Enhanced transcription of rRNA genes by purified androgen receptor complexes in vitro.
Androgens stringently regulate the concentrations of the polyamines, spermine and spermidine, in rat ventral prostate. The intracellular distribution of the polyamines is also dependent on the androgenic milieu; a small proportion of polyamines is associated tightly with the nucleus. Soluble nucleolar chromatin contains RNA polymerase A and protein kinase activities, both of which are regulated by androgens and markedly stimulated by polyamines. It remains to be established whether the polyamines modulate these enzyme moieties directly, or whether the phosphorylation of other nucleolar proteins plays a crucial role in the androgenic regulation of rRNA synthesis. Using a protocol centered on affinity chromatography, the purification of the androgen receptor protein to a state approaching homogeneity has been accomplished. The purified receptor stimulates the RNA polymerase A and protein kinase activities associated with prostate nucleolar chromatin. The significance of these findings in the light of correct concepts of steroid hormone action is discussed.


Yeast Replicator: A High-Throughput Multiplexed Microfluidics Platform for Automated Measurements of Single-Cell Aging.
The yeast Saccharomyces cerevisiae is a model organism for replicative aging studies; however, conventional lifespan measurement platforms have several limitations. Here, we present a microfluidics platform that facilitates simultaneous lifespan and gene expression measurements of aging yeast cells. Our multiplexed high-throughput platform offers the capability to perform independent lifespan experiments using different yeast strains or growth media. Using this platform in minimal media environments containing glucose, we measured the full lifespan of individual yeast cells in wild-type and canonical gene deletion backgrounds. Compared to glucose, in galactose we observed a 16.8% decrease in replicative lifespan accompanied by an ∼2-fold increase in single-cell oxidative stress levels reported by PSOD1-mCherry. Using PGAL1-YFP to measure the activity of the bistable galactose network, we saw that OFF and ON cells are similar in their lifespan. Our work shows that aging cells are committed to a single phenotypic state throughout their lifespan.


Adverse associations between maternal deoxynivalenol exposure and birth outcomes: a prospective cohort study in China.
Deoxynivalenol (DON), one of the most prevalent mycotoxins, has been found to cause fetal growth retardation in animals. However, limited evidence exists regarding its effects on pregnant women. Maternal urinary concentration of total DON (tDON) and free DON (fDON) in the second trimester was measured using liquid chromatography with tandem mass spectrometry. Provisional daily intake (PDI) of DON was calculated based on tDON concentration. Linear and logistic regression models were used to evaluate the association between DON exposure levels and birth weight, birth length, and the risk of small for gestational age (SGA). Among 1538 subjects, the median concentrations of tDON and fDON were 12.1 ng/mL and 5.1 ng/mL, respectively. The PDI values revealed that the median DON intake was 0.7 µg/kg bw, and 35.9% of the total population exceeded the provisional maximum tolerable daily intake (PMTDI) of 1 µg/kg bw. Compared with the lowest tertile, birth weight decreased by 81.11 g (95% CI: -127.00, -35.23) for tDON (P-trend < 0.001) and 63.02 g (95% CI: -108.72, -17.32) for fDON (P-trend = 0.004) in the highest tertile. Each unit increase in Ln-tDON and Ln-fDON was also inversely associated with birth weight. Furthermore, compared to those who did not exceed PMTDI, pregnant women whose PDI exceeded PMTDI had lower birth weight (β = -79.79 g; 95% CI: -119.09, -40.49) and birth length (β = -0.21 cm; 95% CI: -0.34, -0.07), and a higher risk of SGA (OR = 1.48; 95% CI: 1.02, 2.15) in their offspring. Similar associations with birth weight, birth length, and SGA were found when comparing the highest tertile of PDI to the lowest tertile (all P-trend < 0.05). Maternal DON exposure is related to decreased birth weight. Our findings implicate that DON exposure during pregnancy may cause fetal growth faltering, and measures should be taken to reduce DON exposure in pregnant women.


Reliability of the clinical teaching effectiveness instrument.
The Clinical Teaching Effectiveness Instrument (CTEI) was developed to evaluate the quality of the clinical teaching of educators. Its authors reported evidence supporting content and criterion validity and found favourable reliability findings. We tested the validity and reliability of this instrument in a European context and investigated its reliability as an instrument to evaluate the quality of clinical teaching at group level rather than at the level of the individual teacher. Students participating in a surgical clerkship were asked to fill in a questionnaire reflecting a student-teacher encounter with a staff member or a resident. We calculated variance components using the urgenova program. For individual score interpretation of the quality of clinical teaching the standard error of estimate was calculated. For group interpretation we calculated the root mean square error. The results did not differ statistically between staff and residents. The average score was 3.42. The largest variance component was associated with rater variance. For individual score interpretation a reliability of > 0.80 was reached with 7 ratings or more. To reach reliable outcomes at group level, 15 educators or more were needed with a single rater per educator. The required sample size for appraisal of individual teaching is easily achievable. Reliable findings can also be obtained at group level with a feasible sample size. The results provide additional evidence of the reliability of the CTEI in undergraduate medical education in a European setting. The results also showed that the instrument can be used to measure the quality of teaching at group level.


Origin of cyanobacterial thylakoids via a non-vesicular glycolipid phase transition and their impact on the Great Oxygenation Event.
The appearance of oxygenic photosynthesis in cyanobacteria is a major event in evolution. It had an irreversible impact on the Earth, promoting the Great Oxygenation Event (GOE) ~2.4 billion years ago. Ancient cyanobacteria predating the GOE were Gloeobacter-type cells lacking thylakoids, which hosted photosystems in their cytoplasmic membrane. The driver of the GOE was proposed to be the transition from unicellular to filamentous cyanobacteria. However, the appearance of thylakoids expanded the photosynthetic surface to such an extent that it introduced a multiplier effect, which would be more coherent with an impact on the atmosphere. Primitive thylakoids self-organize as concentric parietal uninterrupted multilayers. There is no robust evidence for an origin of thylakoids via a vesicular-based scenario. This review reports studies supporting that hexagonal II-forming glucolipids and galactolipids at the periphery of the cytosolic membrane could be turned, within nanoseconds and without any external source of energy, into membrane multilayers. Comparison of lipid biosynthetic pathways shows that ancient cyanobacteria contained only one anionic lamellar-forming lipid, phosphatidylglycerol. The acquisition of sulfoquinovosyldiacylglycerol biosynthesis correlates with thylakoid emergence, possibly enabling sufficient provision of anionic lipids to trigger a hexagonal II-to-lamellar phase transition. With this non-vesicular lipid-phase transition, a framework is also available to re-examine the role of companion proteins in thylakoid biogenesis.


Loss of heterozygosity (LOH) at 17q and 14q in human lung cancers.
Our recent linkage study of urethane-induced pulmonary adenomas in SMXA RI strains of mouse revealed two host resistance genes, Par1 (chromosome 11) and Par3 (chromosome 12). The map positions of Par1 and Par3 correspond to human 17q11-23 and 14q11-24, based on synteny between mouse and human. In this study, we examined the loss of heterozygosity (LOH) in these two homologous human chromosomal regions in 30 primary lung adenocarcinoma samples with matched normal DNA. Using 15 highly polymorphic markers, two commonly deleted regions were identified on human chromosomes 14 and 17, respectively. At 17q21, nine (53%) of 17 informative tumors showed LOH between D17S588 and D17S518. On the other hand, at 14q11-12, seven (32%) of 22 informative tumors showed LOH at loci between D14S261 and D14S80. Subsequently, we examined 25 squamous cell carcinomas (SQ) and 24 small cell carcinomas (SCC). At 14q11-12, six (38%) of 16 informative SQ and five (42%) of 12 informative SCC showed LOH. In contrast, at 17q11-23, one (7%) of 15 informative SQ and two (14%) of 14 SCC showed LOH. Therefore, the gene on 17q seemed to affect selectively adenocarcinomas, whereas the other gene on 14q, all three types of lung carcinomas. These observations indicate that a comparative genetic analysis provides a promising approach to survey genes involved in multifactorial process of human lung carcinogenesis.


Inferring longitudinal patterns of group B Streptococcus colonization during pregnancy.
Maternal colonization by Group B Streptococcus (GBS) can lead to severe infection in neonates and has also been associated with prematurity and stillbirth. Better quantitative understanding of the trajectories of GBS carriage during pregnancy is essential for the design of informative epidemiological studies. Here, we describe analyses of published longitudinal data using Bayesian hidden Markov models, which involve the estimation of parameters related to the succession of latent states (infection status) and observations (culture positivity). In addition to quantifying infection acquisition and clearance probabilities, the statistical approach also suggests that for some longitudinal patterns of culture results, pregnant women were likely to have been GBS-colonized despite a negative diagnostic result. We believe this method, if used in future longitudinal studies of maternal GBS colonization, would improve our understanding of the pathologies linked to this bacterium and could also inform maternal GBS vaccine trial design.


Warfarin therapy for atrial fibrillation in the elderly.
To evaluate a clinical practice model that addresses special needs for managing anticoagulation in a community-dwelling elderly population with atrial fibrillation and high risk of stroke. Medical records of 18 patients (mean age 82 y) followed by the Geriatric Ambulatory Program over 2 years, with a target international normalized ratio (INR) of 2.0-3.0, were reviewed. Risk factors for stroke, number and results of INR tests, suspected reasons for suboptimal response, and adverse events were analyzed. Patients were defined as having cognitive impairment if they had a Folstein Mini-Mental State Exam score < or = 26. Functional impairment was defined by > or = 2 disabilities in activities of daily living. Eighty-three percent (15/18) had > or = 2 additional stroke risk factors. Fifty-one percent (273/541) of INR responses were therapeutic. Female gender (p = 0.015) and cognitive (p = 0.019) and functional impairment (p = 0.001) were associated with supratherapeutic INR response. All patients with cognitive impairment and 85% of those with functional impairment received caregiver support for medication administration. There were 4 minor bleeding events and no thromboembolic events. The mean number of medications was 9.3 in those with bleeding versus 6.8 in those without bleeding (p = 0.052). Elderly patients with high stroke risk achieved therapeutic INR responses. However, those with significant cognitive or functional impairment require caregiver support and special consideration for anticoagulation management.


