Effects of the antacids in didanosine tablets on dapsone pharmacokinetics.
To investigate 1) the effects of the magnesium-aluminum antacids in didanosine tablets on dapsone absorption in healthy volunteers and 2) the effects of the antacid formulation of active didanosine tablets on dapsone pharmacokinetics in patients seropositive for human immunodeficiency virus (HIV). Two separate, randomized, two-period, two-treatment, crossover studies with a 21-day washout period between treatments. Clinical investigation unit of a university hospital. Six healthy men and six HIV-positive men. Serial dapsone plasma concentrations were measured when dapsone, 100 mg, was administered alone and with either the third of four doses of didanosine placebo tablets (group 1) or the 27th of 28 doses of active didanosine tablets (group 2). In each study, pharmacokinetic variables were determined using noncompartmental methods and compared by analysis of variance. When dapsone was administered alone, pharmacokinetic variables did not significantly differ from those with dapsone given in either combination (P > 0.10 for all comparisons). Didanosine, antacids, and other excipients in the currently used didanosine chewable tablets did not significantly affect plasma concentrations of dapsone.


[Osteomyelitis of the spine and its surgical treatment--personal experience].
Evaluation of surgical treatment of osteomyelitis of the spine, determination of efficient diagnostic and therapeutic criteria. The examined group comprised in total 27 patients. No selection was made, all patients who were operated on and treated at the Department of the authors were followed gradually as they were admitted at the Department. Evaluation was based on the monitoring of the clinical condition of patients including laboratory tests. Evaluated was also the achievement of vertebral body fusion three months after the surgery. The surgical procedure always contained a radical debridement of inflammatory focus, a massive autogenous cancellous bone grafting mixed with Gentamycin foam, either separately or in combination with instrumentation. Approach to the inflammatory focus was in 9 cases transforaminal posterior, in 2 cases transforaminal combined with transversectomy, in 2 cases transforaminal combined with laminectomy, in 4 cases transthoracic, in 1 case transabdominal, in 7 cases retroperitoneal and once percutaneous technique was used. All patients operated on healed without complications. Local pain and temperature subsided rapidly (in terms of days). Also objective inflammatory criteria returned to normal (in terms of weeks). Seven patients show improved neurological status. X-ray examination 3 months after the surgery always shows consolidation and ingrowth of cancellous bone grafts without local relapse or loss of correction. The numbers of patients of the authors' group correspond with the numbers of patients presented in literature. There is also agreement with other authors that a modern treatment of spinal osteomyelitis should include a radical debridement of the focus combined with instrumentation, neurological decompression and application of cancellous bone grafts completed by local antibiotics. As compared to other authors who present a prevalence of abscess the histological examination of our cases shows predominantly inflammatory granuloma. Radical surgical treatment of spinal osteomyelitis including debridement, cancellous bone grafting, internal fixation (instrumentation) and decompression together with a general treatment and orthotic management seems to be an efficient way of the treatment of this serious disease.


Dependency of Experimental Autoimmune Encephalomyelitis Induction on MOG35-55 Properties Modulating Matrix Metalloproteinase-9 and Interleukin-6.
Experimental autoimmune encephalomyelitis (EAE) is commonly induced with myelin oligodendrocyte glycoprotein (MOG)35-55; occasionally, EAE is not well induced despite MOG35-55 immunization. To confirm that EAE induction varies with difference in MOG35-55 properties, we compared three MOG35-55 from different commercial sources, which are MOG-A, MOG-B, and MOG-C. The peptides induced EAE disease with 100, 40, and 20 % incidence, respectively. Compared with others, MOG-A showed higher peptide purity (99.2 %) and content (92.2 %) and presented a sheet shape with additional sodium and chloride chemical elements. In MOG-A-treated group, MMP-9 activity and IL-6 levels were considerably higher than the other groups in CNS tissues, and significantly increased VCAM-1, IFN-γ, and decreased IL-4 were also shown compared to MOG-B- and/or MOG-C-treated group. In conclusion, the immunological and toxicological changes by the difference in MOG35-55 properties modulate EAE induction, and MOG35-55 which affects MMP-9 activity and IL-6 levels may be the most effective EAE-inducing antigen. This study can be potentially applied by researchers using MOG35-55 peptide and manufacturers for MOG35-55 synthesis.


Detection of spatiotemporal changes in ecological quality in the Chinese mainland: Trends and attributes.
Global climate change and revegetation programs have significantly changed the ecological quality (EQ) in the Chinese mainland after 1999. Monitoring and assessing the changes in the regional EQ and analyzing their drivers are crucial for ensuring ecological restoration and rehabilitation. However, it is challenging to carry out a long-term and large-scale quantitative assessment of the EQ of a region based on traditional field investigations and experiment methods alone; notably, in previous studies, the effects of carbon and water cycles and human activities on the variations in EQ have not been studied comprehensively. Therefore, in addition to remote sensing data and principal component analysis, we used the remote sensing-based ecological index (RSEI), to assess the EQ changes in the Chinese mainland during 2000-2021. Additionally, we also analyzed the impacts of carbon and water cycles and anthropological activities on the changes in the RSEI. The main conclusions of this study were: since the beginning of the 21st century, we observed a fluctuating upward trend in the EQ changes in the Chinese mainland and eight climatic regions. From 2000 to 2021, in terms of the EQ, North China (NN) portrayed the highest increase rate (2.02 × 10-3 year-1, P < 0.05). There was a breaking point in 2011, the EQ in the region experienced a change, from a downward trend to an upward one. Northwest China, Northeast China, and NN portrayed an overall significant increasing trend in the RSEI, whereas the southwest part of the Southwest Yungui Plateau (YG) and a part of the plain region of the Changjiang (Yangtze) River (CJ) river region portrayed a significant decreasing trend in the EQ. Overall, the carbon and water cycles and human activities played a pivotal role in determining the spatial patterns and trends of the EQ in the Chinese mainland. In particular, the self-calibrating Palmer Drought Severity Index, actual evapotranspiration (AET), gross primary productivity (GPP), and soil water content (Soil_w) were identified as the key drivers of the RSEI. In the central and western Qinghai-Tibetan Plateau (QZ) and the northwest region of NW, the changes in RSEI were dominated by AET; however, in central NN, southeastern QZ, northern YG, and central NE, the changes were driven by GPP, and in the southeast region of NW, south region of NE, northern region of NN, middle YG region, and a part of the middle CJ region, the changes were driven by Soil_w. The population-density-related change in the RSEI was positive in the northern regions (NN and NW) but negative in the southern regions (SE), whereas the RSEI change related to ecosystem services was positive in the NE, NW, QZ, and YG regions. These results are beneficial for the adaptive management and protection of the environment and the realization of green and sustainable developmental strategies in the Chinese mainland.


Bilateral electrodermal activity in depressive patients.
Integrated bilateral skin conductance response and reactive lateral eye movements were studied in 19 depressive patients and 14 normal subjects during visual and verbal tasks and tone habituation sequence. Distinct right-left differences between depressed and normal subjects in reactive lateral eye movements were revealed. Both endogenous and reactive depression patients were classified as 'left-movers'. Marked bilateral differences were observed in electrodermal activity (EDA) in patients during verbal and visual tasks. In endogenous depression EDA was higher on the left hand compared with the right under all the conditions studied (visual and verbal, and tone habituation sequence). In reactive depression EDA was higher on the left side during the verbal task and tone habituation sequence and on the right side in the visual task. Neither result in reactive depression group attained significance. The results are interpreted as a reflection of a different pathophysiology underlying reactive and endogenous depression.


Expression of gingival crevicular fluid markers during early and late healing of intrabony defects after surgical treatment: a systematic review.
Surgical treatments such as guided tissue regeneration (GTR) and access flap surgery are widely employed for the treatment of intrabony defects. However, little is known regarding the postoperative expression of gingival crevicular fluid (GCF) markers. The aim of this systematic review was to compare the expression of GCF markers following treatment of periodontal intrabony defects with guided tissue regeneration or access surgery. The association of the markers' expression with the clinical outcome was also assessed. An electronic literature search was conducted in MEDLINE, EMBASE, OpenGrey, LILACS and Cochrane Library up to December 2018 complemented by a manual search. Human, prospective clinical studies were identified. The changes from baseline up to 30 days (early healing) and 3 months (late healing) were assessed. A total of 164 publications were identified and reviewed for eligibility. Of these, 10 publications fulfilled the inclusion criteria. The included studies evaluated 15 different GCF markers with a follow-up time between 21 and 360 days postoperatively. PDGF, VEGF and TIMP-1 changes were often investigated in the included studies; however, contrasting results were reported. Two studies agreed that both GTR and OFD lead to similar OPG level changes. TGF-β1 is increased early postoperatively, irrespective of the surgical technique employed. There is limited evidence available on the expression of GCF markers after surgical interventions of intrabony periodontal defects. However, OPG and TGF-β1 tend to increase early post-operatively, irrespective of the surgical technique employed, irrespective of the surgical technique employed. More well-designed, powered studies with sampling periods reflecting the regenerative process are needed, and future research should focus on employing standardised protocols for collecting, storing and analysing GCF markers.


