Comparison of the performance of immunosorbents prepared by site-directed or random coupling of monoclonal antibodies.
The majority of methods used to prepare immunosorbents immobilize antibodies through their reactive amino acid residues. The bound antibody activity of these immunosorbents is low. Hydrazide-based matrices couple antibodies through carbohydrate chains frequently located in the Fc region. This paper reports a comparative study of the performance of immunosorbents prepared by cyanogen bromide or hydrazide immobilization methods. The experiments utilized murine monoclonal antibodies to the human plasma proteins Factor IX or Protein C. The antibodies were immobilized at low densities to beaded agarose matrices which had similar properties. The hydrazide immunosorbents had binding efficiencies which were lower (anti-Factor IX) or up to 1.6-fold higher (anti-Protein C) than comparable cyanogen bromide coupled gels. However, there was no improvement in performance due to lower recoveries of bound protein from the hydrazide gels. Control experiments demonstrated that oxidation of antibody which is required for its coupling to hydrazide gels had no effect on antibody binding to antigen. Our results indicate that, as with cyanogen bromide coupling methods, site-directed immobilization through carbohydrate residues results in a restricted ability to bind to antigen. Both monoclonals were found to contain carbohydrate in their Fab' regions through which coupling may have occurred. The frequency of carbohydrate in the Fab region and the ability to control glycosylation at these sites are factors which may impact the utility of carbohydrate-directed immobilization of antibodies.


The New York State universal newborn hearing screening demonstration project: outpatient outcome measures.
To investigate outpatient outcome measures of a multi-center, state-wide, universal newborn hearing screening project. Eight hospitals participated in a 3-yr, funded project. Each hospital designed its own protocol using common criteria for judging whether an infant passed a hearing screening. Infants were tested in the hospital, and those either failing the in-hospital screening or who were not tested in the hospital (missed) were asked to return 4 to 6 wk after hospital discharge for outpatient rescreening. Those infants failing the outpatient rescreening were referred for diagnostic auditory brain stem response testing. Each hospital used its own audiological equipment and criteria to determine whether a particular infant had a hearing loss. All data were collected and analyzed for individual hospitals, as well as totaled across all hospitals. Data were analyzed in terms of year of program operation, nursery type, and geographic region. Seventy-two percent of infants who failed the in-hospital screening returned for outpatient testing. The percentage of in-hospital fails returning for retesting was significantly higher than the percentage of in-hospital misses returning for retesting. The percentage of infants returning for retesting increased with successive years of program operation. Some differences were noted in the percentage of infants returning for retesting among hospitals and geographic regions of the state. Some differences in outpatient outcome measures also were noted between infants originally born into the neonatal intensive care unit (NICU) and the well-baby nursery (WBN). The percentage of infants from the NICU who returned for retesting was slightly higher than that for infants from the WBN. The percentage of infants from the WBN passing the outpatient rescreening was higher than that for the NICU infants. The overall prevalence of hearing loss was 1.96/1000, with that in the NICU being 8/1000 and that in the WBN being 0.9/1000. Positive predictive value for permanent hearing loss based on inpatient screening was approximately 4% and based on outpatient rescreening was approximately 22%. Several outpatient outcome measures changed with successive years of program operation, suggesting that programs improve over time. Also, some outpatient outcome measures differ between NICU and WBN populations. The differences noted across regions of the state in the percentage of infants returning for outpatient retesting require further research to determine whether differences are due to demographic and/or procedural differences.


Mouse translation elongation factor eEF1A-2 interacts with Prdx-I to protect cells against apoptotic death induced by oxidative stress.
eEF1A-1 and eEF1A-2 are two isoforms of translation elongation factor eEF1A. In adult mammalian tissues, isoform eEF1A-1 is present in all tissues except neurons, cardiomyocytes, and myotubes, where its isoform, eEF1A-2, is the only form expressed. Both forms of eEF1A have been characterized to function in the protein elongation step of translation, and eEF1A-1 is shown to possess additional non-canonical roles in actin binding/bundling, microtubule bundling/severing, and cellular transformation processes. To study whether eEF1A-2 has similar non-canonical functions, we carried out a yeast two-hybrid screening using a full sequence of mouse eEF1A-2 as bait. A total of 78 hits, representing 23 proteins, were identified and validated to be true positives. We have focused on the protein with the highest frequency of hits, peroxiredoxin I (Prdx-I), for in-depth study of its functional implication for eEF1A-2. Here we show that Prdx-I coimmunoprecipitates with eEF1A-2 from extracts of both cultured cells and mouse tissues expressing this protein, but it does not do so with its isoform, eEF1A-1, even though the latter is abundantly present. We also report that an eEF1A-2 and Prdx-I double transfectant increases resistance to peroxide-induced cell death as high as 1 mM peroxide treatment, significantly higher than do single transfectants with either gene alone; this protection is correlated with reduced activation of caspases 3 and 8, and with increased expression of pro-survival factor Akt. Thus, our results suggest that eEF1A-2 interacts with Prdx-I to functionally provide cells with extraordinary resistance to oxidative stress-induced cell death.


Differential diagnosis of pulmonary carcinoma following head and neck cancer by genetic analysis.
Patients with head and neck cancer often develop a lung tumor that can be diagnosed as distant metastasis (DM) or second primary tumor (SPT). In this study, we use TP53 mutation analysis for validation of an allelic loss marker panel and a decision algorithm for distinguishing between DM and SPT. Tumor pairs of 39 patients were analyzed for TP53 mutations, for patterns of allelic loss and immunohistochemical analysis of p53 expression. Results of these three analyses were compared, using mutation analysis as gold standard. Loss of heterozygosity (LOH) analysis indicated DM in 15 and SPT in 23 cases (one inconclusive). TP53 mutation analysis was informative in 15 cases. Based on the p53 mutation status alone, nine tumors were diagnosed as SPT and six as DM. In all 15 cases the LOH analysis was in concordance with the TP53 mutation analysis. Immunostaining for p53 showed promise as a first scan to diagnose lung tumors as SPT but cannot be used to diagnose DM. The TP53 mutation data validate the suitability of the LOH marker panel and decision algorithm for differential diagnosis of DM and SPT in the lung. LOH analysis can theoretically be exploited in almost all cases and is less laborious than TP53 mutation analysis.


Advances in dielectric elastomers for actuators and artificial muscles.
A number of materials have been explored for their use as artificial muscles. Among these, dielectric elastomers (DEs) appear to provide the best combination of properties for true muscle-like actuation. DEs behave as compliant capacitors, expanding in area and shrinking in thickness when a voltage is applied. Materials combining very high energy densities, strains, and efficiencies have been known for some time. To date, however, the widespread adoption of DEs has been hindered by premature breakdown and the requirement for high voltages and bulky support frames. Recent advances seem poised to remove these restrictions and allow for the production of highly reliable, high-performance transducers for artificial muscle applications.


Assignment of the chloramphenicol resistance gene to mitochondrial deoxyribonucleic acid and analysis of its expression in cultured human cells.
The mitochondrial deoxyribonucleic acids (mtDNA's) from human HeLa and HT1080 cells differed in their restriction endonuclease cleavage patterns for HaeII, HaeIII, and HhaI. HaeII digestion yielded a 9-kilobase fragment in HT1080, which was replaced by 4.5-, 2.4-, and 2.1-kilobase fragments in HeLa. HaeIII and HhaI yielded distinctive 1.35- and 0.68-kilobase HeLa fragments. These restriction endonuclease polymorphisms were used as mtDNA markers in HeLa-HT1080 cybrid and hybrid crosses involving the cytoplasmic chloramphenicol resistance mutation was used. mtDNA's were purified and digested with the restriction endonucleases, the fragments were separated on agarose gels, and the bands were detected by ethidium bromide staining and Southern transfer analysis. Three cybrids and four hybrids (four expressing HeLa and three expressing HT1080 chloramphenicol resistance) contained 2- to 10-fold excesses of the mtDNA of the chloramphenicol-resistant parent. One cybrid, which was permitted to segregate chloramphenicol resistance and was then rechallenged with chloramphenicol, had approximately equal proportions of the two mtDNA's. Only one hybrid was discordant. These results indicated that chloramphenicol resistance is encoded in mtDNA and that expression of chloramphenicol resistance is related to the ratio of chloramphenicol-resistant and -sensitive genomes within cells.


Cervical spine bending: a factor confounding whole trunk and lumbar forward bending range of motion.
Knee bending during tests of lumbar forward bending (FB) may introduce confounding variability. Precluding bending at the knees has, therefore, long been standard protocol to produce valid and reproducible results. However, there is limited research on cervical spine bending as a confounding variable in whole trunk and lumbar FB. To examine the role of cervical spine bending on the range of whole trunk and lumbar FB. Participants were recruited from the faculty, staff, and student population of Nova Southeastern University's Health Professions Division. Each participant underwent 4 FB tests with varying cervical starting positions. Range of motion was measured for whole trunk FB and lumbar FB by using the fingertip-to-floor and double digital inclinometer techniques, respectively. Two hundred thirty-six participants met the study criteria. Statistically significant differences were found in both whole trunk (6.96 cm) and lumbar (3.95°) FB range of motion when the cervical spine was backward bent after full spine FB (P<.05). Statistically significant differences were also found in both whole trunk (15.72 cm) and lumbar (7.38°) FB when the cervical spine was backward bent before thoracolumbar spine FB (P<.05). Cervical spine bending influences the ability of the trunk and lumbar spine to bend forward and is, therefore, a confounding variable during tests of whole trunk and lumbar spine FB.


Is a behavioral graded activity program more effective than manual therapy in patients with subacute neck pain? Results of a randomized clinical trial.
A randomized clinical trial. To compare the effectiveness of a behavioral graded activity program with manual therapy in patients with subacute (4-12 weeks) nonspecific neck pain. Neck pain is a common complaint, for which many conservative therapies are available in primary care. There is strong evidence for manual therapy in combination with exercises. Psychosocial factors are also believed to play a role in chronic pain. The evidence of the effectiveness of a program focused on these factors is still unknown. A randomized clinical trial was conducted, involving 146 patients with subacute nonspecific neck pain. The BGA program can be described as a time-contingent increase in activities from baseline toward predetermined goals. Manual therapy consists of specific spinal mobilization techniques and exercises. Primary outcomes were global perceived effect, the Numerical Rating Scale for pain and the Neck Disability Index. Secondary outcomes were the Tampa Scale for Kinesiophobia, the 4 Dimensional Symptom Questionnaire, and the Pain Coping and Cognition List. Measurements were carried out at baseline and 6, 13, 26, and 52 weeks after randomization. Data are analyzed according to the intention-to-treat principle, using multilevel analysis. The success rates at 52 weeks, based on the GPE were 89.4% for the BGA program and 86.5% for MT. This difference was not statistically significant. For pain and disability, a difference was found in favor of the BGA program; mean difference for pain = 0.99 (95% CI 0.15-1.83) and mean difference for NDI = 2.42 (95% CI 0.52-4.32). All other differences between the interventions in the primary and secondary outcomes were not statistically significant. Based on this trial it can be concluded that there are only marginal, but not clinically relevant, differences between a BGA program and MT.