Infracoccygeal sacropexy for uterovaginal prolapse.
To report on the efficacy and safety of infracoccygeal sacropexy for the treatment of uterovaginal prolapse. A retrospective analysis of 87 women who underwent an infracoccygeal sacropexy procedure, 53 receiving a multifilament and 34 receiving a monofilament tape. The mean follow-up was 27 months. Postoperative perineal pain was reported by 7 women (10%), and dyschesia and dyspareunia were observed de novo in 4 (5%) and 5 women (6%), respectively. There were 5 cases (9%) of vaginal extrusion and 9 cases (18%) of prolapse recurrence in the multifilament tape group, and in the monofilament tape group there were no cases of vaginal extrusion and 4 cases (14%) of prolapse recurrence (P=0.79 for prolapse recurrence). The recurrence-free survival curves of the 2 groups were similar. Univariate analysis revealed no independent factors of recurrence. Although infracoccygeal sacropexy was found to be associated with few prolapse recurrences, complications were not rare.


Carboxymethyl functionalization of amylopectin and its evaluation as a nanometric drug carrier.
Carboxymethylation of amylopectin was accomplished by reacting it with monochloroacetic acid. The results of characterization study confirmed the carboxymethyl functionalization with degree of substitution of 1.25. Carboxymethyl amylopectin was found to be slightly more crystalline, appearing morphologically as polyhedral flakes with striated surface. Further, carboxymethylation of amylopectin modified its compression characteristic from fragmentary to plastic. Interaction of carboxymethyl amylopectin (0.001-0.005%, w/v) with chitosan (0.05-0.075% w/v) using ibuprofen (0.05%, w/v) as a model drug provided polyelectrolyte nanoparticles of size 182.7-353.3 nm and 75-92% drug entrapment. The polyelectrolyte nanoparticles showed a sustained release of drug following zero-order kinetics with mechanism of release being combination of matrix erosion and diffusion through the matrix.


Efficient algorithm for calculating two-photon absorption spectra.
We propose an efficient time-dependent algorithm for nonlinear response function that requires CPU time proportional to the system size, and study the size effects in two-photon absorption spectra of Si nanocrystallites by using this algorithm.


Behavioral thermoregulation and critical thermal limits of giant keyhole limpet Megathura crenulata(Sowerby 1825) (Mollusca; Vetigastropoda).
The thermoregulatory behavior of the giant keyhole limpet Megathura crenulata was determined in a horizontal thermal gradient during the day at 18.9 °C and 18.3 °C for the night. The final preferendum determined for giant keyhole limpets was of 18.6±1.2 °C. Limpets' displacement velocity was 10.0±3.9 cm h(-1) during the light phase and 8.4±1.6 cm h(-1) during the dark phase. The thermotolerance (measured as CTMax at 50%) was determined in a keyhole limpet in three acclimation temperatures 17, 20, and 23 °C. Limpets were subjected to water increasing temperatures at a rate of 1 °C every 30 min, until they detached from the substrate. The critical thermal maximum at 50% was 27.2, 27.9 and 28.3 °C respectively.


Transcriptome analysis evinces anti-neoplastic mechanisms of hypericin: A study on U87 glioblastoma cell line.
Hypericin (HYP) from Hypericum perforatum has cytotoxic effects on a variety of malignant cell types, but the pattern of gene expression mediating the effect is largely unknown. Here we sought to analyze the response of U87 glioblastoma (GBM) cell lines in response to HYP. U87 cell line was treated by HYP. Cytotoxicity was assessed using MTT and Annexin V/PI assays. Gene expression profile was obtained using high-throughput sequencing. Enrichment analysis was performed on differentially expressed genes (DEGs). Upstream transcription factors and microRNAs regulating DEGs were predicted. The effects of DEGs on survival of GBM patients were calculated. Protein-protein interaction analysis was conducted to obtain key altered genes. The possible effect of HYP treatment on immunity response was evaluated. The IC50 of HYP on U87 cell line was determined to be 1.5 μg/ml. The main type of cell death was apoptosis. A total of 312 DEGs were found. Affected Gene Ontology terms and pathways were identified. Analysis of upstream modulators of DEGs pointed out to transcription factors that significantly overlap with GBM stem cell transcription factor. Survival analysis suggested that HYP works best for the mesenchymal subtype patients. Tumor infiltration analysis predicted that HYP may affect Treg and macrophage infiltration in vivo. Using expression pattern of GBM patients and HYP-induced DEGs we suggested Fedratinib as a complementary drug to HYP. Our study represents the response of U87 cell line to HYP, with analyses on survival, transcription factors and personalization according to GBM subtype.


Developing and implementing the Active Design Guidelines in New York City.
Physical inactivity is a leading cause of death in the United States and globally and is also associated with several additional leading causes of death, including obesity, high blood pressure and high blood glucose. The built environment plays a critical role in promoting or discouraging physical activity among adults and children. To create a healthier and more physically active city, a working group comprising several New York City agencies, including the Departments of Design and Construction, Health and Mental Hygiene, Transportation and City Planning, and in collaboration with design organizations and academics, published the Active Design Guidelines (ADG; <www.nyc.gov/adg>) in January 2010. The ADG is a manual of evidence-based and best-practice strategies for increasing physical activity in the design and construction of neighborhoods, streets and buildings. The commentary discusses key activities and events leading up to the publication as well as current implementation activities. It also shares the lessons learned that could assist other communities interested in improving their built environments in developing and implementing similar activities and initiatives.


Evaluating the feasibility of ReWork-SCI: a person-centred intervention for return-to-work after spinal cord injury.
To evaluate the feasibility of: (1) ReWork-SCI with regard to adherence and acceptability and (2) a study design for evaluating ReWork-SCI with regard to recruitment, retention and outcome measures. Pre-test and post-test, single group, feasibility study. Spinal cord injury (SCI) unit at a regional rehabilitation centre in Sweden. Two women and five men (n=7). Eligible criteria: (1) sustained traumatic or non-traumatic SCI; (2) completed the first acute care episode in a hospital; (3) between 18 to 65 years of age; (4) assessed by a physician as approachable for participation in the intervention; (5) history of permanent or temporary employment; (6) self-reported desire to return to work; and (7) ability to communicate in English or Swedish. ReWork-SCI is a person-centred intervention for return-to-work (RTW), developed and evaluated using the Medical Research Council's guidelines. ReWork-SCI follows a person-centred, structured and coordinated intervention process led by a coordinator within a SCI rehabilitation team. The feasibility of ReWork-SCI and a study design was evaluated using a set of outcome measurement tools, vocational data, logbooks and semi-structured interviews. All eligible participants accepted enrolment and follow-up. All participants had a plan for RTW after 3 months and four participants had initiated part-time work or work trial 6 months after commencement of intervention. Adherence and acceptability were overall good. Challenges of the intervention related to the person-centred follow-up, staff shortage and rootedness in the SCI team. ReWork-SCI was feasible and can contribute to a systematic design of an individualised plan, facilitate decision-making and build trust in the RTW process after SCI. Core features of the intervention was the systematic structure, use of a person-centred approach and dialogue with the employer. For the effectiveness of ReWork-SCI, modifications and considerations of study design are needed.


Dance as a neurorehabilitation strategy: A systematic review.
This systematic review aimed to demonstrate the benefits of dance-based therapeutic interventions in rehabilitation processes in patients with neurological disorders. Searches on the following databases and electronic search engines were conducted: MEDLINE, LILACS, Science Direct, SCOPUS, PeDro, BVS (virtual health library), and Google Scholar. Data extraction was independently performed by two authors. Twenty-five clinical trial-type studies that included dance and established outcome measures were included, and studies in which a musicalized exercise modality was used without the connotation of dance were excluded. From the results of multiple studies, the short-term motor benefits of rhythmic auditory stimulation on gait parameters were highlighted. Furthermore, there was scientific evidence demonstrating the benefits of the cognitive and social parameters of group dancing as significant improvements in cognitive flexibility and processing speed were highlighted. Recent studies showed that interventions based on exercise and/or rhythmic choreography can reduce the risk of falls in patients with neurological disorders, thereby improving their quality of life. These findings suggest that dance is an innovative and effective method that can be included in therapies as it allows for a promising prognosis in motor, cognitive, and social performances of patients with neurological disorders that alter mobility and quality of life.


Kinematic and Kinetic Gait Characteristics in People with Patellofemoral Pain: A Systematic Review and Meta-analysis.
Patellofemoral pain (PFP) is a prevalent knee condition with many proposed biomechanically orientated etiological factors and treatments. We aimed to systematically review and synthesize the evidence for biomechanical variables (spatiotemporal, kinematic, kinetic) during walking and running in people with PFP compared with pain-free controls, and determine if biomechanical variables contribute to the development of PFP. Systematic review and meta-analysis. We searched Medline, CINAHL, SPORTDiscus, Embase, and Web of Science from inception to October 2021. All study designs (prospective, case-control [± interventional component, provided pre-intervention data were reported for both groups], cross-sectional) comparing spatiotemporal, kinematic, and/or kinetic variables during walking and/or running between people with and without PFP. We identified 55 studies involving 1300 people with PFP and 1393 pain-free controls. Overall pooled analysis identified that people with PFP had slower gait velocity [moderate evidence, standardized mean difference (SMD) - 0.50, 95% confidence interval (CI) - 0.72, - 0.27], lower cadence (limited evidence, SMD - 0.43, 95% CI - 0.74, - 0.12), and shorter stride length (limited evidence, SMD - 0.46, 95% CI - 0.80, - 0.12). People with PFP also had greater peak contralateral pelvic drop (moderate evidence, SMD - 0.46, 95% CI - 0.90, - 0.03), smaller peak knee flexion angles (moderate evidence, SMD - 0.30, 95% CI - 0.52, - 0.08), and smaller peak knee extension moments (limited evidence, SMD - 0.41, 95% CI - 0.75, - 0.07) compared with controls. Females with PFP had greater peak hip flexion (moderate evidence, SMD 0.83, 95% CI 0.30, 1.36) and rearfoot eversion (limited evidence, SMD 0.59, 95% CI 0.03, 1.14) angles compared to pain-free females. No significant between-group differences were identified for all other biomechanical variables. Data pooling was not possible for prospective studies. A limited number of biomechanical differences exist when comparing people with and without PFP, mostly characterized by small-to-moderate effect sizes. People with PFP ambulate slower, with lower cadence and a shortened stride length, greater contralateral pelvic drop, and lower knee flexion angles and knee extension moments. It is unclear whether these features are present prior to PFP onset or occur as pain-compensatory movement strategies given the lack of prospective data. PROSPERO # CRD42019080241.