Adapting the Short Grit Scale with Exploratory Structural Equation Modeling for Portuguese College Students.
The Short Grit Scale (Grit-S) is a self- and informant-report version of the longer Grit Scale, and it retains the 2-factor structure of the original scale. Our purpose in this research was to measure trait-level perseverance and passion for long-term goals by translating and validating the Grit-S for Portuguese respondents. Our participants were 572 college students (135 female, 437 male; age range 18-30 years, M age = 21.47, SD = 2.29 years) from twelve Portuguese universities. Our data confirmed the scale's two-factor structure ("consistency of interests" and "perseverance of effort") and demonstrated appropriate adjustment values (CFI = 0.999, TLI = 0.981, SRMR = 0.017, RMSEA = 0.001, CI90%= 0.000-0.041). We found the adapted scale to be invariant for sex. Use of the scale confirmed an association between grit and well-being. These results imply that other investigators and practitioners interested in this scale may now apply it with Portuguese young adults.


An elastic material for cartilage replacement in an arthritic shoulder joint.
Inevitable complications after total shoulder replacements, mainly due to wear and loosening of glenoid components, have stimulated research on a new glenoid implant design. The aim of this study is to analyse a new design concept of the glenoid component with the articular surface made of artificial cartilage-specifically a poly(vinyl-alcohol)cryogel (PVA-c). Based on the performed experiments and hyperelasticity law the material model of the PVA-c is defined. Mechanical effects of using a cryogel layer with different geometries in glenoid components are analysed using a finite element method. The analysis allows for identification of design solutions with reduced, lower than failure, strains and stresses in the cryogel. The layer shape similar to the glenoid cartilage shows the most favourable mechanical effects. This indicates a high potential of using this soft material for the articular surface. Replacing the polyethylene with a cryogel layer results in a significant reduction in contact stresses together with the growth in contact area. This can promote fluid film lubrication and lower the wear, thus lower implant failure. Moreover, soft cryogel, having shock absorbing properties, may result in lower stresses at the implant-bone, thus lowering the risk of implant loosening. This study indicates a high importance of an elastic implant for restoring full function of the articular surface in a glenoid component.


New insulin sensitivity adjusted beta cell indices: validation according to a genetic study.
The role of hepatocyte nuclear factor-1alpha (HNF1alpha) in the maturity-onset diabetes of the young 3 (MODY3) is well established. A common polymorphism, I27L that is not linked to MODY3, has been demonstrated to affect beta cell function. To facilitate the identification of subjects with a low beta cell reserve, we developed beta cell indices and validated according to a genetic study. Insulin sensitivity index (ISI), 1st phase insulin response (1stIR), and 2nd phase insulin response (2ndIR) were assessed in 60 glucose tolerant subjects using hyperglycemic clamps. Delta1stIR and delta2ndIR were defined as differences between 1stIR (or 2ndIR) and the ISI-adjusted 1stIR (or 2ndIR). The genotypes were determined from genomic DNA. Delta1stIR (P = 0.0130) and delta2ndIR (P = 0.0482) differed among the 3 genotypic groups. Multivariate analysis confirmed the independent influence of the I27L polymorphism on delta1stIR (P = 0.0130) and delta2ndIR (P = 0.0369). Within the LL group, 75% and 63% of the subjects were within the lowest quartile of delta1stIR (P = 0.0011) and delta2ndIR (P = 0.0277), respectively. With new beta cell indices, we demonstrated the independent impact of the I27L polymorphism on beta cell function. The new beta cell indices will facilitate the identification of glucose tolerant subjects with reduced beta cell reserve.


Neonatal hypoxia/ischemia is associated with decreased inflammatory mediators after erythropoietin administration.
Erythropoietin (EPO), a hematopoietic growth factor, has been shown to be neuroprotective when administered as either a pretreatment or posttreatment. This study tested the hypothesis that one of the mechanisms of protection afforded by posttreatment with recombinant human EPO (rh-EPO) is an anti-inflammatory effect via inhibition of interleukin (IL)-1beta. Seven-day-old rat pups were subjected to unilateral carotid artery ligation followed by 90 minutes of hypoxia (8% O2 at 37 degrees C). Pups were divided into the following groups: control, hypoxia/ischemia, and hypoxia/ischemia plus rh-EPO. In the rh-EPO-treated pups, rh-EPO (5 U/g body weight IP) was administered starting 24 hours after the insult and then for 2 additional days. Samples were collected at 3, 7, 14, and 21 days after the insult. IL-1beta mRNA and protein levels were determined by quantitative real-time reverse transcription-polymerase chain reaction and ELISA. Tumor necrosis factor (TNF)-alpha mRNA levels were determined by colorimetric microplate assay. rhEPO attenuated brain injury, as assessed by brain weight, and attenuated both the hypoxia/ischemia-induced increases in IL-1beta mRNA and protein levels. TNF-alpha mRNA levels did not increase at 3 to 14 days after the hypoxic/ischemic insult. Administration of exogenous rh-EPO starting 24 hours after a hypoxic/ischemic insult is neuroprotective in the neonatal rat. This neuroprotective activity prevented the secondary, delayed rise in IL-1beta and attenuated the infiltration of leukocytes into the ipsilateral hemisphere.


Hyperimmunoglobulinemia D and periodic fever syndrome (HIDS) in a child with normal serum IgD, but increased serum IgA concentration.
This report describes a boy with hyperimmunoglobulinemia D and periodic fever syndrome (HIDS). The serum IgD level was normal, but the serum IgA concentration was markedly elevated. In addition, he had a history of orchitis on two occasions, a previously unreported manifestation of HIDS. This report expands the clinical and laboratory features associated with HIDS and serves to emphasize that a normal serum IgD level does not exclude the diagnosis of HIDS.


Teaching periodontics. Past patterns and future prospects.
The article presents a brief history of the development of periodontal concepts and their data base. Because of the rapid change in periodontal science and knowledge, the author pleads for periodontal education based on "show, deliberate, choose, and do" in our dental curricula.


The glycolysis/HIF-1α axis defines the inflammatory role of IL-4-primed macrophages.
T helper type 2 (Th2) cytokine-activated M2 macrophages contribute to inflammation resolution and wound healing. This study shows that IL-4-primed macrophages exhibit a stronger response to lipopolysaccharide stimulation while maintaining M2 signature gene expression. Metabolic divergence between canonical M2 and non-canonical proinflammatory-prone M2 (M2INF) macrophages occurs after the IL-4Rα/Stat6 axis. Glycolysis supports Hif-1α stabilization and proinflammatory phenotype of M2INF macrophages. Inhibiting glycolysis blunts Hif-1α accumulation and M2INF phenotype. Wdr5-dependent H3K4me3 mediates the long-lasting effect of IL-4, with Wdr5 knockdown inhibiting M2INF macrophages. Our results also show that the induction of M2INF macrophages by IL-4 intraperitoneal injection and transferring of M2INF macrophages confer a survival advantage against bacterial infection in vivo. In conclusion, our findings highlight the previously neglected non-canonical role of M2INF macrophages and broaden our understanding of IL-4-mediated physiological changes. These results have immediate implications for how Th2-skewed infections could redirect disease progression in response to pathogen infection.


