Effects of galacto-oligosaccharide prebiotics in blood profile of severely acute malnourished children.
This study assessed the effects of galacto-oligosaccharides (Oligomate) on hematocrit, serum enzymes, total bilirubin levels, and serum electrolytes in controls and severely malnourished infants, with emphasis on gastrointestinal symptoms. Oligomate doses and phases did not affect stools frequency per day, indicating that prebiotic effect on stool may be due to the prebiotic type. The number of vomits per day during phases 2 and 3 were significantly reduced (p<0.05) in response to prebiotics, despite the prebiotic dose effect was not significant (p>0.05). Moreover, prebiotics administration during phases 2 and 3 markedly improved hemoglobin levels (p<0.05), but not the dose. Similarly, hematocrit levels and white blood cells were significantly improved during the last 2 phases, but dose have no effects on blood hematocrit levels. Erythrocyte sedimentation rate significantly decreased (p<0.05) in phases 2 and 3 compared to phase 1. No dose-related effect was stated on erythrocytes sedimentation rate. Regarding the serum enzymes, SGPT significantly decreased (p<0.05) in phases 2 and 3 compared to phase 1, whereas SGOT significantly decreased only in phase 3. Total bilirubin levels increased significantly (p<0.05) in phase 3 when compared to phases 1 or 2. Prebiotics significantly decreased (p<0.05) sodium levels in the treated group, while potassium levels did not change in all groups, excepting during phase 2, where it increased significantly. Thus, our results confirm the hypothesis that prebiotic supplementation improves blood parameters and health status, consequently decreasing the infection risk and number of vomit per day in infants.


Ultrasound-guided lumbar puncture improves success rate and efficiency in overweight patients.
Approximately 400,000 diagnostic lumbar punctures (LPs) are performed by neurologists yearly in the United States. There has been a transition from neurologists performing >40% of LPs 2 decades ago to now <15% of the time. High body mass index (BMI) is one of the key obstacles of LP for neurology residents, and the success rates drop to 58% when BMI is >35 kg/m2. A prospective intervention study was conducted from March 2017 to March 2018 at an academic medical center. Patients were divided into ultrasound-guided LP or conventional LP. The primary outcome was the success rate of LP. The secondary outcome was the parameters regarding LP performance, the patients' feedback after procedures, and the residents' feedback about the learning modules. Eighty-two patients were enrolled with inclusion criteria of age >18 years, BMI ≥25 kg/m2, and able to give written informed consent. The ultrasound-guided LP group showed higher success rate (92.7% vs 68.3%, p < 0.05) and less duration of time (17.02 vs 37.63 minutes, p < 0.05) compared with the conventional LP group. It also showed advantages of decreased trials of needle insertions and redirections and reduced pain levels perceived by patients. This study suggests that using ultrasound to localize before LP could increase the LP success rate and improve other related LP outcomes in the overweight populations. Our study also provides evidence that brief, targeted teaching modules are feasible and effective for ultrasound-guided LP training. This study provides Class III evidence that for patients with BMI ≥25 kg/m2 undergoing LP, ultrasound guidance increases the LP success rate.


ACR Appropriateness Criteria® Sinonasal Disease.
Imaging of sinonasal pathology may occur for assessment of rhinosinusitis or mass lesions. Rhinosinusitis is prevalent in up to 16% of the US population with annual economic burdens estimated at 22 billion dollars. Rhinosinusitis is characterized as acute or chronic based on symptom duration; if four or more episodes occur annually, the term recurrent acute rhinosinusitis (RARS) is used. In acute uncomplicated rhinosinusitis when inflammatory change remains in the paranasal sinuses and nasal cavity, imaging may not be required. Distinction between viral or bacterial rhinosinusitis is a clinical diagnosis, and imaging should be interpreted in conjunction with clinical and endoscopic findings. Sinus CT imaging is appropriate per clinical judgment in associated complications including headache, facial pain, swelling, orbital proptosis, or cranial nerve palsies. In maxillary sinusitis, teeth may require assessment because 20% may be odontogenic in origin. MRI may be complementary in aggressive infections with intraocular/intracranial complications, invasive fungal sinusitis, or sinonasal masses. The American College of Radiology Appropriateness Criteria are evidence-based guidelines for specific clinical conditions that are reviewed annually by a multidisciplinary expert panel. The guideline development and revision include an extensive analysis of current medical literature from peer reviewed journals and the application of well-established methodologies (RAND/UCLA Appropriateness Method and Grading of Recommendations Assessment, Development, and Evaluation or GRADE) to rate the appropriateness of imaging and treatment procedures for specific clinical scenarios. In those instances where evidence is lacking or equivocal, expert opinion may supplement the available evidence to recommend imaging or treatment.


Value redefined for inflammatory bowel disease patients: a choice-based conjoint analysis of patients' preferences.
Value-based healthcare is an upcoming field. The core idea is to evaluate care based on achieved outcomes divided by the costs. Unfortunately, the optimal way to evaluate outcomes is ill-defined. In this study, we aim to develop a single, preference based, outcome metric, which can be used to quantify overall health value in inflammatory bowel disease (IBD). IBD patients filled out a choice-based conjoint (CBC) questionnaire in which patients chose preferable outcome scenarios with different levels of disease control (DC), quality of life (QoL), and productivity (Pr). A CBC analysis was performed to estimate the relative value of DC, QoL, and Pr. A patient-centered composite score was developed which was weighted based on the stated preferences. We included 210 IBD patients. Large differences in stated preferences were observed. Increases from low to intermediate outcome levels were valued more than increases from intermediate to high outcome levels. Overall, QoL was more important to patients than DC or Pr. Individual outcome scores were calculated based on the stated preferences. This score was significantly different from a score not weighted based on patient preferences in patients with active disease. We showed the feasibility of creating a single outcome metric in IBD which incorporates patients' values using a CBC. Because this metric changes significantly when weighted according to patients' values, we propose that success in healthcare should be measured accordingly.


Symptomatic response rates to testosterone therapy and the likelihood of completing 12 months of therapy in clinical practice.
Despite increasing medical interest in testosterone therapy (TTh) for men with testosterone deficiency (TD) there is limited information regarding subjective response rates and acceptability of medium- to long-term TTh in routine clinical practice. To evaluate results in a consecutive series of men in clinical practice treated with TTh. A chart review was performed for a consecutive series of men for whom TTh was initiated over 1 year for a clinical diagnosis of TD. A diagnosis of TD was based on the presence of symptoms and on laboratory evaluation indicating total testosterone (<300 ng/dL) or free testosterone (FT) (<1.5 ng/dL). Presenting symptoms were noted at baseline, and improvement was documented in domains of erectile function, libido, energy,and mood. Percentage of men who completed 12 months of TTh, and symptomatic response rates. There were 127 men included in the evaluation. The most common presenting symptoms were the combination of erectile dysfunction (ED) and reduced libido in 82 (64.6%), ED alone in 29 (22.8%), and reduced libido alone in 13 (10.2%). Initial mode of TTh was injections (testosterone enanthate or cypionate) in 70 (55.1%)and transdermal gel (Androgel, Solvay Pharmaceuticals, Marietta, GA, USA) in the remainder. Improvements in erections, libido, energy, and/or mood were reported by 70% of men by 3 months. Eighty men (63%) completed 12 months of TTh with subjective benefit (responders). Treatment was discontinued in 34 (26.8%) who reported no major benefit (non-responders), and 13 (10.2%) were lost to follow-up. Among men who discontinued TTh, 64.7%failed to report benefits by 3 months. Baseline FT was lower among responders than non-responders. One case(1.25%) of prostate cancer was identified after one year of TTh. Approximately two-thirds of men with TD who begin TTh will experience symptomatic benefit and will complete at least 12 months of treatment. Benefit was noted in a majority by 3 months.


Genetic exchange among natural isolates of bacteria: recombination within the phoA gene of Escherichia coli.
An 1871-nucleotide region including the phoA gene (the structural gene encoding alkaline phosphatase, EC 220.127.116.11) was cloned and sequenced from eight naturally occurring strains of Escherichia coli. Alignment with the sequence from E. coli K-12 made apparent that there were 87 polymorphic nucleotide sites, of which 42 were informative for phylogenetic analysis. Maximum parsimony analysis revealed six equally parsimonious trees with a consistency index of 0.80. Of the 42 informative sites, 22 were inconsistent with each of the maximum parsimony trees. The spatial distribution of the inconsistent sites was highly nonrandom in a manner implying that intragenic recombination has played a major role in determining the evolutionary history of the nine alleles. The implication is that different segments of the phoA gene have different phylogenetic histories.


Controversies in the cellular pharmacology of oligodeoxynucleotides.
Phosphodiester and phosphorothioate oligodeoxynucleotides are polyanions that cannot passively diffuse across cell membranes. Instead, the processes of adsorbtive endocytosis and pinocytosis probably account for the great majority of oligodeoxynucleotide internalization in most cell types. Oligodeoxynucleotides can adsorb to heparin-binding, cell surface proteins. An example of such a protein is the integrin Mac-1 (alpha M beta 2; CR3; CD11b/CD18), a receptor for fibrinogen which is found on neutrophils, macrophages and natural killer cells. Up-regulation of neutrophil cell surface Mac-1 expression by interleukin 8, arachidonic acid or tumour necrosis factor alpha leads to increased cell surface oligodeoxynucleotide binding and internalization. Binding and internalization can be blocked by both fibrinogen and by anti-Mac-1 monoclonal antibodies. Subsequent to internalization, oligodeoxynucleotides reside in subcellular vesicular structures, i.e. endosomes and lysosomes. However, in the absence of permeabilizing agents, these compartments may be sites of sequestration and the oligomers may be unavailable for antisense activity. At present, controversy surrounds the use of guanosine-rich phosphorothioate oligodeoxynucleotides as antisense agents. We examined the ability of the 24mer antisense rel A (p65) phosphorothioate oligodeoxynucleotide to inhibit nuclear translocation of NF kappa B in K-BALB murine fibroblasts. 7-Deaza-2'-deoxyguanosine substitution in the 5' guanosine quartet region demonstrated that inhibition of nuclear translocation could not be due to a Watson-Crick antisense effect. Rather, we favour the explanation that the parent molecule may be a sequence-specific, apatameric decoy.