The high-level tetraplegic: psychological survival and adjustment.
From the date of injury tetraplegics with spinal cord lesions of C4 and above are faced with unparalleled social and psychological adjustment. The individual will be involved in a process of re-establishing his identity by learning to develop and adapt his only remaining resources, his mind and his ability to communicate. This patient population is absolutely dependent in all aspects of meeting basic needs during the initial stages of the injury. If we attempt to return the patient to an active role in society, the rehabilitation team must be aware of the dynamics by which the patient achieves control of both himself and his environment. He will undergo a marked regression of his physical and emotional being and will face the turmoil of realigning ego parameters. The focal point of patient, family and staff interaction is the months of rehabilitation, where the team deals with the problem of social reintegration. Family involvement is essential from the beginning and reaches a high point at discharge and home planning. The scope of the paper will be to trace the psycho-social development of the patient with cervical lesion of C4 and above through the initial, rehabilitation and discharge phases of the injury. Our aim is to develop sufficient insight to the problems facing our patient population that we may aid in their search for meaningful lives.


Etiology of Obesity Over the Life Span: Ecological and Genetic Highlights from Asian Countries.
Obesity is a worldwide pandemic, and the prevalence rate has doubled since the 1980s. Asian countries are also experiencing the global epidemic of obesity with its related health consequences. The prevalence of overweight and obesity are increasing at an alarming rate across all age groups in Asia. These increases are mainly attributed to rapid economic growth, which leads to socio-economic, nutrition and lifestyle transitions, resulting in a positive energy balance. In addition, fat mass and obesity-associated gene variants, copy number variants in chromosomes and epigenetic modifications have shown positive associations with the risk of obesity among Asians. In this review highlights of prevalence and related ecological and genetic factors that could influence the rapid rise in obesity among Asian populations are discussed.


Origin, occurrence, and biodegradation of long-side-chain alkyl compounds in the environment: a review.
Long-side-chain alkyl compounds, such as those present in oil and oil products, either of natural or of anthropogenic origin or released by industrial activities, are a ubiquitous group of chemicals in the environment. Among them, long-side-chain alkylmonocycloalkanes, alkylbenzenes, and alkyl organic sulfur compounds are largely found in fossil fuels while long-side-chain alkylbenzene sulfonates and alkylphenols are released into the environment primarily due to domestic activities. The present article briefly summarizes the advances that have occurred in this field in terms of the identity, abundance, possible origin and significance of these long-side-chain alkyl compounds found in the environment. In addition, the susceptibility to biodegradation and the fate of these chemicals in the environment are discussed.


Global trends and themes in genetic counseling research.
This article seeks to highlight the most recent trends and themes in genetic counseling that are of broad interest. A total of 3505 documents were published between 1952 and 2021, with a trend toward increase in paper/year. The most common documents are original articles (2515, 71.8%), followed by review articles (341, 9.7%). Journal of Genetic Counseling publishes the highest number of genetic counseling articles (587, 16.7%), followed by Clinical Genetics (103, 2.9%) and the South American Journal of Medical Genetics (95, 2.7%). Co-occurrence analysis revealed five research themes: genetic testing, cancer, genetic counselor, prenatal diagnosis, and psychiatry. The genetic counselor theme contained most of the recent keywords, including "covid-19," "underrepresented population," "service delivery models," "workforce," "disparities," "service delivery," "professional development," "cultural competence," "access," "diversity," "telemedicine," and "health literacy." Genetic counseling researchers may use these keywords to find topics pertinent to their future research and practice.


Urine growth hormone determinations compared with other methods in the assessment of growth hormone secretion.
Urinary excretion of hGH was studied in children with short stature using a sensitive sandwich enzyme immunoassay technique. Urinary hGH excretion, in terms of hGH: creatinine ratio, showed excellent correlation with the mean and peak hGH values during physiological and pharmacological tests. It seems that the urinary hGH levels reflect serum hGH profiles during the urine collection period. A border zone for the lower limits of normal hGH levels in the urine was 7.5-13.4 ng/g creatinine for the physiological test at night (from 2000 hours to 0600 hours) and 17.4-35.0 ng/g creatinine for the pharmacological tests. Assessment of hGH secretory status by the urinary hGH levels showed good agreement with the serum hGH response. Measurement of urinary hGH could be used as a diagnostic test for impaired hGH secretion, and the multiple blood drawing required in physiological and pharmacological tests might be replaced by urine sampling.


Persistence of candiduria in ICU catheterized patients is not linked to adherence and proteolytic activities of Candida strains.
In this study, the patients who developed asymptomatic candiduria in the intensive care unit were followed prospectively for the persistence of candiduria after the replacement of indwelling urethral catheter and the correlation between persistence and virulence factors (proteinase enzyme activity and epithelial adhesion) was assessed. Prospective study. Intensive care unit and mycology laboratory at a university hospital. Thirty-four patients with asymptomatic candiduria were included in the study. Candiduria persisted in 19 of 34 patients(56%; group 1) and cleared in 15 of 34 patients(44%; group 2) after urinary catheters were changed. When the virulence factors (epithelial adhesion and proteinase activity) and distribution of Candida spp. were compared between two groups, no statistically significant correlation was found. The host immune response might be more important than virulence factors of Candida spp. for persistence of candiduria.


Pancreatic atrophy and recovery after allogeneic hematopoietic cell transplantation.
Pancreatic atrophy after allogeneic hematopoietic cell transplantation (HCT) is one of the symptoms associated with chronic graft-versus-host disease (GVHD). Although pancreatic atrophy has been considered to cause exocrine insufficiency and weight loss, it is not yet clear what kinds of recipients can be expected to recover their body weight (BW) or pancreatic thickness. In addition, the effect of pancreatic atrophy on the prognosis has not been clarified. We retrospectively analyzed 170 recipients who received allogeneic bone marrow transplantation or peripheral blood stem cell transplantation, and evaluated them using the CT scan images obtained closest to 1, 2, 3, and 4 years after HCT. Fifty-five recipients (32.4%) demonstrated pancreatic atrophy, and 11 (20%) of them recovered their pancreatic thickness. While recipients without pancreatic atrophy gradually recovered their BW (P < 0.001), those with atrophy did not (P = 0.12). Moderate and severe chronic GVHD tended to be slightly more common in the atrophy group (47.3% vs 38.3%), whereas the pancreatic thickness tended to recover in these recipients (30.8% vs 10.3%). HCT from a female donor to a male recipient showed superior pancreatic recovery compared to other donor and recipient sex combinations. Pancreatic atrophy treated as a significantly associated with inferior survival (HR 4.91, P < 0.001) and an increased risk of non-relapse mortality (HR 8.75, P < 0.001). These results suggest that it is important to monitor pancreatic thickness after HCT. Further prospective investigations are warranted to clarify the significance of pancreatic atrophy on clinical outcomes.


Thyroid stimulating hormone exhibits the impact on LDLR/LDL-c via up-regulating hepatic PCSK9 expression.
Thyroid stimulating hormone (TSH) has received increasing attention as being closely associated with increased low-density lipoprotein cholesterol (LDL-c) level and higher atherosclerotic risks. Proprotein convertase subtilisin/kexin type 9 (PCSK9) is known for increasing circulating LDL-c level by inducing LDL receptor degradation. However, whether TSH influences hepatic PCSK9 expression and LDL-c metabolism remains unclear. First, the correlation between TSH and lipid profiles were investigated in euthyroid population and in subclinical hypothyroidism patients. Then, an in vitro study was conducted to validate the effects of TSH on hepatic PCSK9 expression in HepG2 cells. Serum TSH concentrations positively correlated with LDL-c levels in euthyroid subjects. Subclinical hypothyroidism patients with higher serum TSH levels showed significantly increased serum PCSK9 levels than the matched euthyroid participants (151.29 (89.51-293.03) vs. 84.70 (34.98-141.72) ng/ml, P<0.001), along with increased LDL-c concentrations. In HepG2 cells, LDLR expression on the plasma membrane was decreased, and PCSK9 mRNA and protein levels were synchronously upregulated after recombinant human TSH (rhTSH) treatment, while the effects could be blocked by TSH receptor blocking antibody K1-70. Sterol regulatory element binding protein (SREBP) 1c and SREBP2 mRNA expressions were enhanced after rhTSH treatment, and specific siRNAs significantly inhibited the effects of rhTSH. Furthermore, there was a noticeable induction of PCSK9 expression by rhTSH even though HMGCR gene expression was silenced. We conclude a regulating role of TSH on hepatic PCSK9 expression, which further contributing to a higher LDL-c level.


Nitrate tolerance in heart failure: differential venous, pulmonary and systemic arterial effects.
The hemodynamic profile of tolerance to intravenous nitroglycerin was studied in 9 patients with New York Heart Association Class III to IV congestive heart failure. After rapid dosage build-up to the maximal tolerated dose (decrease in pulmonary wedge pressure to 10 mm Hg or systolic blood pressure to 90 mm Hg), nitroglycerin (525 +/- 548 micrograms/min) was administered at a constant continuous intravenous infusion for a total of 24 hours. The extent of nitrate tolerance at 24 hours was calculated as the percentage loss of the benefit achieved at time of peak effect of nitroglycerin. Tolerance had a different time course and magnitude in the venous, arterial and pulmonary circulations. At 24 hours, right atrial pressure and pulmonary vascular resistance returned to control values in most patients, while 40 to 50% of the effect on systemic vascular resistance, cardiac index and pulmonary wedge pressure was maintained. These findings emphasize the importance of precise definitions in studies relating to nitrate tolerance.


Direct Simulation Monte Carlo investigation of fluid characteristics and gas transport in porous microchannels.
The impetus of the current research is to use the direct simulation Monte Carlo (DSMC) algorithm to investigate fluid behaviour and gas transport in porous microchannels. Here, we demonstrate DSMC's capability to simulate porous media up to 40% porosity. In this study, the porous geometry is generated by a random distribution of circular obstacles through the microchannel with no interpenetration between the obstacles. The influence of the morphology along with rarefaction and gas type on the apparent permeability is investigated. Moreover, the effects of porosity, solid particle's diameter and specific surface area are considered. Our results demonstrate that although decreasing porosity intensifies tortuosity in the flow field, the tortuosity reduces at higher Knudsen numbers due to slip flow at solid boundaries. In addition, our study on two different gas species showed that the gas type affects slippage and apparent gas permeability. Finally, comparing different apparent permeability models showed that Beskok and Karniadakis model is valid only up to the early transition regime and at higher Knudsen numbers, the current data matches those models that take Knudsen diffusion into account as well.


The management of priapism in sickle cell anemia.
After assessing the data currently available on the multiple modalities used to manage priapism secondary to sickle cell disease we adopted a specific therapeutic regimen for the treatment of this problem. Early non-surgical measures, including hydration, alkalization and analgesics, rapid red cell infusion to double the hematocrit and late shunting procedures in conjunction with exchange transfusion, offer the soundest approach on pathophysiological grounds. There is currently no evidence to support early surgical intervention as the best approach to conserve potency in this problem.