Myricetin Induces Apoptosis of Human Anaplastic Thyroid Cancer Cells via Mitochondria Dysfunction.
Thyroid cancer is the most common endocrine malignancy, with an increasing incidence worldwide. Most thyroid cancers are well differentiated and have a favorable outcome. However, undifferentiated thyroid cancers are one of the most lethal human malignancies. Anaplastic thyroid cancer (ATC) accounts for 2% of all thyroid cancers, and its median survival rate is low. ATC is responsible for more than one-third of thyroid cancer-related deaths. Myricetin is a flavonol compound found in walnuts, herbs, and various berries and is known to induce apoptotic death of various types of cancer cells. However, an anticancer effect of myricetin against human anaplastic thyroid cancer (HATCs) cells has not been demonstrated. In the present study, the anticancer effects and mechanism of action of myricetin were examined using SNU-80 HATC cells. SNU-80 HATC cells were treated with various concentrations of myricetin and compared with untreated controls. Myricetin significantly reduced HATC cell proliferation, by approximately 70%. A substantial proportion of dead cells exhibited arrest in the sub-G1 phase. Myricetin also exhibited cytotoxicity and induced DNA condensation in SNU-80 HATC cells in a dose-dependent manner. The mechanism of myricetin-induced cell death involved an increase in the activation of caspase cascades and the Bax:Bcl-2 ratio at a concentration of 100 μM. Myricetin also induced the release of apoptosis-inducing factor (AIF) from mitochondria into the cytosol and altered the mitochondrial membrane potential. Our results indicate that myricetin is a potent inducer of HATC cell death and may thus prove useful in the development of therapeutic agents for HATC.


The Association of Health Literacy with Illness and Medication Beliefs among Patients with Chronic Obstructive Pulmonary Disease.
Low health literacy is associated with low adherence to self-management in many chronic diseases. Additionally, health beliefs are thought to be determinants of self-management behaviors. In this study we sought to determine the association, if any, of health literacy and health beliefs among elderly individuals with COPD. We enrolled a cohort of patients with COPD from two academic urban settings in New York, NY and Chicago, IL. Health literacy was measured using the Short Test of Functional Health Literacy in Adults. Using the framework of the Self-Regulation Model, illness and medication beliefs were measured with the Brief Illness Perception Questionnaire (B-IPQ) and Beliefs about Medications Questionnaire (BMQ). Unadjusted analyses, with corresponding Cohen's d effect sizes, and multiple logistic regression were used to assess the relationships between HL and illness and medication beliefs. We enrolled 235 participants, 29% of whom had low health literacy. Patients with low health literacy were more likely to belong to a racial minority group (p<0.001), not be married (p = 0.006), and to have lower income (p<0.001) or education (p<0.001). In unadjusted analyses, patients with low health literacy were less likely to believe they will always have COPD (p = 0.003, Cohen's d = 0.42), and were more likely to be concerned about their illness ((p = 0.04, Cohen's d = 0.17). In analyses adjusted for sociodemographic factors and other health beliefs, patients with low health literacy were less likely to believe that they will always have COPD (odds ratio [OR]: 0.78, 95% confidence interval [CI]: 0.65-0.94). In addition, the association of low health literacy with expressed concern about medications remained significant (OR: 1.20, 95% CI: 1.05-1.37) though the association of low health literacy with belief in the necessity of medications was no longer significant (OR: 0.92, 95% CI: 0.82-1.04). In this cohort of urban individuals with COPD, low health literacy was prevalent, and associated with illness beliefs that predict decreased adherence. Our results suggest that targeted strategies to address low health literacy and related illness and medications beliefs might improve COPD medication adherence and other self-management behaviors.


Retrograde intrarenal surgery in patients with isolated anomaly of kidney rotation.
The purpose of the study was to investigate feasibility of retrograde intrarenal surgery (RIRS) in patients with isolated anomaly of kidney rotation with kidney stones. We retrospectively reviewed the medical records of patients who underwent RIRS due to kidney stones in three referral hospitals. Twenty-four patients who had kidney rotation were included in this study. The patients with horseshoe kidney or pelvic kidney were not included in the study. The patients were evaluated for final success 1 month after surgery. Success was defined as stone-free or fragment smaller than 3 mm. Mean age of patients was 39.5 years (1-71 years) and male/female ratio was 3:1. Mean stone size was 13.46 mm (5-30 mm). Twelve (50 %) patients had the operation on the right side and other 12 (50 %) patients had on the left side. Eighteen (75 %) patients were stone-free after single procedure. And stone-free rate increased to 83.3 % after additional treatment procedures. When we compared the successful and failed procedures, gender, stone side and size, preoperative hydronephrosis, access sheet usage, ureteral double-J stent insertion were not statistically significant in two groups. We did not confront major complication. RIRS seems to be an effective and safe treatment option for renal stones in patients with isolated anomaly of kidney rotation.


A case of Kleine-Levin syndrome examined with SPECT and neuropsychological testing.
A case of Kleine-Levin syndrome with typical periodic hypersomnia and bulimia was diagnosed. On examination with single photo emission tomography (SPECT) (CERETEC) during a relapse period and 2 weeks later there was marked cortical hypoperfusion of the frontal and temporal lobes, especially on the left side as well as in the right parietal lobe. Neuropsychological testing performed 1 week after a relapse showed a reduction in encoding to memory function of verbal learning indicating neocortical damage of the left fronto-temporal region. A follow-up 2 months later after the patient had spontaneously recovered showed only a slight left fronto-temporal disturbance. CT and MRI of the brain were normal although the MRI showed a large and asymmetric mamillary body. Neuropsychological testing 6 years after recovery showed pronounced reduction in short-time verbal and visual memory. Seven years after recovery SPECT demonstrated a normalized frontal perfusion but still a slight hypoperfusion in the left temporal lobe. Our results correlate to autopsy findings in two cases described previously.


Compression of the posterior interosseous nerve secondary to a synovial cyst: Case report and review of the literature.
Posterior interosseous nerve syndrome secondary to compression by a synovial cyst at the elbow is a rare and often unrecognized pathology. Early management relies on complete neurolysis to achieve satisfactory functional recovery. Increasing awareness among the orthopedics will help in the early diagnosis of the disease and in the initiation of early and proper treatment. In this article, we report the case of a 32-year-old patient with posterior interosseous nerve syndrome secondary to compression by a synovial cyst of the elbow. Surgical management combined with post-operative rehabilitation resulted in indolence with good functional recovery. Posterior interosseous nerve syndrome secondary to compression by a synovial cyst at the elbow is a rare entity. Anatomically, the deep branch of the radial nerve or posterior interosseous nerve passes through the Fröhse's arch or arch of the supinator muscle at the elbow, then travels between the two heads of this muscle. Several anatomical structures may compress the NIOP. Clinically, it presents as paralysis or paresis of the extensor muscles of the fingers and the abductor muscle of the thumb. Limitation of the ulnar extensor carpi may be responsible for radial deviation of the carpus in some cases. MRI is the radiological examination of choice. Electromyography plays a contributory role in diagnosis prior to surgical exploration. Surgical excision is the treatment of choice. It may be combined with radial neurolysis for better recovery. Progression after surgical treatment is generally favourable. Ignorance of posterior interosseous nerve palsy syndrome frequently leads to misdiagnosis. Early management relies on complete neurolysis to achieve satisfactory functional recovery.


Translational physiology: porcine models of human coronary artery disease: implications for preclinical trials of therapeutic angiogenesis.
"Therapeutic angiogenesis" describes an emerging field of cardiovascular medicine whereby new blood vessels are induced to grow to supply oxygen and nutrients to ischemic cardiac or skeletal muscle. Various methods of producing therapeutic angiogenesis have been employed, including mechanical means, gene therapy, and the use of growth factors, among others. The use of appropriate large-animal models is essential if these therapies are to be critically evaluated in a preclinical setting before their use in humans, yet little has been written comparing the various available models. Over the past decade, swine have been increasingly used in studies of chronic ischemia because of their numerous similarities to humans, including minimal preexisting coronary collaterals as well as similar coronary anatomy and physiology. Consequently, this review describes the most commonly used swine models of chronic myocardial ischemia with special attention to regional myocardial blood flow and function and critically evaluates the strengths and weaknesses of each model in terms of utility for preclinical trials of angiogenic therapies.


Neonatal resuscitation 3: manometer use in a model of face mask ventilation.
Adequate ventilation is the key to successful neonatal resuscitation. Positive pressure ventilation (PPV) is initiated with manual ventilation devices via face masks. These devices may be used with a manometer to measure airway pressures delivered. The expiratory tidal volume measured at the mask (V(TE(mask))) is a good estimate of the tidal volume delivered during simulated neonatal resuscitation. To assess the effect of viewing a manometer on the peak inspiratory pressures used, the volume delivered, and leakage from the face mask during PPV with two manual ventilation devices in a model of neonatal resuscitation. Participants gave PPV to a modified resuscitation mannequin using a Laerdal infant resuscitator and a Neopuff infant resuscitator at specified pressures ensuring adequate chest wall excursion. Each participant gave PPV to the mannequin with each device twice, viewing the manometer on one occasion and unable to see the manometer on the other. Data from participants were averaged for each device used with the manometer and without the manometer separately. A total of 7767 inflations delivered by the 18 participants were recorded and analysed. Peak inspiratory pressures delivered were lower with the Laerdal device. There were no differences in leakage from the face mask or volumes delivered. Whether or not the manometer was visible made no difference to any measured variable. Viewing a manometer during PPV in this model of neonatal resuscitation does not affect the airway pressure or tidal volumes delivered or the degree of leakage from the face mask.