Current practice of venous thromboembolism prevention in acute trusts: a qualitative study.
To explore the current practice of venous thromboembolism (VTE) prevention in acute trusts. A qualitative research design was used to explore the perceived current practice of thromboprophylaxis, and knowledge and experience of VTE prevention. Data were collected via interviews with personnel from acute trusts and other relevant organisations and charities. Constant comparison was used to generate themes grounded in the data. The UK. 17 participants, sampled due to their expertise and knowledge in the field of VTE, were interviewed for the study. No one felt directly responsible for VTE risk assessment and treatment in acute trusts. There were concerns whether any action takes place based on the risk assessment. Low levels of VTE knowledge existed throughout the system. Our study highlights the importance of continuous training to prevent VTE risk assessment being considered a tick box exercise and for clinicians to understand the significance of the procedure to ensure that VTE preventative measures are administered. It is essential that acute trust staff acknowledge that VTE prevention is the responsibility of everyone involved in a patient's care. Concerns remain around prophylaxis treatment, administration and contraindications.


Cognitive-behavioral treatment with adult alcohol and illicit drug users: a meta-analysis of randomized controlled trials.
This meta-analysis examined 53 controlled trials of cognitive-behavioral treatment (CBT) for adults diagnosed with alcohol- or illicit-drug-use disorders. The aims were to provide an overall picture of CBT treatment efficacy and to identify client or treatment factors predictive of CBT effect magnitude. The inverse variance weighted effect size (Hedges' g) was calculated for each study and pooled using fixed and random effects methods. Potential study-level moderators were assessed in subgroup analyses by primary drug, type of CBT, and type of comparison condition. In addition, seven client and treatment variables were examined in meta-regression analyses. Across studies, CBT produced a small but statistically significant treatment effect (g = 0.154, p < .005). The pooled effect was somewhat lower at 6-9 months (g = 0.1 15, p < .005) and continued to diminish at 12-month follow-up (g = 0.096, p < .05). The effect of CBT was largest in marijuana studies (g = 0.513, p < .005) and in studies with a no-treatment control as the comparison condition (g = 0.796, p < .005). Meta-regression analyses indicated that the percentage of female participants was positively associated and the number of treatment sessions was negatively associated with effect size. The findings demonstrate the utility of CBT across a large and diverse sample of studies and under rigorous conditions for establishing efficacy. CBT effects were strongest with marijuana users, when CBT was compared with no treatment, and may be larger with women than with men and when delivered in a brief format.


[Rapid analysis of genetically modified soybean by a duplex PCR-capillary electrophoresis system with laser-induced fluorescence detection].
To develop a rapid detection method for genetically modified soybean resistant to glyphosate. A duplex PCR was performed with primers designed in this study to simultaneously amplify heterogenous genes in transgenic soybean: CaMV-35S promoter, NOS terminator and CP4-EPSPS gene. And a simple capillary electrophoresis with laser-induced fluorescence detection (CE-LIF) was developed and applied to the rapid analysis of the above PCR products by using a 50 cm length x 100 microm i.d. capillary coated with linear polyacrylamide and an 8 g/L HPMC-4000 sieving buffer under 200 V/cm electric field strength. The proposed method was able to simultaneously detect the three heterogenous genes existing in genetically modified soybean under the optimization conditions of PCR and capillary electrophoresis. The measured sequences of the duplex PCR products were identical with the original genes' sequences. Moreover, the sample volumes required were not more than 5 nl and the detection could be completed in less than 24 min. The relative standard deviations (R. S. D.) of the migration times for the PCR products were < or = 3.2%. In comparison with agarose gels electrophoresis, the duplex PCR-based capillary electrophoretic method with laser-induced fluorescence detection is rapid, sensitive and accurate, and it is suitable for detection of genetically modified soybean.


Microbiome of Nodules and Roots of Soybean and Common Bean: Searching for Differences Associated with Contrasting Performances in Symbiotic Nitrogen Fixation.
Biological nitrogen fixation (BNF) is a key process for the N input in agriculture, with outstanding economic and environmental benefits from the replacement of chemical fertilizers. However, not all symbioses are equally effective in fixing N2, and a major example relies on the high contribution associated with the soybean (Glycine max), contrasting with the low rates reported with the common bean (Phaseolus vulgaris) crop worldwide. Understanding these differences represents a major challenge that can help to design strategies to increase the contribution of BNF, and next-generation sequencing (NGS) analyses of the nodule and root microbiomes may bring new insights to explain differential symbiotic performances. In this study, three treatments evaluated in non-sterile soil conditions were investigated in both legumes: (i) non-inoculated control; (ii) inoculated with host-compatible rhizobia; and (iii) co-inoculated with host-compatible rhizobia and Azospirillum brasilense. In the more efficient and specific symbiosis with soybean, Bradyrhizobium presented a high abundance in nodules, with further increases with inoculation. Contrarily, the abundance of the main Rhizobium symbiont was lower in common bean nodules and did not increase with inoculation, which may explain the often-reported lack of response of this legume to inoculation with elite strains. Co-inoculation with Azospirillum decreased the abundance of the host-compatible rhizobia in nodules, probably because of competitiveness among the species at the rhizosphere, but increased in root microbiomes. The results showed that several other bacteria compose the nodule microbiomes of both legumes, including nitrogen-fixing, growth-promoters, and biocontrol agents, whose contribution to plant growth deserves further investigation. Several genera of bacteria were detected in root microbiomes, and this microbial community might contribute to plant growth through a variety of microbial processes. However, massive inoculation with elite strains should be better investigated, as it may affect the root microbiome, verified by both relative abundance and diversity indices, that might impact the contribution of microbial processes to plant growth.


Spectroscopic evidence for interfacial Fe(II)-Fe(III) electron transfer in a clay mineral.
Interfacial electron transfer has been shown to occur between sorbed Fe(II) and structural Fe(III) in Fe oxides, but it is unknown whether a similar reaction occurs between sorbed Fe(II) and Fe(III)-bearing clay minerals. Here, we used the isotopic specificity of (57)Fe Mössbauer spectroscopy to demonstrate electron transfer between sorbed Fe(II) and structural Fe(III) in an Fe-bearing smectite clay mineral (NAu-2, nontronite). Mössbauer spectra of NAu-2 reacted with aqueous (56)Fe(II) (which is invisible to (57)Fe Mössbauer spectroscopy) showed direct evidence for reduction of NAu-2 by sorbed Fe(II). Mössbauer spectra using aqueous (57)Fe(II) showed that sorbed Fe(II) is oxidized upon sorption to the clay and pXRD patterns indicate that the oxidation product is lepidocrocite. Spectra collected at different temperatures indicate that reduction of structural Fe(III) by sorbed Fe(II) induces electron delocalization in the clay structure. Our results also imply that interpretation of room temperature and 77 K Mössbauer spectra may significantly underestimate the amount of Fe(II) in Fe-bearing clays. These findings provide compelling evidence for abiotic reduction of Fe-bearing clay minerals by sorbed Fe(II), and require us to reframe our conceptual model for interpreting biological reduction of clay minerals, as well as contaminant reduction by reduced clays.


Spread of antibiotic resistance with food-borne pathogens.
This short review summarizes data on antibiotic resistance profiles of common food-borne pathogens like Salmonella sp., Escherichia coli, Campylobacter sp., Listeria monocytogenes, Clostridium perfringens, Staphylococcus aureus, and coagulase-negative staphylococci. As a flashlight on the literature of the last few years, it provides ample evidence that antibiotic resistance traits have entered the microflora of farm animals and the food produced from them. Molecular analysis of the resistance genes, where available, shows that the food microflora is not separated from its human counterpart and conjugative transfer of resistance genes has been demonstrated in vitro and in a few cases in vivo. For example, for Salmonella typhimurium, resistance towards tetracyclines has increased from zero in 1948 to a 98% level in certain epidemic populations of S. typhimurium DT104 in 1998. The high incidence of food-borne pathogens in raw meat and milk together with a high level of therapeutic, prophylactic and nutritional application of antibiotics in agriculture reveals an antibiotic resistance problem of global dimensions. The resistance problem in human medicine will not be solved if there is a constant influx of resistance genes into the human microflora via the food chain.


Abnormal anxiety- and depression-like behaviors in mice lacking both central serotonergic neurons and pancreatic islet cells.
Dysfunction of central serotonin (5-HT) system has been proposed to be one of the underlying mechanisms for anxiety and depression, and the association of diabetes mellitus and psychiatric disorders has been noticed by the high prevalence of anxiety/depression in patients with diabetes mellitus. This promoted us to examine these behaviors in central 5-HT-deficient mice and those also suffering with diabetes mellitus. Mice lacking either 5-HT or central serotonergic neurons were generated by conditional deletion of Tph2 or Lmx1b respectively. Simultaneous depletion of both central serotonergic neurons and pancreatic islet cells was achieved by administration of diphtheria toxin (DT) in Pet1-Cre;Rosa26-DT receptor (DTR) mice. The central 5-HT-deficient mice showed reduced anxiety-like behaviors as they spent more time in and entered more often into the light box in the light/dark box test compared with controls; similar results were observed in the elevated plus maze test. However, they displayed no differences in the immobility time of the forced swimming and tail suspension tests suggesting normal depression-like behaviors in central 5-HT-deficient mice. As expected, DT-treated Pet1-Cre;Rosa26-DTR mice lacking both central serotonergic neurons and pancreatic islet endocrine cells exhibited several classic diabetic symptoms. Interestingly, they displayed increased anxiety-like behaviors but reduced immobility time in the forced swimming and tail suspension tests. Furthermore, the hippocampal neurogenesis was dramatically enhanced in these mice. These results suggest that the deficiency of central 5-HT may not be sufficient to induce anxiety/depression-like behaviors in mice, and the enhanced hippocampal neurogenesis may contribute to the altered depression-like behaviors in the 5-HT-deficient mice with diabetes. Our current investigation provides understanding the relationship between diabetes mellitus and psychiatric disorders.