The low dose gamma ionising radiation impact upon cooperativity of androgen-specific proteins.
The paper deals with effects of the ionising radiation (γ-IR, 0.5 Gy) upon serum testosterone (T), characteristics of testosterone-binding globulin (TeBG) and androgen receptor (AR) in parallel with observation of androgen (A) responsive enzyme activity - hexokinase (HK). The interdependence or relationships of T-levels with parameters of the proteins that provide androgenic regulation are consequently analyzed in post-IR dynamics. The IR-stress adjustment data reveal expediency of TeBG- and AR-cooperativity measurements for more precise assessments of endocrine A-control at appropriate emergencies.


Radiocarbon dates and Bayesian modeling support maritime diffusion model for megaliths in Europe.
There are two competing hypotheses for the origin of megaliths in Europe. The conventional view from the late 19th and early 20th centuries was of a single-source diffusion of megaliths in Europe from the Near East through the Mediterranean and along the Atlantic coast. Following early radiocarbon dating in the 1970s, an alternative hypothesis arose of regional independent developments in Europe. This model has dominated megalith research until today. We applied a Bayesian statistical approach to 2,410 currently available radiocarbon results from megalithic, partly premegalithic, and contemporaneous nonmegalithic contexts in Europe to resolve this long-standing debate. The radiocarbon results suggest that megalithic graves emerged within a brief time interval of 200 y to 300 y in the second half of the fifth millennium calibrated years BC in northwest France, the Mediterranean, and the Atlantic coast of Iberia. We found decisive support for the spread of megaliths along the sea route in three main phases. Thus, a maritime diffusion model is the most likely explanation of their expansion.


The comparison of counterimmunoelectrophoresis with indirect haemagglutination test for detection of antibodies in experimentally infected guinea pigs with Toxocara canis.
The humoral response of guinea pigs infected with doses of 500, 1000, and 1500 infective Toxocara canis ova were measured by counterimmunoelectrophoresis and indirect haemmagglutination tests one to five weeks after infection. The antigens used in these measurements were prepared from adult T. canis. Counterimmunoelectrophoresis showed lower titres than haemagglutination method, but it was performed rapidly and with ease. The possible use of these techniques for immunodiagnosis of human visceral larval migrans is discussed.


Update in allergy and immunology.
Internal medicine has witnessed astounding developments in our understanding of the immune system's role in the pathophysiology of many diseases. The present challenge is to harness this knowledge to develop more effective therapies for immune-related illness. Guidelines for the care of common illnesses such as asthma may encourage appropriate clinical application of advances in our armamentarium against immune disease.


Promotive effects of non-digestible disaccharides on rat mineral absorption depend on the type of saccharide.
We examined the effects of feeding non-digestible disaccharides, difructose anhydride III (DFAIII), maltitol, melibiose and, cellobiose, on calcium, magnesium, and iron absorption in comparison with fructo-oligosaccharide (FOS) in normal and ovariectomized rats. In experiment 1, six groups of male Sprague-Dawley rats were fed a control diet (100 g of cellulose/kg of diet), test diets containing 30 g of FOS, or the four non-digestible disaccharides in place of the cellulose in the control diet for 4 wk. In experiment 2, two groups of female Sprague-Dawley rats (sham or ovariectomized) were assigned to one of four subgroups and fed the control or test diet containing FOS, DFAIII, and melibiose for 5 wk. Feces and cecal contents were collected to evaluate mineral absorption and cecal fermentation. In experiment 1, calcium absorption in all the disaccharides groups except the cellobiose group, magnesium absorption in all test diet groups, and iron absorption in the FOS, DFAIII, and melibiose groups were higher than those in the control group. In ovariectomized rats (experiment 2), calcium absorption in the DFAIII and melibiose groups, magnesium absorption in all test diet groups, and iron absorption in the DFAIII group alone were higher than those in the control group. Cecal organic acids were positively and pH was negatively correlated with the absorption of these minerals, although the effects varied. Non-digestible disaccharides increase calcium, magnesium, and iron absorption in normal and ovariectomized rats; however, the effects depend on the disaccharide tested, a fact that may be partly associated with the cecal fermentation of these disaccharides.


Pardaxin, a new pharmacological tool to stimulate the arachidonic acid cascade in PC12 cells.
The effect of Pardaxin, a neurotoxin that induces neurotransmitter release from neurons, on the arachidonic acid (AA) cascade was studied in PC12 cells. Both native and the synthetic Pardaxin selectively stimulated phospholipase A2 (PLA2) activity (measured by [3H]AA release) in the presence as well as in the absence of extracellular calcium. Pardaxin-stimulated PLA2 activity was also evident in the increased formation of lysophosphatidylcholine. Pardaxin analogs, lacking the alpha-helical structure that is essential for insertion into the plasma membrane, were ineffective in stimulating the AA cascade in PC12 cells. Pardaxin stimulation of PLA2 was markedly inhibited by the nonselective PLA2 inhibitors bromophenacyl bromide and mepacrine, by methyl arachidonyl fluorophosphonate, a dual inhibitor of calcium-dependent cytosolic PLA2 and the calcium-independent PLA2 and by bromoenol lactone[(E)-6-(bromoethylene)tetrahydro-3-(1-naphthalenyl-2H-pyran -2- one], a highly specific inhibitor of calcium-independent PLA2. After Pardaxin treatment, there was increased release of AA metabolites produced by the cyclooxygenase pathway as expressed in an 8-fold increase of PGE2 release. The release of other eicosanoids, such as 6-keto-PGF1alpha and thromboxane B2, was also augmented. Pardaxin-induced PGE2 release was observed in calcium-free medium and in the absence of any increase in cytosolic calcium. Dexamethasone partially inhibited Pardaxin-induced PGE2 release. This effect was reversed by the type II corticosteroid receptor antagonist RU-38486. Our results indicate that Pardaxin stimulates release of AA and eicosanoids, independently of calcium, and suggest that calcium-independent PLA2 plays an important role in Pardaxin stimulation of the AA cascade.


The structure of neuromelanin as studied by chemical degradative methods.
The biosynthesis, structure and function of neuromelanin (NM), the dark brown melanin-like pigment present in the substantia nigra (SN), are not well characterized, in spite of the possible involvement of NM in the etiology and pathogenesis of Parkinson's disease. NM was isolated from the SN of five non-Parkinsonian human brains. NM and synthetic melanins, employed as models, were characterized by chemical analysis. Alkaline hydrogen peroxide (H2O2) oxidation of NM generated four degradation products, pyrrole-2,3-dicarboxylic acid (PDCA), pyrrole-2,3,5-tricarboxylic acid (PTCA), thiazole-4,5-dicarboxylic acid (TDCA) and thiazole-2,3,5-tricarboxylic acid (TTCA), whose ratios, especially the TTCA to PDCA ratio, indicate that NM is derived mostly from dopamine (DA) with 25% incorporation of cysteine (Cys) in the form of a benzothiazine structure. Hydriodic acid (HI) reductive hydrolysis of NM yielded 4-amino-3-hydroxyphenylethylamine (4-AHPEA) as a marker of cysteinyldopamine (CysDA)-derived units. The 4-AHPEA to PDCA ratio indicates a 21% incorporation of CysDA-derived units into NM. These degradative experiments also suggest that DOPA is not incorporated into NM to a significant extent (approximately 6% the level of DA). It is concluded that the TTCA to PDCA ratio is a useful indicator of CysDA-derived units in NM, and NM consists mainly of DA-melanin with some contribution from CysDA-melanin. The involvement of DA and CysDA as building blocks of NM demonstrates the detoxifying role of NM synthesis, since it prevents the intraneuronal accumulation of DA and CysDA, which would cause toxic effects.


A topographical map of spatiotemporal patterns of gene expression.
A recent study by Folkes et al. in Cell generated a 3D atlas of gene expression for the Drosophila blastoderm embryo using a new approach for image registration. This virtual embryo allows in silico multiplexing of in situ hybridizations and lays the groundwork for new insights into gene regulatory networks.


Tonsillar chancre as unusual manifestation of primary syphilis.
Primary syphilis with oropharyngeal manifestations should be kept in mind, though. Lips and tongue ulcers are the most frequently reported lesions and tonsillar ulcers are much more rare. We report the case of a 24-year-old woman with a syphilitic ulcer localized in her left tonsil.


Do certain surgical steps increase postoperative morbidity after cytoreductive surgery and HIPEC- a retrospective analysis.
It has been shown that cytoreductive surgery (CRS) in combination with hyperthermic intraperitoneal chemotherapy (HIPEC) is an effective treatment for patients suffering from peritoneal malignancies. Despite good results, there is an ongoing debate about this treatment due to perioperative morbidity. The aim of this study is to identify relevant risk factors for an unfavorable postoperative outcome after CRS and HIPEC. A retrospective analysis of a prospectively recorded database of all patients undergoing CRS and HIPEC between 2013 and 2020 in the Department of Surgery of the University Hospital Dresden was performed with a special focus on certain surgical steps of multivisceral resection, one- or 2- stage CRS/HIPEC and underlying diagnosis as possible risk factors for worse postoperative course. N = 173 CRS and HIPEC procedures were performed for various diagnoses. Relevant postoperative morbidity was 24% and 30d-mortality 1.2%. Simultaneous liver resections, preoperative hypalbuminemia and 2-staged CRS/HIPEC were significant risk factors for a worse postoperative course in multivariable analysis. Assessment of the association of simultaneous anastomoses and morbidity and mortality was inconclusive. CRS and HIPEC is a safe treatment without relevant intraoperative morbidity and mortality and acceptable postoperative outcome. One-stage CRS/HIPEC should be preferred.


Fracture risk in very young women using combined oral contraceptives.
Little is known on fracture risk in young women using oral contraceptives. Case-control study (64,548 cases, 193,641 controls). Overall, no major association with risk of fracture was present. Only in a few age groups a limited increase (OR 1.1-1.5) was seen in those using low average doses (less than one tablet per day - i.e., intermittent use). Neither 20 nor >or=30 mcg per day of ethinylestradiol was associated with risk of fractures, while changing from 20 to >or=30 mcg per day or vice versa was associated with a limited increase in fracture risk (OR 1.1-1.4) in some age groups. In general, use of oral contraceptives in young women did not seem to be associated with an increased risk of fractures. The isolated increase seen in some strata with low average doses may be linked to factors associated with discontinuation of oral contraceptives rather than pharmacological effects.