Impact of introducing a lactation consultant into a neonatal unit.
Benefits of mothers' own milk (MOM) for premature and sick neonates are well documented. To increase access, many neonatal units have a lactation consultant (LC) on staff. This study aimed to assess the impact of a permanent LC on (i) maternal access to LC support; (ii) staff confidence in providing Breast Feeding (BF) education and (iii) provision of MOM. Study included a staff survey and chart audit. Questions provided feedback on access to lactation support and meeting maternal needs. Audit data included: gestational age, birthweight, intention to breastfeed, documentation of LC appointment, provision of MOM at 12 hours, days 3, 7, 28 and discharge. Student's t-tests were used for numerical data and chi-squared tests for categorical variables. Ninety-one staff surveys were returned, (pre 35/75 (47%), post 56/85 (66%) with staff reporting organising an LC appointment was significantly easier (P < 0.0001). Staff perceived maternal lactation needs and confidence to breastfeed post-discharge had significantly improved post-LC. The chart audit showed a significant increase in maternal access to LC appointments (15% vs. 80%; P < 0.01), breast pump education by day 3 (65% vs. 81%; P < 0.01), and an increase in MOM provision by 12 h (46% vs. 61%; P < 0.01) post-LC but not at days 7, 28 or discharge. A dedicated LC increases staff and maternal access to lactation education and support, improving provision of early MOM. Further research is required to assess the effect of LCs in improving breastfeeding rates in neonatal units.


Complete Genome Sequence of the Clinical Beijing-Like Strain Mycobacterium tuberculosis 323 Using the PacBio Real-Time Sequencing Platform.
We report here the whole-genome sequence of the multidrug-resistant Beijing-like strain Mycobacterium tuberculosis 323, isolated from a 15-year-old female patient who died shortly after the initiation of second-line drug treatment. This strain is representative of the Beijing-like isolates from Colombia, where this lineage is becoming a public health concern.


Molecular mechanism of antibiotic resistance induced by mono- and twin-chained quaternary ammonium compounds.
The usage of quaternary ammonium compounds (QACs) as disinfectants has increased dramatically since the outbreak of COVID-19 pandemic, leading to potentially accelerated emergence of antibiotic resistance. Long-term exposure to subinhibitory level QACs can lead to multidrug resistance, but the contribution of mutagenesis to resistance evolution is obscure. In this study, we subcultured E. coli K-12 under subinhibitory (0.25 × and 0.5 × Minimum Inhibitory Concentration, MIC) or inhibitory (1 × and 2 × MIC) concentrations of benzalkonium chloride (BAC, mono-chained) or didecyldimethylammonium chloride (DDAC, twin-chained) for 60 days. The sensitivity of QAC-adapted cells to five typical antibiotics decreased significantly, and in particular, the MIC of rifampicin increased by 85 times. E. coli adapted faster to BAC but developed 20-167% higher antibiotic resistance with 56% more mutations under DDAC exposure. The broader mutations induced by QACs, including negative regulators (acrR, marR, soxR, and crp), outer membrane proteins and transporters (mipA and sbmA), and RNA polymerase (rpoB and rpoC), potentially contributed to the high multi-drug resistance. After QACs stresses were removed, the phenotypic resistance induced by subinhibitory concentrations of QACs was reversible, whereas that induced by inhibitory concentrations of QACs was irreversible. The different patterns and molecular mechanism of antibiotic resistance induced by BAC and DDAC is informative to estimating the risks of broader QACs present at varied concentrations in the environment.


Effect of the Time to Change anti-stigma campaign on trends in mental-illness-related public stigma among the English population in 2003-13: an analysis of survey data.
Understanding trends and effective mechanisms that are likely to reduce public stigma and discrimination towards people with mental illness is important. We aimed to assess changes in public stigma in England after the introduction of the Time to Change anti-stigma campaign. We used data from the 2003 and 2007-13 national Attitudes to Mental Illness surveys to investigate 10-year trends in public attitudes across England before and during the Time to Change anti-stigma campaign. We present annual mean scores for attitude items related to prejudice and exclusion, and tolerance and support for community care. We also present an extrapolated linear trend line for the years 2009-13 and estimate population attitude scores without the campaign. We present unadjusted and adjusted linear regression models. In addition, we used multivariable linear regression models fitted to data aggregated by region to investigate whether a dose-effect response exists between campaign awareness and regional outcomes related to knowledge, attitudes, and intended behaviour. About 1700 respondents were surveyed each year. Significant increases in positive attitudes related to prejudice and exclusion occurred after the Time to Change campaign. In the multivariable analysis, we noted a significant increase in positive attitudes in relation to prejudice and exclusion after the launch of Time to Change (reverse-coded Z score 0·02, 95% CI 0·01 to 0·05; p=0·01), but not for tolerance and support for community care (Z score 0·01, -0·01 to 0·03; p=0·27). We also found evidence for a dose-effect relation between campaign awareness and regional improvement in knowledge (p=0·004) and attitudes (tolerance and support p<0·0001; prejudice and exclusion p=0·001), but not intended behaviour (p=0·20). The positive effects of Time to Change seem to be significant and moderate. Although attitudes are probably more at risk of deterioration during times of economic hardship, anti-stigma programmes might still play an active part in long-term reduction of stigma and discrimination, especially in relation to prejudice and exclusion of people with mental health problems. UK Department of Health, Comic Relief, Big Lottery.


Synthesis and structure-activity relationships of 1,2,4-triazoles as a novel class of potent tubulin polymerization inhibitors.
A novel triazole-containing chemical series was shown to inhibit tubulin polymerization and cause cell cycle arrest in A431 cancer cells with EC(50) values in the single digit nanomolar range. Binding experiments demonstrated that representative active compounds of this class compete with colchicine for its binding site on tubulin. The syntheses and structure-activity relationship studies for the triazole derivatives are described herein.


Soluble HLA class I antigen secretion by normal lymphocytes: relationship with cell activation and effect of interferon-gamma.
HLA class I antigens are thought to be integral membrane proteins. However, soluble forms of these molecules have been detected. Our laboratory has recently shown that the predominant form of these soluble proteins present in human serum, spleen tissue and culture supernatant of activated lymphocytes exhibits molecular weight and structure similar to classical HLA class I antigens, but lacks HLA A or B polymorphic determinants. In the present study, the secretion of such soluble proteins by lymphocytes has been further explored. Phytohaemagglutinin-stimulated normal lymphocytes secrete considerable quantities of soluble HLA (sHLA) class I proteins. This secretion seems to be a general property of lymphocytes, since activation of T as well as B cells by appropriate mitogens equally induce sHLA I secretion. Lymphocytes require RNA and protein synthesis, but not DNA synthesis, for the secretion to occur. Kinetic studies reveal that maximal sHLA I secretion precedes the peak of DNA synthesis by 24 h. In vitro stimulation with antigens or alloantigens also provokes sHLA I secretion. Moreover, this phenomenon has also been detected for in vivo-activated lymphocytes, as enhanced spontaneous sHLA I secretion was observed in cultures of low-density blastic B and T cells, and of blood lymphocytes obtained from normal subjects who had received a booster immunization 5 days earlier. Interferon-gamma (IFN-gamma) increases the expression of membrane-bound class I antigens but does not induce any sHLA I secretion, suggesting that both molecules are under different regulatory mechanisms. Our results indicate that human lymphocytes, upon stimulation, actively secrete considerable amounts of a soluble form of these biologically relevant proteins.


An equatorial oscillation in Saturn's middle atmosphere.
The middle atmospheres of planets are driven by a combination of radiative heating and cooling, mean meridional motions, and vertically propagating waves (which originate in the deep troposphere). It is very difficult to model these effects and, therefore, observations are essential to advancing our understanding of atmospheres. The equatorial stratospheres of Earth and Jupiter oscillate quasi-periodically on timescales of about two and four years, respectively, driven by wave-induced momentum transport. On Venus and Titan, waves originating from surface-atmosphere interaction and inertial instability are thought to drive the atmosphere to rotate more rapidly than the surface (superrotation). However, the relevant wave modes have not yet been precisely identified. Here we report infrared observations showing that Saturn has an equatorial oscillation like those found on Earth and Jupiter, as well as a mid-latitude subsidence that may be associated with the equatorial motion. The latitudinal extent of Saturn's oscillation shows that it obeys the same basic physics as do those on Earth and Jupiter. Future highly resolved observations of the temperature profile together with modelling of these three different atmospheres will allow us determine the wave mode, the wavelength and the wave amplitude that lead to middle atmosphere oscillation.