Pituitary stalk interruption syndrome and isolated pituitary hypoplasia may be caused by mutations in holoprosencephaly-related genes.
Holoprosencephaly (HPE) is a developmental defect characterized by wide phenotypic variability, ranging from minor midline malformations (eg, single central incisor) to severe deformities. In 10-15% of HPE patients, mutations in specific genes have been identified (eg, SHH, TGIF, SIX3). Pituitary stalk interruption syndrome (PSIS) constitutes a distinct abnormality of unknown pathogenesis, whereas isolated pituitary hypoplasia (IPH) has been linked to various developmental genes. Three of our patients with PSIS had a single central incisor, a malformation encountered in some HPE cases. Based on this observation, we initiated a search for mutations in HPE-associated genes in 30 patients with PSIS or IPH. The entire coding region of the TGIF, SHH, and SIX3 genes was sequenced in patients with combined pituitary hormone deficiency associated with either PSIS or IPH and in healthy controls. Two novel mutations in the HPE-related genes were detected (ie, c.799 C>T, p.Q267X in the TGIF gene, and c.1279G>A, p.G427R in the SHH gene) in 2 of our patients. The overall incidence of HPE-related gene mutations in our nonsyndromic and nonchromosomal patients was 6.6%. No molecular defect in the SIX3 gene was detected in our cohort. The data suggest that HPE-related gene mutations are implicated in the etiology of isolated pituitary defects (PSIS or IPH). Alternatively, PSIS or IPH may constitute mild forms of an expanded HPE spectrum.


Modified broth-disk method for testing the antibiotic susceptibility of anaerobic bacteria.
The most commonly used method for testing the antibiotic susceptibility of aerobic and facultative bacteria is the disk diffusion method. However, some anaerobic bacteria do not grow well enough in anaerobic jars for performance of disk diffusion tests. A modification of the broth-disk method of Schneierson allowed us to determine antibiotic susceptibility in a completely anaerobic environment. Commercial antibiotic disks were added anaerobically to tubes of prereduced brain heart infusion broth to achieve a concentration of each antibiotic approximating that attainable in blood. The tubes were then inoculated and incubated for 18 h. Resistance or susceptibility to each antibiotic was determined according to the amount of growth in each tube as compared with a control culture without the antibiotic. There was good correlation between results obtained by this broth-disk method and minimal inhibitory concentrations.


Development of an ADL instrument targeting elderly persons with age-related macular degeneration.
In order to estimate the needs of early intervention in a health education program, an ADL instrument measuring feelings of (in)security was needed. The instrument was intended to detect early signs of decline and to evaluate ADL performance among persons with AMD. An evaluative instrument must have a high level of responsiveness, which is the ability to detect true changes over time but must also show a high level of intra-individual agreement in a test-retest assessment. The purpose of this study was to develop an ADL instrument for evaluative purposes and to establish the reliability and the responsiveness of the instrument. The target group comprised all persons referred by an ophthalmologist for low vision rehabilitation, 65 years and older, living at home, with aged-related macular degeneration as the primary diagnosis and with distance visual acuity of the better eye with best correction not lower than 0.1. A non-parametric statistical method that is developed for paired ordered parametrical data was applied in order to measure the systematic and occasional intra-rater disagreement separately and to measure the responsiveness. The test-retest study showed that the ADL-instrument had a high level of test-retest stability, which is a condition for responsiveness. The instrument was found to be responsive. It could therefore detect true longitudinal changes and be used for evaluative purposes targeting elderly with AMD.


Very large (CAG)(n) DNA repeat expansions in the sperm of two spinocerebellar ataxia type 7 males.
Genetic anticipation, i.e. increasing disease severity and decreasing age of onset from one generation to the next, is observed in a number of diseases, including myotonic dystrophy type 1, Huntington's disease and several of the spinocerebellar ataxias. All of these disorders are associated with the expansion of a trinucleotide repeat and array length is positively correlated with disease severity and inversely correlated with the age of onset. The expanded repeat is highly unstable and continues to expand from one generation to the next, providing a molecular explanation for anticipation. Spinocerebellar ataxia type 7 (SCA7) is one of the latest additions to the list of triplet repeat diseases and is distinct from the other SCAs in that it is accompanied by retinal degeneration. Pedigree analyses have previously revealed that the SCA7 repeat is highly unstable and liable to expand, in particular when transmitted by a male. Surprisingly, though, an under-representation of male transmission has also been reported. We now demonstrate directly by single molecule analyses that the expanded repeat is extraordinarily unstable in the male germline and biased toward massive increases. Nearly all of the mutant sperm of two SCA7 males contain alleles that are so large that most of the affected offspring would at best have a severe infantile form of the disease. Indeed, the gross under-representation of such very large expanded alleles in patients suggests that a significant proportion of such alleles might be associated with embryonic lethality or dysfunctional sperm.


RhoJ/TCL regulates endothelial motility and tube formation and modulates actomyosin contractility and focal adhesion numbers.
RhoJ/TCL was identified by our group as an endothelial-expressed Rho GTPase. The aim of this study was to determine its tissue distribution, subcellular localization, and function in endothelial migration and tube formation. Using in situ hybridization, RhoJ was localized to endothelial cells in a set of normal and cancerous tissues and in the vasculature of mouse embryos; endogenous RhoJ was localized to focal adhesions by immunofluorescence. The proangiogenic factor vascular endothelial growth factor activated RhoJ in endothelial cells. Using either small interfering (si)RNA-mediated knockdown of RhoJ expression or overexpression of constitutively active RhoJ (daRhoJ), RhoJ was found to positively regulate endothelial motility and tubule formation. Downregulating RhoJ expression increased focal adhesions and stress fibers in migrating cells, whereas daRhoJ overexpression resulted in the converse. RhoJ downregulation resulted in increased contraction of a collagen gel and increased phospho-myosin light chain, indicative of increased actomyosin contractility. Pharmacological inhibition of Rho-kinase (which phosphorylates myosin light chain) or nonmuscle myosin II reversed the defective tube formation and migration of RhoJ knockdown cells. RhoJ is endothelial-expressed in vivo, activated by vascular endothelial growth factor, localizes to focal adhesions, regulates endothelial cell migration and tube formation, and modulates actomyosin contractility and focal adhesion numbers.


Gastro-oesophageal candidiasis.
A prospective search for gastro-oesophageal candidiasis was made by histological examination of all the biopsies taken from 465 patients endoscoped consecutively during a 12 month period. The criterion for diagnosis was the demonstration of infiltration of tissue or ulcer slough by yeasts and hyphae. Nineteen cases of candidiasis were found giving an overall incidence of 4%. There were 12 cases with oesophageal candidiasis, two with both oesophageal and gastric candidiasis, and five with gastric candidiasis. In none of the patients was candidiasis suspected before endoscopy. Symptoms referable to the candidiasis were uncommon and radiology was not helpful in diagnosis. There was associated local pathology (particularly peptic ulceration and carcinoma of the stomach or oesophagus) in all except two patients, which suggests that the candidiasis is usually secondary to mucosal damage. In the series, candidiasis was present in 27% of patients with oesophageal cancer, 20% of patients with gastric cancer, 16% of patients with benign gastric ulcers, and 15% of patients with oesophagitis.


Tone-detection training enhances spectral integration mediated by intracortical pathways in primary auditory cortex.
Auditory-cued behavioral training can alter neural circuits in primary auditory cortex (A1), but the mechanisms and consequences of experience-dependent cortical plasticity are not fully understood. To address this issue, we trained adult rats to detect a 5 kHz target in order to receive a food reward. After 14 days training we identified three locations within A1: (i) the region representing the characteristic frequency (CF) 5 kHz, (ii) a nearby region with CF ∼10 kHz, and (iii) a more distant region with CF ∼20 kHz. In order to compare functional connectivity in A1 near to, vs. far from, the representation of the target frequency, we placed a 16-channel multiprobe in middle- (∼10 kHz) and high- (∼20 kHz) CF regions and obtained current-source density (CSD) profiles evoked by a range of tone stimuli (CF±1-3 octaves in quarter-octave steps). Our aim was to construct "CSD receptive fields" (CSD RFs) in order to determine the laminar and spectral profile of tone-evoked current sinks, and infer changes to thalamocortical and intracortical inputs. Behavioral training altered CSD RFs at the 10 kHz, but not 20 kHz, site relative to CSD RFs in untrained control animals. At the 10 kHz site, current sinks evoked by the target frequency were enhanced in layer 2/3, but the initial current sink in layer 4 was not altered. The results imply training-induced plasticity along intracortical pathways connecting the target representation with nearby cortical regions. Finally, we related behavioral performance (sensitivity index, d') to CSD responses in individual animals, and found a significant correlation between the development of d' over training and the amplitude of the target-evoked current sink in layer 2/3. The results suggest that plasticity along intracortical pathways is important for auditory learning.


Efficacy and safety of electrical stimulation for stress urinary incontinence in women: a systematic review and meta-analysis.
This systematic review and meta-analysis was aimed at investigating the safety and short- and long-term efficacy of electrical stimulation (ES) in women with stress urinary incontinence (SUI). PubMed, Embase, and the Cochrane database were searched for randomized controlled trials (RCTs) conducted up to 2020. Studies comparing ES with sham ES or no intervention were included. Standardized mean differences (SMDs), weighted mean differences (WMDs), relative risks (RR), and 95% confidence intervals (CIs) were calculated. This study included 9 RCTs, involving a total of 982 patients, of whom 520 received ES. Our results showed that in the short term (< 3 months), compared with sham ES or no intervention, ES significantly improved incontinence-specific quality of life (IQOL) (p = 0.003; SMD = 0.90 [95% CI, 0.30 to 1.50]; I2 = 88%) and reduced urine leakage (p < 0.00001; WMD = -6.15 [95% CI, -8.29 to -4.01]; I2 = 0%) but did not significantly reduce the frequency of incontinence episodes (p = 0.34; WMD = -0.98 [95% CI, -2.99 to 1.04]; I2 = 85%). In the long term (3-7.5 months), ES significantly improved IQOL (p = 0.0009; SMD = 1.14 [95% CI, 0.47 to 1.81]; I2 = 91%) and reduced the frequency of incontinence episodes (p = 0.0009; WMD = -2.45 [95% CI, -3.90 to -1.01]; I2 = 79%) but did not significantly reduce urine leakage (p = 0.27; WMD = -9.21 [95% CI, -25.57 to 7.14]; I2 = 71%). There was no significant difference in adverse events between ES and sham ES or no intervention (p = 0.36; RR = 1.34 [95% CI, 0.72 to 2.50]; I2 = 0%). A test for subgroup differences showed that electroacupuncture (EA) improved long-term IQOL to a greater extent than vaginal ES (VES), whereas there was no significant difference in efficacy between EA and VES in short- or long-term reduction of urine leakage, frequency of incontinence episodes, or short-term IQOL improvement. ES may improve short- and long-term IQOL for women with SUI, but it appears to provide only short-term reduction in urine leakage and long-term reduction in frequency of incontinence episodes. However, we cannot draw any conclusion on the safety between ES and sham ES or no intervention because of the rarity of adverse events. It is still uncertain whether EA is comparable or superior to VES owing to an insufficient number of studies and patients. The conclusions should be considered carefully because of the limited quality and quantity of the RCTs included. Further rigorous RCTs with adequate sample sizes and long follow-up are necessary to fully validate our findings.