Acceptability to patients, carers and clinicians of an mHealth platform for the management of Parkinson's disease (PD_Manager): study protocol for a pilot randomised controlled trial.
Parkinson's disease is a degenerative neurological condition causing multiple motor and non-motor symptoms that have a serious adverse effect on quality of life. Management is problematic due to the variable and fluctuating nature of symptoms, often hourly and daily. The PD_Manager mHealth platform aims to provide a continuous feed of data on symptoms to improve clinical understanding of the status of any individual patient and inform care planning. The objectives of this trial are to (1) assess patient (and family carer) perspectives of PD_Manager regarding comfort, acceptability and ease of use; (2) assess clinician views about the utility of the data generated by PD_Manager for clinical decision making and the acceptability of the system in clinical practice. This trial is an unblinded, parallel, two-group, randomised controlled pilot study. A total of 200 persons with Parkinson's disease (Hoehn and Yahr stage 3, experiencing motor fluctuations at least 2 h per day), with primary family carers, in three countries (110 Rome, 50 Venice, Italy; 20 each in Ioannina, Greece and Surrey, England) will be recruited. Following informed consent, baseline information will be gathered, including the following: age, gender, education, attitudes to technology (patient and carer); time since Parkinson's diagnosis, symptom status and comorbidities (patient only). Randomisation will assign participants (1:1 in each country), to PD_Manager vs control, stratifying by age (1 ≤ 70 : 1 > 70) and gender (60% M: 40% F). The PD_Manager system captures continuous data on motor symptoms, sleep, activity, speech quality and emotional state using wearable devices (wristband, insoles) and a smartphone (with apps) for storing and transmitting the information. Control group participants will be asked to keep a symptom diary covering the same elements as PD_Manager records. After a minimum of two weeks, each participant will attend a consultation with a specialist doctor for review of the data gathered (by either means), and changes to management will be initiated as indicated. Patients, carers and clinicians will be asked for feedback on the acceptability and utility of the data collection methods. The PD_Manager intervention, compared to a symptom diary, will be evaluated in a cost-consequences framework. Information gathered will inform further development of the PD_Manager system and a larger effectiveness trial. ISRCTN Registry, ISRCTN17396879 . Registered on 15 March 2017.


Spermatogonial Stem Cells and In Vitro Spermatogenesis: How Far Are We from a Human Testis on a Chip?
An in vitro testis model will provide a superior platform for studying the testis microenvironment and molecular mechanisms that affect male fertility. The ultimate aim is to provide reproductive hope for children diagnosed with cancer who were sterilized by aggressive gonadotoxic therapies.


Anthropometric studies on African athletes who participated in the 1st African University Games.
1. Height, weight, skinfold thicknesses and mid-arm circumference were measured in 540 males and 117 females aged 20-24 years who took part in the 1st African University Games, held at the University of Ghana, Legon. Body fat content, Quetelet's index (weight divided by height X 100; Khosla & Lowe (1967)) and mid-arm muscle circumference were derived from the measurements taken. 2. The physique or body-build of the subjects as assessed by Quetelet's index showed that both male and female subjects from the various countries were of medium body-build. The body fat content for males was between 10 and 12% with the exception of the Egyptians (12.8%) while that of females was between 23-24%. 3. Body measurements of the subjects compared favourably with that of international standards (WHO, 1966) with the exception of the triceps skinfold thickness which was only approximately 60% of the standard value. 4. The low values for triceps skinfold thickness are probably due to differences in the distribution of subcutaneous fat at different sites in the body as found between caucasian and non-caucasian population groups. The results are discussed in relation to the findings of other workers on ethnic differences in skinfold thickness.


Food bolus impaction secondary to reflux strictures: a rural surgical experience.
Benign esophageal strictures at the gastroesophageal (GE) junction secondary to acid reflux have an unknown altered natural history after dilation. We carried out a 7-year retrospective investigation from 2001 to 2007. Sixty-four patients presented with food bolus impaction at the GE junction. Reflux strictures were the cause in 42 (66%), neuromuscular disease or dysmotility in 2 (3%), esophageal cancer in 1 (2%), and no underlying pathology could be identified in 19 (29%). Duration of dilation-induced relief from impaction recurrence decreased with each subsequent dilation: first 30 +/- 21 months, second 27 +/- 15 months, and third 8 +/- 6 months. Patients taking proton pump inhibitors (PPIs) had a longer interval between dilations (25 +/- 9 months) compared with those who were not (20 +/- 12 months, p = 0.06). PPIs also decreased the number of subsequent dilations needed during the course of the study (1 +/- 1 versus 2 +/- 1 dilations, p = 0.02). Reflux strictures were more likely to cause another episode of impaction than no underlying pathology (relative risk, 2.7; 95% CI, 1.8-4.1; p < 0.0001). Mean followup was 38 months (range 2 to 120 months). Reflux strictures are well treated with dilation and PPIs. Although food bolus impaction can occur without an underlying pathology, patients with relux strictures are likely to have a recurrence and should be treated aggressively. Addition of PPIs appears to decrease the number of dilations that a patient will require and lengthen the interval between dilations. Patients requiring more than two dilations after initiation of PPI therapy are unlikely to have durable relief.


The effect of ethanol on cell wall antigens of Saccharomyces cerevisiae and specific isolation of high ethanol producing strains of this yeast, making use of a serological technique.
Generally, natural isolates of high ethanol producing Saccharomyces cerevisiae obtained by screening are used in alcoholic industries. The methods involved in their isolation and identification are elaborate. Antigenic analysis using antibodies raised against whole Saccharomyces cells indicated species specificity of cell wall surface thermostable antigens. By affinity purification, the specific antibodies could be obtained and used for specific isolation of S. cerevisiae. Antigenic studies using antibodies raised against isolated cell walls of fermentatively grown S. cerevisiae indicated the occurrence of thermolabile antigens common to Saccharomyces species. Higher concentrations of these antigens could be detected in those S. cerevisiae that had the ability for high ethanol production. The concentrations of these cell wall common antigens increased with increasing culture age and ethanol accumulation in culture broths. In younger yeast cells, the concentration could be increased by growing the cells in a medium containing added ethanol. Using dilutions of cross absorbed antibody specific for common antigens and Ouchterlony test, high ethanol producing S. cerevisiae could be identified.


Electrophoretic Approach for the Simultaneous Deposition and Functionalization of Reduced Graphene Oxide Nanosheets with Diazonium Compounds: Application for Lysozyme Sensing in Serum.
Electrophoretic deposition (EPD) of reduced graphene oxide nanosheets (rGO) offers several advantages over other surface coating approaches, including process simplicity, uniformity of the deposited films, and good control of the film thickness. The EPD conditions might also be of interest for the reduction of diazonium salts, which upon the release of N2 molecules and generation of radicals, can form covalent bonds with the sp2 hybridized carbon lattice atoms of rGO films. In this work, we report on the coating of gold electrodes in one step with rGO/polyethylenimine (PEI) thin films and their simultaneous modification using different phenyl (Ph) diazonium salt precursors bearing various functionalities such as -B(OH)2, -COOH, and -C≡CH. We show further the interest of such interfaces for designing highly sensitive sensing platforms. Azide-terminated lysozyme aptamers were clicked onto the rGO/PEI/Ph-alkynyl matrix and used for the sensing of lysozyme levels in patients suffering from inflammatory bowel disease (IBD), where lysozyme levels are up-regulated. The approach attained the required demand for the determination of lysozyme level in patients suffering from IBD with a 200 fM detection limit and a linear range up to 20 pM without signal amplification.


Hereditary pancreatitis caused by a double gain-of-function trypsinogen mutation.
Hereditary pancreatitis, an autosomal dominant disease with approximately 80% penetrance, can be caused by both 'gain-of-function' missense and copy number mutations in the cationic trypsinogen gene (PRSS1). Here we demonstrate a heterozygous hybrid PRSS2 (encoding anionic trypsinogen)/PRSS1 gene in a French white family with hereditary pancreatitis, by means of quantitative fluorescent multiplex PCR and RT-PCR analyses. The hybrid gene, in which exons 1 and 2 are derived from PRSS2 and exons 3-5 from PRSS1, apparently resulted from a non-allelic homologous recombination (NAHR) event between the chromosome 7 homologs or sister chromatids during meiosis. Interestingly, this hybrid gene causes the disease through a combination of its inherent 'double gain-of-function' effect, acting simultaneously as a 'quantitative' copy number mutation and a 'qualitative' missense mutation (i.e. the known disease-causing p.N29I mutation). Our finding reveals a previously unknown mechanism causing human inherited disease, enriches the lexicon of human genetic variation and goes beyond the known interaction between copy number variations (CNVs) and single nucleotide substitutions in health and disease. Our finding should also stimulate more interest in analyzing both types of genetic variation whenever one tries to determine the contribution of a specific locus to a given disease phenotype.


Biochar accelerates PAHs biodegradation in petroleum-polluted soil by biostimulation strategy.
Sawdust and wheat straw biochars prepared at 300°C and 500°C were applied to petroleum-polluted soil for an 84-day incubation to estimate their effectiveness on polycyclic aromatic hydrocarbons (PAHs) removal. Biochars alone were most effective at reducing PAHs contents. However, adding biochar to soils in company with NaN3 solution resulted in a decreasing trend in terms of PAHs removal, which was even lower than treatment CK without biochar. Moreover, it was discovered by PCR-DGGE files and sequencing analysis that the predominant bacterial diversity slightly decreased but the abundance of some specific taxa, including PAHs degraders, was promoted with biochar input. These results highlighted the potential of biochar application on accelerating PAHs biodegradation, which could be attributed to the properties of biochars that benefit for making the amended soil a better habitat for microbes. The impacts of biochar preparation and pollutants nature on PAHs removal were also determined. Significant reduction in the PAHs contents was detected when adding biochar prepared at a high temperature (500°C), while the feedstocks of biochar showed little effect on PAHs removal. Due to the high hydrophobicity of aromatic rings, high-molecular weight PAHs were found much more resistant to microbial degradation in comparison with low-molecular weight PAHs.


A decision support system for telemedicine through the mobile telecommunications platform.
In this paper we have discussed the application of artificial intelligence in telemedicine using mobile device. The main goal of our research is to develop methods and systems to collect, analyze, distribute and use medical diagnostics information from multiple knowledge sources and areas of expertise. Physicians may collect and analyze information obtained from experts worldwide with the help of a medical decision support system. In this information retrieval system, modern communication tools such as computers and mobile phones can be used efficiently. In this work we propose a medical decision support system using the general packet radio service (GPRS). GPRS, a data extension of the mobile telephony standard Global system for mobile communications (GSM) is emerging as the first true packet-switched architecture to allow mobile subscribers to benefit from high-speed transmission rates and run JAVA based applications from their mobile terminals. An academic prototype of a medical decision support system using mobile device was implemented. The results reveal that the system could find acceptance from the medical community and it could be an effective means of providing quality health care in developing countries.