Mapping the road to elimination: a 5-year evaluation of implementation strategies associated with hepatitis C treatment in the veterans health administration.
While few countries and healthcare systems are on track to meet the World Health Organization's hepatitis C virus (HCV) elimination goals, the US Veterans Health Administration (VHA) has been a leader in these efforts. We aimed to determine which implementation strategies were associated with successful national viral elimination implementation within the VHA. We conducted a five-year, longitudinal cohort study of the VHA Hepatic Innovation Team (HIT) Collaborative between October 2015 and September 2019. Participants from 130 VHA medical centers treating HCV were sent annual electronic surveys about their use of 73 implementation strategies, organized into nine clusters as described by the Expert Recommendations for Implementing Change taxonomy. Descriptive and nonparametric analyses assessed strategy use over time, strategy attribution to the HIT, and strategy associations with site HCV treatment volume and rate of adoption, following the Theory of Diffusion of Innovations. Between 58 and 109 medical centers provided responses in each year, including 127 (98%) responding at least once, and 54 (42%) responding in all four implementation years. A median of 13-27 strategies were endorsed per year, and 8-36 individual strategies were significantly associated with treatment volume per year. Data warehousing, tailoring, and patient-facing strategies were most commonly endorsed. One strategy-"identify early adopters to learn from their experiences"-was significantly associated with HCV treatment volume in each year. Peak implementation year was associated with revising professional roles, providing local technical assistance, using data warehousing (i.e., dashboard population management), and identifying and preparing champions. Many of the strategies were driven by a national learning collaborative, which was instrumental in successful HCV elimination. VHA's tremendous success in rapidly treating nearly all Veterans with HCV can provide a roadmap for other HCV elimination initiatives.


[Testing of some grapevine cultivars for the presence of crown gall disease agent and fanleaf and leafroll viruses].
Some rootstocks and scion cultivars grown in Ukraine have been tested for the presence of tumorigenic agrobacteria, grapevine fanleaf and grapevine leafroll viruses of the 1st and 3rd serotypes by polymerase chain reaction and enzyme-linked immunosorbent assay. The conditions of polymerase chain reaction were modified in the course of investigations. It was shown that the certified material as well as regular planting material may be infected by the agent of crown gall disease. Certified grapevine material was free from grapevine fanleaf and grapevine leafroll viruses; at the same time the regular planting material was infected by both viruses. Our data indicate the necessity of including Agrobacterium vitis in the list of pathogens that should be tested in the process of certified planting material production.


Post-Transcriptional Dysregulation of microRNA and Alternative Polyadenylation in Colorectal Cancer.
Colorectal cancer (CRC) is one of the leading causes of cancer death worldwide. microRNAs (miRNAs) repress gene expression by binding to complementary sequences in the 3' untranslated region (3'UTR) of target mRNAs. Alternative polyadenylation (APA) are relevant to the variability of the 3'UTR of mRNA. However, the posttranscriptional dysregulation of miRNAs and APA in CRC are poorly understood. In this study, we conducted small RNA sequencing to identify differentially expressed miRNAs (DERs) and their target genes. Function analysis on DER-target genes can explain the regulation roles of miRNAs in CRC. The mutual regulation of miRNAs and APA was analyzed by combining miRNA data to 3'UTR alteration using 3' termini of polyadenylated RNAs sequencing (3T-seq) technique, and this was validated using TCGA gene expression data. Our results showed 64 significant differentially expressed miRNAs (DERs) in CRC patients. Their target genes were related to cell adhesion and transcription regulation and were prevailingly involved in the CRC-related pathway. Integrative analysis of the miRNA and APA profile revealed 16 DERs were correlated with 12 polyadenylation factors, and six of them were significantly differently expressed in CRC. We also found four DERs that lost binding sites due to APA and showed a positive correlation between the miRNA and gene expression. Our study found that miRNAs regulated APA by modulating key polyadenylation factors, and several miRNAs lost their suppression on mRNA due to APA. Associating this with gene expression may provide some important clues for a deeper study of posttranscriptional cellular regulation and biomarker research in CRC. Our data provided the first evidence that the interaction between miRNAs and APA associated with gene expression could serve as biomarkers for CRC, suggesting that hsa-miR-133a-3p and MLEC, hsa-miR-145-5p and SET, hsa-miR-1-3p and PPIA, and hsa-miR-378d and YY1 might be novel and potential biomarkers in improving the diagnosis of CRC.


Analysis of Psychological and Gut Microbiome Characteristics in Patients With Non-erosive Reflux Disease.
Objective: To assess the correlation between the incidence of non-erosive reflux disease (NERD) and psychological factors, especially somatoform disorders. To investigate the characteristics of gut microbiome in NERD patients. Methods: We enrolled 24 NERD patients and 24 healthy controls. All patients were evaluated via GerdQ, SOMS-7, SAS, HAMA, and HAMD. Fecal samples were collected and 16S rRNA sequencing was performed to evaluate the gut microbiome composition. Results: The main symptoms of the NERD patients were regurgitation (87.5%), belching (66.7%), pharyngeal discomfort (50%), and heartburn (37.5%). The average score of GerdQ was 13.42 ± 3.41. In 15 patients (62.5%), the total score of the last two items was <3 points, while the average score of 24 patients was 3.63 ± 2.32. NERD patients with somatoform disorders accounted for 50%. There were 17 patients without anxiety, 6 patients with mild anxiety (25%), 1 patient with moderate anxiety (4.2%), and no patient with severe anxiety. There were 22 patients (91.7%) without depression, 2 patients (8.3%) with mild depression, and no patient with moderate or severe depression. The alpha diversity of NERD group was higher than HC, which showed significant difference (P < 0.05). The beta-diversity was significantly different between HC and NERD patients (P = 0.026), male and female patients (P = 0.009). The beta-diversity was also significantly different between male and female patients (P = 0.009). There were several bacteria with significant differences between HC and NERD group, and NERD patients with or without somatoform disorders, such as Firmicutes, TM7 were enriched in the NERD group compared with the healthy control group, while Bacteroidetes were enriched in the healthy controls. Conclusions: NERD symptoms overlap with somatoform disorders. NERD symptoms have an impact on the daily life quality of patients. Some of them are accompanied by anxiety and depression of different degrees, and the two are significantly correlated. The diversity of gut microbiome in patients with NERD is significantly higher than healthy controls, which has its characteristics. The predominant bacteria in gut microbiome of patients with NERD are similar to the healthy population, with Firmicutes and Bacteroidetes as the main ones. The composition of gut microbiome in NERD patients with or without somatoform disorder is significantly different, which may be related to the interaction of microbiome-brain-gut axis.


Reliability and validity of a Swedish version of the Revised Leeds Disability Questionnaire for patients with ankylosing spondylitis.
The aim of the study was to investigate the reliability and the validity of a Swedish version of the Revised Leeds Disability Questionnaire (RLDQ). Forty-two patients, 36 men and 6 women, median age 52.5 years, median symptom duration 24 years, with spondylarthropathy were assessed with the RLDQ, three questions on the content of the questionnaire, and range-of-motion measures. The results indicated satisfactory test-retest stability and internal consistency, and only minor internal redundancy. RLDQ items were generally considered relevant and suggested additions were similar to items already included in the questionnaire. Patients with low disability scores tended to be older and have better joint mobility than those with higher scores. Physiotherapists observed and scored disability significantly higher than patients. Correlations between subscores of the RLDQ and range-of-motion measures were in the main weak. In conclusion the Swedish version of the RLDQ may be considered as reliable and valid.


The Role of Apparent Diffusion Coefficient Quantification in Differentiating Benign and Malignant Renal Masses by 3 Tesla Magnetic Resonance Imaging.
Diffusion-weighted magnetic resonance imaging (DWI) is a widely-accepted diagnostic modality whose efficacy has been investigated by numerous past studies in the differentiation of malignant lesions from benign entities. The aim of this study was to evaluate the efficiency of diffusion-weighted magnetic resonance imaging in the characterization of renal lesions. Diagnostic accuracy study. A total of 137 patients with renal lesions were included in this study. The median apparent diffusion coefficient (ADC) values as well as the b 800 and b 1600 signal intensities of normal kidneys, solid components of mixed renal masses, and total cystic lesions were evaluated. There were significant differences between the ADC values of lesions and normal renal parenchyma, and between the ADC values of benign and malignant renal lesions on DWIs at b values of 800 and 1600 s/mm(2) (p<0.001 and p<0.001, respectively). There were significant differences between the ADC values of Bosniak Category 1 and 2 cysts and the ADC values of Bosniak Category 1 and 3 cysts on DWIs at b values of 800 s/mm(2) (p<0.001) and 1600 s/mm(2) (p<0.001). A cutoff value of 1.902 × 10(-3) mm(2)/s for the ADC with a b value of 800 s/mm(2) provided 88% sensitivity and 96% specificity for differentiation between benign and malignant renal lesions. A cutoff value of 1.623 × 10(-3) mm(2)/s for the ADC with a b value of 1600 s/mm(2) provided 79% sensitivity and 96% specificity (p<0.001) for the differentiation between benign and malignant renal lesions. Accurate assessment of renal masses is important for determining the necessity for surgical intervention. DWI provides additional value by differentiating benign from malignant renal tumors and can be added to routine kidney MRI protocols.


Effectiveness of magnetic resonance angiography in the evaluation of lung perfusion in constrictive bronchiolitis obliterans.
Childhood constrictive bronchiolitis obliterans (CBO) is characterized clinically by the persistence of obstructive respiratory symptoms following a prolonged episode of lower respiratory viral infection. The diagnosis depends on the clinical, radiological and laboratory findings. Perfusion defects on scintigraphy may predict the severity. This report describes a 2-year-old girl who presented with a 15-day history of respiratory distress despite treatment. Auscultation of the lungs revealed prolonged expiration and bilateral crepitant rales. CXR revealed bilateral paracardiac infiltration. She improved with a treatment regimen of bronchodilators, systemic steroids and antibiotherapy; however, rales were detected again 10 days after discharge. HRCT of the lung revealed an oligaemic-mosaic pattern, more prominent in the left lung. The diagnosis of CBO was made on the basis of these clinical and radiological findings. Perfusion was normal in the right lung but diffusely decreased in the left lung on perfusion scintigraphy by Tc99m-MAA. Magnetic resonance (MR) angiography source images of the lungs revealed significant but not diffuse perfusion defects in the left lung, in contrast to radionuclide scintigraphy. There were areas of low perfusion in the right lung, which were not evident in radionuclide scintigraphy. The use of source images of MRI angiography of the lungs may be more sensitive and specific for the detection of perfusion defects in patients with CBO and may assist in determining the prognosis.