Corifollitropin alfa: a new option to treat female infertility.
Corifollitropin alfa (Elonva®) is the first hybrid follicle-stimulating hormone (FSH) molecule with demonstrated sustained follicle-stimulating activity. The β subunit of this molecule contains the carboxy-terminal peptide of human chorionic gonadotropin, which alters the pharmacokinetic profile of the molecule. It demonstrates a longer circulation half-life and extended time to peak levels when compared with recombinant FSH (rFSH). Like rFSH, it lacks luteinizing hormone activity and binds specifically to the FSH receptor in vitro. Clinical trials show that corifollitropin alfa is able to sustain multiple follicular growth for a week, with a similar ovarian response and safety profile as rFSH. A single injection of corifollitropin alfa can replace 7 daily injections of rFSH during the first week of ovarian stimulation in gonadotropin-releasing hormone antagonist protocols. Therefore, corifollitropin alfa addresses the need for a simplified treatment approach to lessen the burden of multiple daily injections for in vitro fertilization.


Interacting influences of gender and chronic pain status on parasympathetically mediated heart rate variability in adolescents and young adults.
Considerable research links chronic pain to autonomic nervous system (ANS) dysfunction, specifically low heart rate variability (HRV) mediated by reduced parasympathetic activity. However, little is known about factors that influence ANS function in chronic pain. The ANS is the primary pathway for brain-gut communication, making it of particular interest in gastrointestinal disorders, such as irritable bowel syndrome, characterized by functional abdominal pain (FAP). We evaluated the relation of sex, pain severity, and psychological stress to ANS function in adolescents/young adults from a database of pediatric FAP and control participants enrolled 8 years earlier in a prospective study of pain. At follow-up in adolescence/young adulthood (Mean age = 19.46, SD = 3.48), we classified participants as Pain-Remit (n = 130), Pain-Persist (n = 96), and pain-free controls (n = 123). We recorded electrocardiogram data at rest and during laboratory stressors. Results demonstrated significantly lower HRV in Pain-Persist females compared with Pain-Remit females, female controls, and all males regardless of pain category. Spectral analysis of electrocardiogram showed that Pain-Persist females had reduced power in the high frequency domain of cardiac activity, ie, reduced parasympathetic "braking" of sympathetic activity, both at rest and during stress. Pain-Remit females exhibited levels of autonomic imbalance intermediate between those of females with persistent FAP and all other participants. Parasympathetically mediated low HRV in young women with persistent FAP may reflect a peripheral mechanism (eg, gut dysfunction) or a central nervous system mechanism (eg, pain amplification or poor emotion self-regulation) involving prolonged sympathetic activation.


Infertility treatment: lack of consensus plagues an unregulated field.
Each year, in an attempt to stimulate journalism students' interest in medical writing, CMAJ sponsors the Amy Chouinard Memorial Essay Prize. The $750 award is in memory of Amy Chouinard, a longtime and valued contributor to CMAJ and the Canadian Journal of Surgery. Students from any recognized journalism program at a Canadian college or university are eligible to enter, and the deadline for 1998 entries is June 1. The 1997 winner, Megan Easton, presents a well-written and thorough account of the issues surrounding infertility treatment. Interest in the topic came naturally enough--her father, Dr. William Easton, is a urogynecologist in private practice in Scarborough, Ont.


Exercise: prescriptions for the elderly.
It has been said that those who cannot find time for exercise will most certainly find time for illness. This is especially true for the elderly. Physicians need to provide encouragement and instruction to help the older patient to begin and to continue an exercise program that is designed to avoid injury.


Our second genome-human metagenome: how next-generation sequencer changes our life through microbiology.
Next-generation sequencing has greatly expanded our ability to query the identity and genetic composition of entire communities of microbial organisms. This area of research, known as metagenomics, does not rely upon culturing the individual organisms. Rather, the genetic material from the entire community is processed and sequenced simultaneously. From this sequence data, researchers are able to determine the relative population of organisms within the community as well as determine which genes and metabolic pathways are present and expressed in the microbial community. While these techniques have been applied to a wide range of environmental samples, metagenomics is also the focus of intensive research on human-associated microbial communities. The scope of these human metagenomics studies are quite varied, but all have a common goal of attempting to understand the important role that human commensal microbial communities play in health and disease. The early results from studying the human metagenome indicate a vital role that microbial communities play in immunity, health, and disease. Going forward, human metagenomics is a wide open field of research with many unanswered questions such as which factors are responsible for the variation of composition of an individual's microbiome, how does the microbiome respond to disturbance, and what beneficial functions are the microorganisms performing?


The contribution of K(+) channels to human thoracic duct contractility.
In smooth muscle cells, K(+) permeability is high, and this highly influences the resting membrane potential. Lymph propulsion is dependent on phasic contractions generated by smooth muscle cells of lymphatic vessels, and it is likely that K(+) channels play a critical role in regulating contractility in this tissue. The aim of this study was to investigate the contribution of distinct K(+) channels to human lymphatic vessel contractility. Thoracic ducts were harvested from 43 patients and mounted in a wire myograph for isometric force measurements or membrane potential recordings with an intracellular microelectrode. Using K(+) channel blockers and activators, we demonstrate a functional contribution to human lymphatic vessel contractility from all the major classes of K(+) channels [ATP-sensitive K(+) (KATP), Ca(2+)-activated K(+), inward rectifier K(+), and voltage-dependent K(+) channels], and this was confirmed at the mRNA level. Contraction amplitude, frequency, and baseline tension were altered depending on which channel was blocked or activated. Microelectrode impalements of lymphatic vessels determined an average resting membrane potential of -43.1 ± 3.7 mV. We observed that membrane potential changes of <5 mV could have large functional effects with contraction frequencies increasing threefold. In general, KATP channels appeared to be constitutively open since incubation with glibenclamide increased contraction frequency in spontaneously active vessels and depolarized and initiated contractions in previously quiescent vessels. The largest change in membrane voltage was observed with the KATP opener pinacidil, which caused 24 ± 3 mV hyperpolarization. We conclude that K(+) channels are important modulators of human lymphatic contractility.


Risk Stratification to Guide Prevention and Control Strategies for Arboviruses Transmitted by Aedes aegypti.
Strategies for the prevention of arboviral diseases transmitted by Aedes aegypti have traditionally focused on vector control. This remains the same to this day, despite a lack of documented evidence on its efficacy due to a lack of coverage and sustainability. The continuous growth of urban areas and generally unplanned urbanization, which favor the presence of Ae. aegypti, demand resources, both material and human, as well as logistics to effectively lower the population's risk of infection. These considerations have motivated the development of tools to identify areas with a recurrent concentration of arboviral cases during an outbreak to be able to prioritize preventive actions and optimize available resources. This study explores the existence of spatial patterns of dengue incidence in the locality of Tartagal, in northeastern Argentina, during the outbreaks that occurred between 2010 and 2020. Approximately half (50.8%) of the cases recorded during this period were concentrated in 35.9% of the urban area. Additionally, an important overlap was found between hotspot areas of dengue and chikungunya (Kendall's W = 0.92; p-value < 0.001) during the 2016 outbreak. Moreover, 65.9% of the cases recorded in 2022 were geolocalized within the hotspot areas detected between 2010 and 2020. These results can be used to generate a risk map to implement timely preventive control strategies that prioritize these areas to reduce their vulnerability while optimizing the available resources and increasing the scope of action.


Use of the hyperinsulinemic euglycemic clamp to assess insulin sensitivity in guinea pigs: dose response, partitioned glucose metabolism, and species comparisons.
The guinea pig is an alternate small animal model for the study of metabolism, including insulin sensitivity. However, only one study to date has reported the use of the hyperinsulinemic euglycemic clamp in anesthetized animals in this species, and the dose response has not been reported. We therefore characterized the dose-response curve for whole body glucose uptake using recombinant human insulin in the adult guinea pig. Interspecies comparisons with published data showed species differences in maximal whole body responses (guinea pig ≈ human < rat < mouse) and the insulin concentrations at which half-maximal insulin responses occurred (guinea pig > human ≈ rat > mouse). In subsequent studies, we used concomitant d-[3-3H]glucose infusion to characterize insulin sensitivities of whole body glucose uptake, utilization, production, storage, and glycolysis in young adult guinea pigs at human insulin doses that produced approximately half-maximal (7.5 mU·min-1·kg-1) and near-maximal whole body responses (30 mU·min-1·kg-1). Although human insulin infusion increased rates of glucose utilization (up to 68%) and storage and, at high concentrations, increased rates of glycolysis in females, glucose production was only partially suppressed (~23%), even at high insulin doses. Fasting glucose, metabolic clearance of insulin, and rates of glucose utilization, storage, and production during insulin stimulation were higher in female than in male guinea pigs (P < 0.05), but insulin sensitivity of these and whole body glucose uptake did not differ between sexes. This study establishes a method for measuring partitioned glucose metabolism in chronically catheterized conscious guinea pigs, allowing studies of regulation of insulin sensitivity in this species.