Spectral and temporal response patterns of single units in the chinchilla dorsal cochlear nucleus.
Spectral and temporal response patterns to pure-tone stimuli were collected from single units in the dorsal cochlear nucleus of anesthetized chinchillas. The spectral response profiles were divisible into groups based on the balance of excitation and inhibition. Temporal responses were characterized in chloralose-anesthetized animals by collecting PST-histograms. There appeared to be no simple one-to-one relationship between a unit's spectral and its temporal response pattern. Excitatory spectral responses were generally sharply tuned areas resembling those of auditory nerve fibers. However, unlike the latter, the majority of these had chopper or pauser/buildup temporal responses. Inhibitory spectral responses were of two distinct types: one included lateral inhibitory areas flanking the tuned excitatory areas which occasionally invaded the latter creating a nonmonotonic excitatory response at the unit's characteristic frequency. The other included sharply tuned inhibitory areas. The characteristic frequencies of these units were found to be in close correspondence with those of sharply tuned excitatory units from the same penetration suggesting that these inhibitory units were tonotopically mapped in the same register as tuned excitatory units. The spectral response patterns were studied with three types of anesthesia: ketamine/xylazine, dial/urethane, and chloralose. In each of these groups the patterns were similar. However, the proportions of units showing inhibition was strongly dependent on the choice of anesthetic agent with chloralose yielding the highest proportions (59%) and ketamine/xylazine yielding the lowest (29%).


Myocardial perfusion scintigraphy dosimetry: optimal use of SPECT and SPECT/CT technologies in stress-first imaging protocol.
Over the past decade, nuclear medicine experts have been seeking to minimize patient exposure to radiation in myocardial perfusion scintigraphy (MPS). This review describes the latest technological innovations in MPS, particularly with regard to dose reduction. We searched in PubMed for original clinical papers in English, published after 2008, using the following research criteria: (dose) and ((reduction) or (reducing)) and ((myocardial) or (cardiac) or (heart)) and ((nuclear medicine) or (nuclear imaging) or (radionuclide) or (scintigraphy) or (SPET) or (SPECT)). Thereafter, recent reviews on the topic were considered and other relevant clinical papers were added to the results. Of 202 non-duplicate articles, 17 were included. To these, another eight papers cited in recent reviews were added. By optimizing the features of software, i.e., through algorithms for iterative reconstruction with resolution recovery (IRRs), and hardware, i.e., scanners and collimators, and by preferring, unless otherwise indicated, the use of stress-first imaging protocols, it has become possible to reduce the effective dose by at least 50% in stress/rest protocols, and by up to 89% in patients undergoing a diagnostic stress-only study with new technology. With today's SPECT/CT systems, the use of a stress-first protocol can conveniently be performed, resulting in an overall dose reduction of about 35% if two-thirds of stress-first examinations were considered definitively normal. Using innovative gamma cameras, collimators and software, as well as, unless otherwise indicated, stress-first imaging protocols, it has become possible to reduce significantly the effective dose in a high percentage of patients, even when X-ray CT scanning is performed for attenuation correction.


Activation of synaptic and extrasynaptic glycine receptors by taurine in preoptic hypothalamic neurons.
Taurine is an essential amino-sulfonic acid having a fundamental function in the brain, participating in both cell volume regulation and neurotransmission. Using a whole cell voltage patch clamp technique, the taurine-activated neurotransmitter receptors in the preoptic hypothalamic area (PHA) neurons were investigated. In the first set of experiments, different concentrations of taurine were applied on PHA neurons. Taurine-induced responses were concentration-dependent. Taurine-induced currents were action potential-independent and sensitive to strychnine, suggesting the involvement of glycine receptors. In addition, taurine activated not only α-homomeric, but also αβ-heteromeric glycine receptors in PHA neurons. Interestingly, a low concentration of taurine (0.5mM) activated glycine receptors, whereas a higher concentration (3mM) activated both glycine and gamma-aminobutyric acid A (GABAA) receptors in PHA neurons. These results suggest that PHA neurons are influenced by taurine and respond via glycine and GABAA receptors.


Sleep Measurement in Children-Are We on the Right Track?
Sleep plays a critical role in the development of healthy children. Detecting sleep and sleep disorders and the effectiveness of interventions for improving sleep in children require valid sleep measures. Assessment of sleep in children, in particular infants and young children, can be a quite challenging task. Many subjective and objective methods are available to evaluate various aspects of sleep in childhood, each with their strengths and limitations. None can, however, replace the importance of thorough clinical interview with detailed history and clinical examination by a sleep specialist.


Records management guidance and its implications for research.
The UK Department of Health's code of practice for managing records has important implications for nurse researchers. Naomi Reay and Jill Firth consider the steps researchers need to take to comply with the code.


Influence of the Conditioning Method for Pre-Sintered Zirconia on the Shear Bond Strength of Bilayered Porcelain/Zirconia.
This study evaluated the bond strength of veneering porcelain with an experimental conditioner-coated zirconia. Pre-sintered Y-TZP specimens (n = 44) were divided in two groups based on conditioning type. After sintering, all sample surfaces were sandblasted and layered with veneering porcelain. Additionally, half of the specimens in each group underwent thermal cycling (10,000 cycles, 5-55 °C), and all shear bond strengths were measured. After testing, the failure mode of each fractured specimen was determined. Differences were tested by parametric and Fisher's exact tests (α = 0.05). The differences in bond strength were not statistically significant. Adhesive fractures were dominantly observed for the non-thermal cycled specimens. After thermal cycling, the conditioner-coated group showed cohesive and mixed fractures (p = 0.0021), whereas the uncoated group showed more adhesive fractures (p = 0.0021). Conditioning of the pre-sintered Y-TZP did not change the shear bond strength of the veneering porcelain, but did improve the failure mode after thermal cycling.


Area patterning of the mammalian cortex.
Here we describe mechanisms regulating area patterning of developing mammalian neocortex, referred to as arealization. Current findings indicate an interplay between intrinsic genetic mechanisms and extrinsic information relayed to cortex by thalamocortical input. Intrinsic mechanisms are based on morphogens and signaling molecules secreted by patterning centers, positioned at the perimeter of dorsal telencephalon, that generate across nascent cortex the graded expression of transcription factors in cortical progenitors. Two major patterning centers are the commissural plate, which expresses Fgf8 and Fgf17, and the cortical hem, which expresses Bmps and Wnts. Four transcription factors, COUP-TFI, Emx2, Pax6, and Sp8, with graded expression across the embryonic cortical axes, are shown to determine sizes and positions of cortical areas by specifying or repressing area identities within cortical progenitors. They also interact to modify their expression, as well as expression of Fgf8. We review these mechanisms of arealization and discuss models and concepts of cortical area patterning.


Obstructive sleep apnea improves with non-invasive hypoglossal nerve stimulation using temporal interference.
Peripheral nerve stimulation is used in both clinical and fundamental research for therapy and exploration. At present, non-invasive peripheral nerve stimulation still lacks the penetration depth to reach deep nerve targets and the stimulation focality to offer selectivity. It is therefore rarely employed as the primary selected nerve stimulation method. We have previously demonstrated that a new stimulation technique, temporal interference stimulation, can overcome depth and focality issues. Here, we implement a novel form of temporal interference, bilateral temporal interference stimulation, for bilateral hypoglossal nerve stimulation in rodents and humans. Pairs of electrodes are placed alongside both hypoglossal nerves to stimulate them synchronously and thus decrease the stimulation amplitude required to activate hypoglossal-nerve-controlled tongue movement. Comparing bilateral temporal interference stimulation with unilateral temporal interference stimulation, we show that it can elicit the same behavioral and electrophysiological responses at a reduced stimulation amplitude. Traditional transcutaneous stimulation evokes no response with equivalent amplitudes of stimulation. During first-in-man studies, temporal interference stimulation was found to be well-tolerated, and to clinically reduce apnea-hypopnea events in a subgroup of female patients with obstructive sleep apnea. These results suggest a high clinical potential for the use of temporal interference in the treatment of obstructive sleep apnea and other diseases as a safe, effective, and patient-friendly approach. The protocol was conducted with the agreement of the International Conference on Harmonisation Good Clinical Practice (ICH GCP), applicable United States Code of Federal Regulations (CFR) and followed the approved BRANY IRB File # 22-02-636-1279.


Iodine content of human tissues after administration of iodine containing drugs or contrast media.
Total iodine contents were determined in 209 bioptic or autoptic specimens of various extrathyroidal tissues. Fifty-two of the 80 subjects examined had no previous exposure to excessive iodine, 24 were tested after administration of x-ray contrast media and 4 after treatment with various iodine containing drugs. Unexposed subjects had tissue iodine contents (mean +/- SD) ranging from 0.85 +/- 0.17 microgram/100 g in the brain and 9.78 +/- 2.75 microgram/100 g in the liver of adults. A significantly lower iodine concentration was found in the liver of newborns (2.79 +/- 1.00 microgram, p less than 0.001). Most of the other tissues had iodine concentrations of 2-4 microgram/100 g. Subjects with previous exposure to iodine containing drugs or x-ray contrast media showed increased iodine contents of various degree in all examined tissues, including adipose tissue, bone, brain, kidney, liver, lung, skeletal muscle, skin and spleen. Accumulation of iodine in adipose tissue was still demonstrable more than two years after cholecystography.


NADH diferric transferrin reductase in liver plasma membrane.
Evidence is presented that rat liver plasma membranes contain a distinct NADH diferric transferrin reductase. Three different assay procedures for demonstration of the activity are described. The enzyme activity is highest in isolated plasma membrane, and activity in other internal membranes is one-eighth or less than in plasma membrane. The activity is inhibited by apotransferrin and antitransferrin antibodies. Trypsin treatment of the membranes leads to rapid loss of the transferrin reductase activity as compared with NADH ferricyanide reductase activity. Erythrocyte plasma membranes, which lack transferrin receptors, show no diferric transferrin reductase activity, although NADH ferricyanide reductase is present. The transferrin reductase is inhibited by agents that inhibit diferric transferrin reduction by intact cells and is activated by CHAPS (3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfate) detergent. Inhibitors of mitochondrial electron transport have no effect on the activity. We propose that the NADH diferric transferrin reductase in plasma membranes measures the activity of the enzyme that causes the reduction of diferric transferrin by intact cells. This transmembrane electron transport system requires the transferrin receptor for diferric transferrin reduction. Because the transmembrane electron transport has been shown to stimulate cell growth, the reduction of diferric transferrin at the cell surface may be an important function for diferric transferrin in stimulation of cell growth, in addition to its role in iron transport.