Uniform Selection And Representation Matching: A Framework For Classifying Wound Healing Stage.
We propose the Uniform Selection and Representation Matching framework, an image classification framework that leverages co-teaching, contrastive learning, representation matching, and uniform selection to perform accurate wound stage classification with limited and noisy-labeled data. Given that descriptors of wound stages are under-specified, making accurate recognition difficult, images that generate low classification confidence are identified using an entropy-based selection process. Pseudo-labels are assigned to the low-confidence images through the representation matching process, where images are embedded into latent space and labels are assigned through majority voting. The Uniform Selection and Representation Matching framework demonstrates high accuracy in classifying wound-stage images by achieving a classification accuracy of 90.0%, a significant improvement over conventional convolutional neural networks.Clinical relevance- This work proposes a wound-stage classification algorithm trained with minimal data and noisy labels. Applications include remotely monitoring wound healing, recommending treatments, and incorporating intelligent bandage devices.


E-Textiles for Healthy Ageing.
The ageing population has grown quickly in the last half century with increased longevity and declining birth rate. This presents challenges to health services and the wider society. This review paper considers different aspects (e.g., physical, mental, and social well-being) of healthy ageing and how health devices can help people to monitor health conditions, treat diseases and promote social interactions. Existing technologies for addressing non-physical (e.g., Alzheimer's, loneliness) and physical (e.g., stroke, bedsores, and fall) related challenges are presented together with the drivers and constraints of using e-textiles for these applications. E-textiles provide a platform that enables unobtrusive and ubiquitous deployment of sensors and actuators for healthy ageing applications. However, constraints remain on battery, integration, data accuracy, manufacturing, durability, ethics/privacy issues, and regulations. These challenges can only effectively be met by interdisciplinary teams sharing expertise and methods, and involving end users and other key stakeholders at an early stage in the research.


Epitopes and structural domains of a RNA-binding nuclear protein, SS-B/La: similarities with adenovirus DNA-binding protein.
SS-B/La is a conserved cellular phosphoprotein of 46/48 KD molecular weight which is the target antigen of autoantibodies in sera of patients with Sjögren's syndrome. Two relatively protease-resistant domains (X and Y) were defined in the SS-B antigen from HeLa cells. Human autoantibodies to SS-B were used as reagents. Domain X is a methionine-containing, non-phosphorylated 28 KD polypeptide and is resistant to partial digestion with six different proteases. Domain Y is a 23 KD polypeptide which contains little if any methionine, but all the detectable phosphorylated amino acids. These results demonstrate that there are at least two distinct antigenic epitopes on the 46/48 KD SS-B protein, each located on a separate structural domain. The asymmetric distribution of methionine and phosphorylated amino acid residues in SS-B show striking similarity to two reported domains of the adenovirus 72 KD DNA-binding protein, which also have separate methionine and phosphorylated amino acid distributions.


Comparison of CO2 capture by ex-situ accelerated carbonation and in in-situ naturally weathered coal fly ash.
Natural weathering at coal power plants ash dams occurs via processes such as carbonation, dissolution, co-precipitation and fluid transport mechanisms which are responsible for the long-term chemical, physical and geochemical changes in the ash. Very little information is available on the natural carbon capture potential of wet or dry ash dams. This study investigated the extent of carbon capture in a wet-dumped ash dam and the mineralogical changes promoting CO2 capture, comparing this natural phenomenon with accelerated ex-situ mineral carbonation of fresh fly ash (FA). Significant levels of trace elements of Sr, Ba and Zr were present in both fresh and weathered ash. However Nb, Y, Sr, Th and Ba were found to be enriched in weathered ash compared to fresh ash. Mineralogically, fresh ash is made up of quartz, mullite, hematite, magnetite and lime while weathered and carbonated ashes contained additional phases such as calcite and aragonite. Up to 6.5 wt % CO2 was captured by the fresh FA with a 60% conversion of calcium to CaCO3 via accelerated carbonation (carried out at 2 h, 4Mpa, 90 °C, bulk ash and a S/L ratio of 1). On the other hand 6.8 wt % CO2 was found to have been captured by natural carbonation over a period of 20 years of wet disposed ash. Thus natural carbonation in the ash dumps is significant and may be effective in capturing CO2.


Sentinel lymph node biopsy in cases of skin melanoma: initial experiences at a center in northeastern Brazil.
The incidence of skin melanoma is increasing worldwide. The presence of lymph node metastasis is the most important prognostic factor of this disease, the thicker the lesion the greater the likelihood of lymph node involvement. Approximately 20% of patients with Breslow depth 1-4 mm have lymph node metastasis. Sentinel lymph node biopsy is standard procedure in the management of patients with skin melanoma, reflecting progress in the treatment of this disease since this procedure avoids an unnecessary radical lymphadenectomy, thus reducing the morbidity of treatment. This paper describes a series of cases involving sentinel lymph node biopsy in melanoma patients in Teresina, Piauí, Brazil between 2008 and 2009.


Global protocol for healthcare administration.
Managers and administrators wanting to improve their leadership and influence should carefully analyze the factors which comprise their success. The first three factors described in this article are often learned easily through seminars and study. The final factor, developing a sense of the appropriate, though, is more difficult. It requires a willingness to recognize the importance of the unwritten rules of society which are placed upon leaders and then dictates that the leader continually engage in self-evaluation and correction of personal habits, characteristics and idiosyncrasies. Practicing appropriate protocols necessitates suppression of the personal ego and an openness to the suggestions of others.


Characterization of β-lactamase and porin mutants of Enterobacteriaceae selected with ceftaroline + avibactam (NXL104).
Ceftaroline + avibactam (NXL104) is a novel inhibitor combination active against Enterobacteriaceae with class A and C β-lactamases. We investigated its risk of mutational resistance. Single- and multi-step mutants were sought and characterized from Enterobacteriaceae with extended-spectrum β-lactamases (ESBLs), AmpC β-lactamases and KPC β-lactamases. Overgrowth occurred on agar with low MIC multiples of ceftaroline + avibactam, but frequencies for single-step mutants were <10(-9). Most mutants were unstable, with only three remaining resistant on subculture. For one, from an CTX-M-15-positive Escherichia coli, the ceftaroline + avibactam MIC was raised, but the organism had reduced resistance to ceftaroline and lost resistance to other oxyimino-cephalosporins, with this profile retained when the mutant bla(CTX-M-15) was cloned into E. coli DH5α. Sequencing identified a Lys237Gln substitution in the CTX-M-15 variant. The other two stable single-step mutants were from an AmpC-derepressed Enterobacter cloacae strain; these had unaltered or slightly reduced resistance to other β-lactams. Both had amino acids 213-226 deleted from the Ω loop of AmpC. Further stable mutants were obtained from AmpC-inducible and -derepressed E. cloacae in multi-step selection, and these variously had reduced expression of OmpC and OmpF, and/or Asn366His/Ile substitutions in AmpC. Stable resistant mutants were difficult to select. Those from AmpC-derepressed E. cloacae had porin loss or AmpC changes, including Ω loop deletions. A Lys237Gln substitution in CTX-M-15 conferred resistance, but largely abolished ESBL activity.


[The morphological and karyological characteristics of MDCK and vero (B) cells cultures on plant hydrolyzate-based nutrient media].
MDCK and Vero (B) cell cultures were propagated during 10 passages in the experimental nutrient media containing the soybean powder hydrolyzate prepared using trypsin and bromelain enzymes and the rice powder hydrolysate prepared with trypsin and in the control DMEM and SFM4 MegaVir media. The karyological, morphological, and proliferative characteristics of continuous cultures were examined and compared. The experimental media supplied with 3% fetal bovine serum (FBS) (Gibco, U.S.A.) showed high growth-enhancing properties and failed to affect their morphology. After propagated during 10 passages in the experimental media preserved a stable karyotype. MDCK cell cultures in the nutrient media based on rice and soybean powder hydrolyzates low (2%) in FBS caused no substantial changes in the proliferation indices and morphological and karyological characteristics of cell cultures.


Nonparametric Maximum Likelihood Estimators of Time-Dependent Accuracy Measures for Survival Outcome Under Two-Stage Sampling Designs.
Large prospective cohort studies of rare chronic diseases require thoughtful planning of study designs, especially for biomarker studies when measurements are based on stored tissue or blood specimens. Two-phase designs, including nested case-control (Thomas, 1977) and case-cohort (Prentice, 1986) sampling designs, provide cost-effective strategies for conducting biomarker evaluation studies. Existing literature for biomarker assessment under two-phase designs largely focuses on simple inverse probability weighting (IPW) estimators (Cai and Zheng, 2011; Liu et al., 2012). Drawing on recent theoretical development on the maximum likelihood estimators for relative risk parameters in two-phase studies (Scheike and Martinussen, 2004; Zeng et al., 2006), we propose nonparametric maximum likelihood based estimators to evaluate the accuracy and predictiveness of a risk prediction biomarker under both types of two-phase designs. In addition, hybrid estimators that combine IPW estimators and maximum likelihood estimation procedure are proposed to improve efficiency and alleviate computational burden. We derive large sample properties of proposed estimators and evaluate their finite sample performance using numerical studies. We illustrate new procedures using a two-phase biomarker study aiming to evaluate the accuracy of a novel biomarker, des-γ-carboxy prothrombin, for early detection of hepatocellular carcinoma (Lok et al., 2010).


A case of erythromycin-induced carbamazepine toxicity.
A case of serious carbamazepine toxicity as a result of concomitant erythromycin therapy is presented. Carbamazepine competes with erythromycin for metabolism by hepatic microsomal enzymes, resulting in decreased clearance of carbamazepine. Carbamazepine drug interactions will become increasingly important to psychiatrists as this anticonvulsant becomes more widely used to treat affective disorders.


Diagnosing the health of organizations.
The concept of a healthy organization is idealistic. Organizations are never fully stable, they never maximize their potential, and they never achieve complete harmony. It is a common myth that the leader determines an organization's health. It is how the psychodynamics of an organization are managed and the capacity of leaders and followers to repair broken relationships that influence an organization's health.


