In vitro activity of flurithromycin, a novel macrolide antibiotic.
Flurithromycin is an (8,S)-8-fluoroerythromycin isolated from the fermentation broth of Streptomyces erythraeus ATCC 31772, a blocked mutant of a strain producer of erythromycin. Its in vitro antibacterial activity has been determined on recent clinical isolates of respiratory pathogens. The range of MIC for Streptococcus pneumoniae and Streptococcus beta-haemolyticus group A is from 0.0015 to 0.006 microgram/ml, for Haemophilus influenzae from 0.012 to 0.4 microgram/ml, for Staphylococcus aureus from 0.1 to 3.1 micrograms/ml. Its action is bacteriostatic and increases at alkaline pH. Among anaerobes Clostridium perfringens, Bacteroides fragilis, other species of Bacteroides and Peptostreptococcus are particularly susceptible. Flurithromycin also showed some activity on Mycobacterium bovis, M. scrofulaceum and M. phley. The determination of killing curves indicated that in most cases a killing effect was obtained at 4 X MIC. A combination of flurithromycin with ampicillin or doxycycline sometimes was synergic, but more often additive and never antagonistic. The possible interference of flurithromycin on some parameters of the natural system of defense was determined. At concentrations equal to therapeutic levels in blood and tissues, flurithromycin did not influence chemotaxis, phagocytosis, metabolic activation and the killing activity of neutrophils.


Crystallization and preliminary X-ray crystallographic studies of Escherichia coli xanthine phosphoribosyltransferase.
Xanthine phosphoribosyltransferase (XPRT; EC 18.104.22.168) from Escherichia coli is a purine salvage enzyme which synthesizes the nucleotides GMP, XMP, and IMP. A mutant C59A, which is more stable than wild-type XPRT while retaining high activity, has been prepared and crystallized to give three different crystal forms (A, B, and C). Form A crystals are orthorhombic (P21212), with unit cell dimensions a = 59.2 A, b = 92.9 A, c = 53.2 A. Form B crystals are monoclinic (C2) with unit cell dimensions a = 84.4 A, b = 70.8 A, c = 54.1 A, and beta = 113.4 degrees, and form C crystals are tetragonal (P41212 or P43212) with unit cell dimensions a,b = 94 A, c = 167.5 A. Wild-type XPRT and a selenomethionine derivative of C59A XPRT have also been crystallized in the orthorhombic form. The selenomethionine derivative was prepared by expressing XPRT in the usual E. coli strain without the need for a methionine auxotroph. Cells were grown in a methionine-deficient medium supplemented with selenomethionine which gave >95% incorporation. Both the wild-type and selenomethionine C59A XPRT crystals are isomorphous with C59A form A crystals.


Impact of COVID-19 lockdown measures on oncological surgical activity: Analysis of the surgical pathology caseload of a tertiary referral hospital in Northwestern Italy.
Italy was severely affected by the severe acute respiratory syndrome coronavirus 2 pandemic. Our Institution, Piedmont's largest tertiary referral center, was designated as a non-COVID-19 hospital and activities were reorganized to prioritize critical services like oncological care. The aim of this study was to investigate the efficacy in preserving the oncological surgical practice at our Institution during the most critical months of the COVID-19 epidemic by analyzing the surgical pathology activity. The number of oncological surgical resections submitted to histopathological examination from 9th March 2020 to 8th May 2020 were collected as well staging/grading data and compared with the previous three pre-COVID-19 years (2017-2019). Overall, no decrease was observed for most tumor sites (5/9) while breast resections showed the largest drop (109 vs. 160; -31.9%), although a full recovery was already noticed during the second half of the period. Conversely, the selected control benchmarks showed a sharp decrease (-80.4%). Distribution of pathological TNM stages (or tumor grades for central nervous system tumors) showed no significant differences during the lockdown compared with previous years (p > .05). The present data suggest the possibility of preserving this cornerstone oncological activity during an evolving public health emergency thanks to a prompt workflow reorganization.


Vocabulary Repetition Following Multisensory Instruction Is Ineffective on L2 Sentence Comprehension: Evidence From the N400.
Putting the principles of multisensory teaching into practice, this study investigated the effect of audio-visual vocabulary repetition on L2 sentence comprehension. Forty participants were randomly assigned to experimental and control groups. A sensory-based model of instruction (i.e., emotioncy) was used to teach a list of unfamiliar vocabularies to the two groups. Following the instruction, the experimental group repeated the instructed words twice, while the control group received no vocabulary repetition. Afterward, their electrophysiological neural activities were recorded through electroencephalography while doing a sentence acceptability judgment task with 216 sentences under acceptable (correct) and unacceptable (pragmatically violated) conditions. A one-way analysis of variance (ANOVA), a multivariate analysis of variance (MANOVA), and a Bayesian repeated-measures ANOVA were used to compare the behavioral and neurocognitive responses [N400 as the main language-related event-related brain potential (ERP) effect] of the two groups. The results showed no significant N400 amplitude difference in favor of any of the groups. The findings corroborated the ineffectiveness of two repetitions preceded by multisensory instruction on L2 sentence comprehension.


Recent Research Trends on the Enzymatic Synthesis of Structured Lipids.
Structured lipids (SLs) are lipids that have been chemically or enzymatically modified from their natural biosynthetic form. Because SLs are made to possess desired nutritional, physicochemical, or textural properties for various applications in the food industry, many research activities have been aimed at their commercialization. The production of SLs by enzymatic procedures has a great potential in the future market because of the specificity of lipases and phospholipases used as the biocatalysts. The aim of this review is to provide concise information on the recent research trends on the enzymatic synthesis of SLs of commercial interest, such as medium- and long-chain triacylglycerols, human milk fat substitutes, cocoa butter equivalents, trans-free or low-trans plastic fats (such as margarines and shortenings), low-calorie fats/oils, health-beneficial fatty acid-rich fats/oils, mono- or diacylglycerols, and structurally modified phospholipids. This limited review covers 108 research articles published between 2010 and 2014 which were searched in Web of Science.


Warfarin therapy: evolving strategies in anticoagulation.
Warfarin is the oral anticoagulant most frequently used to control and prevent thromboembolic disorders. Prescribing the dose that both avoids hemorrhagic complications and achieves sufficient suppression of thrombosis requires a thorough understanding of the drug's unique pharmacology. Warfarin has a complex dose-response relationship that makes safe and effective use a challenge. For most indications, the dose is adjusted to maintain the patient's International Normalized Ratio (INR) at 2 to 3. Because of the delay in factor II (prothrombin) suppression, heparin is administered concurrently for four to five days to prevent thrombus propagation. Loading doses of warfarin are not warranted and may result in bleeding complications. Interactions with other drugs must be considered, and therapy in elderly patients requires careful management. Current dosing recommendations are reviewed, and practical guidelines for the optimal use of warfarin are provided.


[Effect of a synthetic analogue of bacterial anabiosis autoinducers hexylresorcinol on the stability of membrane structures].
The effect of hexylresorcinol (HR), a chemical analogue of microbial anabiosis autoinducers of the alkylhydroxybenzene (AHB) group, on the stability of biological membranes and monolamellar liposomes formed of egg phosphatidylcholine (ePC) was studied. According to spectrophotometry and electron microscopy studying of HR-loaded liposomes in the presence of a surface-active agent Tween 20, the critical ratio between HR and ePC for liposome preservation was found to be close to equimolar. The trends in HR influence on membrane structural organization and stability confirmed in experiments on liposomes were also reproduced on intact bacterial cells explaining non-species-specific effect of AHBs. The demonstrated high efficiency of AHB biocides may be used in material and equipment protection against biocorrosion.


One hundred and ten sports related facial fractures.
In a 1 year period (1993) 332 facial fractures were treated in Cork Regional Hospital. Of these 110 (33%) were sports related. Males accounted for 94.5% of patients and 77% of patients were aged below 30 years. Nasal fractures were the most common fractures seen (56%). Almost half (49%) of sports related facial fractures occurred while playing Gaelic games. The incidence of hurling fractures has been more than halved over the last 20 years. Seventy eight percent of patients required operative treatment.


Crystallography and Microstructure of 7M Martensite in Ni-Mn-Ga Thin Films Epitaxially Grown on (1 1 2¯ 0)-Oriented Al2O3 Substrate.
Epitaxial Ni-Mn-Ga thin films have been extensively investigated, due to their potential applications in magnetic micro-electro-mechanical systems. It has been proposed that the martensitic phase in the <1 1 0>A-oriented film is much more stable than that in the <1 0 0>A-oriented film. Nevertheless, the magnetic properties, microstructural features, and crystal structures of martensite in such films have not been fully revealed. In this work, the <1 1 0>A-oriented Ni51.0Mn27.5Ga21.5 films with different thicknesses were prepared by epitaxially growing on Al2O3(1 1 2¯ 0) substrate by magnetron sputtering. The characterization by X-ray diffraction technique and transmission electron microscopy revealed that all the Ni51.0Mn27.5Ga21.5 films are of 7M martensite at the ambient temperature, with their Type-I and Type-II twinning interfaces nearly parallel to the substrate surface.


To exploit the tumor microenvironment: Passive and active tumor targeting of nanocarriers for anti-cancer drug delivery.
Because of the particular characteristics of the tumor microenvironment and tumor angiogenesis, it is possible to design drug delivery systems that specifically target anti-cancer drugs to tumors. Most of the conventional chemotherapeutic agents have poor pharmacokinetics profiles and are distributed non-specifically in the body leading to systemic toxicity associated with serious side effects. Therefore, the development of drug delivery systems able to target the tumor site is becoming a real challenge that is currently addressed. Nanomedicine can reach tumor passively through the leaky vasculature surrounding the tumors by the Enhanced Permeability and Retention effect whereas ligands grafted at the surface of nanocarriers allow active targeting by binding to the receptors overexpressed by cancer cells or angiogenic endothelial cells. This review is divided into two parts: the first one describes the tumor microenvironment and the second one focuses on the exploitation and the understanding of these characteristics to design new drug delivery systems targeting the tumor. Delivery of conventional chemotherapeutic anti-cancer drugs is mainly discussed.