Acute ischemia causes axonal stasis, swelling, attenuation, and secondary demyelination.
Microsphere embolization of rat sciatic nerve capillaries results in a central fascicular ischemic core. Twenty-four hours after microembolization, the pathological alterations along the length of 55 myelinated fibers were reconstructed by computer imaging of 2,000 serial semi-thin epoxy sections of a tissue block that extended from just above and into an ischemic core. From proximal to distal, the typical sequence of pathological alterations was: normal----swollen dark axons + thin myelin or demyelination----attenuated axons----axon cytolysis (46 fibers) or normal axons (9 fibers). Because organelle accumulation and axonal swelling were the earliest and most proximal pathological lesions, we infer that regional hypoxia causes axonal stasis as a primary event. Demyelination was found in fibers showing swollen dark and attenuated axons. These findings suggest that axons are selectively vulnerable to acute ischemia and that, depending on severity, the fibers either undergo axonal degeneration or transitory structural alterations without axonal degeneration, the latter consisting of axonal changes and secondary demyelination.


Cloning a part of the ochratoxin A biosynthetic gene cluster of Penicillium nordicum and characterization of the ochratoxin polyketide synthase gene.
Penicillium nordicum is a fungal species able to produce high amounts of ochratoxin A. A 10kb genomic DNA fragment of P. nordicumn has been cloned which carries three long open reading frames. One open reading frame (otapksPN) has homology to fungal polyketide synthases. The second open reading frame (npsPN) has homology to non-ribosomal peptide synthetases and the third open reading frame (aspPN) has homology to fungal alkaline serine proteinases. The non-ribosomal peptide synthetase and the polyketide synthase are convergently transcribed. Interestingly, the polyketide synthase can be identified by PCR only in P. nordicum strains and not in the related species Penicillium verrucosum or in ochratoxigenic Aspergillus species, indicating that the ochratoxin polyketide synthases are different in the important ochratoxigenic species. In contrast, the non-ribosomal peptide synthetase can be identified in P. nordicum and P. verrucosum, but not in other species. An inactivation of the polyketide synthase resulted in strains with abolished capacity to produce ochratoxin A. Expression of the polyketide synthase correlates with ochratoxin A biosynthesis.


Mild calorie restriction does not affect testosterone levels and testicular gene expression in mutant mice.
The hypothalamic-pituitary-gonadal (HPG) axis and the somatotropic axis are influenced by nutritional factors. Calorie restriction (CR) extends lifespan but suppresses both the HPG and the somatotropic axes. Since most CR studies use a fairly severe (40%-60%) reduction of calorie intake, we hypothesized that a milder CR (20%) might not be deleterious to reproduction in male mice. To test this hypothesis, we evaluated the effects of 20% CR on testicular testosterone content and on testicular expression of genes that are relevant to testicular function and reproductive competence, including insulin-like growth factor-I, cytochrome P450 aromatase (Cyp19a1), androgen receptor, luteinizing hormone receptor, follicle-stimulating hormone receptor, cytochrome P450c17 and 3-beta-hydroxysteroid dehydrogenase/isomerase. To relate CR effects to the activity of the somatotropic axis, we have used growth hormone-resistant GHR knockout mice as well as transgenic mice overexpressing GH. Mild CR did not affect testosterone levels in testis homogenates and had little effect on expression of the examined genes in the reproductive organs. Altered activity of the GH/insulin-like growth factor-1 axis had a major impact on the parameters analyzed. The results also suggest that expression of several key genes involved in the control of testicular function is preserved under conditions of mild CR and encourage speculation that mild regimens of CR can produce longevity benefits without impairing reproduction.


A novel amidase from Brevibacterium epidermidis ZJB-07021: gene cloning, refolding and application in butyrylhydroxamic acid synthesis.
A novel amidase gene (bami) was cloned from Brevibacterium epidermidis ZJB-07021 by combination of degenerate PCR and high-efficiency thermal asymmetric interlaced PCR (hiTAIL-PCR). The deduced amino acid sequence showed low identity (≤55 %) with other reported amidases. The bami gene was overexpressed in Escherichia coli, and the resultant inclusion bodies were refolded and purified to homogeneity with a recovery of 22.6 %. Bami exhibited a broad substrate spectrum towards aliphatic, aromatic and heterocyclic amides, and showed the highest acyl transfer activity towards butyramide with specific activity of 1331.0 ± 24.0 U mg(-1). Kinetic analysis demonstrated that purified Bami exhibited high catalytic efficiency (414.9 mM(-1) s(-1)) for acyl transfer of butyramide, with turnover number (K cat) of 3569.0 s(-1). Key parameters including pH, substrate/co-substrate concentration, reaction temperature and catalyst loading were investigated and the Bami showed maximum acyl transfer activity at 50 °C, pH 7.5. Enzymatic catalysis of 200 mM butyramide with 15 μg mL(-1) purified Bami was completed in 15 min with a BHA yield of 88.1 % under optimized conditions. The results demonstrated the great potential of Bami for the production of a variety of hydroxamic acids.


Differentiation-dependent sialylation of individual neural cell adhesion molecule polypeptides during postnatal development.
The postnatal sialylation of individual neural cell adhesion molecule (N-CAM) polypeptides by a developmentally regulated sialyltransferase in Golgi-enriched fractions isolated from rat brain is described. The 120-kilodalton polypeptide of N-CAM was found to be sialylated at each developmental age examined. This was in contrast to the 140- and 180-kilodalton N-CAM polypeptides which were only sialylated until postnatal day 10 and from postnatal day 12, respectively. Immunoblotting procedures demonstrated that all N-CAM polypeptides were expressed in the Golgi fractions at each developmental stage examined. The heavily sialylated "embryonic" form of N-CAM was found to be reexpressed at postnatal days 10 and 12, a time coincident with extensive fibre outgrowth. The "embryonic" form of N-CAM incorporated similar amounts of [14C]sialic acid into its constituent polypeptides reflecting the difference in sialic acid to protein ratio, as this form of N-CAM was virtually undetectable in the immunoblots of postnatal material.


Characterization of the human Glvr-1 phosphate transporter/retrovirus receptor gene and promoter region.
The cell surface receptor for gibbon ape leukemia virus (Glvr-1) belongs to the type III sodium-dependent phosphate transporter/retrovirus receptor gene family. Several observations have suggested an important role for Glvr-1 in regulated Pi handling in bone forming cells and prompted us to investigate further the molecular mechanisms regulating Glvr-1 gene expression. In addition, the regulation of Glvr-1 gene expression also has potential applications to gene therapy, since retroviral vectors carrying gibbon ape leukemia virus envelope proteins are used for gene delivery into different cell types. The aim of this study was thus to clone the human Glvr-1 gene in order to describe its structure and its promoter region. Our results indicate that the Glvr-1 gene consists of 11 exons and 10 introns spread over 18kb of genomic DNA. The translation initiation site is located within exon II and the translation stop codon within exon XI. Rapid amplification of cDNA ends (5'-RACE) suggests that, in human SaOS-2 osteoblast-like cells, transcription of Glvr-1 is initiated at multiple sites, mostly located between bp 32 and 50 of the published cDNA sequence, which was initially obtained from HL-60 cells. The 5'-flanking region of the gene is characterized by a very high GC content. Reporter gene assays demonstrate the presence of a functional promoter upstream of exon I and indicate that a GC-rich region, containing two potential SP1 binding sites, is required for high promoter activity in transiently transfected SaOS-2 cells. The description of the human Glvr-1 gene structure, as well as the analysis of some structural and functional characteristics of its promoter region, provide a basis for more detailed investigation of the molecular mechanisms controlling expression of the Glvr-1 gene in bone forming cells and in other cell types.


Validation of the 3M™ Molecular Detection Assay 2 - STEC Gene Screen (stx) for the Detection of Shiga Toxin Gene (stx1 and/or stx2) in Fresh Raw Ground Beef and Fresh Spinach: AOAC Performance Tested MethodSM 071903.
The 3M™ Molecular Detection Assay 2 - STEC Gene Screen (stx) method is based on gene amplification by the use of real time loop-mediated isothermal amplification when used with the 3M Molecular Detection System for the rapid and specific detection of Shiga toxin gene (stx1 and/or stx2) from Shiga toxin-producing Escherichia coli (STEC) in enriched foods. The stx assay does not differentiate between stx1 and stx2 but detects the presence of stx1 and/or stx2. The 3M Molecular Detection Assay 2 - STEC Gene Screen (stx) method was evaluated for AOAC® Performance Tested MethodsSM certification. Matrix studies, inclusivity/exclusivity, robustness testing, product stability, and lot-to-lot variability testing were conducted to assess the method's performance. The 3M Molecular Detection Assay 2 - STEC Gene Screen (stx) demonstrated equivalent results to the United States Department of Agriculture/Food Safety and Inspection Service Microbiology Laboratory Guidebook Chapter 5C.00 reference method for fresh raw ground beef, and the U.S. Food and Drug Administration Bacteriological Analytical Manual Chapter 4A reference method for fresh spinach. The 3M Molecular Detection Assay 2 - STEC Gene Screen (stx) detected all STEC E. coli strains (E. coli strains with stx1 and/or stx2 genes) and did not detect any of the 45 strains from the exclusivity panel. Robustness testing indicated that small variations in critical test parameters did not adversely affect the assay's performance. Product consistency and stability testing demonstrated no differences between the lots evaluated. The data collected in these studies demonstrate that the 3M Molecular Detection Assay 2 - STEC Gene Screen (stx) is a reliable method for the rapid and specific detection of Shiga toxin-producing E. coli in raw ground beef and spinach. The 3M Molecular Detection Assay 2 - STEC Gene Screen (stx) method is suitable for the rapid and specific detection of Shiga toxin-producing E. coli in fresh raw ground beef, and spinach.