OCTN1 (SLC22A4) displays two different transport pathways for organic cations or zwitterions.
OCTN1 belongs to the SLC22 family, which includes transporters for cationic, zwitterionic, and anionic substrates. OCTN1 function and role in cells are still poorly understood. Not only cations, such as TEA, but also zwitterions, such as carnitine and ergothioneine, figure among transported molecules. In this work, we carried out transport assays measuring [14C]-TEA and [3H]-carnitine in proteoliposomes reconstituted with the recombinant human OCTN1 in the presence of Na+ or other cations. The homology model of OCTN1 was built using the structure of OCT3 as a template for docking analysis. TEA and carnitine did not inhibit each other. Moreover, carnitine uptake was not affected by the presence of Na+ and TEBA, whereas TEA was strongly inhibited by both compounds. Computational data revealed that TEA, Na+, and Carnitine can interact with E381 in the OCTN1 substrate site. Differently from TEA, in the presence of Na+, carnitine is still able to interact with the binding site via R469. The lack of mutual inhibition of the two prototype substrates, the different effect of Na+ and TEBA on their transport reaction, together with the computational analysis supports the existence of two transport pathways for cations and zwitterions. The results shed new light on the transport mechanisms of OCTN1, helping to get further insights into the structure/function relationships. The described results correlate well with previous and very recent findings on the polyspecificity of the OCT group of transporters belonging to the same family.


Class II Young Adult Treatment with Twin Force Bite Corrector: 10-Year Follow-up.
One of the most preferred compliance free fixed functional appliances in nongrowing patients is Twin Force Bite corrector (TFBC). The aim of this case report is to evaluate the effectiveness of TFBC in the treatment of an adult Class II case. A 16-year 1-month-old boy having skeletal and dental Class II relationship was selected. Roth 0.018 × 0.025 inch slots brackets were attached; a fixed lingual arch in the mandibular dental arch and a Nance appliance in the maxillary dental arch were used to increase anchorage. The TFBC therapy used for sagittal activation and stimulation of forward mandibular growth lasted for 3 months. The post-TFBC treatment lasted 6 months and the total treatment time was 9 months. Treatment of a young adult Class II malocclusion with TFBC resulted in a Class I molar occlusion, an ideal overjet, overbite, and incisor angulation in a short time and maintained in the 10-year follow-up.


Intraluminal Arterial Filling Defects Misdiagnosed as Pulmonary Emboli: What Else Could They Be?
Pulmonary artery filling defects can be observed in various pathologic processes other than pulmonary embolism, for example, nonthrombotic pulmonary embolism with biological and nonbiological materials and intrinsic pulmonary artery lesions. They have also been described in rare conditions, such as fibrosing mediastinitis and congenital absence or stenosis of pulmonary artery, and some pulmonary parenchymal and airway malignancies. Misdiagnosis is common owing to the relative rarity of these conditions. Correct diagnosis is based on the appropriate clinical suspicion considering the unique clinical features, laboratory findings, and additional radiologic clues inferring a pathology other than pulmonary embolism.


Development of the corpus callosum in childhood, adolescence and early adulthood.
The corpus callosum (CC) is the major commissure connecting the cerebral hemispheres and there is evidence of its continuing development into young adulthood [Ann. Neurol. 34 (1993) 71]. Yet, little is known about changes in the size and tissue characteristics of its sub-regions. The sub-regions of the CC (genu, body, isthmus and splenium) are topographically organized to carry interhemispheric fibres representing heteromodal and unimodal cortical brain regions. Studies of the development of each of these sub-regions can therefore provide insights into the time course of brain development. We assessed age-related changes in the size and the signal intensities (SI) of the subregions of the corpus callosum in the Magnetic Resonance Imaging (MRI) scans of a cross-sectional sample of 109 healthy young individuals aged 7-32 years. Age was significantly positively correlated with the size of the callosal sub-regions (with the exception of the isthmus). On the other hand, there was an age-related decrease in SI across all the CC sub-regions. The rates of CC regional size increases appeared to be most pronounced in childhood. By contrast, SI decreases occurred during childhood and adolescence but reached an asymptote during young adulthood. Finally, the observed size and SI changes were similar across CC sub-regions. The observed increases in CC size in conjunction with the decreases in signal intensity reflect continued maturation of the structure from childhood through young adulthood. An increase in axonal size may underlie growth in the size of the CC during childhood. The continued decrease in the CC signal intensity during adolescence may in addition be related to ongoing maturation of the axonal cytoskeleton. CC maturational changes appeared synchronous across sub-regions suggesting parallel maturation of diverse brain regions during childhood and adolescence.


Recurrence of subglandular breast implant capsular contracture: anterior versus total capsulectomy.
The objective of this study was to determine whether the type of capsulectomy, anterior or total, affects the recurrence of capsular contracture around subglandular silicone-gel breast implants. A retrospective analysis was performed of patients who underwent either anterior or total capsulectomy for Baker grade 3 or 4 subglandular capsular contracture in our unit. All patients were invited to a review clinic where their capsular status was assessed. There were 100 anterior- disc capsulectomies in 60 patients between 1988 and 1997 and 99 total capsulectomies in 60 patients between 1990 and 1998. The follow-up in the former group was a median of 7 years and mean 6.9 years, compared with median 2.5 and mean 3.1 years in the latter group. Eighty-six percent of the implants removed from both groups at capsulectomy were smooth-walled gel-filled implants. Sixty-nine breasts in the anterior group received textured gel implants at capsulectomy; the remaining 31 received polyurethane-coated Meme implants. In the total capsulectomy group, all but two breasts (one patient) received textured gel implants. After review, the capsular status was known in 80 percent of the anterior and 92 percent of the total capsulectomy group. The review clinic found eight new contractures in five patients to have developed in the anterior compared with none in the total group. Recurrent contractures affected 50 percent of patients (46 percent of breasts) in the anterior and 11 percent of patients (10 percent of breasts) in the total capsulectomy group. Kaplan-Meier survival analysis was applied to the data. By including only patients who received textured gel implants at capsulectomy, the Logrank found a statistical difference between the two treatment groups (0.01 < p < 0.5). We believe that this study provides some evidence that total capsulectomy for subglandular silicone breast implant capsular contracture results in a lower capsular recurrence than anterior- disc capsulectomy. The pattern and risk of recurrence after total capsulectomy and exchange for a modern textured prosthesis appear to approach those following primary augmentation.


Structure of the type III pantothenate kinase from Bacillus anthracis at 2.0 A resolution: implications for coenzyme A-dependent redox biology.
Coenzyme A (CoASH) is the major low-molecular weight thiol in Staphylococcus aureus and a number of other bacteria; the crystal structure of the S. aureus coenzyme A-disulfide reductase (CoADR), which maintains the reduced intracellular state of CoASH, has recently been reported [Mallett, T.C., Wallen, J.R., Karplus, P.A., Sakai, H., Tsukihara, T., and Claiborne, A. (2006) Biochemistry 45, 11278-89]. In this report we demonstrate that CoASH is the major thiol in Bacillus anthracis; a bioinformatics analysis indicates that three of the four proteins responsible for the conversion of pantothenate (Pan) to CoASH in Escherichia coli are conserved in B. anthracis. In contrast, a novel type III pantothenate kinase (PanK) catalyzes the first committed step in the biosynthetic pathway in B. anthracis; unlike the E. coli type I PanK, this enzyme is not subject to feedback inhibition by CoASH. The crystal structure of B. anthracis PanK (BaPanK), solved using multiwavelength anomalous dispersion data and refined at a resolution of 2.0 A, demonstrates that BaPanK is a new member of the Acetate and Sugar Kinase/Hsc70/Actin (ASKHA) superfamily. The Pan and ATP substrates have been modeled into the active-site cleft; in addition to providing a clear rationale for the absence of CoASH inhibition, analysis of the Pan-binding pocket has led to the development of two new structure-based motifs (the PAN and INTERFACE motifs). Our analyses also suggest that the type III PanK in the spore-forming B. anthracis plays an essential role in the novel thiol/disulfide redox biology of this category A biodefense pathogen.


Clozapine pharmacokinetics and pharmacodynamics studied with Cyp1A2-null mice.
The aim of this study was to use the CYP1A2-null mouse to investigate the in-vivo contribution of CYP1A2 to clozapine pharmacokinetics and pharmacodynamics. An intraperitoneal injection of 10 mg/kg clozapine was administered to four male CYP1A2 -/- mice and four male wild-type mice. Clozapine, desmethylclozapine, and clozapine N-oxide concentrations in sequential tail blood samples were measured by HPLC with UV detection. Behavioural parameters were recorded at each time point. The area under the curve (AUC) of clozapine was 2.6 times greater, the clearance of clozapine was 2.6 times slower, and the half-life was 1.2 times longer in the CYP1A2 -/- mice (p = 0.0143) as compared to the wild-type mice. Sixty-one percent of the clozapine clearance in wild-type mice was calculated to be mediated by CYP1A2. The AUC of desmethylclozapine was 1.6 times lower in the CYP1A2 -/- mice compared to the wild-type mice (p = 0.0286), while there was a trend for the AUC of clozapine N-oxide to be greater in the CYP1A2 -/- mice (p = 0.0571). The CYP1A2 -/- mice were significantly more drowsy and showed more motor impairment (p = 0.0145) and myoclonus than the wild-type mice. Our results indicate that, in vivo, CYP1A2 is the major determinant of clozapine clearance, contributes significantly to the demethylation of clozapine, and has a negligible contribution to the N-oxidation. Our data also indicate that CYP1A2 poor metabolizers might be more susceptible than extensive metabolizers to dose-related adverse effects of clozapine, such as sedation, myoclonus and seizures.


Anatomical and behavioral comparison of unilateral dopamine-rich grafts implanted into the striatum of neonatal and adult rats.
The anatomical and functional characteristics of dopamine neuron-rich grafts implanted into rat pups were compared with those of identical grafts implanted into adult hosts. The host nigrostriatal dopaminergic pathway was unilaterally destroyed by an intrahypothalamic injection of 6-hydroxydopamine. This was followed five days later by the implantation of a cellular suspension obtained from rat embryonic mesencephali. Identical operations were carried out on adult and infant (PD3) rats. The survival rate of implanted tyrosine hydroxylase-positive cells was lower in the neonates. On the other hand, in the neonate hosts, surviving immunoreactive cells migrated extensively throughout the host striatum coursing preferentially below the corpus callosum and towards the subependymal zone. The structural integrity of the host parenchyma was well maintained after the neonatal implantation, in contrast to that observed in the adults. Despite a difference in the cell survival rate, there was no major difference in reinnervation density between the two types of host. The functional capacities of the implants were evaluated by measuring the rotational responses of the animals to dopaminergic agonists. The implants compensated the lesion-induced contralateral rotational response to the mixed agonist apomorphine and the D1 agonist SCH-38393 in both neonates and adults. However, the response to the D2 agonist LY-171555 was not significantly attenuated by the implant. The ipsilateral rotational response to amphetamine observed in the lesioned animals was also compensated and even reversed by the graft. It is concluded that with respect to rotational behavior, similar functional benefits were observed following adult stage or neonatal implantation, despite differences in their anatomical development.