The great mimicker: Phenotypic overlap between constitutional mismatch repair deficiency and Tuberous Sclerosis complex.
Constitutional mismatch repair deficiency is a rare cancer predisposition syndrome caused by biallelic mutations in one of the four mismatch repair genes. Patients are predisposed to various tumors including hematological malignancies, brain tumors and colorectal carcinomas. Phenotypic overlap with Neurofibromatosis-1 is well known, with most patients presenting with café-au-lait macules. Other common features include axillary and/or inguinal freckling and intracranial MRI foci of high T2W/FLAIR signal intensity similar to the typical FASI seen in Neurofibromatosis-1. In this cohort of eight patients with constitutional mismatch repair deficiency we describe overlapping phenotypical features with Tuberous Sclerosis complex. In addition to "ash-leaf like" hypomelanotic macules (five patients), we detected intracranial tuber-like lesions (three patients), renal cysts (three patients) and renal angiomyolipomas (two patients). All our patients also had Neurofibromatosis-1 like features, mainly café-au-lait macules. This study suggests that features of Tuberous sclerosis especially when overlapping with those of Neurofibromatosis 1 or malignancies atypical for these syndromes should raise the possibility of constitutional mismatch repair deficiency. Correct diagnosis is essential for appropriate genetic counseling and pre-emptive cancer surveillance.


Effect of intermediate dose dexamethasone on post-operative pain in lumbar spine surgery: A randomized, triple-blind, placebo-controlled trial.
Dexamethasone has demonstrated analgesic properties and is used as an adjunctive pain agent for many procedures. We evaluated the efficacy of a single, intermediate dose of dexamethasone on post-operative analgesic consumption, and pain scores for lumbar spine surgery. Eighty patients aged between 18 and 70 scheduled for lumbar decompressive laminectomy were randomly allocated into two groups to receive either intravenous 0.2 mg/kg dexamethasone (group D = 40) or normal saline (group P = 40) before anesthetic induction. Post-operative total morphine consumption and the respective pain score at the PACU, 4, 6, 12, 24 and 48 h were evaluated. In addition, any adverse events were recorded. Total post-operative morphine consumption within 48 h was significantly lower in group D (34.5 vs. 42.5 mg, p = 0.031); however, the respective morphine consumption at each assessment was similar between groups. The respective NRS pain score at rest and upon movement in both groups was not significantly different for any time comparison. The average NRS pain score at rest and upon movement within 48 h was similar in both groups (i.e., NRS at rest Group D 3.6 vs. Group P 3.8, p = 0.936, and NRS for movement Group D 6.2 vs. Group P 6.3, p = 0.791). The adverse events within 48 h were also similar and serious complications (i.e., respiratory depression or surgical infection) were not found in either group. A single, intermediate dose of dexamethasone before anesthetic induction could minimally decrease post-operative morphine consumption within 48 h after lumbar decompressive laminectomy without any effect on the pain score.


Nurse prescribing of medicines in Western European and Anglo-Saxon countries: a survey on forces, conditions and jurisdictional control.
The number of Western European and Anglo-Saxon countries where nurses are legally allowed to prescribe medicines is growing. As the prescribing of medicines has traditionally been the task of the medical profession, nurse prescribing is changing the relationship between the medical and nursing professions. To gain more insight into the forces that led to the introduction of nurse prescribing of medicines in Western European and Anglo-Saxon countries, as well as into the legal, educational and organizational conditions under which nurses prescribe in these countries. Moreover, this study sought to determine which consequences nurse prescribing has for the division of jurisdictional control over prescribing between the nursing and medical professions. International survey. An email survey was sent to 60 stakeholders of professional nursing or medical associations or government bodies, at national, state or provincial level across ten Western European and Anglo-Saxon countries, namely Australia, Canada, Finland, Ireland, the Netherlands, New Zealand, Spain, Sweden, the United Kingdom and the United States of America. The survey addressed the reasons for the introduction of nurse prescribing and the conditions under which nurses are or will be prescribing medicines. The response rate was 65% (n=39). It was shown that a diversity of forces led to the introduction of nurse prescribing, and respondents from nursing and medical associations and government bodies cited different forces as being important for the introduction of nurse prescribing. Representatives of nurses' associations oftentimes emphasized the medication needs of patients living in remote geographical areas, while representatives of medical associations more often pointed to workforce shortages within the health care service. The conditions under which nurses prescribe medicines vary considerably, from countries where nurses prescribe independently to countries in which prescribing by nurses is only allowed under strict conditions and the supervision of physicians. Citing different forces as being important in the introduction of nurse prescribing can be conceived as a professional 'problem construction' in order to gain jurisdiction over the prescribing task. In most countries, nurses prescribe in a subordinate position and the jurisdiction over prescribing remains predominantly with the medical profession.


Modeling and assessing water and nutrient balances in a tile-drained agricultural watershed in the U.S. Corn Belt.
Identifying the key processes and primary sources of water and nutrient losses is essential for water quantity and quality management in watersheds. This is especially true in the U.S. Corn Belt, which has been recognized as the primary region contributing nutrient loads to the Great Lakes and the Gulf of Mexico. A SWAT (Soil and Water Assessment Tool) model simulation was set up in an agricultural watershed with about 50% tile drainage area in the U.S. Corn Belt to study the water and nutrient balance components for the whole watershed and the corn-soybean rotation system. The SWAT model was improved to consider additional nitrogen and phosphorus loss paths from the soil. The model was comprehensively calibrated and validated for simulating monthly stream flow, total suspended solids (TSS), nutrient loads (including total Kjeldahl nitrogen (TKN), nitrate and nitrite nitrogen (NOx-N), total phosphorus (TP) and orthophosphate phosphorus (orthoP)), actual evapotranspiration (ETa), leaf area index (LAI) and annual crop yields in the watershed from 2011 to 2019. Results showed the model performance was very good for simulating the stream flow, TSS and ETa, and acceptable for nutrient loads, LAI and crop yields. ETa, surface runoff, lateral soil flow, tile drainage and percolation respectively accounted for 65%, 15%, 2%, 8% and 9% of the precipitation. Fertilizer was the main source of nitrogen and phosphorus input to the watershed, and harvested crops were the main paths removing nutrients. Surface runoff, tile drainage and percolation each contributed about 30% of total nitrogen losses to water, with surface runoff being dominated by organic nitrogen while tile drainage and percolation were dominated by nitrate nitrogen. Phosphorus losses were mainly through surface runoff, which resulted in 66% of the total losses and was dominated by organic phosphorus and soluble phosphorus. Representing about 49% of the watershed area, the corn-soybean rotation system contributed 83% and 88% of the total nitrogen and phosphorus inputs, respectively, to the watershed, as well as 64% and 46% of the nitrogen and phosphorus losses to the water system, respectively. The non-growing season (October to the next April) was identified as the critical period resulting in water and nutrient losses due to low evapotranspiration and plant uptake. Targeted management strategies for reducing nutrient loads in key hydrological paths were suggested.


Detection of acute Epstein Barr virus cerebellitis using sequential brain HMPAO-SPECT imaging.
We used Technetium-99m hexamethyl propylene amine oxime single-photon emission computed tomography (HMPAO-SPECT) and magnetic resonance imaging (MRI) to evaluate a 30-year-old woman who developed cerebellar dysfunction after infection with Epstein Barr virus. Although no abnormal findings were evident from a brain MRI, the HMPAO-SPECT revealed increased perfusion in both cerebellar hemispheres. The patient improved without specific treatment. Two sequential HMPAO-SPECTs showed gradually reduced hyperperfusion in the cerebellum, which was correlated with her clinical recovery. We suggest that brain HMPAO-SPECT may thus be useful for identifying acute cerebellitis and predicting its clinical outcome.