Positive aspect of caregiving among primary informal dementia caregivers in Singapore.
The present study investigated the factor structure of positive aspects of caregiving (PAC) scale among primary informal caregivers providing care to persons with dementia (PWD) in Singapore. 282 primary informal caregivers of PWD were recruited from the Institute of Mental Health, and Changi General Hospital and administered the 9-item PAC scale. A confirmatory factor analyses (CFA) was conducted to test the model fit of the 9-item PAC proposed by the scale developer and multiple linear regression was used to investigate the significant socio-demographic correlates. CFA showed that the 2-factor structure including 'Self-Affirmation' and 'Outlook on Life' had an acceptable model fit. After controlling for confounding variables, Malay caregivers were associated with higher scores on PAC and 'Self-Affirmation' compared to caregivers of other ethnicities. Caregivers with Secondary or below education level had higher PAC and 'Outlook on Life' scores. Caregivers who had received formal training scored higher in PAC, 'Self-Affirmation' and 'Outlook on Life'. The present study confirmed that the 2-factor structure of the 9-item PAC was suitable for informal caregivers of PWD in Singapore. The findings have important implications for locally available interventions to enhance caregiver's psychological well-being and reduce burden of care.


Increased serum lactate dehydrongenase should be included among the variables that define very-high-risk multiple myeloma.
In patients who have symptomatic multiple myeloma (MM), a high serum lactate dehydrogenase (LDH) level is associated with features of advanced disease, adds prognostic value to the international staging system (ISS) and predicts for inferior survival. However, it has not been clearly defined what the impact of this abnormality is for patients treated upfront with novel agent-based regimens. To address this issue we analyzed 203 consecutive unselected patients with symptomatic MM who received upfront treatment with novel agents in a single center. The median overall survival for patients with normal LDH was 54 months but in patients with increased LDH levels it was 21 months (P = .003), whereas increased serum LDH was associated with a higher probability of early death. Multivariate analysis confirmed that an increased LDH level is independently associated with poor survival. Furthermore, increased LDH levels could identify subgroups of patients within ISS-2 and ISS-3 with even worse outcome. We conclude that serum LDH is a simple, inexpensive, and readily available blood test that may be included among the variables that define very-high-risk MM.


Roles of hippocampal NMDA receptors and nucleus accumbens D1 receptors in the amphetamine-produced conditioned place preference in rats.
There are glutamatergic projections from the hippocampus to the nucleus accumbens (NAc), which regulate DA transmission in this structure. To be precise, the ventral hippocampal (VH) glutamatergic neurons project to the nucleus accumbens shell region (NAcSh), whereas the dorsal hippocampus (DH) sends glutamatergic projections to the nucleus accumbens core region (NAcC). This study investigates the roles of hippocampal N-methyl-D-aspartate (NMDA) glutamate receptors and NAc type 1 dopamine receptor (D1) in amphetamine-produced conditioned place preference (AMPH-CPP) in rats. Our earlier reports showed that AMPH-CPP results in the enhancement of hippocampal CaMKII activity and it can be impaired by NMDA antagonist (AP5). In this study AMPH-CPP did not alter the NAc CaMKII activity, although AMPH-CPP was impaired by a blockade of D1 receptors (SCH23390) during conditioning. Moreover, inactivation of hippocampal area (dorsal hippocampus or ventral hippocampus) impaired AMPH-CPP, but its effect was diminished by the activation of D1 receptors in accumbal region (NAc core or NAc shell). By inactivating both DH and NAc core resulted in the disruption of rat's CPP expression. However, the impaired CPP expression was recovered during the next testing session, suggesting the disruption of CPP expression was a short term effect. Moreover, the disruption of CPP expression was not exhibited if NAc core was not inactivated. Interestingly, the rats that received activation in VH but an inactivation in NAc shell before testing show impaired CPP expression compared to those received inactivation in both VH and NAc shell. DH activation plus an inactivation in NAc core before testing show a significantly higher rate of the weakening of AMPH-CPP expression. Similarly, an activation of VH plus an inactivation of NAc shell before testing also show a statistically significant lower CPP score on tests 3 and 4. These results, taken together, indicate that NMDA receptor activation in DH and VH have different enhancing effects on the AMPH-CPP as their innervations onto the different NAc regions are essential for AMPH-CPP establishment. If the deterioration of AMPH-CPP expression (or extinction process) resembles the formation of new learning, then this active process might have been facilitated by the hippocampal NMDA receptor activations during testing.


The mitochondrial toxin 3-nitropropionic acid aggravates reserpine-induced oral dyskinesia in rats.
The effects of a previous long-term administration of the mitochondrial toxin 3-nitropropionic acid were studied on an animal model of tardive dyskinesia, i.e., the frequency of spontaneous tongue protrusions in rats repeatedly treated with reserpine. 3-Nitropropionic acid (10 or 15 mg/kg i.p., every other day for 17 days) potentiated the increase in tongue-protrusion frequency induced by reserpine (1 mg/kg, s.c., every other day for 3 days) but did not modify reserpine-induced increase in immobility duration and decrease in locomotion frequency. These results support the notion that neurotoxic events are associated with the development of tardive dyskinesia.


Evaluation of 294 psychiatric consultations involving in-patients above 70 years of age in somatic departments in a university hospital.
The integrated geropsychiatric service of the Psychiatric Department of the University Hospital in Linköping consists of an observation ward, a day unit and an out-patient unit. One of the main tasks of the last-mentioned unit is to provide geropsychiatric consultations for in-patients in the non-psychiatric departments of the hospital. During the first 2 years from October 1982 to October 1984 300 consultations were performed. The consultations included registration of social, medical and psychiatric data, a psychiatric examination, discussions with the staff and a psychiatric report. Out of the total number of 300 consultations 294 could be evaluated completely. They comprised 247 individuals. The evaluation was performed after the first 2 years by screening the patients' psychiatric records and reports and by computer-analysis of 89 variables covering medical as well as social data. The patients' mean age was 78 years and 52% were treated in the Department of Internal Medicine. Fifty-three percent had had previous psychiatric contacts. Many were suffering from serious somatic diseases and 22% from depressive states. Only 9% showed no signs of psychiatric disorders. Twenty-three percent of the patients were transferred to the geropsychiatric observation ward or day unit and only 3% were recommended to be certified and transferred to a mental hospital. Pertinent findings are discussed in the light of international data on geropsychiatric consultations and liaison psychiatry. The study shows the importance of providing special geropsychiatric consultations for in-patients in somatic departments as a part of an integrated geropsychiatric service.


Effect of siRNA pre-Exposure on Subsequent Response to siRNA Therapy.
An alternative cancer therapy based on RNA interference (RNAi) has shown considerable promise but the possibility of resistance development is not known. This study explored the possibility of therapeutic resistance against siRNA nanoparticles in human cancer cells. Two approaches to siRNA treatment were undertaken using lipid-modified polyethylenimines, a single high concentration (shock) and repeated increasing concentrations (gradual). The targets were Mcl-1, RPS6KA5 and KSP in MDA-MB-435 cells. There was no evidence of resistance development in shock-treated cells, while the decrease in mRNA levels of targeted proteins was not as robust in naïve cells in gradual treatment. However, silencing efficiency was restored after a 7-day recovery period when expression of suppressed proteins returned to normal levels. Cellular uptake of siRNA was not affected by pre-treatments. Other mediators involved in cell survival and proliferation were altered in siRNA-treated cells, but only JUN silencing led to a heightened loss of viability. In vivo experiments demonstrated similar silencing efficiency at mRNA level after repeat doses. Human cancer cells responded to repeat siRNA nanoparticles in a similar fashion after a temporary initial alteration and little, if any, resistance was evident against repeated siRNA treatments.


Human leucocyte migration inhibitory factor (LIF). V. Specific regulation of the lymphokine serine protease activity by 3',5'-cGMP and, possibly, 2',3'-cCMP.
The cyclic nucleotides guanosine 3',5'-cyclic monophosphoric acid (3',5'-cGMP) and cytidine 2',3'-cyclic monophosphoric acid (2',3'-cCMP) but not cyclic phosphodiesters derived from the bases adenine and uracil preserved LIF activity against the blocking effect of the serine protease inhibitor phenylmethylsulphonyl fluoride (PMSF). Phosphomonoesters derived from guanosine and cytidine as well as 2',3'-cGMP and 3',5'-cCMP were all inactive, indicating specificity for phosphodiester bonds and their respective positions in the two active nucleotides. The protection afforded by 3',5'-cGMP and 2',3'-cCMP was dose dependent. Thus, using 10(-3) M PMSF, 3',5'-cGMP was active at concentrations higher than 10(-5) to 10(-4) M, and 2',3'-cCMP at concentrations higher than 3 X 10(-4) to 10(-3) M. The more pronounced LIF-inhibitory effect obtained by increased concentrations of PMSF could be overcome by raising the levels of the nucleotides, indicating that the interactions between PMSF and the nucleotides with LIF were mutally exclusive. The possibility that 3',5'-cGMP and perhaps 2',3'-cCMP function as modulators of LIF is discussed, and models for the function of this lymphokine are proposed.


HBVsvp-Pulsed Dendritic Cell Immunotherapy Induces Th1 Polarization and Hepatitis B Virus-Specific Cytotoxic T Lymphocytes Production.
In chronic hepatitis B virus (CHB) patients, both dendritic cells (DCs) and T cells are functionally impaired and consequently the HBV-specific cellular immune responses are downregulated. The present study aims to investigate whether monocyte-derived DC (MoDCs)-pulsed-HBV subviral particles (HBVsvp) can polarize Th1 cells to induce HBV-specific cytotoxic T-lymphocytes (CTL) responses in CHB patients. To this end, the human hepatoma HepG2.2.15 cell line was used to produce HBVsvp as a culturing system, and HBVsvp were concentrated for highly virus titer using the polyethylene glycol protocol. Peripheral blood mononuclear cells (PBMCs), collected from CHB patients and healthy donors, were differentiated into MoDCs and T cells. PBMCs-derived MoDCs were first pulsed with HBVsvp and then cultured with PBMCs-derived T cells. MoDCs and/or T subsets cells were identified for phenotypic activation by FACS analysis. The cytokine secretion of IL-4, IL-12, and IFN-γ in the culture supernatants was detected. The MoDCs were restored for their activation upon pulsing with HBVsvp in vitro, as identified by significantly overexpression of both CD86 and HLA-DR, and overproduction of IL-4 and IL-12. Furthermore, MoDCs-pulsed-HBVsvp induced Th1 frequencies and activated HBV-specific CTL to produce significantly highest amount of IFN-γ. Enhanced HBV-specific CTL led to strong cytolytic capacity against HepG2.2.15. Overall, our data suggest that in vitro activation of MoDCs with HBVsvp overcomes the functionally impaired DCs and T cells in CHB patients offering a promising tool for therapeutic or vaccine-based approaches against HBV.