[Intestinal duplication in the adult. A case report of colonic duplication and a review of the literature].
Alimentary tract duplications are uncommon congenital anomalies that are usually present during the first decade of life. However, a smaller number of cases may remain unsuspected until adulthood. They are most common in the ileum but can occur anywhere along the alimentary tract. Duplications may be cystic or tubular in appearance and characteristically arise from the mesenteric aspect of the intestine. Abdominal pain, nausea and/or vomiting, palpable mass, weight loss, and bleeding are the most common symptoms. This paper reports the case of a 24-year-old male with a cystic duplication of the ascending colon not correctly diagnosed prior to operation. When faced with such an abdominal cystic lesion, differential diagnoses versus duplication cyst, mesenteric cyst, choledochal cyst, giant diverticulum, and cystic tumour of the pancreas should be considered. Ultrasound and computerized tomography are good tools for diagnosis and for delineating the relationship between the cyst and peripheral structures. Surgery provides treatment and allows a final pathological diagnosis. Removal of the cyst could be considered satisfactory but resection of the duplication and the adjacent bowel is recommended because of the possibility of malignant degeneration and the risk of gastrointestinal ulceration and haemorrhage due to ectopic gastric mucosa.


[Inter-observer agreement and validity of photographic dietary assessment].
Estimation of nutritional intake from photographs of served meals has been proposed. We examined the inter-observer agreement and validity of this method for assessment of the daily diet. Twenty-five family members (13 men and 12 women, mean age +/- SD: 47.3 +/- 5.6 years) of students in a dietetic course performed both meal photography and traditional weighed dietary records for four days. For the photo method, two observers independently identified foods and estimated their portion sizes from the photographs, converting them into energy and nutrient intake with a food composition table. The inter-observer agreement with the photo method was assessed in terms of inter-observer ratios, correlations, and coefficients of variation (CV), and the method was validated using the weighed dietary records as the reference. Its running cost was also calculated. The inter-observer ratios for estimation of mean daily nutritional intake ranged from 0.89 (magnesium) to 1.14 (retinol) with a median of 1.03. The correlation coefficients between observers varied from 0.65 (saturated fatty acids [SFA]) to 0.92 (vitamin C) on a daily basis (median, 0.79), and from 0.65 (SFA) to 0.96 (vitamin C) on an individual basis (median, 0.78). The CVs ranged from 7.9% (energy) to 23.8% (carotenc) (median, 13.3%), and from 5.2% (energy and magnesium) to 17.8% (carotene) (median, 8.8%) for daily and individual intake, respectively. Regarding validity, the ratios between methods (the photo method/weighed dietary records) ranged from 0.96 (potassium and SFA) to 1.11 (retinal and salt) with a median of 1.00. Correlations between the two methods were distributed from 0.40 (salt) to 0.82 (vitamin C and retinol) on a daily basis (median, 0.67), and from 0.47 (salt) to 0.90 (vitamin C) on an individual basis (median, 0.74). The CVs ranged from 10.5% (energy) to 39.6% (carotene) (median, 16.9%), and from 6.1% (protein) to 20.6% (carotene) (median, 11.2%), respectively. It was calculated to cost 105 Japanese yen per meal when using a lens-attached film for 25 shots. Inter-observer agreement and validity of the photo method, though varying with the nutrient, were generally acceptable. Although some modifications are needed for diners-out, the method appears to be useful as a dietary recording tool.


Characterization of Shiga toxin gene (stx)-positive and intimin gene (eae)-positive Escherichia coli isolates from wastewater of slaughterhouses in France.
Wastewater samples from 12 slaughterhouses located in different regions in France were tested for the presence of stx-positive and eae-positive Escherichia coli isolates, and characteristics of the isolates obtained were determined. A total of 224 wastewater samples were collected in wastewater treatment plants at different stages of wastewater processing. Altogether, 5,001 E. coli isolates were obtained by colony counting and screened for the presence of stx and eae genes by multiplex PCR. stx-positive and eae-positive E. coli isolates were detected in 25% of the samples collected; they were found in 13% and 3% of the samples obtained from treated effluent and sludge, respectively, suggesting that they could be spread into the environment. Screening of the samples collected by immunomagnetic separation allowed us to isolate 31 additional E. coli serogroup O157 isolates. Four of these isolates harbored stx and eae genes. All stx-positive and eae-positive E. coli isolates were analyzed for eae and stx genetic variants, as well as for additional virulence factors and serotypes. Our results suggest that the majority of the stx- and eae-positive E. coli isolates from wastewater have low virulence for humans. However, the diversity of the enterohemorrhagic E. coli-associated virulence factors in the strains indicates that the environment may play an important role in the emergence of new pathogenic enterohemorrhagic E. coli strains.


Optical Excitation of Carbon Nanotubes Drives Localized Diazonium Reactions.
Covalent chemistries have been widely used to modify carbon nanomaterials; however, they typically lack the precision and efficiency required to directly engineer their optical and electronic properties. Here, we show, for the first time, that visible light which is tuned into resonance with carbon nanotubes can be used to drive their functionalization by aryldiazonium salts. The optical excitation accelerates the reaction rate 154-fold (±13) and makes it possible to significantly improve the efficiency of covalent bonding to the sp(2) carbon lattice. Control experiments suggest that the reaction is dominated by a localized photothermal effect. This light-driven reaction paves the way for precise nanochemistry that can directly tailor carbon nanomaterials at the optical and electronic levels.


Children value objects with distinctive histories.
From ancient objects in museums to souvenirs obtained on vacation, we often value objects for their distinctive histories. The present experiments investigate the developmental origins of people's feelings that objects with distinctive histories are special. In each of four experiments, 4- to 7-year-olds (total N = 400) saw pairs of identical-looking objects in which one object was new and the other had a history that was either distinctive or mundane. In the first experiment, the histories did not involve people; in the remaining experiments, the histories were personal and related the objects to particular people. Distinctive histories affected children's valuations of regular objects (all experiments), but not their valuations of stuffed animals. Both older and younger children viewed regular objects with distinctive histories as more special than those with mundane histories. Older children mostly viewed objects with distinctive histories as more special than new objects, and younger children showed similar judgments when judging which object a person cared about more. Together, the findings reveal a novel way that information about the past influences children's judgments about the present and suggest that young children's valuations of objects depend on objects' histories. (PsycINFO Database Record (c) 2019 APA, all rights reserved).


Tumour responses following a steroid switch from prednisone to dexamethasone in castration-resistant prostate cancer patients progressing on abiraterone.
Abiraterone is a CYP17A1 inhibitor that improves survival in castration-resistant prostate cancer (CRPC). Abiraterone is licensed in combination with prednisone 5 mg twice daily to prevent a syndrome of secondary mineralocorticoid excess. We hypothesised that a 'steroid switch' from prednisone to dexamethasone would induce secondary responses in patients progressing on abiraterone and prednisone 5 mg b.i.d. We performed a 'steroid switch' in patients with CRPC at PSA progression on abiraterone and prednisolone. Patients were monitored for secondary declines in PSA, radiological tumour regression and toxicity. A retrospective analysis of 30 CRPC patients who underwent a steroid switch from prednisolone to dexamethasone while on abiraterone was performed. A total of six patients (20%) had a ⩾50% PSA decline that was confirmed by a second PSA level at least 3 weeks later. In all, 11 patients (39.2%) had a confirmed ⩾30% PSA decline. Median time to PSA progression on abiraterone and dexamethasone was 11.7 weeks (95% CI: 8.6-14.8 weeks) in the whole cohort and 27.6 weeks (95% CI: 14.5-40.7 weeks) in patients who achieved a confirmed 50% PSA decline. Nine patients had RECIST evaluable disease: two of these patients had RECIST partial response, six patients had stable disease and one patient had progressive disease at the first imaging assessment. Treatment was well tolerated, with no grade 3 and grade 4 adverse events. One patient had to be reverted to prednisolone because of grade 2 hypotension. Durable PSA responses occur in up to 40% of patients following a 'steroid switch' for PSA progression on abiraterone and prednisone. Studies are ongoing to elucidate the mechanisms underlying this response.


Association between vitamin D and risk of colorectal cancer: a systematic review of prospective studies.
To conduct a systematic review of prospective studies assessing the association of vitamin D intake or blood levels of 25-hydroxyvitamin D [25(OH)D] with the risk of colorectal cancer using meta-analysis. Relevant studies were identified by a search of MEDLINE and EMBASE databases before October 2010 with no restrictions. We included prospective studies that reported relative risk (RR) estimates with 95% CIs for the association between vitamin D intake or blood 25(OH)D levels and the risk of colorectal, colon, or rectal cancer. Approximately 1,000,000 participants from several countries were included in this analysis. Nine studies on vitamin D intake and nine studies on blood 25(OH)D levels were included in the meta-analysis. The pooled RRs of colorectal cancer for the highest versus lowest categories of vitamin D intake and blood 25(OH)D levels were 0.88 (95% CI, 0.80 to 0.96) and 0.67 (95% CI, 0.54 to 0.80), respectively. There was no heterogeneity among studies of vitamin D intake (P = .19) or among studies of blood 25(OH)D levels (P = .96). A 10 ng/mL increment in blood 25(OH)D level conferred an RR of 0.74 (95% CI, 0.63 to 0.89). Vitamin D intake and blood 25(OH)D levels were inversely associated with the risk of colorectal cancer in this meta-analysis.


Modified semilunar coronally advanced flap: A case series.
Dentists traditionally think of periodontal treatment as a means of saving the teeth while leaving the patient with an esthetic problem. The goal of gingival esthetics is to maintain normal healthy gingival appearance around teeth that must be restored. Gingival recession represents a significant concern for patients and a therapeutic problem for the clinician. Root coverage is the goal of periodontal plastic surgery when treating gingival recessions in the esthetic zone. Correction of mucogingival recession deformities with a variety of periodontal plastic surgical procedures have been described each demonstrating a variable degree of success. This case report presents to you the treatment outcomes and predictability of modified semilunar coronally advanced flap (Kamran Haghighat) techniques described for the treatment of recession defects on single and multiple adjacent teeth, respectively.