Systemic absorption of adrenaline after aerosol, eye-drop and subcutaneous administration to healthy volunteers.
Adrenaline is the drug of choice for management of the anaphylactic reaction. The objective of this study was to compare systemic absorption of adrenaline after administration by different routes to healthy volunteers. Ten puffs (1.5 mg as adrenaline base) with 10-15 s intervals between them followed 2 h later by 20 puffs (3 mg) of adrenaline from a pressurized aerosol (Medihaler-Epi, 3M Riker, 14.0 mg/ml adrenaline acid tartrate) were sprayed into the cheek pouch or inhaled through the mouth or the nostrils. Adrenaline was also administered to the eyes by giving 2 drops (1 mg) of Isopto-Epinal (Alcon, 10 mg X ml-1). Finally, 0.5 ml (0.5 mg) of adrenaline was given subcutaneously in the upper arm of the same individuals. The systemic absorption was determined by measuring plasma adrenaline levels and effects on blood pressure, heart rate and finger tremor before and 5, 15, 30, 60, 90, and 120 min after adrenaline administration. Adrenaline given as eye-drops did not have any significant effect on these parameters. Subcutaneously administered adrenaline caused within 5 min a significant increase of plasma adrenaline level (from 1.0 +/- 0.2 to peak of 6.5 +/- 1.2 nM) which gradually decreased during 2 h. This mode of adrenaline administration increased the systolic blood pressure by a maximum of 11 +/- 3.5 mmHg, heart rate by 9 +/- 2.2 beats X min-1, tremor ratio by 4 +/- 0.6 and reduced the diastolic blood pressure by 18 +/- 4.7 mmHg. The cardiovascular effects were approximately maximum 15 min after administration and lasted almost 90 min.(ABSTRACT TRUNCATED AT 250 WORDS)


Ultrastructural and diffusion tensor imaging studies reveal axon abnormalities in Pompe disease mice.
Pompe disease (PD) is caused by lysosomal glycogen accumulation in tissues, including muscles and the central nervous system (CNS). The intravenous infusion of recombinant human acid alpha-glucosidase (rhGAA) rescues the muscle pathologies in PD but does not treat the CNS because rhGAA does not cross the blood-brain barrier (BBB). To understand the CNS pathologies in PD, control and PD mice were followed and analyzed at 9 and 18 months with brain structural and ultrastructural studies. T2-weighted brain magnetic resonance imaging studies revealed the progressive dilatation of the lateral ventricles and thinning of the corpus callosum in PD mice. Electron microscopy (EM) studies at the genu of the corpus callosum revealed glycogen accumulation, an increase in nerve fiber size variation, a decrease in the g-ratio (axon diameter/total fiber diameter), and myelin sheath decompaction. The morphology of oligodendrocytes was normal. Diffusion tensor imaging (DTI) studies at the corpus callosum revealed an increase in axial diffusivity (AD) and mean diffusivity (MD) more significantly in 9-month-old PD mice. The current study suggests that axon degeneration and axon loss occur in aged PD mice and are probably caused by glycogen accumulation in neurons. A drug crossing the BBB or a treatment for directly targeting the brain might be necessary in PD.


IFNL4 genetic variant can predispose to COVID-19.
Interferon lambda 4 (IFNλ4) has shown antiviral activity against RNA viruses, including some coronaviruses. Besides, genetic variants of IFNL4 can be predictive of the clearance of RNA viruses. However, little is known about the effect of these genetic variants on SARS-CoV-2 infection. In this study, we investigated whether there was a relationship of the rs12979860 polymorphism of IFNL4 with COVID-19. We found that the T allele of rs12979860 was overexpressed in COVID-19 patients with regard to the general population without this disease (36.16% vs. 26.40%, p = 6.4 × 10-4; OR 0.633 C vs T; 95% CI 0.487, 0.824), suggesting that this allele could be a risk factor for COVID-19. Accordingly, the CC genotype was significantly lower in COVID-19 patients compared to controls (37.85% vs. 55.51%, p = 8 × 10-5; OR 0.488; 95% CI 0.342, 0.698). These results were not affected by sex, age, and disease severity in patients with COVID-19. Our findings suggest that, like other infectious diseases caused by RNA viruses, genetic variants of IFNL4 can predispose to COVID-19. Confirmation of our results may contribute to better understanding the mechanisms of this disease.


Brief report: The effect of delayed matching to sample on stimulus over-selectivity.
Stimulus over-selectivity occurs when one aspect of the environment controls behavior at the expense of other equally salient aspects. Participants were trained on a match-to-sample (MTS) discrimination task. Levels of over-selectivity in a group of children (4-18 years) with Autism Spectrum Disorders (ASD) were compared with a mental-aged matched typically-developing group. There was more over-selectivity in the ASD group. When retention intervals were added between the sample and comparisons in the MTS task, both groups showed an increased level of over-selectivity, with the ASD group showing a more pronounced effect.


Quantitation of plasma and urine 3-hydroxyglutaric acid, after separation from 2-hydroxyglutaric acid and other compounds of similar ion transition, by liquid chromatography-tandem mass spectrometry for the confirmation of glutaric aciduria type 1.
Glutaric aciduria type 1, a deficiency of glutaryl-CoA dehydrogenase, causes an accumulation of neurotoxic metabolites glutaric acid and 3-hydroxyglutaric acid (3-HGA). Testing of these analytes is routinely done by GC-MS but seldom account for interference from isomers or compounds with similar ion transitions. We developed a liquid chromatography tandem mass spectrometry method that accurately measures 3-HGA in urine and plasma specimens, while utilizing similar reagents and instrumentation used for the routine performance of amino acid and acylcarnitine analysis in determining the diagnosis of several metabolic disorders. Plasma and urine samples were added aliquots of the deuterated 3-HGA internal standard and acetonitrile. The protein-free supernatant was brought to dryness, and the residue derivatized using 3 M HCL in 1-butanol with heating. The dried derivative was then reconstituted in 50% methanol-water solution and aliquot transferred to an HPLC vial for analysis by LC-MS/MS. Separation was performed using a C8 HPLC column under flow gradient conditions of 0.2% formic acid in water and methanol, respectively. Ionization was by ESI and detection of selected precursor-product ion transitions by multiple reaction monitoring (MRM) in positive mode. The butyl-ester derivative of 3-HGA eluted at 7.82 min while 2-hydroxyglutaric acid (2-HGA) eluted at 8.21 min. This was equivalent to a separation factor of 1.05 and a resolution of 1.03, respectively. The 3-HGA calibration curve was linear over the range 6.20-319 ng mL-1 (r2 = 0.9996), and the reportable range determined by the linearity was found to be 1.54-384 ng mL-1. The calculated limits of detection and quantitation were 0.348 and 1.56 ng mL-1, respectively. Intra- and Inter-assay %CVs for quality control plasma and urine samples ranged from 2 to 18%, with recoveries of 66-115%. The method correlated to the gold standard GC-MS method for both serum (r2 ≥ 0.996) and urine analysis (r2 ≥ 0.949). The concentration of 3-HGA in normal, non-GA1 individuals was ≤25.2 ng mL-1 (in plasma) and ≤ 35.0 μmol mmol-1 of creatinine (in urine). This LC-MS/MS method accurately quantified plasma and urine 3-HGA concentration after successful resolution from 2-HGA and other compounds with similar ion transitions. This method is suitable for confirmatory testing of 3-HGA, as a follow-up to an abnormal newborn screen test result, with concern for GA type 1.


5-Methyltetrahydrofolic acid and folic acid measured in plasma with liquid chromatography tandem mass spectrometry: applications to folate absorption and metabolism.
We describe a liquid chromatography (LC) tandem mass spectrometry (MS-MS) method for the determination of 5-methyltetrahydrofolic acid (5-methylTHF) and folic acid concentrations and enrichments in human plasma. It was used to study absorption and initial metabolism in five volunteers with two simultaneously administered oral test doses ([(13)C(6)]folic acid in capsules and [(2)H(2)]folic acid in a drink). [(13)C(5)]5-methylTHF and [(2)H(4)]folic acid were used as internal standards. Plasma samples (2 ml) were purified using folate binding protein affinity columns, followed by a concentration step. After LC separation, folates were detected using positive electrospray ionization MS-MS under multiple reaction monitoring conditions. Calibrations were linear for 5-methylTHF over the range 1.2 x 10(-11) (=limit of detection) to 3.2 x 10(-7)mol/L and for folic acid over the range 5 x 10(-10) (=limit of detection) to 4.5 x 10(-8)mol/L. For 5-methylTHF concentration in plasma, intraassay coefficient of variation was within 8.6% (and for unlabeled 5-methylTHF it was within 2.8%) and interassay coefficient of variation was within 9.0%. For folic acid concentrations these coefficient of variations were within 7.5% and within 6.5%, respectively. The [(13)C(6)] and [(2)H(2)] isotopomers of folic acid and 5-methylTHF were measured in the plasma of each volunteer for 8h. After accounting for the time delay due to capsule opening, the modeling results showed no significant differences in absorption time, first pass effect, and elimination rate in the folic acid test doses in capsule or drink. We conclude that LC-MS-MS offers increased sensitivity for quantification of plasma concentrations and enrichments of 5-methylTHF and folic acid and is applicable to stable-isotope studies in humans.