Porphyromonas Gingivalis Elevated High-Mobility Group Box 1 Levels After Myocardial Infarction in Mice.
High mobility group box 1 (HMGB1) is a nuclear protein released from necrotic cells, inducing inflammatory responses. Epidemiological studies suggested a possible association between periodontitis and cardiovascular diseases (CVDs). Due to tissue damage and necrosis of cardiac cells following myocardial infarction (MI), HMGB1 is released, activating an inflammatory reaction. However, it remains unclear whether periodontitis is also involved in myocardial damage. The purpose of this study was to determine the effect of the periodontal pathogen Porphyromonas gingivalis (P.g.) after MI in mice.C57BL/6J wild type mice in post-MI were inoculated with P.g. in the infected group (P.g.-inoculated MI group) and with phosphate buffer saline (PBS) in the control group (PBS-injected MI group). Plasma samples and twelve tissue samples from mice hearts after MI were obtained. We determined the expression of HMGB1 by ELISA and immunohistochemistry.The level of HMGB1 protein in the P.g.-inoculated MI group was significantly higher than in the PBS-injected MI group on day 5, but not on day 14. Immunohistochemistry analysis revealed that HMGB1 was mainly expressed in cardiomyocytes, immune cells, and vascular endothelial cells in the PBS-injected MI group, while HMGB1 was seen broadly in degenerated cardiomyocytes, extracellular fields, immune cells, and vascular endothelial cells in the P.g.-inoculated MI group. A significant increase in the number of HMGB1 positive cells was observed in the P.g.-inoculated MI group compared to the PBS-injected MI group.Infection with P.g. after MI enhanced myocardial HMGB1 expression. There is a possible relationship between periodontitis and post-infarction myocardial inflammation through HMGB-1.


Emerging treatments in acute lymphoblastic leukemia.
Acute lymphoblastic leukemia (ALL) is a clonal proliferation of early B- and T-lymphocyte progenitors and results in the accumulation of leukemic blasts in the bone marrow and various extramedullary sites. It affects both children and adults, with peak prevalence between the ages of 2 to 5 years. Despite current treatment protocols achieving rapid cytoreduction in the vast majority of patients, serious acute and late complications are frequent and resistance to chemotherapy often develops. In contrast to the successes obtained with pediatric patients, treatment outcomes for adults remain poor with only 40% of patients being long-term survivors. Extensive research in the field of ALL has helped understand the mechanisms that control leukemic cells, facilitating the design of new drugs that specifically interfere with leukemic pathways and overcome chemo-resistance induced by common treatment regimens. Herein, we review the current status of the development of novel anti-leukemic agents, with emphasis on small molecular inhibitors that have already translated into clinical trials and are in the advanced stages of preclinical development. Challenges to successful development of each strategy are discussed.


Over-expression of the cucumber expansin gene (Cs-EXPA1) in transgenic maize seed for cellulose deconstruction.
Plant cell wall degradation into fermentable sugars by cellulases is one of the greatest barriers to biofuel production. Expansin protein loosens the plant cell wall by opening up the complex of cellulose microfibrils and polysaccharide matrix components thereby increasing its accessibility to cellulases. We over-expressed cucumber expansin in maize kernels to produce enough protein to assess its potential to serve as an industrial enzyme for applications particularly in biomass conversion. We used the globulin-1 embryo-preferred promoter to express the cucumber expansin gene in maize seed. Expansin protein was targeted to one of three sub-cellular locations: the cell wall, the vacuole, or the endoplasmic reticulum (ER). To assess the level of expansin accumulation in seeds of transgenic kernels, a high throughput expansin assay was developed. The highest expressing plants were chosen and enriched crude expansin extract from those plants was tested for synergistic effects with cellulase on several lignocellulosic substrates. Activity of recombinant cucumber expansin from transgenic kernels was confirmed on these pretreated substrates. The best transgenic lines (ER-targeted) can now be used for breeding to increase expansin expression for use in the biomass conversion industry. Results of these experiments show the success of expansin over-expression and accumulation in transgenic maize seed without negative impact on growth and development and confirm its synergistic effect with cellulase on deconstruction of complex cell wall substrates.


Lack of angiotensin II conversion to angiotensin III increases water but not alcohol consumption in aminopeptidase A-deficient mice.
Elevated central concentrations of the vasopressor octapeptide angiotensin (Ang) II increase the water intake in mammals. Recently, we showed that central AngII is also crucial in alcohol-consuming behavior. Since the heptapeptide AngIII, an AngII metabolite, is discussed to mediate AngII-related effects, we investigated water and alcohol consumption in mice, genetically deficient in aminopeptidase A (APA), a peptidase responsible for AngII conversion to AngIII. Sixteen male APA-deficient mice and their age matched wild-type controls were monitored on their water intake under basal conditions and total fluid and alcohol intake before and after social stress in a two-bottle free-choice paradigm. Alterations were connected to the regulation in activity of Ang-related peptidases (APA, ACE; ACE2) in brain regions involved in alcohol intake and peripheral organs. In comparison to their wild-type controls, APA-deficient mice drank significantly more water but not more alcohol at all investigated time points. A reduction in water intake, as observed in wild-type animals after social stress, did not occur in knockout mice. However, the reduction in alcohol consumption after social stress was significantly reduced in both strains. Alcohol consumption upregulated all three peptidases in the kidney, but not in lung. Notable, renal ACE2 activity was significantly higher in APA-deficient mice under basal condition. While the inhibition of AngII metabolism to AngIII does not influence the alcohol intake, water consumption in mice deficient for APA was significantly elevated. These differences induced by an altered AngII/AngIII ratio oppose the hypothesis that central AngII and AngIII act in a congruent pattern.


An increase in intercellular crosstalk and electrotonic coupling between cardiomyocytes and nonmyocytes reshapes the electrical conduction in the metabolic heart characterized by short QT intervals in ECGs.
Cardiac conduction abnormalities are disorders in metabolic syndrome (MetS), however, their mechanisms are unknown. Although ventricular arrhythmia reflects the changes in QT-interval of electrocardiograms associated with the changes in cardiomyocyte action potential durations (APDs), recent studies emphasize role of intercellular crosstalk between cardiomyocytes and nonmyocytes via passive (electrotonic)-conduction. Therefore, considering the possible increase in intercellular interactions of nonmyocytes with cardiomyocytes, we hypothesized an early-cardiac-remodeling characterized by short QT-interval via contributions and modulations of changes by nonmyocytes to the ventricular APs in an early-stage MetS hearts. Following the feeding of 8-week-old rats with a high-sucrose diet (32%; MetS rats) and validation of insulin resistance, there was a significant increase in heart rate and changes in the electrical characteristics of the hearts, especially a shortening in action potential (AP) duration of the papillary muscles. The patch-clamp analysis of ventricular cardiomyocytes showed an increase in the Na+ -channel currents while there were decreases in l-type Ca2+ -channel (LTCC) currents with unchanged K+ -channel currents. There was an increase in the phosphorylated form of connexin 43 (pCx43), mostly with lateral localization on sarcolemma, while its unphosphorylated form (Cx43) exhibited a high degree of localization within intercalated discs. A high-level positively-stained α-SMA and CD68 cells were prominently localized and distributed in interfibrillar spaces of the heart, implying the possible contributions of myofibroblasts and macrophages to both shortened APDs and abnormal electrical conduction in MetS hearts. Our data propose a previously unrecognized pathway for SQT induction in the heart. This pathway includes not only the contribution of short ventricular-APDs via ionic mechanisms but also increasing contributions of the electrotonic-cardiomyocyte depolarization, spontaneous electrical activity-associated fast heterogeneous impulse conduction in the heart via increased interactions and relocations between cardiomyocytes and nonmyocytes, which may be an explanation for the development of an SQT in early-cardiac-remodeling.


Influence of hyperhomocysteinemia on the cellular redox state--impact on homocysteine-induced endothelial dysfunction.
Hyperhomocysteinemia is an independent risk factor for the development of atherosclerosis. An increasing body of evidence has implicated oxidative stress as being contributory to homocysteine's deleterious effects on the vasculature. Elevated levels of homocysteine may lead to increased generation of superoxide by a biochemical mechanism involving nitric oxide synthase, and, to a lesser extent, by an increase in the chemical oxidation of homocysteine and other aminothiols in the circulation. The resultant increase in superoxide levels is further amplified by homocysteine-dependent alterations in the function of cellular antioxidant enzymes such as cellular glutathione peroxidase or extracellular superoxide dismutase. One direct clinical consequence of elevated vascular superoxide levels is the inactivation of the vasorelaxant messenger nitric oxide, leading to endothelial dysfunction. Scavenging of superoxide anion by either superoxide dismutase or 4,5-dihydroxybenzene 1,3-disulfonate (Tiron) reverses endothelial dysfunction in hyperhomocysteinemic animal models and in isolated aortic rings incubated with homocysteine. Similarly, homocysteine-induced endothelial dysfunction is also reversed by increasing the concentration of the endogenous antioxidant glutathione or overexpressing cellular glutathione peroxidase in animal models of mild hyperhomocysteinemia. Taken together, these findings strongly suggest that the adverse vascular effects of homocysteine are at least partly mediated by oxidative inactivation of nitric oxide.


Uterine serous carcinoma: a comparison of therapy for advanced-stage disease.
Management of advanced-stage uterine serous carcinoma (USC) is uncertain, and postsurgical therapeutic options swing between radiation and chemotherapy. The aim of this study is to evaluate the utility of radiotherapy compared to platinum-based chemotherapy in women with advanced-stage USC. We retrospectively identified cases of USC at our institution. Survival distributions were calculated by the Kaplan-Meier method. Two-tailed t-tests were used to compare time to progression and time to death. We identified 24 women diagnosed with either stage III or IV USC. Time to progression for women receiving radiotherapy was 5.3 months as compared with 12.4 months for women receiving chemotherapy (P = 0.01). Mean time to death for the radiotherapy group was 8 months compared to 18 months in the chemotherapy group (P = 0.04). Kaplan-Meier survival curves were significantly different between the two groups (P = 0.01). While radiotherapy appears to control USC recurrences in the pelvis, the disease often recurs distantly. When compared to radiotherapy, platinum-based chemotherapy appears to increase disease-free survival and time to death in women with advanced-stage USC.


Reflections on being therapeutic and reflection.
This article offers a reflective account of an incident that occurred between a nurse tutor and a patient on a cardiology ward. It highlights the importance of interpersonal skills in creating a therapeutic relationship, in particular those of self-awareness, empathy and intuition. The author's difficulties in running reflective practice sessions for pre-registration students are discussed and insights are offered into why these difficulties arose.