Rapid, transient, and dose-dependent expression of hsp70 messenger RNA in the rat brain after morphine treatment.
Induction of Hsp70 in the brain has been reported after intake of drugs of abuse like amphetamine and lysergic acid diethylamide. In this investigation, gene expression of Hsp70 and other heat shock genes in the rat brain was studied in response to morphine. Twenty milligrams per kilogram morphine intraperitoneally resulted in a marked induction of Hsp70 messenger RNA (mRNA) expression in the frontal cortex with a maximum increase of 13.2-fold after 2 hours. A moderate increase of Hsp27 mRNA expression (6.7-fold) could be observed after 4 hours, whereas mRNA expression of Hsp90 and of the constitutive Hsc70 did not exceed a mean factor of 1.8-fold during the 24 hours interval. The increase in Hsp70 mRNA was dose dependent, showing a significant elevation after doses ranging from 10 to 50 mg/kg morphine. In situ hybridization revealed enhanced Hsp70 mRNA expression mainly in cortical areas, in the hippocampus, in the paraventricular and supraoptic nuclei of the hypothalamus, in the locus coeruleus, as well in the pineal body. The double in situ hybridization technique revealed increased Hsp70 mRNA expression mainly in VGLUT1-positive neurons and to a lesser extent in olig1-positive oligodendroglia. Immunohistochemistry revealed a marked increase of Hsp70 protein in neuronal cells and blood vessels after 12 hours. In contrast to animal experiments, morphine did not increase Hsp70 mRNA expression in vitro in micro-opioid receptor (MOR1)-expressing human embryonic kidney 293 cells, suggesting no direct MOR1-mediated cellular effect. To exclude a body temperature-related morphine effect on Hsp70 mRNA expression, the temperature was recorded. Five to 20 mg/kg resulted in hyperthermia (maximum 40.6 degrees), whereas a high dose (50 mg/kg) that produced the highest mRNA induction, showed a clear hypothermia (minimum 37.2 degrees C). These findings argue against the possibility that Hsp70 induction by morphine is caused by its effect on body temperature. It may be speculated that increased expression of Hsp70 after morphine application protects brain structures against potentially hazardous effects of opiates.


Blood pressure regulation, peripheral renin activity and aldosterone in patients with pyelonephritic renal scarring.
Patients with renal scarring due to previous upper urinary tract infections (UTI) are at risk of developing hypertension and renal insufficiency. In this study glomerular filtration rate (GFR), systolic (SBP) and diastolic (DBP) blood pressure, peripheral renin activity (PRA), plasma (p-Aldo) and urine aldosterone (u-Aldo), the urinary excretion of sodium (UNa) and potassium (UK) and the fractional sodium (CNa/CIn) and potassium (CK/CIn) excretion were determined in 22 female patients with verified renal scarring and a history of febrile UTI and in nine age-matched healthy women with normal i.v. urograms. The patients had significantly lower GFR, higher SBP, higher PRA and higher CK/CIn than the healthy controls. A significant correlation between CNa/CIn and SBP (r = 0.51, P less than 0.05), DBP (r = 0.50, P less than 0.05) and PRA (r = -0.47, P less than 0.05) was found. The ratio of UK/UNa was significantly correlated to SBP (r = -0.43, P less than 0.05), DBP (r = -0.44, P less than 0.05), PRA (r = 0.65, P less than 0.01) and p-Aldo (r = 0.68, P less than 0.01). It is concluded that the renin-aldosterone system probably is involved in the pathogenesis of hypertension in patients with pyelonephritic renal scarring and that screening determinations of UNa and UK may prove useful for detection of individuals with increased PRA and p-Aldo.


Droplet digital PCR of viral DNA/RNA, current progress, challenges, and future perspectives.
High-throughput droplet-based digital PCR (ddPCR) is a refinement of the conventional polymerase chain reaction (PCR) methods. In ddPCR, DNA/RNA is encapsulated stochastically inside the microdroplets as reaction chambers. A small percentage of the reaction chamber contains one or fewer copies of the DNA or RNA. After PCR amplification, concentrations are determined based on the proportion of nonfluorescent partitions through the Poisson distribution. Some of the main features of ddPCR include high sensitivity and specificity, absolute quantification without a standard curve, high reproducibility, good tolerance to PCR inhibitor, and high efficacy compared to conventional molecular methods. These advantages make ddPCR a valuable addition to the virologist's toolbox. The following review outlines the recent technological advances in ddPCR methods and their applications in viral identification.


Syndromic Craniosynostosis: Complexities of Clinical Care.
Patients with syndromic craniosynostosis have a molecularly identified genetic cause for the premature closure of their cranial sutures and associated facial and extra-cranial features. Their clinical complexity demands comprehensive management by an extensive multidisciplinary team. This review aims to marry genotypic and phenotypic knowledge with clinical presentation and management of the craniofacial syndromes presenting most frequently to the craniofacial unit at Great Ormond Street Hospital for Children NHS Foundation Trust.


Rationalized polytherapy for epilepsy.
In the past, epilepsy was usually treated with polytherapy, but with little knowledge as to the interactions and side-effects of the combinations of the anti-epileptic drugs used. Adverse events and sparse clinical knowledge led to monotherapy becoming the treatment regime of choice. A new generation of drugs, which are well-tolerated and have few or predictable interactions, have enabled the reassessment of polytherapy for the treatment of epilepsy. Extensive clinical trials of these drugs are allowing the emergence of a new, rationalized approach to polytherapy. In our study, 19 patients with refractory partial epilepsy, and who were 'socially active and integrated into society', received vigabatrin as add-on therapy. Patients were taking a mean of 1.5 drugs, and five patients were taking small doses of drugs which lead to tolerance, such as barbiturates and benzodiazepines. With vigabatrin as add-on therapy, 14 patients (73%) had a greater than 50% reduction in seizure frequency, and 10 (52%) had a greater than 70% reduction in seizure frequency. In one patient, seizure frequency increased, and two patients developed myoclonic jerks. Vigabatrin was not shown to have any harmful effects in extensive laboratory, EEG and cognitive function tests. In fact, a minor improvement occurred in visual memory, which was probably related to the reduction in seizures. Addition of vigabatrin may, therefore, be of benefit to patients with partial epilepsy refractory to monotherapy with standard anti-epilepsy drugs.


A Retrospective Study From a Single Center in Romania of 36 Patients Aged Between 37 and 59 Years Who Presented With Locally Advanced Colorectal Cancer.
Background The incidence of colorectal carcinoma is increasing in younger adults. This retrospective study was conducted at a single center in Romania and included 36 patients aged between 37 and 59 years who presented with locally advanced colorectal cancer. The purpose of this study is to show the importance of colorectal cancer screening in young patients. Materials and methods The study included 36 patients with histologically proven colorectal cancer evaluated in OncoFort Hospital. Disease staging was based on surgical findings and pre or post-operative abdominal CT or MRI of the abdomen and pelvis. The inclusion criteria were defined as a history of adjuvant chemotherapy plus radiotherapy and whether one had locally advanced colorectal cancer or recurrent or metastatic disease. Results Of the 36 patients, 13 (36.11%) were women, and 23 (63.8%) were men. The mean age was 47.4 years (range: 37-59 years). The colon cancers were more frequent than tumours of the rectum (n = 19, 52.77% versus n = 17, 47.23%). A total of 44.44% of patients were classified as stage III-IV. We found no significant correlation between mutation status or histologic grade and age. Conclusion This real-world study from a single center in Romania highlights that colorectal carcinoma may present in advanced stages in younger patients and may support consideration of a need to perform further studies to determine if the current age recommendations for screening should be lowered.


Nanowire-based sensors.
Nanowires are important potential candidates for the realization of the next generation of sensors. They offer many advantages such as high surface-to-volume ratios, Debye lengths comparable to the target molecule, minimum power consumption, and they can be relatively easily incorporated into microelectronic devices. Accordingly, there has been an intensified search for novel nanowire materials and corresponding platforms for realizing single-molecule detection with superior sensing performance. In this work, progress made towards the use of nanowires for achieving better sensing performance is critically reviewed. In particular, various nanowires types (metallic, semiconducting, and insulating) and their employment either as a sensor material or as a template material are discussed. Major obstacles and future steps towards the ultimate nanosensors based on nanowires are addressed.


Effect of increased abdominal pressure on cytokines (IL1 beta, IL6, TNFalpha), C-reactive protein (CRP), free radicals (NO, MDA), and histology.
It is generally accepted that proinflammatory mediators, including cytokines, are responsible for the metabolic changes associated with injury. Recent clinical and experimental studies have also shown that the laparoscopic procedures actually produce ischemia-reperfusion injury in the organs by oxygen-derived free radicals. This study aimed to assess the effect of different insufflation pressures and laparotomy on tissue response by comparing the proinflammatory cytokines, C-reactive protein, and serum and tissue levels of oxygen-derived free radicals. Forty mature New Zealand white rabbits were assigned to 4 groups of 10 animals. In groups 1 to 3, CO2 pneumoperitoneum was created using an automatic insufflator to the designated pressure of 10, 15, and 20 mm Hg, respectively. The remaining 10 animals underwent laparotomy using 10 cm midline incision (group 4). Blood samples were collected before (0 min) and at the end of the procedure (60 min). After the collection of last blood samples, all animals were killed and samples from liver and gut were obtained for measurements of tissue malondialdehyde levels and histology. The proinflammatory cytokine levels were increased significantly in groups 1 to 3, but did not change in the laparotomy group. Serum C-reactive protein levels were elevated in all groups. The comparison of the results between the laparotomy and laparoscopy groups showed that serum interleukin 6 and nitric oxide levels were significantly elevated in relation the intra-abdominal pressure, and serum interleukin 6 and nitric oxide levels peaked in group 3. Tissue malondialdehyde levels were significantly higher in groups 1 and 2 than in groups 3 and 4. The findings of our experiment suggest that the elevated intra-abdominal pressure is responsible for ischemia, free radical production, and proinflammatory cytokine response-mediated cell damage during laparoscopic surgery.


A sheep preparation for studying interactions between blood flow and drug disposition. I: Physiological profile.
A sheep preparation has been developed which allows repeated measurements of regional blood flow, oxygen consumption and drug disposition in awake, unrestrained animals. This allows systematic studies of both acute changes, such as haemodynamic disturbances, and of chronic changes, such as enzyme induction, to be carried out. Good agreement was shown between the values for cardiac output and regional blood flow obtained by the Fick and indicator dilution methods, and those obtained by others using microspheres. Significant day-to-day fluctuations in haemodynamic indices were shown to occur; assumptions that hepatic or renal blood flows are constant fractions of cardiac output, or that renal or hepatic flow indicator extraction ratios remain unchanged from day-to-day, will lead to significant errors. Thus, control measurements for each experiment are necessary. It is proposed that physiological models for studying drug disposition based on data from awake, unrestrained animals may provide insight into some mechanisms of changes in drug disposition that cannot be obtained using the traditional compartmental method.