Dynamic bacterial assembly driven by Streptomyces griseorubens JSD-1 inoculants correspond to composting performance in swine manure and rice straw co-composting.
The effect of Streptomyces griseorubens JSD-1 inoculant on composting performance and bacterial community assembly during the swine manure and rice straw co-composting was studied by a high-throughput pyrosequencing technology. The JSD-1 inoculant contributed to a higher temperature (maximum 66.8 °C), a longer thermophilic phase (46 days), and a lower bacterial diversity in JSD-1 compost. The principle component analysis confirmed that JSD-1 inoculant significantly reshaped the microbial communities. The difference in genera significantly increased during both composting processes. The predominant biomarkers were members of Bacteroidetes in JSD-1 composting. The network analysis also showed different chief "connecting" genera in both composts. Moreover, JSD-1 inoculant increased the total nitrogen, phosphorus, and potassium content in composts. The redundancy analysis showed that the bacterial community was mainly influenced by temperature; additionally, the nutrient contents were positively correlated with temperature. These results demonstrated that JSD-1 inoculant drove the bacterial assembly to induce physicochemical property changes in co-composting.


Preoperative evaluation of Klatskin tumor: accuracy of spiral CT in determining vascular invasion as a sign of unresectability.
To assess the accuracy of spiral computed tomography (CT) in predicting the resectability of Klatskin tumor as determined by vascular invasion. Twenty-one consecutive patients with Klatskin tumor who had undergone laparotomy were included in this study. The preoperative thin-section (5-mm-thick) spiral CT scans of these patients were assessed for the surgical resectability of tumor by evaluating the vascular invasion. The criterion for vascular invasion indicating unresectability was the tumoral invasion of the proper hepatic artery or main portal vein or simultaneous invasion of one side of the hepatic artery and the other side of the portal vein. All nine patients with tumors thought to be unresectable on the basis of CT findings had tumors that were unresectable at surgery (positive predictive value, 100%). Of 12 patients with tumors thought to be resectable, six had resectable tumors (negative predictive value, 50%). Spiral CT failed to detect small hepatic metastasis (n = 1), lymph node metastasis (n = 1), extensive tumor (n = 2) and variation of bile duct (n = 2), which precluded surgical resection. Spiral CT is a reliable method for detecting vascular invasion and unresectable tumors. However, it has limitations in detecting variations of the bile duct or the intraductal extent of tumor.


Mechanism of cholera toxin activation by a guanine nucleotide-dependent 19 kDa protein.
Cholera toxin causes the devastating diarrheal syndrome characteristic of cholera by catalyzing the ADP-ribosylation of Gs alpha, a GTP-binding regulatory protein, resulting in activation of adenylyl cyclase. ADP-ribosylation of Gs alpha is enhanced by 19 kDa guanine nucleotide-binding proteins known as ADP-ribosylation factors or ARFs. We investigated the effects of agents known to alter toxin-catalyzed activation of adenylyl cyclase on the stimulation of toxin- and toxin subunit-catalyzed ADP-ribosylation of Gs alpha and other substrates by an ADP-ribosylation factor purified from a soluble fraction of bovine brain (sARF II). In the presence of GTP, sARF II enhanced activity of both the toxin catalytic unit and a reduced and alkylated fragment ('A1'), as a result of an increase in substrate affinity with no significant effects on Vmax. Activation of toxin was independent of Gs alpha and was stimulated 4-fold by sodium dodecyl sulfate, but abolished by Triton X-100. sARF II therefore serves as a direct allosteric activator of the A1 protein and may thus amplify the pathological effects of cholera toxin.


Facial nerve paralysis following cochlear implant surgery.
Facial nerve paralysis is a rare but devastating complication of cochlear implant surgery. The aims of the study were to define the incidence of facial nerve paralysis in our series and understand possible mechanisms of injury. Retrospective chart review and case reports. Charts were reviewed of all 705 patients implanted between 1980 and 2002 at the authors' institutions to identify those with postoperative facial nerve weakness and determine incidence. For patients with facial nerve weakness, onset, degree, and timing of paralysis were noted; clinical findings were correlated to operative report findings. The method of treatment was noted, and the final facial nerve function outcome was recorded. Five patients (one child and four adults) were found to have postoperative facial nerve weakness, for an incidence of 0.71%. This complication was delayed in all cases, ranging from 18 hours to 19 days postoperatively. All patients were treated with steroids or steroids combined with antiviral medication, and all ultimately recovered normal facial function. In the study series, the incidence of facial nerve paralysis following cochlear implant surgery was 0.71%. Possible mechanisms of injury included heating injury and viral reactivation. All patients presented with a delayed facial nerve paralysis and did recover normal facial nerve function.


Protective effects of N-acetylcysteine on 3, 4-methylenedioxymethamphetamine-induced neurotoxicity in male Sprague-Dawley rats.
Exposure to 3, 4-methylenedioxymethamphetamine (MDMA) leads to spatial memory impairment and hippocampal cell death. In the present study we have examined the protective effects of N-acetyl-L-cysteine (NAC) on MDMA-induced neurotoxicity. A total of 56 male Sprague Dawley rats (200-250 g) received twice daily intraperitoneal (IP) injections of 5, 10 or 20 mg/kg MDMA plus NAC (100 mg/kg). Rectal temperatures were recorded before and after daily treatment. We used a Morris water maze (MWM) to assess spatial learning and memory. At the end of the study rats' brains were removed, cells were counted and the level of Bcl-2, Bax and caspase-3 expression in the hippocampi were measured. NAC pretreatment significantly reduced MDMA-induced hyperthermia. In the MWM, NAC significantly attenuated the MDMA-induced increase in distance traveled; however the observed increase in escape latency was not significant. The decrease in time spent in the target quadrant in MDMA animals was significantly attenuated (p < 0.001, all groups). NAC protected against MDMA-induced cell death and the up -regulation of Bax and Caspase-3, in addition to the down-regulation of Bcl-2. This data suggested a possible benefit of NAC in the treatment of neurotoxicity among those who use MDMA.


Combining thin-film microextraction and surface enhanced Raman spectroscopy to sensitively detect thiram based on 3D silver nanonetworks.
By coupling thin-film microextraction (TFME) with surface enhanced Raman scattering (SERS), a facile method was developed for the determination of thiram in the complex matrix (orange juice or grape peel). The substrate of TFME was made by self-assembling silver sol on the silicon wafer to form a three-dimensional (3D) silver nanonetwork structure, without adding any template, which was used for TFME and SERS detection, respectively. The substrate exhibits high reproducibility with a relative standard deviation of about 7.32 % in spot and spot SERS intensity. The SERS signal intensity at a shift of 1384 cm-1 and the thiram concentration showed good linearity in the range of 0.01-5 µg/L and the linear correlation coefficient was 0.9912. The detection limit for thiram was found to be 0.01 µg/L. The TFME-SERS method was applied for the determination of thiram in fruit juice and the results were obtained very well. Therefore, this method is expected to play a role in the detection of trace pollutants.


Mechanistic insights into the size-dependent effects of nanoparticles on inhibiting and accelerating amyloid fibril formation.
The aggregation of peptides into amyloid fibrils has been linked to ageing-related diseases, such as Alzheimer's and type 2 diabetes. Interfaces, particularly those with large nanostructured surfaces, can affect the kinetics of peptide aggregation, which ranges from complete inhibition to strong acceleration. While a number of physiochemical parameters determine interfacial effects, we focus here on the role of nanoparticle (NP) size and curvature. We used thioflavin T (ThT) fluorescence assays to demonstrate the size-dependent effects of NPs on amyloid fibril formation for the peptides Aβ40, NNFGAIL, GNNQQNY and VQIYVK. While 5 nm gold NPs (AuNP-5) retarded or inhibited the aggregation of all peptides except NNFGAIL, larger 20 nm gold NPs (AuNP-20) tended to accelerate or not influence peptide aggregation. Differences in the NP effects for the peptides resulted from the different peptide properties (size, tendency to aggregate) and associated surface binding affinities. Additional dynamic light scattering (DLS), electron microscopy, and atomic force microscopy (AFM) experiments with the Aβ40 peptide confirmed size-dependent NP effects on peptide aggregation, and also suggested a structural influence on the formed fibrils. NPs can serve as a surface for the adsorption of peptide monomers and enable nucleation to oligomers and fibril formation. However, molecular dynamics (MD) simulations showed that peptide oligomers were less stable at smaller NPs. High surface curvatures destabilized prefibrillar structures, which provides a possible explanation for inhibitory effects on fibril growth, provided that peptide-NP surface binding was relevant for fibril formation. These mechanistic insights can support the design of future nanostructured materials.