Physician-initiated clinical study of limb ulcers treated with a functional peptide, SR-0379: from discovery to a randomized, double-blind, placebo-controlled trial.
SR-0379 is a functional peptide that has wound healing effect with anti-microbial action, making it an ideal drug to prevent infection. To evaluate the safety, efficacy, and pharmacokinetics of SR-0379 for the treatment of leg ulcers, a physician-initiated, phase I/IIa, first-in-patient clinical study was designed. A multi-center, double-blind, randomized clinical study was conducted from October 2015 to September 2016. The inclusion criteria for leg ulcers were (1) diabetes or critical limb ischemia and (2) wound size <6 cm in diameter. Twelve patients were randomized into four groups and administered 0.02%, 0.1%, or 0.5% SR-0379 or placebo treatment on skin ulcers once per day for 28 days. Efficiency was evaluated by determining the rate of wound size reduction as a primary endpoint at 4 weeks after the first treatment compared with the pre-treatment wound size. As a secondary endpoint, the DESIGN-R score index, time to wound closure, and the 50% wound size reduction ratio were also evaluated. The safety of SR-0379 was evaluated during the study period. In the evaluation of efficiency, the skin ulcer reduction rates at the last evaluation were 44.73% for the 0.02% SR-0379 group, 68.25% for the 0.1% group, and 71.61% for the 0.5% group, compared with 9.95% for the placebo group. Six adverse events were reported in four patients, of which one occurred in the placebo group, and causal relationships to study drugs were denied for all six events. Treatment with SR-0379 for chronic leg ulcers was safe, well tolerated, and effective.


Random Migration Assays of Mammalian Cells and Quantitative Analyses of Single Cell Trajectories.
Cell migration is essential to many biological processes such as embryonic development, immune surveillance and wound healing. Random cell migration refers to the intrinsic ability of cells to migrate, often called cell motility. This basal condition contrasts with directed cell migration, where cells migrate toward a chemical or physical cue. Unlike Brownian particles, however, randomly migrating cells exhibit a directional persistence, i.e., they are more likely to sustain the movement in the direction they previously took than to change, even if this direction is randomly chosen in an isotropic environment. Here we describe how to set up time-lapse recording of mammalian cells freely moving on a two-dimensional surface coated with extracellular matrix proteins, how to acquire single cell trajectories from movies and how to extract key parameters that characterize cell motility, such as cell speed, directionality, mean square displacement, and directional persistence.


[Computed tomography in the comprehensive diagnosis of acute diseases and injuries].
The author presents the results of a CT use during combined investigation on 2500 patients and the wounded who were admitted to hospital on emergency as well as patients who developed serious complications during treatment, especially in the postoperative period. CT findings were verified during operation, at autopsy or during a follow-up. They were indicative of a high effectiveness of emergency CT of different organs and systems in the diagnosis of lesions, acute diseases and their complications. The use of CT permitted considerable reduction of a diagnostic (preoperative) period, a decrease in use of routine invasive x-ray and surgical methods, and the improvement of therapeutic results.


A platform for interrogating cancer-associated p53 alleles.
p53 is the most frequently mutated gene in human cancer. Compelling evidence argues that full transformation involves loss of growth suppression encoded by wild-type p53 together with poorly understood oncogenic activity encoded by missense mutations. Furthermore, distinguishing disease alleles from natural polymorphisms is an important clinical challenge. To interrogate the genetic activity of human p53 variants, we leveraged the Drosophila model as an in vivo platform. We engineered strains that replace the fly p53 gene with human alleles, producing a collection of stocks that are, in effect, 'humanized' for p53 variants. Like the fly counterpart, human p53 transcriptionally activated a biosensor and induced apoptosis after DNA damage. However, all humanized strains representing common alleles found in cancer patients failed to complement in these assays. Surprisingly, stimulus-dependent activation of hp53 occurred without stabilization, demonstrating that these two processes can be uncoupled. Like its fly counterpart, hp53 formed prominent nuclear foci in germline cells but cancer-associated p53 variants did not. Moreover, these same mutant alleles disrupted hp53 foci and inhibited biosensor activity, suggesting that these properties are functionally linked. Together these findings establish a functional platform for interrogating human p53 alleles and suggest that simple phenotypes could be used to stratify disease variants.


CagA-positive strains of Helicobacter pylori may protect against Barrett's esophagus.
Helicobacter pylori (H. pylori) colonization is associated with chronic gastritis, peptic ulcer disease, and adenocarcinoma of the distal stomach. However, the role of H. pylori strain variation in complicated gastroesophageal reflux disease, especially Barrett's esophagus, is unknown. Therefore, the aim of this study was to evaluate the prevalence of colonization by cagA+ and cagA- H. pylori strains in the spectrum of gastroesophageal reflux disease, including Barrett's esophagus. A total of 251 patients undergoing endoscopy were categorized into four groups: controls, patients with gastroesophageal reflux disease alone, and patients with short- and long-segment Barrett's esophagus. All patients underwent upper endoscopies with biopsies and serum collections. H. pylori and degree of mucosal inflammation in gastric biopsies were assessed and serological assessment made for H. pylori and cagA status. The overall prevalence of H. pylori colonization in the study population was 35% (95% confidence interval = 29.5-41.4%) which did not differ significantly among the groups. However, colonization by cagA+ H. pylori strains was significantly more prevalent among controls (11/25; 44%) and patients with gastroesophageal reflux disease (13/36; 36%) than in patients with short-segment (2/10; 20%) or long-segment Barrett's esophagus (0/18; 0%). Patients with Barrett's esophagus were less likely to be colonized by cagA+ H. pylori strains than reflux patients without Barrett's esophagus (odds ratio = 0.27, 95% confidence interval = 0.11-0.67, p = 0.004). Colonization by cagA+ H. pylori strains may be protective against the formation of short- and long-segment Barrett's esophagus and its malignant complications.


Regional deposition of inhaled particles in human lungs: comparison between men and women.
We measured detailed regional deposition patterns of inhaled particles in healthy adult male (n = 11; 25 +/- 4 yr of age) and female (n = 11; 25 +/- 3 yr of age) subjects by means of a serial bolus aerosol delivery technique for monodisperse fine [particle diameter (Dp) = 1 micron] and coarse aerosols (Dp = 3 and 5 micron). The bolus aerosol (40 ml half-width) was delivered to a specific volumetric depth (Vp) of the lung ranging from 100 to 500 ml with a 50-ml increment, and local deposition fraction (LDF) was assessed for each of the 10 local volumetric regions. In all subjects, the deposition distribution pattern was very uneven with respect to Vp, showing characteristic unimodal curves with respect to particle size and flow rate. However, the unevenness was more pronounced in women. LDF tended to be greater in all regions of the lung in women than in men for Dp = 1 micron. For Dp = 3 and 5 micron, LDF showed a marked enhancement in the shallow region of Vp </= 200 ml in women compared with men (P < 0.05). LDF in women was comparable to or smaller than those of men in deep lung regions of Vp > 200 ml. Total lung deposition was comparable between men and women for fine particles but was consistently greater in women than men for coarse particles regardless of flow rates used: the difference ranged from 9 to 31% and was greater with higher flow rates (P < 0.05). The results indicate that 1) particle deposition characteristics differ between healthy men and women under controlled breathing conditions and 2) deposition in women is greater than that in men.


Long-term results of the radial artery used for myocardial revascularization.
No information is available on the long-term results of radial artery (RA) grafts used as coronary artery bypass conduits. In this report, we describe the long-term (105+/-9 months) angiographic results of a series of 90 consecutive patients in whom the RA was used as a coronary artery bypass conduit directly anastomosed to the ascending aorta. The long-term patency and perfect patency rates of the RA were 91.6% and 88%, respectively, versus 97.5% and 96.3% for internal thoracic artery grafts. The severity of stenosis of the target vessel clearly influenced long-term RA patency, whereas location of the target vessel and long-term use of calcium channel blockers did not influence angiographic results. Preserved endothelial function and absence of flow-limiting, fibrous, intimal hyperplasia were also documented. Ten years after surgery, RA grafts have excellent patency and perfect patency rates. Appropriate surgical technique and correct indication are the key factors for long-term RA patency.


Response to Wolf et al.: Furthering Debate over the Suitability of Trap-Neuter-Return for Stray Cat Management.
To continue dialogue over proposed Australian trials of Trap-Neuter-Return (TNR), we applied a framework requiring identification of areas of agreement, areas of disagreement, and identification of empirical data collection required to resolve disagreements. There is agreement that Australia has a problem with stray cats, causing problems of impacts on wildlife, nuisance, disease transmission (including public health issues and exchange of diseases between stray cat and pet cat populations), poor welfare outcomes for stray cats, and an emotional burden on staff euthanising healthy stray cats. There is disagreement on whether (i) current measures are failing, leading to unacceptably high euthanasia levels, (ii) some contributors to the debate misunderstand TNR, (iii) TNR trials will reduce urban cat populations and associated problems, (iv) TNR is an ethical solution to cat overpopulation, and (v) some contributors to the debate promulgated misinformation. Although not everyone agrees that TNR trials should proceed, as a hypothetical exploration, we propose an experimental approach explicitly comparing TNR to alternatives. Trials could only be considered if other detailed and well-funded attempts at stray cat control focusing across an entire Local Government Area (LGA) prove ineffective.


A tissue culture model of the hypophysiotrophic CRF producing neuronal system.
To investigate functional and chemical properties of anatomically characterized corticotropin-releasing factor-41 (CRF-41) producing neurons in vitro, hypothalamic slices of 6-day-old rats were maintained in culture for up to 6 weeks using a modified roller culture technique. This technique yields thick (100 microns) slices that contained an average of 300-400 CRF-41-immunostained neurons. The majority of CRF-41-positive cells were of small size (12-15 microns in diameter), and contained CRF-41-labeled dense core vesicles of 100 nm diameter as detected by electron microscopic postembedding immunocytochemistry. These cells represented the only CRF-41-positive cell population in the culture. Light microscope double immunolabelling of colchicine-treated cultures kept in a serum-containing media (SCM) indicated that about 60% of these CRF-41-positive neurons contains detectable levels of vasopressin-associated neurophysin (VP-NP). Culturing slices in serum-free, chemically defined media (SFM) resulted in an increased VP-NP immunostaining: parvicellular neurons labeled for both CRF-41 and VP-NP could be detected without colchicine treatment, and practically all CRF-41-positive neurons expressed VP-NP immunoreactivity. At the electron microscopic level there was a significant increase in VP-NP labeling density in the dense core vesicle compartment of CRF-41-positive varicosities. Adding dexamethasone (10 nM) to the SFM restored the staining pattern originally observed in SCM. Hence, the increased VP-NP and CRF-41 immunostaining after culturing CRF-41 neurons in SFM is most likely due to the absence of inhibitory glucocorticoids. The capacity of cultured paraventricular cells to release CRF-41 was assessed using an immunoassay. Unstimulated (basal) secretion of CRF-41 was not altered by five successive samplings at 2-hour intervals and stimulation of the same culture with 56 mmol K+ significantly increased (2-3 times) the CRF-41 content in the medium. The presence of dexamethasone (10 nM) in SFM induced a 6-fold reduction of K(+)-stimulated CRF-41 release and a 5 times reduction in tissue content in relation to cultures maintained in SFM without dexamethasone. In summary, we have demonstrated that cultured CRF-41 cells display morphological and biochemical features, as well as responsiveness to glucocorticoids, that is reminiscent to the situation in vivo. Thus, the model is well suited for studies of hypophysiotrophic CRF-41 cell functions.