Physical activity profile of students in Mansoura University, Egypt.
A cross-sectional study was carried out on 1708 students from Mansoura University, Egypt, to describe the pattern of physical activity, predictors of physical inactivity and perceived barriers to and benefits of physical activity. The International Physical Activity Questionnaire (IPAQ) was used to measure physical activity. Data were analysed according to the guidelines for data processing and analysis of the IPAQ. Perceived barriers to and potential benefits of physical activity were asked for. More than 11% of students were physically inactive. On logistic regression analysis, the independent predictors of physical inactivity were high socioeconomic standard of the family (OR 2.1), female sex (OR 1.8), medical education (OR 1.8) and non-membership in sports clubs OR 1.6). The most frequent barriers to physical activity were time limitation and lack of accessible and suitable sporting places. More than 70% of the participants stated that physical activity promotes and maintains health.


Nanofibers for Biomedical and Healthcare Applications.
Unique features of nanofibers provide enormous potential in the field of biomedical and healthcare applications. Many studies have proven the extreme potential of nanofibers in front of current challenges in the medical and healthcare field. This review highlights the nanofiber technologies, unique properties, fabrication techniques (i.e., physical, chemical, and biological methods), and emerging applications in biomedical and healthcare fields. It summarizes the recent researches on nanofibers for drug delivery systems and controlled drug release, tissue-engineered scaffolds, dressings for wound healing, biosensors, biomedical devices, medical implants, skin care, as well as air, water, and blood purification systems. Attention is given to different types of fibers (e.g., mesoporous, hollow, core-shell nanofibers) fabricated from various materials and their potential biomedical applications.


[Effects of melatonin on the voltage-gated delayed rectifier potassium channels of cultured hippocampal neurons of new-born rats].
To investigate the effects of melatonin on voltage-gated delayed rectifier potassium channels. Hippocampus neurons were obtained from newborn Wistar rat and cultured. Primary cultured for 7 to 12 days of new-born Wistar rat were selected as objectives. Patch clamp whole-cell recording technique was used on the hippocampus neurons cultured for 7 to 12 day. to record the delayed rectifier potassium current to analyze the basic electrophysiological characteristics. The effects of melatonin of the concentrations of 1 nmol/L, 10 nmol/L, 100 nmol/L, 1 mol/L, 10 mol/L, 100 mol/L, and 1 mmol/L on the amplitudes and kinetics of delayed rectifier potassium currents were investigated. With different voltage protocols and specific blockers of potassium channel (4-AP and TEA) a delayed rectifier potassium current that activated and inactivated slowly and had the outward rectifying characteristics (Ik) from the outward potassium currents in cultured new-born hippocampus neurons was separated. The effect of melatonin on the delayed rectifier channel was rapid, reversible and voltage-dependent Melatonin had no effect on the kinetic characteristics of the I -V curve. Melatonin increased the potassium current concentration-dependently. 1 - 100 nmol/L melatonin increased the amplitude of potassium current gradually; the effects of 1 - 100 micromol/L melatonin on the potassium current increased concentration-dependently, while the action of 1 mmol/L melatonin decreased. Melatonin reversibly increases the rectifier delayed potassium currents of the cultured hippocampus neurons of new-born rat. This may be involved in some aspects of physiological and pathological significance of potassium currents.


Giant electron-phonon coupling detected under surface plasmon resonance in Au film.
Surface plasmon resonance (SPR) is a powerful tool to amplify coherent phonon signals in metal films. In a 40 nm Au film excited with a 400 nm pump, we observed an abnormally large electron-phonon coupling constant of about 8×1017 W/(m3K), almost 40× larger than those reported in noble metals, and even comparable to transition metals. We attribute this phenomenon to quantum confinement and interband excitation. With SPR, we also observed two coherent phonon modes in a GaAs/AlAs quantum well thin film. Our finding provides a new approach to generate high-frequency acoustic phonons in noble metals and to study their nonlinear nature of propagation.


Arterial baroreflex control of sympathetic nerve activity and heart rate in patients with type 2 diabetes.
Despite greater blood pressure reactivity to acute cardiovascular stressors and a higher prevalence of hypertension in type 2 diabetes (T2D) patients, limited information is available regarding arterial baroreflex (ABR) control in T2D. We hypothesized that ABR control of muscle sympathetic nerve activity (MSNA) and heart rate (HR) are attenuated in T2D patients. Seventeen T2D patients (50 ± 2 yr; 31 ± 1 kg/m2), 9 weight-matched controls (WM-CON, 46 ± 2 yr; 32 ± 2 kg/m2) and 10 lean controls (Lean-CON, 49 ± 3 yr; 23 ± 1 kg/m2), underwent bolus infusions of sodium nitroprusside (100 μg) followed 60 s later by phenylephrine (150 μg) and weighted linear regression performed. No group differences in overall sympathetic baroreflex gain were observed (T2D: -2.5 ± 0.3 vs. WM-CON: -2.6 ± 0.2 vs. Lean-CON: -2.7 ± 0.4 arbitrary units·beat·mmHg-1, P > 0.05) or in sympathetic baroreflex gain when derived separately during blood pressure (BP) falls (nitroprusside) and BP rises (phenylephrine). In contrast, overall cardiac baroreflex gain was reduced in T2D patients compared with Lean-CON (T2D: 8.2 ± 1.5 vs. Lean-CON: 15.6 ± 2.9 ms·mmHg-1, P < 0.05) and also tended to be reduced in WM-CON (9.3 ± 1.9 ms·mmHg-1) compared with Lean-CON (P = 0.059). Likewise, during BP rises, cardiac baroreflex gain was reduced in T2D patients and weight-matched controls compared with lean controls (P < 0.05), whereas no group differences were found during BP falls (P > 0.05). Sympathetic and cardiac ABR gains were comparable between normotensive and hypertensive T2D patients (P > 0.05). These findings suggest preserved ABR control of MSNA in T2D patients compared with both obese and lean age-matched counterparts, with a selective impairment in ABR HR control in T2D that may be related to obesity.


Association between decreased disc signal intensity in preoperative T2-weighted MRI and a 5-year outcome after lumbar minimally invasive discectomy.
A total of 39 patients who had undergone microdiscectomy or percutaneous nucleotomy for lumbar disc herniation were examined after a follow-up of 5 years. The overall outcome was satisfactory in 80% of the patients treated, and only 1 (3%) patient had been reoperated during the follow-up. Clinical signs and symptoms of lumbar instability were detected in 10 (26%) patients. All these 39 patients had been examined with lumbar magnetic resonance imaging (MRI) on the day preceding the operation; the presence of disc degeneration was graded as severe, mild or non-existent depending on the visual brightness of the discs on T2-weighted images, as compared to the signal intensity of the lumbar vertebrae. None of the 12 patients with no preoperative disc degeneration in MRI suffered from postoperative clinical signs and symptoms of instability as compared to 10 (37%) of the 27 patients with mild or severe disc degeneration suffering from instability (p = 0.04). Thus, the results of the present study imply that the grade of the disc degeneration in preoperative T2-weighted MR images significantly predicted the occurrence of postoperative clinical instability.


Peptide nucleic acid (PNA) conformation and polymorphism in PNA-DNA and PNA-RNA hybrids.
Two hydrogen-bonding motifs have been proposed to account for the extraordinary stability of polyamide "peptide" nucleic acid (PNA) hybrids with nucleic acids. These interresidue- and intraresidue-hydrogen-bond motifs were investigated by molecular mechanics calculations. Energy-minimized structures of Watson-Crick base-paired decameric duplexes of PNA with A-, B-, and Z-DNA and A-RNA polymorphs indicate that the inherent stability of the complementary PNA helical structures is derived from interresidue, rather than from intraresidue, hydrogen bonds in all hybrids studied. Intraresidue-hydrogen-bond lengths are consistently longer than interresidue hydrogen bonds. Helical strand stability with interresidue hydrogen bond stabilization follows the order: B-(DNA.PNA) > A-(DNA.PNA) congruent to A-RNA.PNA > Z-(DNA.PNA). In the triplex hybrids A-(RNA.PNA2) and B-(DNA.PNA2), differences between stabilities of the two decamers of thyminyl PNA with lysine amide attached to the C terminus (pnaT)10 strands are small. The Hoogsteen (pnaT)10 strands are of slightly higher potential energy than are the Watson-Crick (pnaT)10 strands. Antiparallel arrangement of PNAs in the triplex is slightly favored over the parallel arrangement based on the calculations. Examination by molecular mechanics of the PNA.DNA analogue of the NMR-derived structure for the B-double-stranded DNA dodecamer d(CG-CAAATTTGCG)2 in solution suggests that use of all bases of the genetic alphabet should be possible without loss of the specific interresidue-hydrogen-bonding pattern within the PNA strand.


Comparative Transcriptomics Reveals the Role of the Toll-Like Receptor Signaling Pathway in Fluoride-Induced Cardiotoxicity.
Many studies have shown that fluorosis due to long-term fluoride intake has damaging effects on the heart. However, the mechanisms underlying cardiac fluorosis have not been illuminated in detail. We performed high-throughput transcriptome sequencing (RNA-Seq) on rat cardiac tissue to explore the molecular effects of NaF exposure. In total, 372 and 254 differentially expressed genes (DEGs) were identified between a group given 30 mg/L NaF and control and between a group given 90 mg/L NaF and control, respectively. The transcript levels of most of these genes were significantly down-regulated and many were distributed in the Toll-like receptor signaling pathway. Transcriptome analysis revealed that herpes simplex infection, ECM-receptor interaction, influenza A, cytokine-cytokine receptor interaction, apoptosis, and Toll-like receptor signaling pathway were significantly affected. IL-6 and IL-10 may play a crucial role in the cardiac damage caused by NaF as external stimuli according to protein-protein interaction (PPI) network analysis. The results of qRT-PCR and Western blotting showed a marked decreased mRNA and protein levels of IL-1, IL-6, and IL-10 in the low concentration fluoride (LF) and high concentration fluoride (HF) groups, which was in agreement with RNA-Seq results. This is the first study to investigate NaF-induced cardiotoxicity at a transcriptome level.