Analgesic treatment after laparoscopic cholecystectomy: a critical assessment of the evidence.
Acute pain after laparoscopic cholecystectomy is complex in nature. The pain pattern does not resemble pain after other laparoscopic procedures, suggesting that analgesic treatment might be procedure specific and multimodal. Randomized trials of analgesia after laparoscopic cholecystectomy were identified by systematic electronic literature searches (1985 to June 2005) supplemented with manual searching. The trials were categorized by well-defined criteria into high, moderate, or poor methodologic quality. Conclusions were based on trials of high and moderate methodologic quality. In total, 64 randomized analgesic trials were identified, comprising a total of 5,018 evaluated patients. The literature suggests a multimodal analgesic regimen consisting of a preoperative single dose of dexamethasone, incisional local anesthetics (at the beginning or at the end of surgery, depending on preference), and continuous treatment with nonsteroidal antiinflammatory drugs (or cyclooxygenase-2 inhibitors) during the first 3-4 days. Opioids should be used only when other analgesic techniques fail.


Does multivariate analysis of post-thaw sperm characteristics accurately estimate in vitro fertility of boar individual ejaculates?
The objective of this study was to determine if a multivariate pattern analysis of frozen-thawed sperm characteristics of boar semen of unknown fertility, thus identifying groups of ejaculates as "good" or "bad" freezers, would estimate their fertilizing potential in an in vitro embryo production (IVP) system. Frozen-thawed spermatozoa from a single ejaculate collected from 46 boars were evaluated for sperm motility and kinematic patterns, for sperm viability and for early changes in sperm membrane stability. All data generated were used for a multivariate pattern analysis (PATN; CSIRO, Canberra, Australia) which objectively classified all ejaculates within a data set in to one of two groups, categorised as "good" (n = 25) or "bad" (n = 21) according with their freezability. In vitro matured oocytes were exposed to 2000 or 4000 frozen-thawed spermatozoa per oocyte for 6h and then cultured in embryo culture medium for either 6h (assurance of fertilization) or 7 days (to collect data on embryo development). Rates of sperm oocyte penetration and of embryo development significantly (p < 0.05) increased in a sperm:oocyte ratio-dependent manner. A similar pattern was observed when sperm characteristics were grouped. Indeed, ejaculates classified as "good" showed significantly (p < 0.05) higher rates of oocyte penetration, cleavage and of blastocyst formation than those classified as "bad". However, variation was still present among individuals (ejaculates, boars) in their ability to produce blastocysts in vitro. It is therefore concluded that despite the presence of a relationship for ejaculates with good semen quality post-thaw (thus grouped as "good") to higher IVP-results, the presence of individual variation does not allow for an accurate estimation of in vitro fertility based solely on the frozen-thawed semen quality parameters of a single ejaculate from a given boar.


Monte Carlo based protocol for cell survival and tumour control probability in BNCT.
A mathematical model to calculate the theoretical cell survival probability (nominally, the cell survival fraction) is developed to evaluate preclinical treatment conditions for boron neutron capture therapy (BNCT). A treatment condition is characterized by the neutron beam spectra, single or bilateral exposure, and the choice of boron carrier drug (boronophenylalanine (BPA) or boron sulfhydryl hydride (BSH)). The cell survival probability defined from Poisson statistics is expressed with the cell-killing yield, the 10B(n,alpha)7Li reaction density, and the tolerable neutron fluence. The radiation transport calculation from the neutron source to tumours is carried out using Monte Carlo methods: (i) reactor-based BNCT facility modelling to yield the neutron beam library at an irradiation port; (ii) dosimetry to limit the neutron fluence below a tolerance dose (10.5 Gy-Eq); (iii) calculation of the 10B(n,alpha)7Li reaction density in tumours. A shallow surface tumour could be effectively treated by single exposure producing an average cell survival probability of 10(-3)-10(-5) for probable ranges of the cell-killing yield for the two drugs, while a deep tumour will require bilateral exposure to achieve comparable cell kills at depth. With very pure epithermal beams eliminating thermal, low epithermal and fast neutrons, the cell survival can be decreased by factors of 2-10 compared with the unmodified neutron spectrum. A dominant effect of cell-killing yield on tumour cell survival demonstrates the importance of choice of boron carrier drug. However, these calculations do not indicate an unambiguous preference for one drug, due to the large overlap of tumour cell survival in the probable ranges of the cell-killing yield for the two drugs. The cell survival value averaged over a bulky tumour volume is used to predict the overall BNCT therapeutic efficacy, using a simple model of tumour control probability (TCP).


Motion and shape in common fate.
We determined how much motion coherence was needed to detect a target group of four moving dots in a dynamic visual noise (DVN) background. The lifetimes of the trajectories of the target and that of the noise dots were the same. In addition to parallel trajectories and collinear dot arrangements, divergent, convergent, or crossing trajectories and non-collinear dot arrangements were also tested. Performance saturated at a lifetime of approximately 600 ms. It was best for parallel trajectories and collinear dots, and worse for crossed trajectories with non-collinear dots, where it approached performance in a no-motion, form-only control experiment. Results illustrate the importance of common fate in motion perception in DVN, when other factors are equated.


[Psychophysiological factors and their role in assessing the work capacity of arterial hypertension patients].
The homeostatic and information tests were used for the first time ever in addition to the MMPI method to evaluate the effects of psychophysiological factors on working capacity of 250 patients with arterial hypertensions and 127 normal subjects. Optimum performance in the solving of a homeostatic search problem was shown to be twice as rare in hypertensive patients, as compared to the normal subjects. A clear-cut relationship was established between the quality of information processing as evidenced by the information test, and the patients' psychological characteristics, BP values and the extent of cerebral hemodynamic changes. An index capable of quantitative characterization of the working capacity of hypertensive patients is proposed.


Effect of Bacillus Calmette-Guérin infection on delayed footpad reaction to Listeria monocytogenes.
The role of peritoneal macrophages induced by Bacillus Calmette-Guérin (BCG) in the induction of immune responses to Listeria monocytogenes was studied in mice. The peritoneal macrophages from mice treated with BCG 14 days previously contained a high proportion of Ia-bearing macrophages (approximately 56%) and the cells showed not only a high level of listericidal activity but also a strong ability for presentation of listerial antigen to Listeria-immune T cells. An intraperitoneal inoculation with a low dose of Listeria, which can induce the maximal level of delayed footpad reaction (DFR) and positive migration inhibitory activity of macrophages in untreated mice, did not induce a detectable level of such responses in BCG-treated mice. The bacterial growth at an early stage of infection was suppressed by scavenger macrophages in these mice. On the other hand, BCG-treated mice showed the early development of DFR and macrophage migration inhibitory activity after an inoculation with a high dose of Listeria. It is revealed in transfer experiments that Listeria-pulsed peritoneal exudate cells induced by BCG elicited the highest level of DFR and positive migration inhibition of macrophages in normal mice at the earlier period of injection compared with Listeria-pulsed resident peritoneal cells. These results suggested that the increased activities of macrophages acting as scavenger cells and as antigen-presenting cells play important roles in the modification of immune responses to Listeria in BCG-treated mice.


Sestrin2 aggravates oxidative stress of neurons by decreasing the expression of Nrf2.
Oxidative injury is an essential part of the pathological changes of cerebral ischemia-reperfusion. Sestrin2 (Sesn2), a conserved antioxidant protein, is activated under stress to protect cells against oxidative stress. This study mainly explored the function and mechanism of Sesn2 during cerebral ischemia-reperfusion injury in rats. Oxygen-glucose deprivation/reoxygenation (OGD/R) model of primary neurons was established. MTS and LDH (lactate dehydrogenase) kit were used to detect cell viability and cell damage by colorimetric method. The superoxide dismutase (SOD) kit and malondialdehyde (MDA) kit were used to access the level of SOD and MDA in neurons. To establish a model of middle cerebral artery occlusion (MCAO) in adult male Sprague-Dawley (SD) rats, the process of cerebral ischemia-reperfusion injury in animals was simulated. Western blot and immunofluorescence were utilized to further examine the relationship between the expression of Sesn2 and Nrf2 (nuclear factor E2 related factor 2). Sesn2 aggravated neuronal damage and enhanced cell viability in OGD/R model. Intraventricular injection of Sesn2 siRNA lentivirus aggravated nerve function damage in MCAO model, increased cerebral infarction area and water content. Sesn2 overexpression resulted in the increase of total, nuclear levels of Nrf2, as well as the downstream proteins of Nrf2, sulfiredoxin1 (Srx1) and thioredoxin1 (Trx1). On the contrary, after knockdown of Sesn2, we obtained the opposite result. Knockdown of Sesn2 reduced Nrf2, Srx1 and Trx1 levels in rat cerebral cortex in MCAO model. Sesn2 promoted the transfer of nuclear Nrf2 to the cytoplasm, it decreased the expressions of Nrf2 and its downstream proteins, Srx1 and Trx1. Meanwhile, it increased the cerebral ischemia-reperfusion injury by changing the distribution of Nrf2.


Assessment of walking features from foot inertial sensing.
An ambulatory monitoring system is developed for the estimation of spatio-temporal gait parameters. The inertial measurement unit embedded in the system is composed of one biaxial accelerometer and one rate gyroscope, and it reconstructs the sagittal trajectory of a sensed point on the instep of the foot. A gait phase segmentation procedure is devised to determine temporal gait parameters, including stride time and relative stance; the procedure allows to define the time intervals needed for carrying an efficient implementation of the strapdown integration, which allows to estimate stride length, walking speed, and incline. The measurement accuracy of walking speed and inclines assessments is evaluated by experiments carried on adult healthy subjects walking on a motorized treadmill. Root-mean-square errors less than 0.18 km/h (speed) and 1.52% (incline) are obtained for tested speeds and inclines varying in the intervals [3, 6] km/h and [-5, + 15]%, respectively. Based on the results of these experiments, it is concluded that foot inertial sensing is a promising tool for the reliable identification of subsequent gait cycles and the accurate assessment of walking speed and incline.