Augmenting cognitive architectures to support diagrammatic imagination.
Diagrams are a form of spatial representation that supports reasoning and problem solving. Even when diagrams are external, not to mention when there are no external representations, problem solving often calls for internal representations, that is, representations in cognition, of diagrammatic elements and internal perceptions on them. General cognitive architectures--Soar and ACT-R, to name the most prominent--do not have representations and operations to support diagrammatic reasoning. In this article, we examine some requirements for such internal representations and processes in cognitive architectures. We discuss the degree to which DRS, our earlier proposal for such an internal representation for diagrams, meets these requirements. In DRS, the diagrams are not raw images, but a composition of objects that can be individuated and thus symbolized, while, unlike traditional symbols, the referent of the symbol is an object that retains its perceptual essence, namely, its spatiality. This duality provides a way to resolve what anti-imagists thought was a contradiction in mental imagery: the compositionality of mental images that seemed to be unique to symbol systems, and their support of a perceptual experience of images and some types of perception on them. We briefly review the use of DRS to augment Soar and ACT-R with a diagrammatic representation component. We identify issues for further research.


Effect of powder-to-liquid ratio on the distortion of repaired denture acrylic resin.
The aim of this study was to determine the distortion of repaired autopolymerising poly(methylmethacrylate) (PMMA) resin. Eighteen rectangular test specimens with a 45-degree edge profile repair surface were repaired with autopolymerising PMMA using different powder-to-liquid (PL) ratios. None of the test specimens distorted during repair, although microscopical examination showed porosities which may have been due to polymerisation shrinkage. The results suggest that autopolymerising PMMA with various PL ratios can be used to repair fractured pieces of PMMA without affecting the distortion of the specimen.


Regulation of the number of functional voltage-sensitive Ca++ channels on PC12 cells by chronic changes in membrane potential.
The properties of the various types of voltage-sensitive Ca++ channels (VSCC) are becoming increasingly well characterized, but the mechanisms which control the number and types of channels expressed by cells are virtually unknown. To study the regulation of VSCC in neuronal cells we have used PC12 pheochromocytoma cells. Binding of [3H]nitrendipine was used to determine the number of dihydropyridine-sensitive channels, and the uptake of 45Ca++ was used to determine the functional state of VSCC on the cell surface. Prolonged depolarization by elevation of extracellular K+ caused concomitant time and concentration-dependent decreases in both [3H]nitrendipine binding and depolarization-dependent uptake of 45Ca++. Changes in binding and ion flux plateaued at about a 50% decrease with 3 days of depolarization and an extracellular K+ concentration of 50 mM. Return of the cells to normal K+ caused the recovery of both [3H]nitrendipine binding and 45Ca++ uptake within 24 hr. Measurements of the intracellular free Ca++ concentration determined that it remained elevated for several hours with K+ depolarization, but returned to normal within 15 hr. Growth of the cells with a concentration of ionomycin, which caused a similar increase in intracellular free Ca++, also caused a loss of [3H]nitrendipine binding sites. Thus, it appears that the number of functional VSCC can be regulated by changes in intracellular Ca++ such as those associated with prolonged depolarization. However, because Ca++ channel number remained depressed while intracellular free Ca++ returned to normal, other mechanisms controlling channel number also must be involved.


The reliability of the American Community Survey for injury surveillance.
To evaluate the reliability and predictability of 49 socioeconomic indicators constructed from the annual and multiyear American Community Survey (ACS) data cycles for monitoring injury inequalities across the USA. Cross-sectional analysis of the 2006-2013 annual and multiyear county-level ACS data cycles. Indicator reliability was assessed using the margin of error and coefficient of variation (CV). Overlapping multiyear data cycles were assessed for statistical dependence in the estimates. Negative binomial regression models were constructed from a selection of the most reliable indicators over time and across all data cycles using all-cause unintentional and homicide-related mortality records from the National Center for Health Statistics (NCHS). Fewer than half of all indicators for each data cycle generated 'high reliability' CV estimates for at least 95% of all census counties. Indicator reliability did not linearly improve with increasing sample size afforded from the multiyear surveys. On average, changes in socioeconomic conditions for the same geographic areas were statistically significantly different (p<0.05) in 14% (rage 0-99%) to 16% (rage 0-93%) of all overlapping multiyear data cycles. ACS indicators that were among the most reliable across data cycles corroborated variable relationships derived using estimates from the 2000 decennial census and corresponding NCHS records for that year. Few of the socioeconomic indicators previously used to measure injury disparities are consistently reliable across all ACS data cycles. Researchers should be judicious when selecting consecutive multiyear data cycles to approximate changes in annual socioeconomic conditions. Among the indicators that are reliable, it is advisable to use estimates from the annual ACS data cycle as a crude barometer of injury inequalities and the multiyear files to confirm and add precedence to national trends every three and five years.


cor, a novel carbon monoxide resistance gene, is essential for Mycobacterium tuberculosis pathogenesis.
Tuberculosis, caused by Mycobacterium tuberculosis, remains a devastating human infectious disease, causing two million deaths annually. We previously demonstrated that M. tuberculosis induces an enzyme, heme oxygenase (HO1), that produces carbon monoxide (CO) gas and that M. tuberculosis adapts its transcriptome during CO exposure. We now demonstrate that M. tuberculosis carries a novel resistance gene to combat CO toxicity. We screened an M. tuberculosis transposon library for CO-susceptible mutants and found that disruption of Rv1829 (carbon monoxide resistance, Cor) leads to marked CO sensitivity. Heterologous expression of Cor in Escherichia coli rescued it from CO toxicity. Importantly, the virulence of the cor mutant is attenuated in a mouse model of tuberculosis. Thus, Cor is necessary and sufficient to protect bacteria from host-derived CO. Taken together, this represents the first report of a role for HO1-derived CO in controlling infection of an intracellular pathogen and the first identification of a CO resistance gene in a pathogenic organism. Macrophages produce a variety of antimicrobial molecules, including nitric oxide (NO), hydrogen peroxide (H2O2), and acid (H+), that serve to kill engulfed bacteria. In addition to these molecules, human and mouse macrophages also produce carbon monoxide (CO) gas by the heme oxygenase (HO1) enzyme. We observed that, in contrast to other bacteria, mycobacteria are resistant to CO, suggesting that this might be an evolutionary adaptation of mycobacteria for survival within macrophages. We screened a panel of ~2,500 M. tuberculosis mutants to determine which genes are required for survival of M. tuberculosis in the presence of CO. Within this panel, we identified one such gene, cor, that specifically confers CO resistance. Importantly, we found that the ability of M. tuberculosis cells carrying a mutated copy of this gene to cause tuberculosis in a mouse disease model is significantly attenuated. This indicates that CO resistance is essential for mycobacterial survival in vivo.


A silicon implementation of the fly's optomotor control system.
Flies are capable of stabilizing their body during free flight by using visual motion information to estimate self-rotation. We have built a hardware model of this optomotor control system in a standard CMOS VLSI process. The result is a small, low-power chip that receives input directly from the real world through on-board photoreceptors and generates motor commands in real time. The chip was tested under closed-loop conditions typically used for insect studies. The silicon system exhibited stable control sufficiently analogous to the biological system to allow for quantitative comparisons.


Diagnostic value of immunoperoxidase staining and immunofluorescence in the study of kidney biopsy specimens.
This study aimed to determine diagnostic value of immunoperoxidase in comparison with immunofluorescence in the diagnostic assessment of kidney biopsy specimens. Forty-eight kidney biopsy specimens were used to compare a direct immunofluorescence technique with immunoperoxidase techniques on paraffin sections. The sensitivity and specificity were calculated. The kappa statistic for agreement between the two tests was categorized as poor (zero to 0.2), moderate (0.21 to 0.45), good (0.46 to 0.75), and almost perfect concordance (0.76 to 1.0). Compared with immunofluorescence, the immunoperoxidase technique presented a sensitivity of 88.55% and a specificity of 69.22%. Its sensitivity in the staining for IgG, IgM, and IgA was 93.75%, 95.45%, and 76.47%, respectively. The specificity of this test in the staining for IgG, IgM, and IgA was 54.54%, 57.14%, and 96.00%, respectively. The overall kappa value was 0.60 and it was 0.60 for assessing staining intensity. There was a moderate agreement between immunoperoxidase and immunofluorescence in the positive or negative staining for IgG and IgM, as well as a good agreement in the positive or negative staining for IgA. For the staining intensity, the two tests had a good concordance for IgG and IgA and a moderate concordance for IgM. Although immunoperoxidase method has a lower overall diagnostic performance as compared to immunofluorescence, given the good concordance between the two techniques, it can be an alternative method for immunofluorescence study of kidney biopsy specimens, particularly where immunofluorescence fails or is not available.