Multinuclear Solid-State Magnetic Resonance as a Sensitive Probe of Structural Changes upon the Occurrence of Halogen Bonding in Co-crystals.
Although the understanding of intermolecular interactions, such as hydrogen bonding, is relatively well-developed, many additional weak interactions work both in tandem and competitively to stabilize a given crystal structure. Due to a wide array of potential applications, a substantial effort has been invested in understanding the halogen bond. Here, we explore the utility of multinuclear ((13)C, (14/15)N, (19)F, and (127)I) solid-state magnetic resonance experiments in characterizing the electronic and structural changes which take place upon the formation of five halogen-bonded co-crystalline product materials. Single-crystal X-ray diffraction (XRD) structures of three novel co-crystals which exhibit a 1:1 stoichiometry between decamethonium diiodide (i.e., [(CH3)3N(+)(CH2)10N(+)(CH3)3][2 I(-)]) and different para-dihalogen-substituted benzene moieties (i.e., p-C6X2Y4, X=Br, I; Y=H, F) are presented. (13)C and (15)N NMR experiments carried out on these and related systems validate sample purity, but also serve as indirect probes of the formation of a halogen bond in the co-crystal complexes in the solid state. Long-range changes in the electronic environment, which manifest through changes in the electric field gradient (EFG) tensor, are quantitatively measured using (14)N NMR spectroscopy, with a systematic decrease in the (14)N quadrupolar coupling constant (CQ) observed upon halogen bond formation. Attempts at (127)I solid-state NMR spectroscopy experiments are presented and variable-temperature (19)F NMR experiments are used to distinguish between dynamic and static disorder in selected product materials, which could not be conclusively established using solely XRD. Quantum chemical calculations using the gauge-including projector augmented-wave (GIPAW) or relativistic zeroth-order regular approximation (ZORA) density functional theory (DFT) approaches complement the experimental NMR measurements and provide theoretical corroboration for the changes in NMR parameters observed upon the formation of a halogen bond.


Sorption Kinetics on Open Carbon Nanohorn Aggregates: The Effect of Molecular Diameter.
We present the results of a study of the kinetics of adsorption on aggregates of open carbon nanohorns using argon and CF₄ sorbates. We measured the equilibration times for each value of the sorbent loading along eight adsorption isotherms (four isotherms for each sorbate species). We found that: the equilibration times decrease as the sorbent loading (and the equilibrium pressure of the coexisting gas) increases, for a given temperature; and, that, for a given value of the sorbent loading, the equilibration times decrease with increasing temperature. When considering the effect of scaling of the temperatures by the respective critical temperatures we found that, at the same scaled temperature and at comparable loadings, the equilibration times for CF₄ were longer than those for argon. We discuss a possible explanation for this result.


Prodrug delivery of novel PTP1B inhibitors to enhance insulin signalling.
A growing percentage of the population is resistant to two key hormones - insulin and leptin - as a result of increased obesity, often leading to significant health consequences such as type 2 diabetes. Protein tyrosine phosphatase 1B (PTP1B) is a key negative regulator of signalling by both of these hormones, so that inhibitors of this enzyme may provide promise for correcting endocrine abnormalities in both diabetes and obesity. As with other tyrosine phosphatases, identification of viable drug candidates targeting PTP1B has been elusive because of the nature of its active site. Beginning with novel phosphotyrosine mimetics, we have designed some of the most potent PTP1B inhibitors. However, their highly acidic structures limit intrinsic permeability and pharmacokinetics. Ester prodrugs of these inhibitors improve their drug-like properties with the goal of delivering these nanomolar inhibitors to the cytoplasm of cells within target tissues. In addition to identifying prodrugs that is able to deliver active drugs into cells to inhibit PTP1B and increase insulin signalling, these compounds were further modified to gain a variety of cleavage properties for targeting activity in vivo. One such prodrug candidate improved insulin sensitivity in ob/ob mice, with lowered fasting blood glucose levels seen in the context of lowered fasting insulin levels following 4 days of intraperitoneal dosing. The results presented in this study highlight the potential for design of orally active drug candidates targeting PTP1B, while also delineating the considerable challenges remaining.


Exploring the impact of postponing core clerkships on future performance.
Despite the many clerkship models of medical education, all can be considered a form of experiential learning. Experiential learning is a complex pedagogical approach involving the development of cognitive skills in an environment with a unique culture with multiple stakeholders, which may impact learner motivation, confidence, and other noncognitive drivers of success. Students may delay the transition to the clerkship year for myriad reasons, and the intricate nature of experiential learning suggested this may impact student performance. This retrospective, observational study investigated the impact of clerkship postponement by measuring subsequent clerkship performance. Pre-clerkship and third-year clerkship performance were analyzed for three cohorts of students (classes of 2018, 2019, and 2020, N = 274) where students had the option to delay the start of their clerkship year. A mixed analysis of variance (ANOVA) and paired t-tests were conducted to compare academic performance over time among students who did and did not delay. Across three cohorts of students, 12% delayed the start of the clerkship year (N = 33). Regardless of prior academic performance, these students experienced a significant reduction in clerkship grades compared to their non-delaying peers. Delaying the start of the clerkship year may have negative durable effects on future academic performance. This information should be kept in mind for student advisement.


Two approaches to linking census and hospital data.
This study compares registry and non-registry approaches to linking 2006 Census of Population data for Manitoba and Ontario to hospital data from the Discharge Abstract Database (DAD). Using a probabilistic linkage, the registry approach linked the census data to provincial health insurance registries, followed by a deterministic linkage to the DAD based on health insurance number (HIN). The non-registry approach used hierarchical deterministic exact matching based on three variables common to both files to link census data to the DAD. The approaches were compared in terms of linkage and coverage rates, sensitivity and specificity, and consistency of HINs on the linked records. Results of the registry and non-registry linkage approaches were similar. In Manitoba, 7% and 6% of census long-form respondents linked to the DAD with the registry and non-registry linkage approaches, respectively; in Ontario, the linkage rate was 5% for both approaches. With the registry approach, the linked census-DAD data represented 84% (weighted) of hospital admissions in the 2006/2007 DAD in both provinces, compared with 82% in Manitoba and Ontario with the non-registry approach. In the absence of access to provincial health insurance registries with which census data can be linked, a non-registry approach can be used to create a research-quality dataset.


In vivo aging of orthodontic alloys: implications for corrosion potential, nickel release, and biocompatibility.
Despite the large number of studies investigating nickel release from orthodontic stainless steel and nickel-titanium alloys, there is a lack of conclusive evidence with respect to the composition and kinetics of the corrosive products released. The objective of this review is to address the critical issues of corrosion potential and nickel leaching from alloys by investigating the effect of intraoral conditions on the surface reactivity of the materials. After an overview of fundamentals of metallurgical structure of orthodontic alloys, we provide an analysis of corrosion processes occurring in vivo. We present recent evidence suggesting the formation of a proteinaceous biofilm on retrieved orthodontic materials that later undergoes calcification. We illustrate the vastly irrelevant surface structure of in vivo- vs in vitro-aged alloys and discuss the potential implications of this pattern in the reactivity of the materials. Finally, we present a comprehensive review of the issue of nickel release, based on three perspectives: its biologic effects, the methods used for studying its release, and nickel-induced hypersensitivity in orthodontic patients.


[Measurement of adenosine deaminase activity in bronchoalveolar lavage fluids as a tool for diagnosing miliary tuberculosis].
Measurement of the activity of adenosine deaminase (ADA) in pleural effusions has been reported to be useful in the diagnosis of tuberculous pleuritis. To determine whether ADA activity can also be used to diagnose miliary tuberculosis, it was measured in bronchoalveolar lavage fluid from 64 subjects: 6 patients with miliary tuberculosis, 21 patients with sarcoidosis, 15 patients with idiopathic interstitial pneumonia, 15 patients with other diseases, and 7 healthy controls. Mean ADA activity was 5.02+/-3.75 IU/L (mean+/-SD) in patients with miliary tuberculosis; 1.06+/-0.99 IU/L in patients with sarcoidosis; 0.21+/-0.43 in patients with idiopathic interstitial pneumonia; and 0.30+/-0.51 IU/L in healthy controls. The level in patients with miliary tuberculosis differed significantly from the others (p<01.01). All 6 patients with miliary tuberculosis, 3 patients with sarcoidosis, 1 patient with Wegener's granulomatosis, and 1 patient with lymphangitic carcinomatosis had levels of ADA activity in bronchoalveolar lavage fluid that exceeded 2.0 IU/L. With a cut-off value of 2.0 IU/L the sensitivity of this test to miliary tuberculosis is higher than that of other diagnostic aids. Measurement of ADA activity in bronchoalveolar lavage fluid may be of great value in the early diagnosis of miliary tuberculosis.


Characterization of the surface-active components of sugar beet pectin and the hydrodynamic thickness of the adsorbed pectin layer.
The fraction of sugar beet pectin (SBP) adsorbed onto limonene oil droplets during emulsification has been isolated, and its chemical and physicochemical characteristics have been determined. While the SBP sample itself was found to contain 2.67 and 1.06% protein and ferulic acid, respectively, the adsorbed fraction contained 11.10% protein and 2.16% ferulic acid. The adsorbed fraction was also found to have a higher degree of acetylation, notably at the C2 position on the galacturonic acid residues, and was also found to contain a higher proportion of neutral sugars, which are present in the ramified side chains of the pectin molecules. The thickness of the layer of SBP adsorbed onto polystyrene latex particles was studied by dynamic light scattering and was found to increase with increasing surface coverage. It was found to have a value of approximately 140 nm at plateau coverage, which closely corresponded to the hydrodynamic diameter of the pectin chains. The adsorbed layer thickness was found to be sensitive to pH and the presence of electrolyte. The thickness at a surface coverage of approximately 20 mg/m(2) in the absence of electrolyte at pH approximately 4 was 107 nm and at pH 8.8 was 70 nm, while at pH approximately 4 in the presence of 10 mM NaCl the thickness was found to be 70 nm. It was concluded that the SBP molecules form multilayers at the surface due to electrostatic interaction between the positively charged protein moieties and the galacturonic acid residues. The removal of calcium from the SBP had no effect on the adsorbed layer thickness; hence, multilayer formation due to calcium ion cross-linking was considered unlikely.


Circulating and disseminated tumour cells in head and neck cancer.
Multimodal treatment options in head and neck squamous cell carcinoma have allowed for greater control of locoregional disease, but this has not translated into a significant overall survival advantage for patients. This is partially because these treatment modalities have no influence over the rate of development of distant metastases. This article summarises the current methods of detecting circulating and disseminated tumour cells. It also discusses how these cells can offer prognostic value in head and neck squamous cell carcinoma, and considers questions posed by the identification of these cells. A literature search of relevant journal articles was performed using ScienceDirect and PubMed databases, and a general article search was conducted using the online search engine Google. The evidence presented in this article indicates that circulating tumour cells and disseminated tumour cells may be clinically useful as prognostic markers or in the assessment of response to treatment in head and neck squamous cell carcinoma.