Targeting Mitochondrial Dysfunction and Oxidative Stress in Activated Microglia using Dendrimer-Based Therapeutics.
Mitochondrial oxidative stress is associated with many neurodegenerative diseases, such as traumatic brain injury (TBI). Targeted delivery of antioxidants to mitochondria has failed to translate into clinical success due to their nonspecific cellular localization, poor transport properties across multiple biological barriers, and associated side effects. These challenges, coupled with the complex function of the mitochondria, create the need for innovative delivery strategies. Methods: Neutral hydroxyl-terminated polyamidoamine (PAMAM) dendrimers have shown significant potential as nanocarriers in multiple brain injury models. N-acetyl cysteine (NAC) is a clinically used antioxidant and anti-inflammatory agent which has shown significant potency when delivered in a targeted manner. Here we present a mitochondrial targeting hydroxyl PAMAM dendrimer-drug construct (TPP-D-NAC) with triphenyl-phosphonium (TPP) for mitochondrial targeting and NAC for targeted delivery to mitochondria in injured glia. Co-localization and mitochondrial content of mitochondria-targeted and unmodified dendrimer were assessed in microglia and macrophages in vitro via immunohistochemistry and fluorescence quantification. Therapeutic improvements of TPP-D-NAC over dendrimer-NAC conjugate (D-NAC) and free NAC were evaluated in vitro in microglia under oxidative stress challenge. In vivo neuroinflammation targeting was confirmed in a rabbit model of TBI. Results: TPP-conjugated dendrimer co-localized significantly more with mitochondria than unmodified dendrimer without altering overall levels of cellular internalization. This targeting capability translated to significant improvements in the attenuation of oxidative stress by TPP-D-NAC compared to D-NAC and free NAC. Upon systemic administration in a rabbit TBI model, TPP-conjugated dendrimer co-localized specifically with mitochondria in activated microglia and macrophages in the white matter of the ipsilateral/injured hemisphere, confirming its BBB penetration and glial targeting capabilities. Conclusion: D-NAC has shown promising efficacy in many animal models of neurodegeneration, and this work provides evidence that modification for mitochondrial targeting can further enhance its therapeutic efficacy, particularly in diseases where oxidative stress-induced glial cell death plays a significant role in disease progression.


Increased apoptosis of peripheral blood T cells following allogeneic hematopoietic cell transplantation.
Lymphopenia and immune deficiency are significant problems following allogeneic hematopoietic cell transplantation (HCT). It is largely assumed that delayed immune reconstruction is due to a profound decrease in thymus-dependent lymphopoiesis, especially in older patients, but apoptosis is also known to play a significant role in lymphocyte homeostasis. Peripheral T cells from patients who received HCT were studied for evidence of increased cell death. Spontaneous apoptosis was measured in CD3(+) T cells following a 24-hour incubation using 7-amino-actinomycin D in conjunction with the dual staining of cell surface antigens. Apoptosis was significantly greater among CD3(+) T cells taken from patients 19-23 days after transplantation (30.4% +/- 12.5%, P <.05), and 1 year after transplantation (9.7% +/- 2.8%, P <.05) compared with healthy controls (4.0% +/- 1.5%). Increased apoptosis occurred preferentially in HLA (human leukocyte antigen)-DR positive cells and in both CD3(+)/CD4(+) and CD3(+)/CD8(+) T-cell subsets, while CD56(+)/CD3(-) natural killer cells were relatively resistant to apoptosis. The extent of CD4(+) T-cell apoptosis was greater in patients with grade II-IV acute graft-versus-host disease (GVHD) (33. 9% +/- 11.3%) compared with grade 0-I GVHD (14.6 +/- 6.5%, P <.05). T-cell apoptosis was also greater in patients who received transplantations from HLA-mismatched donors (39.5% +/- 10.4%, P <. 05) or HLA-matched unrelated donors (32.1% +/- 11.4%, P <.05) compared with patients who received transplantations from HLA-identical siblings (19.6% +/- 6.7%). The intensity of apoptosis among CD4(+) T cells was significantly correlated with a lower CD4(+) T-cell count. Together, these observations suggest that activation of T cells in vivo, presumably by alloantigens, predisposes the cells to spontaneous apoptosis, and this phenomenon is associated with lymphopenia. Activation-induced T-cell apoptosis may contribute to delayed immune reconstitution following HCT. (Blood. 2000;95:3832-3839)


Experimental enterotoxin-induced Escherichia coli diarrhea and protection induced by previous infection with bacteria of the same adhesin or enterotoxin type.
The diarrheal response to an initial and a second infection with Escherichia coli expressing various enterotoxins (the heat-stable toxin [ST] alone or in combination with the heat-labile toxin [LT]) and colonization factor antigens (CFA/I, CFA/II, or E8775-type) was studied in the reversible tie adult rabbit diarrhea model. An initial infection with high doses (1 X 10(10) to 5 X 10(11) bacteria) of the various strains regularly induced diarrhea which was usually self-limiting (only 7 of 85 animals died). The diarrheal response to equally effective doses of different strains producing both ST and LT (ST/LT) did not differ significantly with serotype or colonization factor antigen. ST/LT-producing strains appeared to induce severe disease more regularly than ST-producing strains carrying the same adhesin. Previous infection with CFA/I-carrying, ST/LT-producing E. coli protected all animals reinfected with an otherwise highly diarrheogenic dose of the same strain as well as against challenge with a CFA/I-carrying, ST/LT-producing strain with different O-, K-, and H-antigens. Fecal excretion of bacteria was also significantly reduced in the protected animals, although not completely eliminated. When only one of the two antigens, CFA/I and LT, was shared by the immunizing and rechallenge strains, partial protection was evident consistent with independent antibacterial (anti-CFA) and antitoxic (anti-LT) immune mechanisms. Oral immunization with purified CFA/I significantly reduced fluid secretion in intestinal loops infected with CFA/I-carrying enterotoxigenic bacteria.


RASA4 undergoes DNA hypermethylation in resistant juvenile myelomonocytic leukemia.
Aberrant DNA methylation at specific genetic loci is a key molecular feature of juvenile myelomonocytic leukemia (JMML) with poor prognosis. Using quantitative high-resolution mass spectrometry, we identified RASA4 isoform 2, which maps to chromosome 7 and encodes a member of the GAP1 family of GTPase-activating proteins for small G proteins, as a recurrent target of isoform-specific DNA hypermethylation in JMML (51% of 125 patients analyzed). RASA4 isoform 2 promoter methylation correlated with clinical parameters predicting poor prognosis (older age, elevated fetal hemoglobin), with higher risk of relapse after hematopoietic stem cell transplantation, and with PTPN11 mutation. The level of isoform 2 methylation increased in relapsed cases after transplantation. Interestingly, most JMML cases with monosomy 7 exhibited hypermethylation on the remaining RASA4 allele. The results corroborate the significance of epigenetic modifications in the phenotype of aggressive JMML.


In vitro and in vivo studies on ocular vitamin A palmitate cationic liposomal in situ gels.
N-trimethyl chitosan (TMC) with different degree of quaternization (DQ) as the coating materials, vitamin A palmitate (VAP)-loaded cationic liposomes dispersed in thermo-sensitive in situ gels (ISG) with poloxamer 407 (P407) as the base were prepared in this study. VAP-loaded liposomes (VAPL) were prepared using a film dispersion method followed by TMC-coating with DQ of 20%, 40% and 60% (TMC20, TMC40 and TMC60), respectively, then dispersed in P407 solution to obtain TMC-coated VAPL ISG. The in vitro properties of the system including morphology, size, zeta potential, entrapment efficiency, drug release and ocular retention were investigated. The ocular retention in vivo, eye irritation and pharmacokinetics in aqueous humor of the system were also studied in rabbits. VAPL revealed a spherical surface with mean size below 100 nm and negative zeta potential. After TMC-coating, the morphology and entrapment efficiency showed no significant changes, while the mean size was increased, zeta potential was changed into positive, and drug release was further sustained, and above all was influenced by DQ of TMC. TMC-coated VAPL exhibited little effect on the gelation temperature of P407 solution, and at the P407 concentration of 25% (w/v), TMC-coated VAPL ISG had the gelation temperature closest to the eye surface (34 °C) after diluted by the artificial tears. Compared with uncoated VAPL ISG and the marketed Oculotect gels, TMC-coated VAPL ISG displayed more retarded drug release and gel corrosion with a good linear relationship between them, and the higher DQ was, the slower drug release and gel corrosion. The ocular retention time in vitro and in vivo of TMC-coated VAPL ISG were both notable prolonged with a positive correlation with DQ of TMC. Compared with the marketed gels, TMC60-coated VAPL ISG showed delayed Tmax, improved Cmax and AUC(0–24) in rabbit aqueous humor, suggesting the sustained drug release and better corneal penetration and absorption. The local irritation of TMC-coated VAPL ISG was proved to be negligible. TMC-coated VAPL ISG with the properties of sustained drug release, prolonged ocular retention, improved corneal penetration and promising bio-safety is valuable to be studied further.


Focusing characteristics of a planar solid-immersion mirror.
The focusing characteristics of a planar waveguide solid-immersion mirror with parabolic design have been investigated. The solid-immersion mirror is integrated into an optical waveguide, and light focusing is achieved with a parabolic mirror parallel to the waveguide plane and waveguide mode confinement normal to the waveguide plane. Optical-quality tantala silica planar waveguides can be obtained by evaporation. The parabolic sidewall reflects over 50% of the incident waveguide mode and generates a diffraction-limited focus. The measured spot size for the solid-immersion mirror described here is less than one third of the wavelength. Polarization analysis shows that the electric field near the focal region has components parallel and normal to the polarization state of the incident beam. The planar solid-immersion mirror is essentially free of chromatic aberration, and the alignment of the illumination beam is within a fraction of degrees.


[Rhabdomyolysis syndrome and hyperthermia secondary to cocaine and/or ecstasy consumption. Report of 2 new cases and review of the problem].
We describe two cases of hyperthermia and rhabdomyolysis related to cocaine and/or ecstasy consumption. The clinical and laboratory findings are characteristic of hyperthermia syndrome in both cases. The first patient consumed cocaine, ecstasy and alcohol and afterwards suffers from hyperthermia and rhabdomyolysis associated with decreased conscious level, labile blood pressure, severe renal insufficiency (needing hemodialysis) and important liver failure. The second patient consumed ecstasy and alcohol with intense physical activity and shows the same but moderate clinical picture rapidly responding to intravenous fluid therapy. The incidence of this drug-induced hyperthermia is low compared to the important use of these illicit drugs. Nevertheless its increasing consumption will lead physicians to attend more patients with this syndrome and other related complications. For this reason we have reviewed and up-dated this problematic subject.


