)


Current Response in Ca V 1.3-/- Mouse Vestibular and Cochlear Hair Cells.
Signal transmission by sensory auditory and vestibular hair cells relies upon Ca2+-dependent exocytosis of glutamate. The Ca2+ current in mammalian inner ear hair cells is predominantly carried through Ca V 1.3 voltage-gated Ca2+ channels. Despite this, Ca V 1.3 deficient mice (Ca V 1.3-/- ) are deaf but do not show any obvious vestibular phenotype. Here, we compared the Ca2+ current (I Ca ) in auditory and vestibular hair cells from wild-type and Ca V 1.3-/- mice, to assess whether differences in the size of the residual I Ca could explain, at least in part, the two phenotypes. Using 5 mM extracellular Ca2+ and near-body temperature conditions, we investigated the cochlear primary sensory receptors inner hair cells (IHCs) and both type I and type II hair cells of the semicircular canals. We found that the residual I Ca in both auditory and vestibular hair cells from Ca V 1.3-/- mice was less than 20% (12-19%, depending on the hair cell type and age investigated) compared to controls, indicating a comparable expression of Ca V 1.3 Ca2+ channels in both sensory organs. We also showed that, different from IHCs, type I and type II hair cells from Ca V 1.3-/- mice were able to acquire the adult-like K+ current profile in their basolateral membrane. Intercellular K+ accumulation was still present in Ca V 1.3-/- mice during I K,L activation, suggesting that the K+-based, non-exocytotic, afferent transmission is still functional in these mice. This non-vesicular mechanism might contribute to the apparent normal vestibular functions in Ca V 1.3-/- mice.


A comparison of lidocaine 2% with levobupivacaine 0.75% for sub-Tenon's block.
To compare the onset of action, and quality of block, of lidocaine 2% with levobupivacaine 0.75% for sub-Tenon's block in patients undergoing cataract surgery. We performed a two-centre trial in 91 patients who were randomized to receive 4 mL of lidocaine 2% (n = 44) or levobupivacaine 0.75% (n = 47) for sub-Tenon's block, both with hyaluronidase 15 IUmL(-1). Onset of akinesia was assessed every 2 min for 10 min. Numbers of patients requiring supplementary injections to achieve clinically satisfactory akinesia or rescue analgesia were recorded. Data were analyzed with Fisher's exact test, U-test and t-test where appropriate. Results were considered significant when P < 0.05. The speed of onset was statistically significantly faster for lidocaine compared to levobupivacaine (3.02 vs. 5.06 min, P < 0.001). There was no statistical difference in number of patients requiring a supplementary injection of local anaesthetic (levobupivacaine 3 vs. lidocaine 0, P = 0.24), rescue analgesia with topical tetracaine (levobupivacaine 0 vs. lidocaine 2, P = 0.5), or ocular akinesia scores at the completion of surgery (lidocaine 1.4 vs. levobupivacaine 1.6, P = 0.12). Pain scores measured by a verbal analogue scale were not significantly different for injection, perioperatively or postoperatively. Both agents produce a rapid onset of anaesthesia when used for sub-Tenon's block. The difference between the two agents, although statistically significant, is not clinically important.


Life-threatening alterations in heart rate after the use of adenosine in atrial flutter.
Adenosine has become the preferred treatment for common types of supraventricular tachycardia because it is extremely effective and rarely associated with with serious side effects. It has also been advocated as an intervention for diagnostic use to assess uncommon types of tachycardia. Evidence is shown in this report that adenosine was associated with dangerous worsening of arrhythmia in patients with atrial flutter. In two patients, adenosine precipitated acceleration of ventricular response, in one case necessitating emergent cardioversion. Both patients had atrial flutter with 2 to 1 atrioventricular block that evolved into 1 to 1 atrioventricular conduction. In three other patients, adenosine was associated with prolonged bradyasystole and hypotension. In each of the five patients, adenosine was given in a standard fashion (6 or 12 mg). In summary, adenosine should be recognized as a potentially dangerous intervention in patients with atrial flutter. If it is used for diagnostic purposes, resuscitative equipment should be readily available.