The use of pedicle screw-rod system for the posterior fixation in cervico-thoracic junction.
In cervico-thoracic junction (CTJ), the use of strong fixation device such as pedicle screw-rod system is often required. Purpose of this study is to analyze the anatomical features of C7 and T1 pedicles related to screw insertion and to evaluate the safety of pedicle screw insertion at these levels. Nineteen patients underwent posterior CTJ fixation with C7 and/or T1 included in fixation levels. Seventeen patients had tumorous conditions and two with post-laminectomy kyphosis. The anatomical features were analyzed for C7 and T1 pedicles in 19 patients using computerized tomography (CT). Pedicle screw and rod fixation system was used in 16 patients. Pedicle violation by screws was evaluated with postoperative CT scan. The mean values of the width, height, stable depth, safety angle, transverse angle, and sagittal angle of C7 pedicles were 6.9 +/- 1.34 mm, 8.23 +/- 1.18 mm, 30.93 +/- 4.65 mm, 26.42 +/- 7.91 degrees, 25.9 +/- 4.83 degrees, and 10.6 +/- 3.39 degrees. At T1 pedicles, anatomic parameters were similar to those of C7. The pedicle violation revealed that 64.1% showed grade I violation and 35.9% showed grade II violation, overall. As for C7 pedicle screw insertion, grade I was 61.5% and grade II 38.5%. At T1 level, grade I was 65.0% and grade II 35.0%. There was no significant difference in violation rate between the whole group, C7, and T1 group. C7 pedicles can withstand pedicle screw insertion. C7 pedicle and T1 pedicle are anatomically very similar. With the use of adequate fluoroscopic oblique view, pedicle screw can be safely inserted at C7 and T1 levels.


High-rate continuous n-butanol production by Clostridium acetobutylicum from glucose and butyric acid in a single-pass fibrous-bed bioreactor.
Biobutanol produced in acetone-butanol-ethanol (ABE) fermentation at batch mode cannot compete with chemically derived butanol because of the low reactor productivity. Continuous fermentation can dramatically enhance productivity and lower capital and operating costs, but are rarely used in industrial fermentation because of increased risks of culture degeneration, cell washout, and contamination. In this study, cells of the asporogenous Clostridium acetobutylicum ATCC55025 were immobilized in a single-pass fibrous-bed bioreactor (FBB) for continuous production of butanol from glucose and butyrate at various dilution rates. Butyric acid in the feed medium helped maintaining cells in the solventogenic phase for stable continuous butanol production. At a dilution rate of 1.88 h-1 , butanol was produced at 9.55 g/L, with a yield of 0.24 g/g and productivity of 16.8 g/L/h, which was the highest productivity ever achieved for biobutanol fermentation and an 80-fold improvement over the conventional ABE fermentation. The extremely high productivity was attributed to the high density of viable cells (~100 g/L at >70% viability) immobilized in the fibrous matrix, which also enabled the cells to better tolerate butanol and butyric acid. The FBB was stable for continuous operation for an extended period of over 1 month.


Indirect Effects of Social Skills on Health Through Stress and Loneliness.
The social skills deficit vulnerability model predicts that people with inadequate social skills are at risk for a range of psychosocial problems, especially when confronted with stress. People with poor social skills often experience stress and loneliness and these two constructs were tested as potential pathways by which the poor social skills confer a risk for compromised mental and physical health. An online survey was completed by 775 adults, aged 18-91. The sample matched national demographics for race/ethnicity and age, among those over 18. Structural equation modeling revealed indirect effects of social skills on both mental and physical health through both stress and loneliness. The models showed that poor social skills were associated with poor mental and physical health through elevated stress and increased loneliness. The findings reveal that social skills deficits are associated with physical as well as mental health problems.


Photosynthetic rates of citronella and lemongrass.
Ten selections of citronella (Cymbopogon nardus [L.] Rendle) were grown at 32/27, 27/21, or 15/10 C day/night temperatures, and plants from three populations of lemongrass (Cymbopogon citratus [D.C.] Stapf from Japan or Sri Lanka and Cymbopogon flexuosus [D.C.] Stapf from India) were grown at 8- or 15-hour photoperiods. Net photosynthetic rates of mature leaves were measured in a controlled environment at 25 C and 260 microeinsteins per meter(2) per second. Rates declined with increasing leaf age, and from the tip to the base of the leaf blade. Rates for citronella leaves grown at 15/10 C were extremely low for all selections. Highest rates of net photosynthesis were recorded for four selections grown at 27/21 C and for two selections grown at 32/27 C. Lemongrass grown at 8-hour photoperiod had higher photosynthetic rates than that grown at 15-hour photoperiod.


[Indoor air studies of mould fungus contamination of homes of selected patients with bronchial asthma (with special regard to evaluation problems)].
Investigations of indoor air of the homes of seven patients with asthma bronchiale who showed up with positive reactions following intracutaneous application of fungal allergens revealed that their places of residence were contaminated by fungal and bacterial spores. The number of colony forming units of mesophilic fungal spores of the indoor air ranged from 100 to 1000 CFU/m3 and this was much higher than the mould flora of the outdoor air determined simultaneously. The major fungi species found by the indoor investigation were: Penicillium sp. > Aspergillus sp. > Cladosporium sp., Mucor sp., Chrysonilia sp., Verticillium sp. > Geotrichum sp., Trichoderma sp. In two cases Thermoactinomyces species could be detected in the indoor air. The main cause of fungal contamination were moist building materials on room walls, insufficient air ventilation, bad maintenance of the circulating air-machines and insufficient room hygiene (e.g. biological garbage in the kitchen).


A Randomized Parallel Study to Assess the Effect of Three Tongue Cleaning Modalities on Oral Malodor.
The objective of this study was to compare the effects of three tongue hygiene regimens on oral malodor. This was a single-center, randomized, parallel design study with three treatment groups. Subjects were randomly assigned to perform tongue hygiene with either the Philips Sonicare TongueCare+ BreathRx regimen (STC), Listerine Cool Mint antiseptic rinse (LCM), or tongue brushing with an ADA reference manual toothbrush (MTB). Tooth brushing was standardized for all subjects during the study period, and no other oral or breath hygiene measures were allowed. Eligible subjects met the following criteria: aged 18-70 years, in good general and oral health, non-smoker, with an organoleptic score between 2.7 and 4.5 following a 12-18 hour oral hygiene abstention period. Subjects who had oral appliances or who had periodontal disease or excessive recession were not eligible. The primary endpoint analysis was to evaluate oral malodor based on an organoleptic (OL) score. Additional surrogate measures for oral malodor included quantification of oral hydrogen sulfide (H2S) level and counts of oral bacteria in secondary analyses. At Day 1, all three malodor endpoints were assessed prior to product use, immediately after use, and four and eight hours after use. Subjects were then provided with instructions on product use at home. Subjects returned to the clinic on Day 8 and the assessments for malodor were repeated for each of the three endpoints, i.e., prior to in-clinic use of the products, immediately after use, and four and eight hours after use. One hundred sixty-eight (168) subjects were randomized to three groups, with 56 per treatment group. Of these, 165 completed all study visits. Randomized subjects were comparable for baseline characteristics (OL score, age, race, and ethnicity). Overall, oral malodor based on the organoleptic score decreased for all treatment groups at all timepoints. For the primary endpoint, reduction of OL score eight hours following a single product use, the STC regimen reduced malodor per OL score by 46.67% (SE = 2.28%), the LCM value was 22.83% (SE = 2.29%), and MTB was 26.19% (SE = 2.29%). The pair-wise comparisons between STC and each of the treatment groups were statistically significant (p-values < 0.0001). Statistically significant differences were also observed between STC and both LCM and MTB groups in pair-wise comparisons at Day 8 (p-values < 0.0001). Reductions in malodor were evident following a single use of each product, and also following a seven-day repeat use period. The STC regimen, however, was statistically significantly superior to both LCM and MTB at improving malodor eight hours following the first use. Statistically significant differences in OL scores were sustained between STC and LCM, and STC and MTB at each efficacy timepoint following the seven-day home use period.


CMAX3: A Robust Statistical Test for Genetic Association Accounting for Covariates.
The additive genetic model as implemented in logistic regression has been widely used in genome-wide association studies (GWASs) for binary outcomes. Unfortunately, for many complex diseases, the underlying genetic models are generally unknown and a mis-specification of the genetic model can result in a substantial loss of power. To address this issue, the MAX3 test (the maximum of three separate test statistics) has been proposed as a robust test that performs plausibly regardless of the underlying genetic model. However, the original implementation of MAX3 utilizes the trend test so it cannot adjust for any covariates such as age and gender. This drawback has significantly limited the application of the MAX3 in GWASs, as covariates account for a considerable amount of variability in these disorders. In this paper, we extended the MAX3 and proposed the CMAX3 (covariate-adjusted MAX3) based on logistic regression. The proposed test yielded a similar robust efficiency as the original MAX3 while easily adjusting for any covariate based on the likelihood framework. The asymptotic formula to calculate the p-value of the proposed test was also developed in this paper. The simulation results showed that the proposed test performed desirably under both the null and alternative hypotheses. For the purpose of illustration, we applied the proposed test to re-analyze a case-control GWAS dataset from the Collaborative Studies on Genetics of Alcoholism (COGA). The R code to implement the proposed test is also introduced in this paper and is available for free download.


Tipifarnib-mediated suppression of T-bet-dependent signaling pathways.
Large granular lymphocyte (LGL) leukemia is a chronic lymphoproliferative disease in which T-bet [T-box transcription factor 21 gene (tbx21)] overexpression may play a pathogenic role. T-bet orchestrates the differentiation of mature peripheral T-cells into interferon-γ (IFN-γ) and tumor necrosis factor-α producing CD4+ T-helper type I (Th1) and CD8+ T cytotoxic cells that are necessary for antiviral responses. When IL-12 is produced by antigen-presenting cells, T-bet expression is induced, causing direct stimulation of ifng gene transcription while simultaneously acting as a transcriptional repressor of the IL4 gene, which then leads to Th1 dominance and T-helper type 2 differentiation blockade. Additionally, T-bet has been shown to regulate histone acetylation of the ifng promoter and enhancer to loosen condensed DNA, creating greater accessibility for other transcription factor binding, which further amplifies IFNγ production. We found that treatment with a farnesyltransferase inhibitor tipifarnib reduced Th1 cytokines in LGL leukemia patient T-cells and blocked T-bet protein expression and IL-12 responsiveness in T-cells from healthy donors. The mechanism of suppression was based on modulation of histone acetylation of the ifng gene, which culminated in Th1 blockade.