Terbinafine pharmacokinetics after single dose oral administration in the dog.
Terbinafine is an allylamine antifungal prescribed for the treatment of mycoses in humans. It is increasingly being used in veterinary patients. The purpose of this study was to evaluate the pharmacokinetic properties of terbinafine in dogs after a single oral dose. Ten healthy adult dogs were included in the study. A single dose of terbinafine (30-35 mg/kg) was administered orally, and blood samples were periodically collected over a 24 h period during which dogs were monitored for adverse effects. Two of 10 dogs developed transient ocular changes. A high-performance liquid chromatography assay was developed and used to determine plasma terbinafine concentrations. Pharmacokinetic analysis was performed using PK Solutions(®) computer software. Area under the curve (AUC) from time 0 to 24 h was 15.4 μg·h/mL (range 5-27), maximal plasma concentration (C(max) ) was 3.5 μg/mL (range 3-4.9 μg/mL) and time to C(max) (T(max) ) was 3.6 h (range 2-6 h). The time above minimal inhibitory concentration (T > MIC) as well as AUC/MIC was calculated for important invasive fungal pathogens and dermatophytes. The T > MIC was 17-18 h for Blastomyces dermatitidis, Histoplasma capsulatum and dermatophytes (Microsporum spp. and Trichophyton mentagrophytes), while the MIC for Sporothrix schenckii and Coccidioides immitis was exceeded for 9.5-11 h. The AUC/MIC values ranged from 9 to 13 μg h/mL for these fungi. Our results provide evidence supporting the use of terbinafine as an oral therapeutic agent for treating systemic and subcutaneous mycoses in dogs.


Gut Microbiota Composition Associated With Clostridium difficile-Positive Diarrhea and C. difficile Type in ICU Patients.
The gut microbiota composition of intensive care unit (ICU) patients suffering from Clostridium difficile-positive diarrhea (CDpD) is poorly understood. This prospective study aims to use 16S rDNA (and metagenome) sequencing to compare the microbiota composition of 58 (and 5) ICU patients with CDpD (CDpD group), 33 (and 4) ICU patients with C. difficile-negative diarrhea (CDnD group), and 21 (and 5) healthy control subjects (control group), as well as CDpD patients in the A+B+ (N = 34; A/B: C. difficile TcdA/B), A-B+ (N = 7), and A-B- (N = 17) subgroups. For 16S rDNA data, OTU clustering (tool: UPARSE), taxonomic assignment (tool: RDP classifier), α-diversity, and β-diversity analyses (tool: QIIME) were conducted. For metagenome data, metagenome assembly (tool: SOAPdenovo), gene calling (tools: MetaGeneMark, CD-HIT, and SoapAligner), unigene alignment (tool: DIAMOND), taxon difference analysis (tool: Metastats), and gene annotation (tool: DIAMOND) were performed. The microbial diversity of the CDpD group was lower than that of the CDnD and control groups. The abundances of 10 taxa (e.g., Deferribacteres, Cryptomycota, Acetothermia) were significantly higher in the CDpD group than in the CDnD group. The abundances of Saccharomycetes and Clostridia were significantly lower in CDpD in comparison with control. Some taxa were significantly different between the A+B+ and A-B- subgroups. CDpD might relate to a decrease in beneficial taxa (i.e., Saccharomycetes and Clostridia) and an increase in harmful taxa (e.g., Deferribacteres, Cryptomycota, Acetothermia) in gut microbiota of ICU patients. C. difficile toxin type might be slightly associated with gut microbiota composition.


Genetic and biochemical characterization of periplasmic-leaky mutants of Escherichia coli K-12.
Periplasmic-leaky mutants of Escherichia coli K-12 were isolated after nitrosoguanidine-induced mutagenesis. They released periplasmic enzymes into the extracellular medium. Excretion of alkaline phosphatase, which started immediately in the early exponential phase of growth, could reach up to 90% of the total enzyme production in the stationary phase. Leaky mutants were sensitive to ethylenediaminetetraacetic acid, cholic acid, and the antibiotics rifampin, chloramphenicol, mitomycin C, and ampicillin. Furthermore, they were resistant to colicin E1 and partially resistant to phage TuLa. Their genetic characterization showed that the lky mutations mapped between the suc and gal markers, near or in the tolPAB locus. A biochemical analysis of cell envelope components showed that periplasmic-leaky mutants contained reduced amounts of major outer membrane protein OmpF and increased amounts of a 16,000-dalton outer membrane protein.


Global trends and predictors of face mask usage during the COVID-19 pandemic.
Guidelines and recommendations from public health authorities related to face masks have been essential in containing the COVID-19 pandemic. We assessed the prevalence and correlates of mask usage during the pandemic. We examined a total of 13,723,810 responses to a daily cross-sectional online survey in 38 countries of people who completed from April 23, 2020 to October 31, 2020 and reported having been in public at least once during the last 7 days. The outcome was individual face mask usage in public settings, and the predictors were country fixed effects, country-level mask policy stringency, calendar time, individual sociodemographic factors, and health prevention behaviors. Associations were modeled using survey-weighted multivariable logistic regression. Mask-wearing varied over time and across the 38 countries. While some countries consistently showed high prevalence throughout, in other countries mask usage increased gradually, and a few other countries remained at low prevalence. Controlling for time and country fixed effects, sociodemographic factors (older age, female gender, education, urbanicity) and stricter mask-related policies were significantly associated with higher mask usage in public settings. Crucially, social behaviors considered risky in the context of the pandemic (going out to large events, restaurants, shopping centers, and socializing outside of the household) were associated with lower mask use. The decision to wear a face mask in public settings is significantly associated with sociodemographic factors, risky social behaviors, and mask policies. This has important implications for health prevention policies and messaging, including the potential need for more targeted policy and messaging design.


Gene expression in the developing embryo and fetus.
Determining the stage- and tissue-specific patterns of gene expression shown by the embryo and fetus will provide information about the control of normal development. Identification of alterations in these patterns associated with specific abnormal phenotypes will also be informative regarding the underlying molecular mechanisms. In addition, qualitative and quantitative changes in gene expression that deviate from the norm may provide a potential marker system for predicting future developmental defects, a system that would be particularly useful in preimplantation embryo technologies before recipient transfer. However, there are a number of important issues regarding the interpretation and relevance of many gene expression studies currently undertaken that are often not considered or are ignored. Even when rigorous methodology is applied to detect differences in gene expression, their functional significance is rarely defined. This review discusses the relevance of gene expression changes as diagnostic markers in relation to protein and epigenetic changes and indicates that gene expression studies should be rigorously designed and interpreted to yield meaningful results.


Control beliefs and engagement in hygienic and safety behaviours: the case of foodborne illness.
Foodborne illness is an ever-growing concern in public health. Studies found that conventional training is not enough to cause employees to apply the hygiene and safety measures. The present study explores control and fatalistic beliefs as potential factors for explaining engagement in preventive actions. Two-hundred and seventeen employees of a fast-food restaurant (75 % of all staff) answered a questionnaire assessing their control beliefs, fatalistic beliefs, risk perception, and engagement in hygienic and safety behaviours. The results validated our hypotheses. Control beliefs were positively related to engagement in hygienic and safety behaviours (b = 0.43, p < 0.001). The inverse relation was observed for fatalistic beliefs (b = - 0.24, p < 0.001). The perceived effectiveness of the prescribed measures seems to be the best predictor of engagement in preventive behaviours, followed by perceived self-efficacy. To increase adherence to preventive measures, it is recommended to enhance staff's self-efficacy and perceived effectiveness of these measures.


Amplitude of pupillary contraction as a function of intensity of illumination in schizophrenia.
When the amplitude of pupillary contraction of normal, healthy adults was binocularly recorded as a function of increasing intensity of light-stimulation, a linear relationship was found between the amplitude and log intensity. Schizophrenic patients deviated significantly from this systematic response pattern in two ways. One group of patients (35%) manifested the same pupillomotor threshold as the normals, however, as the intensity of light was systematically increased mean amplitude and rate of pupillary contraction fell below that evidenced by normals. Another group of patients (65%) were distinguished by an abnormal pupillomotor threshold, specifically requiring an increase of one log intensity unit for the elicitation of a recordable pupillary contraction. Moreover, as the light intensity increased, the mean amplitude of contraction was significantly attenuated at all intensities below that of the normals and of the schizophrenics who manifested a normal pupillomotor threshold. In addition, it was found that the average diameter of the dark-adapted pupil was smaller in both groups of patients as compared to the healthy adults, although there was no difference between the patient groups on the variable. As pupillary reactivity to stress in normals is characterized by an increase in the diameter of the dark-adapted pupil representing increased sympathetic outflow and by a reciprocal increase in the diameter of the light-adapted pupil representing increased supranuclear inhibition, the results of the study do not support the assertion that schizophrenics are hyperaroused. It is suggested schizophrenia may be considered a tonic autonomic dysfunctional state of mock-arousal characterized by either abnormally high levels of central supranuclear inhibition or by defective sympathetic outflow, or both.


Serum insulin-like growth factor 1 and late-life depression: a population-based study.
Serum insulin-like growth factor 1 (IGF-1) concentration decreases, while the prevalence of depressive symptoms increases with advancing age. Although basic research indicates a link between low IGF-1 concentration and depression, this has scarcely been investigated in humans. This study investigates whether lower IGF-1 concentrations are associated with prevalent and incident late-life depression over a 3-year period. The study included 1188 participants, aged ≥ 65 years, from the Longitudinal Aging Study Amsterdam (LASA), an ongoing, population-based cohort study. Depression was assessed at baseline and after three years using the Center for Epidemiological Studies-Depression Scale (CES-D) and the Diagnostic Interview Schedule (DIS), and categorized into minor depression and major depression (MDD). Serum IGF-1 concentration was determined at baseline. Associations were adjusted for relevant confounders. Serum IGF-1 concentrations were within the normal range (mean 13.9 nmol/l, standard deviation 5.3 nmol/l). At baseline, in men, as compared to high concentrations, mid concentrations decreased the probability of prevalent minor depression (odds ratio [OR] = 0.35, 95% confidence interval [CI] = 0.15-0.82). In women, as compared to high concentrations, low concentrations tended to increase the probability of prevalent MDD (OR = 2.66, 95% CI = 0.89-7.89). At three-year follow-up, in men, no significant prospective associations were detected. In women, as compared to high concentrations, mid concentrations decreased the probability of incident minor depression (OR = 0.43, 95% CI = 0.19-0.95). Several associations, which differed across the genders, were observed between IGF-1 and depression. Cross-sectional findings were not supported by longitudinal findings, which suggest that IGF-1 may not play an important predictive role in the development of depression in older persons over time. However, a more acute role of IGF-1 in current depression, as indicated by the cross-sectional results, may be possible. Further studies are needed to elucidate the complex relation between IGF-1 and late-life depression.