Oxidative stress is present in atherosclerotic peripheral arterial disease and further increased by diabetes mellitus.
Atherosclerosis, which is associated with increased oxidative stress, may be widespread and more advanced in both complicated and uncomplicated diabetics than in healthy, age-matched non diabetic controls. The development of microangiopathy and the early development of microangiopathy and the early development of macroangiopathy which occurs in diabetes have unclear aetiology. We have previously shown an increase in the products of free radical activity in conventional atherosclerosis and this study has considered whether there us a further increase in this oxidative stress in diabetics. We measured oxidation by free radicals as lipid peroxides (malondialdehyde) and plasma thiols in 19 insulin dependent diabetic patients with retinopathy or microproteinuria but no clinical evidence of large vessel disease, 19 non diabetic patients with obstructive peripheral arterial disease confirmed by angiography and 19 asymptomatic matched healthy control subjects. The patients were recruited from either the local diabetic or vascular surgery clinics. There were significant increases in oxidative stress in diabetic patients compared to control subjects with increased malondialdehyde 9.2 mumol/l (7.1-11.2) versus 7.65 mumol/l (5.4-9.6, p < 0.01 Mann Whitney test) and reduced plasma thiols 458 mumol/l (294-595) versus 506 mumol/l (433-589, p < 0.05). Those non-diabetic patients with obstructive peripheral arterial disease also had significantly increased malondialdehyde levels at 8.4 mumol/l (6.5-10.1, p < 0.05) and reduced plasma thiols 435 mumol/l (361-462, p < 0.05) compared to controls. The difference between patients with conventional atherosclerotic peripheral arterial disease and those with additional diabetes was significant for malondialdehyde (p < 0.05). These data confirm the oxidative stress that exists in atherosclerotic peripheral arterial disease and suggest that diabetic patients with microangiopathy have an increase in this oxidative stress which may contribute to their early development of atherosclerosis.


Histone deacetylase inhibitors and anticancer therapy.
Recent reports have shown that pharmacological manipulation of chromatin remodeling by histone deacetylase (HDAC) inhibitors, might develop into a potent and specific strategy for the treatment of cancer. Alterations in histone acetylation may lead to changes in chromatin structure and transcriptional dysregulation of genes that are implicated in controlling either cell cycle progression or pathways regulating cell differentiation and/or apoptosis. Dimethyl sulphoxide was one of the first chemicals to be identified as an inducer of transformed cell differentiation. In the class of HDAC inhibitors, now included a short-chain fatty acids, such as 4-phenylbutyrate and valporic acid, hydroxamic acids, such as suberoylanilide hydroxamic acid (SAHA), pyroxamide, trichostatin A, oxamflatin and CHAPSs, cyclic tetrapeptides, such as trapoxin, apicidin and depsipeptide-also known as FK-228 or FR 901228, and benzamides, such as MS-275. First clinical studies have shown that histone hyperacetylation can be achieved safely in humans and that treatment of cancer with such agents seems to become possible. Thus, HDAC inhibitors remains one of the most promising class of new anticancer agents. Further studies are needed in order to delineate the optimal dosage, the duration of therapy and possibly the efficacy of other agents able to synergize with HDAC inhibitors in the fight against cancer.


Degenerative spondylolisthesis. Predisposing factors.
Study of twenty skeletons and two hundred patients with degenerative spondylolisthesis established that it occurred four times more frequently in females, six to nine times more frequently at the interspace between the fourth and fifth lumbar vertebrae than at adjoining levels, three times more frequently in blacks than in whites, and four times more frequently the fifth lumbar vertebra was sacralized. It did not occur before the fifth decade or in conjunction with spina bifida or isthmic spondylolisthesis. The slipping occurs as a result of degenerative disease of the articular processes, but it never exceeds 30 per cent. When symptoms are severe and unrelieved by conservative treatment (10 per cent of our patients), decompression laminectomy and excision of the medial portion of the articular processes affords relief of pain. The predisposing factor is a straight, stable lumbosacral joint which puts abnormal stress on the intervertebral joint between the fourth and fifth lumbar vertebrae, leading to decompensation of disc and ligaments, hypermobility, and degeneration of the articular processes allowing forward slipping.


Streptozocin diabetes alters immunoreactive beta-endorphin levels and pain perception after 8 wk in female rats.
Plasma, pituitary, and hypothalamic levels of the endogenous opioid peptide beta-endorphin were measured by radioimmunoassay and column chromatography in female rats 8 wk after the induction of diabetes with streptozocin (STZ) and in control female rats. In addition, pain perception was determined by measuring the latency to paw lick or jump after being placed on a hot plate. Plasma levels of immunoreactive beta-endorphin (IR-BE) were significantly reduced in STZ-induced diabetic female rats, as were the content and concentration of IR-BE in the neurointermediate lobe of the pituitary (NIL) and the content of IR-BE in the hypothalamus. The concentration but not the content of IR-BE in the anterior pituitary (AP) of the STZ-induced diabetic rats was increased significantly. Streptozocin-induced diabetes also resulted in a significant reduction in the total protein content of the AP, NIL, and hypothalamus. Column chromatography indicated that the decrease in IR-BE in the plasma, NIL, and hypothalamus represented a decrease in beta-endorphin, whereas the increase in IR-BE in the AP represented an increase in both beta-endorphin and beta-lipotropin. Diabetic animals consistently showed decreased latencies to paw lick or jump when subjected to hot-plate testing after 7 wk. These findings suggest that in female rats, central and peripheral endogenous opiate levels and tolerance to nociceptive thermal stimulation were diminished by 8 wk of chemically induced diabetes.


Live visualization of chromatin dynamics with fluorescent TALEs.
The spatiotemporal organization of genomes in the nucleus is an emerging key player to regulate genome function. Live imaging of nuclear organization dynamics would be a breakthrough toward uncovering the functional relevance and mechanisms regulating genome architecture. Here, we used transcription activator-like effector (TALE) technology to visualize endogenous repetitive genomic sequences. We established TALE-mediated genome visualization (TGV) to label genomic sequences and follow nuclear positioning and chromatin dynamics in cultured mouse cells and in the living organism. TGV is highly specific, thus allowing differential labeling of parental chromosomes by distinguishing between single-nucleotide polymorphisms (SNPs). Our findings provide a framework to address the function of genome architecture through visualization of nuclear dynamics in vivo.


Viral vectors for gene transfer into antigen presenting cells.
Crucial insights for vaccine development have come from examining how the immune system responds to antimicrobial vaccines, as well as to viral vectors employed for gene therapy. The effectiveness of a vaccine depends upon both the method of antigen delivery and the presentation of antigen to lymphocytes. Much focus has turned to delivering antigens to dendritic cells, to promote clinically beneficial T- and B-cell responses. Recombinant viral vectors represent a powerful vehicle to deliver genes encoding microbial- or tumor-derived antigens to generate clinically beneficial immunity. Dendritic cell-based and viral vector-based vaccines are currently being evaluated in clinical trials as a means of inducing antitumor immunity.


A sociological study of adolescent health in Ankara, Turkey.
This study aims to explore the associations between socio-demographic factors and subjective health complaints of adolescents in Turkey. A sample of 248 students in the 8th-grade from four primary schools with different socio-economic family backgrounds completed a questionnaire on their socio-demographic characteristics and health complaints. Findings revealed that adolescents are not homogeneous in terms of their socio-economical backgrounds and infra-structural living conditions. The distribution of illnesses/complaints compared among schools groups, only catching cold was significantly different. Factor analysis results showed that there are three groups of diseases that differ from one another namely psychological, physical and other. Adolescents' perception about their general health varied among the four schools. According to the regression analysis between three factor groups of illness/complains and socio-demographic and personal characteristics of students, only number of children and working status of fathers showed a significant difference among the socio-demographic characteristics, while weight and height among the personal characteristics. Although the first findings suggested that respondents' physical characteristics affected their physical complaints, it seemed that several complaints could also be affected by social and cultural context.


Longitudinal analysis of the electroencephalogram and sleep phenotype in the R6/2 mouse model of Huntington's disease.
Deficits in sleep and circadian organization have been identified as common early features in patients with Huntington's disease that correlate with symptom severity and may be instrumental in disease progression. Studies in Huntington's disease gene carriers suggest that alterations in the electroencephalogram may reflect underlying neuronal dysfunction that is present in the premanifest stage. We conducted a longitudinal characterization of sleep/wake and electroencephalographic activity in the R6/2 mouse model of Huntington's disease to determine whether analogous electroencephalographic 'signatures' could be identified early in disease progression. R6/2 and wild-type mice were implanted for electroencephalographic recordings along with telemetry for the continuous recording of activity and body temperature. Diurnal patterns of activity and core body temperature were progressively disrupted in R6/2 mice, with a large reduction in the amplitude of these rhythms apparent by 13 weeks of age. The diurnal variation in sleep/wake states was gradually attenuated as sleep became more fragmented and total sleep time was reduced relative to wild-type mice. These genotypic differences were augmented at 17 weeks and evident across the entire 24-h period. Quantitative electroencephalogram analysis revealed anomalous increases in high beta and gamma activity (25-60 Hz) in all sleep/wake states in R6/2 mice, along with increases in theta activity during both non-rapid eye movement and rapid eye movement sleep and a reduction of delta power in non-rapid eye movement sleep. These dramatic alterations in quantitative electroencephalographic measures were apparent from our earliest recording (9 weeks), before any major differences in diurnal physiology or sleep/wake behaviour occurred. In addition, the homeostatic response to sleep deprivation was greatly attenuated with disease progression. These findings demonstrate the sensitivity of quantitative electroencephalographic analysis to identify early pathophysiological alterations in the R6/2 model of Huntington's disease and suggest longitudinal studies in other preclinical Huntington's disease models are needed to determine the generality of these observations as a potential adjunct in therapeutic development.


Blockchain-Empowered Multi-Robot Collaboration to Fight COVID-19 and Future Pandemics.
This conceptual paper overviews how blockchain technology is involving the operation of multi-robot collaboration for combating COVID-19 and future pandemics. Robots are a promising technology for providing many tasks such as spraying, disinfection, cleaning, treating, detecting high body temperature/mask absence, and delivering goods and medical supplies experiencing an epidemic COVID-19. For combating COVID-19, many heterogeneous and homogenous robots are required to perform different tasks for supporting different purposes in the quarantine area. Managmnt and decentralizing multi-robot play a vital role in combating COVID-19 by reducing human interaction, monitoring, delivering goods. Blockchain technology can manage multi-robot collaboration in a decentralized fashion, improve the interaction among them to exchange information, share representation, share goals, and trust. We highlight the challenges and provide the tactical solutions enabled by integrating blockchain and multi-robot collaboration to combat the COVID-19 pandemic. The proposed conceptual framework can increase the intelligence, decentralization, and autonomous operations of connected multi-robot collaboration in the blockchain network. We overview blockchain potential benefits to defining a framework of multi-robot collaboration applications to combat COVID-19 epidemics such as monitoring and outdoor and hospital End to End (E2E) delivery systems. Furthermore, we discuss the challenges and opportunities of integrated blockchain, multi-robot collaboration, and the Internet of Things (IoT) for combating COVID-19 and future pandemics.