A herd-level study on colostrum management factors associated with the prevalence of adequate transfer of passive immunity in Québec dairy herds.
The objective of this study was to identify herd-level colostrum management factors associated with the adequate transfer of passive immunity (TPI; defined as serum Brix refractance ≥8.4% in the first week of life). A total of 59 commercial Holstein dairy farms were included in this observational cross-sectional study. In every participating herd, a minimum of 14 Holstein calves were sampled to measure their TPI using a digital Brix refractometer. Colostrum samples fed to each of these calves were collected to estimate IgG concentration (colostrum quality) using a digital Brix refractometer and bacterial contamination using the Petrifilm (3M, St. Paul, MN) culture system. Dairy producers completed a questionnaire on colostrum management to assess on-farm practices. The study outcome was the prevalence of adequate TPI calculated based on the proportion of adequate TPI (defined with an individual threshold ≥8.4% Brix) on the total samples tested within each herd. According to the threshold determined in a previous study investigating the influencing colostrum management factors to achieve adequate TPI at the calf level, the prevalence of an adequate colostrum volume fed at first meal (≥2.5 L), the prevalence of adequate colostrum quality (≥24.5% Brix), the prevalence of an adequate time to first feeding (delay between birth and the first colostrum meal, ≤3 h), the prevalence of low aerobic bacterial contamination (≤20,000 cfu/mL), the prevalence of low coliform contamination (≤1,000 cfu/mL), and the prevalence of females were calculated. The herd-level prevalence of adequate TPI ranged from 24% to 100%, with a median of 68%. The median herd prevalences of an adequate colostrum volume fed at first meal, of adequate colostrum quality, of an adequate time to first feeding, of low aerobic bacterial contamination, of low coliform contamination, and of females, were 71, 42, 41, 64, 88, and 61%, respectively. In the final model, the prevalence of adequate TPI was associated with the prevalence of an adequate colostrum volume fed at first meal and the prevalence of an adequate time to first feeding. In summary, management practices varied greatly between farms and influenced the prevalence of adequate TPI.


Formation of a spiraling line defect and its meandering transition in a period-2 medium.
The instability of a period-1 spiral wave resulting in a period-2 spiral wave with a line defect is investigated for the first time in a laboratory system. At the very onset the transition proceeds by an emergence of a spiraling line defect, "breathing" intermittently while retaining its symmetry of a period-1 spiral wave. With a further change in a control parameter, the line defect undergoes a meandering transition producing a compound tip trajectory, following a dynamic shape transition. The observed transitions have a strong analogy to the phase synchronization transition of two coupled nonlinear oscillators and the meandering transition of a period-1 spiral wave.


[Genetics and molecular biology in cardiology (IV). Myocardial response to biomechanical stress].
Biomechanical stress of the myocardium is the situation resulting from hypoxia, hypertension, and other forms of myocardial injury, that invariably lead to increased demands for cardiac work and/or loss of functional myocardium. As a consequence of biomechanical stress a number of responses develop involving all the myocardial cells, namely cardiomyocytes. As a result some myocardial phenotypic changes develop that are initially compensatory (i.e., hypertrophy) but which may mediate the eventual decline in myocardial function that occurs with the transition from hypertrophy to failure in conditions of persistent stress (i.e., apoptosis and fibrosis). This review focuses on the steps involved in the response of the myocardium to biomechanical stress and highlights the most recent developments in the molecular mechanisms involved in the development of heart failure.


[Effects of different tillage and fertilization modes on the soil physical and chemical properties and crop yield under winter wheat/spring corn rotation on dryland of east Gansu, Northwest China].
Based on the 7-year field experiment on the dryland of east Gansu of Northwest China in 2005-2011, this paper analyzed the variations of soil moisture content, bulk density, and nutrients content at harvest time of winter wheat and of the grain yield under no-tillage and conventional tillage and five fertilization modes, and approached the effects of different tillage and fertilization modes on the soil water storage and conservation, soil fertility, and grain yield under winter wheat/ spring corn rotation. In 2011, the soil moisture content in 0-200 cm layer and the soil bulk density and soil organic matter and available nitrogen and phosphorus contents in 0-20 cm and 20-40 cm layers under different fertilization modes were higher under no-tillage than under conventional tillage. Under the same tillage modes, the contents of soil organic matter and available nitrogen and available phosphorus were higher under the combined application of organic and inorganic fertilizers, as compared with other fertilization modes. The soil available potassium content under different tillage and fertilization modes decreased with years. The grain yield under conventional tillage was higher than that under no-tillage. Under the same tillage modes, the grain yield was the highest under the combined application of organic and inorganic fertilizers, and the lowest under no fertilization. In sum, no-tillage had the superiority than conventional tillage in improving the soil water storage and conservation and soil fertility, and the combined application of organic and inorganic fertilizers under conventional tillage could obtain the best grain yield.


Use of CMFDA and CMTMR fluorescent dyes in FACS-based antibody screening.
Cell-based immunizations are often used when membrane antigens are difficult to purify. To confirm that an antibody binding to the surface of a cell line is, in fact, binding to the desired antigen, FACS can be performed independently on two cell lines, a transfected cell line expressing the antigen of interest and a control cell line not expressing the antigen. Antibodies binding only to the transfected cell line are then selected for further analysis. This approach can be challenging if a large number of antibodies need to be screened and the antibody quantities are limited. Here we describe a novel method that combines the above two steps of FACS screening into a single step, based on the use of two fluorochromes, CMFDA and CMTMR, to stain transfected and control cell lines, respectively. Antibodies conjugated to a thirdfluorochrome are then added to the combined cells, followed by three-color FACS analysis. The newly modified FACS method is simple, sensitive, and high throughput. It can be used for antibody screening in multiple cell lines simultaneously.


Association of nutritional status and pulmonary function in children with cystic fibrosis.
Multiple studies have shown that nutritional status is a strong predictor of morbidity and mortality in patients with cystic fibrosis (CF). Since CF is characterized by progressive lung disease, it could be argued that the underlying lung disease is what determines the nutritional failure seen in most patients. This review will summarize the data available from studies that have attempted to better define this relation and also present a review of the possible mechanisms involved taken from both observational and interventional studies. Longitudinal studies with sufficiently large follow-up times have demonstrated that young underweight patients have worst pulmonary function outcomes. More importantly, these studies concur in that the yearly change in growth parameters has a significant effect on the rate at which pulmonary function develops. Although the mechanisms behind this important association are yet unclear, there is some suggestion from interventional studies that the accrual of lean body mass is the factor that is involved in the preservation of lung function. Nutritional status strongly influences pulmonary health among CF patients. Therefore, aggressive nutritional support aiming at achieving normal growth patterns should lead to adequate development of lung function and maintenance of pulmonary health. However, more research is required with long-term longitudinal studies to better identify the most critical nutritional characteristics influencing this process as well as the most effective nutritional interventions.


Altered signal transduction pathways and induction of autophagy in human myotonic dystrophy type 1 myoblasts.
Congenital myotonic dystrophy type 1 (CDM1) affects patients from birth and is associated with mental retardation and impaired muscle development. CDM1 patients carry 1000-3000 CTG repeats in the DMPK gene and display defective skeletal muscles differentiation, resulting in reduced size of myotubes and decreased number of satellite cells. In this study, human myoblasts in culture deriving from control and DM1 embryos (3200 CTG repeats) were analyzed using both a biochemical and electron microscopic approach, in order to provide new insights into the molecular mechanisms underlying such alteration. Interestingly, electron microscopy analysis showed not only ultrastructural features of abnormal differentiation but also revealed the presence of autophagic vacuoles in DM1 myoblasts not undergoing differentiation. In accordance with the electron microscopic findings, the autophagic markers LC3 and ATG5, but not apoptotic markers, were significantly up regulated in DM1 myoblasts after differentiating medium addition. The induction of autophagic processes in DM1 myoblasts was concomitant to p53 over-expression and inhibition of the mTOR-S6K1 pathway, causatively involved in autophagy. Moreover biochemical alterations of the two main signal transduction pathways involved in differentiation were observed in DM1 myoblasts, in particular decreased activation of p38MAPK and persistent activation of the MEK-ERK pathway. This work, while demonstrating that major signaling pathways regulating myoblasts differentiation are profoundly deranged in DM1 myoblasts, for the first time provides evidence of autophagy induction, possibly mediated by p53 activation in response to metabolic stress which might contribute to the dystrophic alterations observed in the muscles of congenital DM1 patients.


Mobile phone-based e-diary for assessment and enhancement of medications adherence among patients with multiple sclerosis.
Adherence to multiple sclerosis (MS) disease-modifying drugs (DMDs) is essential for realization of their optimal effectiveness and benefits. To evaluate the usefulness and validity of a smartphone-based e-diary as a tool for adherence assessment as well as its effectiveness as a promoter of adherence to DMDs. An MS tailored e-diary (MyMS&Me) reminded patients to take their DMDs on time. DMD intake was self-recorded in the e-diary by the participants. Three methods of adherence evaluation were compared: e-diary derived, retrospective self-reported, and the medication possession rate (MPR). The proportion of patients with poor adherence to DMDs (defined as MPR <80%) among e-diary users was compared with a control group without intervention. Sixty-two patients downloaded the e-diary (Female: 41 (66%), Expanded Disability Status Scale 3.2 ± 2.2) and 55 controls were enrolled. The median difference between e-diary-derived adherence and the MPR was -3% (95% limits of agreement: -53% to 12%). The median difference between retrospective self-reported adherence and the MPR was 0.3% (95% limits of agreement: -20% to 42%). The proportion of participants with poor adherence to DMDs was similar in the e-diary and control groups (10% vs. 13%, p = 0.6). Substantial and clinically important disagreement between methods of medication adherence evaluation was noted. Smartphone reminders did not significantly improve the MPR of DMDs.


Weight-dependent susceptibility of tilapia to tilapia lake virus infection.
The emergence of tilapia lake virus (TiLV) has had a severely negative impact on global tilapia aquaculture. TiLV infection has been reported at different life stages of tilapia, with more emphasis on fry and fingerlings; however, the virulence and pathology of TiLV at different tilapia size remains unexplored. In this study, tilapias from a single spawning were raised to 5 g, 25 g, and 65 g, and subsequently challenged by the intraperitoneal injection and cohabitation of a virulent strain of TiLV. The cumulative mortality, viral load, and histopathology of the fish were determined until 22 days post-infection (dpi). The cumulative mortality of the 5 g, 25 g, and 65 g fish was 85% (±1.67), 55% (±2.89), and 51.67% (±7.49), respectively. At 14 dpi, the mean TiLV load in the liver of the 5 g fish was significantly higher than in the 25 g and 65 g fish. All the weight groups showed severe pathological changes in the liver, spleen, and intestine after TiLV infection, but no particular difference was otherwise noted during the study with the exception of higher pathological scores in the liver of the small fish at 14 dpi. Overall, this study indicated that small fish are more susceptible to TiLV infection than large fish. Although multiple factors, including environmental factors, farm management practices, strains of virus could contribute to different susceptibility of fish to viral infection, the present study provides the evidence to support that fish weight affects the mortality and clinical outcome during TiLV infection. More intensive measures such as strict biosecurity and disease surveillance during the susceptible weight should therefore be emphasized to reduce the impact of this virus.