Cytokine production in a model of wound healing: the appearance of MIP-1, MIP-2, cachectin/TNF and IL-1.
Macrophages are essential for normal wound repair and many of their effects on healing wounds are likely to be mediated by the secretion of cytokines. This study examines the appearance of messenger RNA (mRNA) for cachectin/tumor necrosis factor (TNF), IL 1, and macrophage inflammatory proteins 1 and 2 (MIP-1 and MIP-2), as well as the mature peptides, in a model of wound healing using wound chambers. RNA for all four cytokines can be detected in wound inflammatory cells by polymerase chain reaction amplification throughout the first 7 days. Cachectin/TNF and IL 1 protein levels peaked on the first day after wound chamber implantation, and MIP-1 and MIP-2 were detected only on day 3. The data suggest that these cytokines participate in the early inflammatory response to wounding.


Heparin-independent release of lipoprotein lipase activity from perfused rat hearts.
Heparin-independent release of lipoprotein lipase activity from isolated perfused rat hearts was measured and related to the rapid turnover of the enzyme. Hearts consistently released lipoprotein lipase activity (2.1 +/- 0.2 U/g released per min) during 60 min of nonrecirculating perfusion without heparin. This rate of release did not significantly differ from that measured in heparin-perfused hearts after the first 10 min of perfusion (2.2 +/- 0.2 U/g released per min). The fractional release rate of lipoprotein lipase activity during nonheparin perfusion was 1.3% per min, which was higher than that calculated for alkaline phosphatase (0.002%) and creatine kinase (0.03%) activities. The lipase activity released was activated 4-fold by serum and inhibited 94 and 88% by 0.5 M NaCl and 3 mg/ml protamine sulfate, respectively. Lipoprotein lipase activity in the 1-min heparin-releasable (extracellular) and residual (intracellular) compartment remained stable during the last 40 min of nonheparin perfusion. During this period total heart, intracellular and extracellular enzyme t1/2 were calculated to be 52, 42 and 10 min, respectively. The results are consistent with the postulate that continuous release of lipoprotein lipase into the vascular compartment may be an important determinant of its rapid turnover in the heart, and possibly other tissues.


TcBat a bat-exclusive lineage of Trypanosoma cruzi in the Panama Canal Zone, with comments on its classification and the use of the 18S rRNA gene for lineage identification.
We report TcBat, a recently described genetic lineage of Trypanosoma cruzi, in fruit-eating bats Artibeus from Panama. Infections were common (11.6% prevalence), but no other T. cruzi cruzi genotypes were detected. Phylogenetic analyses show an unambiguous association with Brazilian TcBat, but raise questions about the phylogenetic placement of this genotype using the 18S rRNA gene alone. However, analyses with three concatenated genes (18S rRNA, cytb, and H2B) moderately support TcBat as sister to the discrete typing unit (DTU) TcI. We demonstrate that short fragments (>500 bp) of the 18S rRNA gene are useful for identification of DTUs of T. cruzi, and provide reliable phylogenetic signal as long as they are analyzed within a matrix with reference taxa containing additional informative genes. TcBat forms a very distinctive monophyletic group that may be recognized as an additional DTU within T. cruzi cruzi.


Persistent increase in myofibroblasts in Helicobacter heilmannii-infected mice but not in Helicobacter pylori-infected Mongolian gerbils: colocalization of COX-2 and bFGF immunoreactivity.
The clinical significance of Helicobacter heilmannii infection remains uncertain, owing to the lack of a specific detection method. Recently, we reported a marked increase in myofibroblasts in the early stage of Helicobacter pylori infection in Monglian gerbils. The present study was designed to clarify changes in myofibroblasts, and in the immunoeactivities of basic fibroblast growth factor, cyclooxygenase-2 and inducible nitric oxide synthase after H. pylori infection in Monglian gerbils and H. heilmannii infection in mice. After oral inoculation, changes in the location of bacteria and the immunoreactivity of myofibroblasts, basic fibroblast growth factor, cyclooxygenase-2 and inducible nitric oxide synthase were stained with the indirect immunofluorescent method and observed by confocal laser microscopy. In H. heilmannii-infected mice, the increases in myofibroblasts and in immunoreactivities of these three markers were sustained 12 months after infection. In H. pylori-infected Monglian gerbils, however, these increases were significant at 3 months but had returned to control levels at 12 months. Two types of Helicobacter infection showed different patterns of myofibroblast proliferation, coinciding with the extent of inflammation. These findings suggest that this difference may be related to the consequences of the infection.


Na+, K(+)-adenosine triphosphatase regulation in hypertrophied vascular smooth muscle cells.
Vascular smooth muscle cell hypertrophy is a normal compensatory state that may play a pathogenic role in hypertension. Angiotensin II stimulates a hypertrophic response in cultured vascular smooth muscle cells. As part of the growth response, angiotensin II rapidly activates the Na(+)-H+ exchanger, increasing Na+ influx. Because Na+, K(+)-ATPase is the major cellular mechanism for regulating intracellular Na+, we studied the effects of angiotensin II-induced hypertrophy on Na+, K(+)-ATPase expression and activity. Angiotensin II caused rapid increases in both steady-state Na+, K(+)-ATPase activity (ouabain-sensitive 86Rb uptake) and intracellular [Na+]. Angiotensin II also caused a sustained increase in Na+, K(+)-ATPase at 24 hours with a 73% increase in maximal 86Rb uptake per milligram protein and a fourfold increase in Na+, K(+)-ATPase alpha-1 messenger RNA levels. Thus, angiotensin II hypertrophy was associated with rapid increases in Na+, K(+)-ATPase activity due to increased Na+ entry and sustained increases due to a specific increase in Na+, K(+)-ATPase expression. These data demonstrate dynamic regulation of Na+, K(+)-ATPase at the functional and molecular level and suggest that similar compensatory mechanisms should be present in vivo. Alterations in such compensatory pathways may be fundamental to the pathogenesis of hypertension.


Prenatal diagnosis of spinal muscular atrophy: Indian scenario.
To study the psychosocial issues associated with prenatal diagnosis of SMA in India and the use of SMN1 copy number analysis for carrier detection prior to offering prenatal diagnosis. Homozygous deletion of SMN1 gene was done by PCR-RFLP. Copy number analysis of SMN1 gene was performed by quantitative PCR. We report our experience of eight cases of prenatal diagnosis for SMA and the use of carrier detection prior to offering prenatal diagnosis. Quantitative PCR results show that SMN1 copy number analysis is useful to identify couples at risk. Case analyses depict unique psychosocial issues associated with prenatal diagnosis of SMA from India.


Toxicity of emestrin, a new macrocyclic dithiodioxopiperazine mycotoxin, to mitochondrial function.
The effect of emestrin, a new macrocyclic epidithiodioxopiperazine mycotoxin from severalEmericella species, on mitochondrial reactions was studied using isolated rat liver mitochondria to gain insight into the molecular mechanism for itsin vivo toxicity to rat and mouse. Emestrin was found to inhibit ATP synthesis in mitochondria causing an uncoupling of oxidative phosphorylation and a depression of respiration in isolated mitochondria. In addition to these effects on mitochondrial respiration, emestrin elicited a dratsic structural alteration (swelling) of mitochondria as observed in thein vivo system. The mitochondrial swelling was significantly enhanced by the subsequent addition of calcium ion. Emestrin B, in which dithio group is replaced by trithio group, exerted an uncoupling effect on oxidative phosphorylation without accompanying such depressive effect on state 3 respiration as observed for emestrin.


Immunological characterization of the Marin County strain of astrovirus.
Marin County virus (MCV) was isolated from a stool suspension and serially propagated in human embryonic kidney cell cultures. MCV particles in stool and cell-propagated virus stocks showed reactivity by immune electron microscopy (IEM) with rabbit antiserum to astrovirus type 5. MCV antigen was also detected in two MCV stool samples by enzyme immunoassay (EIA) with an astrovirus group-specific monoclonal antibody. Acute and convalescent sera from 3 of 3 MCV-infected patients showed seroconversion to cell-propagated MCV by EIA. Immunofluorescence of MCV propagated in cell culture showed positive reactivity with an astrovirus group specific monoclonal antibody and astrovirus type 5 antiserum, with some cross-reactivity with astrovirus type 1. Similar results were obtained with the prototype strain of astrovirus type 5. However, in plaque-reduction assays, both the prototype astrovirus type 5 and MCV were neutralized by type 5 antiserum only. We conclude that MCV can be serially propagated by techniques used for previously described astroviruses and is serotypically an astrovirus type 5.