Younger but Not Older Adults Benefit from Salient Feedback during Learning.
Older adults are impaired in reinforcement learning (RL) when feedback is partially ambiguous (e.g., Eppinger and Kray, 2011). In this study we examined whether older adults benefit from salient feedback information during learning. We used an electrophysiological approach and investigated 15 younger and 15 older adults with a RL task in which they had to learn stimulus-response associations under two learning conditions. In the positive learning conditions, participants could gain 50 Cents for a correct response but did not gain or lose money (*00 Cent) for an incorrect response. In negative learning conditions, they could lose 50 Cents for an incorrect response but did not gain or lose money (*00 Cent) for a correct response. As the identical outcome "00 Cent" is either better or worse than the alternative outcome depending on the learning condition, this feedback type is ambiguous. To examine the influence of feedback salience we compared this condition with a condition in which positive and negative outcomes were color-coded and thereby clearly separable. The behavioral results indicated that younger adults reached higher accuracy levels under salient feedback conditions. Moreover, the error-related negativity and the feedback-related negativity for losses were larger if the good-bad dimension of feedback was salient. Hence, in younger adults salient feedback facilitates the rapid evaluation of outcomes on a good-bad dimension and by this supports learning. In contrast, for older adults we obtained neither behavioral nor electrophysiological effects of feedback salience. The older adults' performance monitoring system therefore appears less flexible in integrating additional information in this evaluation process.


Inhibition of influenza A virus infection in vitro by peptides designed in silico.
Influenza A viruses are enveloped, segmented negative single-stranded RNA viruses, capable of causing severe human respiratory infections. Currently, only two types of drugs are used to treat influenza A infections, the M2 H(+) ion channel blockers (amantadine and rimantadine) and the neuraminidase inhibitors (NAI) (oseltamivir and zanamivir). Moreover, the emergence of drug-resistant influenza A virus strains has emphasized the need to develop new antiviral agents to complement or replace the existing drugs. Influenza A virus has on the surface a glycoprotein named hemagglutinin (HA) which due to its important role in the initial stage of infection: receptor binding and fusion activities of viral and endosomal membranes, is a potential target for new antiviral drugs. In this work we designed nine peptides using several bioinformatics tools. These peptides were derived from the HA1 and HA2 subunits of influenza A HA with the aim to inhibit influenza A virus infection. The peptides were synthetized and their antiviral activity was tested in vitro against several influenza A viral strains: Puerto Rico/916/34 (H1N1), (H1N1)pdm09, swine (H1N1) and avian (H5N2). We found these peptides were able to inhibit the influenza A viral strains tested, without showing any cytotoxic effect. By docking studies we found evidence that all the peptides were capable to bind to the viral HA, principally to important regions on the viral HA stalk, thus could prevent the HA conformational changes required to carry out its membranes fusion activity.


An Extra Amino Acid Residue in Transmembrane Domain 10 of the γ-Aminobutyric Acid (GABA) Transporter GAT-1 Is Required for Efficient Ion-coupled Transport.
The GABA transporter GAT-1 mediates electrogenic transport of its substrate together with sodium and chloride. It is a member of the neurotransmitter:sodium:symporters, which are crucial for synaptic transmission. Compared with all other neurotransmitter:sodium:symporters, GAT-1 and other members of the GABA transporter subfamily all contain an extra amino acid residue at or near a conserved glycine in transmembrane segment 10. Therefore, we studied the functional impact of deletion and replacement mutants of Gly-457 and its two adjacent residues in GAT-1. The glycine replacement mutants were devoid of transport activity, but remarkably the deletion mutant was active, as were mutants obtained by deleting positions on either side of Gly-457. However, the inward rectification of GABA-induced transport currents by all three deletion mutants was diminished, and the charge-to-flux ratio was increased by more than 2.5-fold, both of which indicate substantial uncoupled transport. These observations suggest that the deletions render the transporters less tightly packed. Consistent with this interpretation, the inactive G457A mutant was partially rescued by removing the adjacent serine residue. Moreover, the activity of several gating mutants was also partially rescued upon deletion of Gly-457. Structural modeling showed that the stretch surrounding Gly-457 is likely to form a π-helix. Our data indicate that the "extra" residue in transmembrane domain 10 of the GABA transporter GAT-1 provides extra bulk, probably in the form of a π-helix, which is required for stringent gating and tight coupling of ion and substrate fluxes in the GABA transporter family.


Vitamin A status of children in the urban slums of Karachi, Pakistan, assessed by clinical, dietary, and biochemical methods.
We assessed the vitamin A status of 532 children with an age range of 6-60 months who were living in slum areas of Karachi, Pakistan, using three methodologies: clinical eye examination, dietary vitamin A intake, and serum retinol level. No definite clinical signs of xerophthalmia were observed in any of these children. The mean +/- SD vitamin A intake estimated from a food frequency questionnaire for the group with inadequate (low and deficient) serum retinol levels (< 20 micrograms/dl) was 362 +/- 332 retinol equivalents (RE) compared with 431 +/- 332) RE in the group with adequate serum levels (P < 0.005). Deficient serum retinol levels (< 10 micrograms/dl) were present in 12 children (2%); two of these had a healed corneal scar. Low serum retinol levels (10-19 micrograms/dl) were present in 46%, while 51% children had adequate levels (> or = 20 micrograms/dl). The mean +/- SD serum retinol level for the inadequate (< 20 micrograms/dl) and adequate groups were 15.3 +/- 2.8 and 26.6 +/- 6.7 micrograms/dl, respectively. These results suggest that a significant number of children in these communities have low vitamin A levels and thus may constitute an at risk group. These results also suggest that the dietary intake method may be a simple and inexpensive screening tool for assessment of vitamin A status in communities.


5'-Bis-conjugation of oligonucleotides by amidative oxidation and click chemistry.
A pent-4-ynyl tert-butyl N,N-diisopropyl phosphoramidite was coupled at the 5'-end of oligonucleotides to give a phosphite triester linkage, which forms an H-phosphonate diester linkage during treatment with dichloroacetic acid. Then an amidative oxidation with CCl(4) in the presence of an amine and a 1,3-dipolar cycloaddition with an azide under copper(I) catalysis afforded the bis-conjugated oligonucleotides with high efficiency. The introduction of a bromoalkyl group as a precursor of azidoalkyl by amidative oxidation allowed the performance of two selective 1,3-dipolar cycloadditions.


MUSI: an integrated system for identifying multiple specificity from very large peptide or nucleic acid data sets.
Peptide recognition domains and transcription factors play crucial roles in cellular signaling. They bind linear stretches of amino acids or nucleotides, respectively, with high specificity. Experimental techniques that assess the binding specificity of these domains, such as microarrays or phage display, can retrieve thousands of distinct ligands, providing detailed insight into binding specificity. In particular, the advent of next-generation sequencing has recently increased the throughput of such methods by several orders of magnitude. These advances have helped reveal the presence of distinct binding specificity classes that co-exist within a set of ligands interacting with the same target. Here, we introduce a software system called MUSI that can rapidly analyze very large data sets of binding sequences to determine the relevant binding specificity patterns. Our pipeline provides two major advances. First, it can detect previously unrecognized multiple specificity patterns in any data set. Second, it offers integrated processing of very large data sets from next-generation sequencing machines. The results are visualized as multiple sequence logos describing the different binding preferences of the protein under investigation. We demonstrate the performance of MUSI by analyzing recent phage display data for human SH3 domains as well as microarray data for mouse transcription factors.


Studies of the modulation of MHC antigen and cell adhesion molecule expression on human dermal microvascular endothelial cells.
Interactions between leukocytes and endothelial cells, particularly in the microvasculature, are important for the initiation and regulation of tissue inflammation. These interactions are regulated by the recognition of specific cell adhesion molecules (CAM) on both leukocytes and endothelial cells. In this study, we examined the modulation of cell surface expression of MHC antigens and the CAM intercellular adhesion molecule 1 (ICAM-1), lymphocyte function antigen 3 (LFA-3), and CD44 on human dermal microvascular endothelial cells (HDMEC) both grown in monolayers and differentiated into capillary-like structures on the basement membrane-like substrate matrigel. HDMEC grown in monolayers or differentiated on matrigel express comparable cell surface MHC class I, LFA-3, CD44, and ICAM-1. ICAM-1, but not LFA-3 or CD44, was increased in expression in a dose- and time-dependent manner by interleukin 1 (IL-1) alpha, tumor necrosis factor (TNF) alpha, lipopolysaccharide (LPS), or interferon (IFN) gamma. Comparable upregulation was observed both in cells grown in monolayers and cells differentiated on matrigel. IL-1 alpha, TNF alpha, and LPS increased ICAM-1 expression on average 100-200% whereas IFN gamma was somewhat less potent. Comparative studies with human umbilical vein endothelial cells (HUVEC) demonstrated consistently lower levels of ICAM-1 expression on HUVEC, but greater increases after cytokine stimulation. Pretreatment with dexamethasone or transforming growth factor (TGF) beta did not affect baseline expression of ICAM-1 or inhibit upregulation of ICAM-1 on HDMEC by IL-1 alpha, TNF alpha, LPS, or IFN gamma. Both IFN gamma and TNF alpha, but not IL-1 alpha increased MHC class I expression, whereas only IFN gamma induced the expression of HLA-DR on HDMEC. The effect of IL-1 alpha, TNF alpha, or IFN gamma was inhibited by antibody to the specific cytokine, but was unaffected by antibody to other cytokines. Additionally, IFN alpha or beta inhibited upregulation of HLA-DR by IFN gamma, but had no effect on the increased MHC class I or ICAM-1 expression mediated by this cytokine. These data demonstrate that the expression of CAM and MHC antigens on small vessel-derived endothelial cells is different from that observed on large-vessel HUVEC, is regulated by the presence of multiple cytokines operating via distinct pathways, and the expression and regulation of these proteins appear to be similar on cells that have been grown in monolayers to those morphologically differentiated into blood vessel-like structures.