Pain-free injection in infants. Use of a lignocaine-prilocaine cream to prevent pain at intravenous induction of general anaesthesia in 1-5-year-old children.
A randomised, placebo-controlled, double-blind study was undertaken in 111 children between the ages of 1 and 5 years to assess the efficacy of EMLA 5% cream in the alleviation of venepuncture pain at intravenous induction of general anaesthesia using 27-gauge needles. Pain assessment was made by an operating department assistant using both verbal rating scale and visual analogue scale methods. Seventy-five children, of whom 24 were premedicated, were treated with EMLA cream and 36 with placebo, 14 of whom were premedicated. Significantly lower pain scores were recorded in the children treated with EMLA cream (verbal rating scale: premedicated p less than 0.05, unpremedicated p less than 0.001; visual analogue scale: premedicated p less than 0.0005, unpremedicated p less than 0.0002). No variation in analgesia was found for application times between 30 and 300 minutes and there were no serious side effects.


Do the right precordial leads during exercise testing contribute to detection of coronary artery disease?
It is still unknown whether or not the additional right precordial leads (RPL) during exercise testing contribute to detection of coronary artery disease (CAD). The aim of this study was to evaluate the RPL during exercise testing for the detection of CAD. The study included 157 consecutive patients (116 men and 41 women, mean age 66 years) suspected of having CAD, who underwent conclusive treadmill exercise testing (heart rate reached at least 85% of the predicted maximum or positive electrocardiogram [ECG] changes were exhibited) and coronary angiography. During exercise testing, the ECG was recorded with the standard 12 leads and 4 RPL (V3R, V4R, V5R, V6R). Of the 157 patients, 67 had CAD (> 75% stenosis in at least one major coronary artery), and 64 had positive ST changes in the standard ECG leads during exercise testing. Using the conventional 12-lead method, sensitivity and specificity were 76 and 86%, respectively. Only three patients exhibited positive changes in the RPL leads; all had > 0.1 mV ST elevation in one of the RPL leads with > 0.1 mV ST elevation in aVR. Two of these patients had significant right coronary artery lesions and the other had a lesion of the left anterior descending artery which perfused the inferior as well as the anteroseptal area. In the standard 12 leads, one of the patients with an abnormal RPL and a right coronary lesion was negative, while the other two patients were positive. Combining RPL with the conventional 12-lead method, sensitivity and specificity were 78 and 86%, respectively. Therefore, RPL did not improve the accuracy of the exercise ECG. The use of RPL during exercise testing may contribute to the detection of ischemia perfused by the right coronary artery; however, it does not improve the diagnostic accuracy of the exercise test.


Predictors of the prevalence of bacterial STI among young disadvantaged Indigenous people in north Queensland, Australia.
To identify sexually transmitted infections in rural and remote Indigenous communities in north eastern Australia and examine factors that may influence prevalence. A cross sectional survey of 26 Aboriginal and Torres Strait Islander communities in northern Queensland was carried out. 3313 people (2862 Indigenous) aged 15 years and over resident in participating communities during the period March 1998 to December 2000. The main outcome measures were community and population prevalence of chlamydia and gonorrhoea and independently associated risk factors. A total of 238 cases of chlamydia, 66 cases of gonorrhoea, and 37 cases of co-infection were detected among Indigenous participants. Prevalence of chlamydia and/or gonorrhoea ranged from 23.0% among 15-19 year olds to 3.5% among those 40 years and older. In the adjusted analysis younger age, female sex, lower socioeconomic status, the use of alcohol and tobacco, and the structure of community health services were independently associated with a higher prevalence of bacterial STI. This study highlights the need for improved STI control in north Queensland Indigenous communities through strategies to improve the reach and accessibility of primary healthcare services.


Genicular Artery Embolisation in Patients with Osteoarthritis of the Knee (GENESIS 2): Protocol for a Double-Blind Randomised Sham-Controlled Trial.
Knee osteoarthritis is a leading cause of chronic disability and economic burden. In many patients who are not surgical candidates, existing treatment options are insufficient. Clinical evidence for a new treatment approach, genicular artery embolisation (GAE), is currently limited to single arm cohort, or small population randomised studies. This trial will investigate the use of a permanent embolic agent for embolisation of abnormal genicular arterial vasculature to reduce pain in patients with mild to moderate knee osteoarthritis. Up to 110 participants, 45 years or older, with knee pain for ≥ 3 months resistant to conservative treatment will be randomised (1:1) to GAE or a sham procedure. The treatment group will receive embolisation using 100-micron Embozene™ microspheres (Varian, a Siemens Healthineers Company) (investigational use for this indication in the UK), and the sham group will receive 0.9% saline in an otherwise identical procedure. Patients will be followed for 24 months. At 6 months, sham participants will be offered crossover to GAE. The primary endpoint is change of 4 Knee Injury and OA Outcome Score subscales (KOOS4) at 6 months post-randomisation. The study will also evaluate quality of life, health economics, imaging findings, and psychosocial pain outcomes. The primary manuscript will be submitted for publication after all participants complete 6 months of follow-up. The trial is expected to run for 3.5 years. Trial Registration: ClinicalTrials.gov, Identifier: NCT05423587.


Quantitative assessment of the association between +61A>G polymorphism of epidermal growth factor gene and susceptibility to glioma.
Numerous studies have investigated the risk of cancer associated with the polymorphism of epidermal growth factor (EGF) 61A>G, but results have been inconsistent. We performed this meta-analysis to drive a more precise estimation of the association between this polymorphism and risk of glioma. A comprehensive search was conducted to identify all case-control studies on the EGF +61A>G polymorphism and glioma risk. Odds ratios (ORs) and 95% confidence intervals (95% CIs) were calculated to assess the strength of the association. Statistical analysis was performed with the software program Stata (version 12.0). A total of ten eligible studies, including 1,888 cases and 2,836 controls were included in this work. Overall, there was a significant association between EGF +61A>G polymorphism and glioma risk in the allele model (OR = 1.419, 95% CI = 1.144-1.759, P = 0.001). In the subgroup analysis by ethnicity, significant associations were also found in Asian populations under all different genetic models (homozygote model: OR = 1.727, 95% CI = 1.310-2.275, P = 0.000; heterozygote model: OR = 1.202, 95% CI = 1.023-1.413, P = 0.025; dominant model: OR = 1.279, 95% CI = 1.096-1.491, P = 0.002; recessive model: OR = 1.590, 95% CI = 1.221-2.070, P = 0.001; and A-allele versus G-allele OR = 1.600, 95% CI = 1.145-2.236, P = 0.006). However, no significant associations were found among Caucasians in all comparison models. In conclusion, the results suggest that there is a significant association between EGF +61A>G polymorphism and glioma risk among Asians.


Hierarchy in the assembly of HLA-B27 and HLA-Cw3 molecules in transgenic mice.
Cell surface expression of human class I molecules in transgenic mice is dependent upon the available pool of beta 2-microglobulin (beta 2m) and the affinity between mouse beta 2m and human class I molecules. HLA-B27 and HLA-Cw3 transgenes can be expressed in mouse strains of the H-2 haplotypes b, f, k, and s which encode two endogenous class I genes mapping to H-2K and H-2D. The human class I genes cannot be expressed on H-2d and H-2q haplotypes which encode three endogenous class I molecules (K,D,L). This suggests that there may be only enough mouse beta 2m molecules to support three class I molecules. When both the HLA-B27 and HLA-Cw3 genes are introduced into H-2b mice, only HLA-Cw3 reaches the cell surface. This suggests that HLA-Cw3 has a higher affinity than HLA-B27 for mouse beta 2m. The possible implications of our findings regarding the assembly, transport, and expression of class I MHC molecules in vivo are discussed.


Comparison of the antiarrhythmic effects of acebutolol and propranolol in the treatment of ventricular arrhythmias.
This study was designed to assess the relative antiarrhythmic activity of fixed doses of 2 beta-adrenergic blocking agents, propranolol and acebutolol, in a prospective double-blind crossover trial. Twenty-one patients who had at least 30 premature ventricular contractions (PVCs) per hour while receiving placebo were entered into the study. Ten patients were randomized to initially receive propranolol, 40 mg every 8 hours, and 11 were assigned to receive acebutolol, 300 mg every 8 hours. After 6 weeks of treatment, patients were weaned off medication for 1 week and then placed on placebo for 1 week. Eighteen patients were available and eligible for crossover to the alternative regimen for an additional 6 weeks. All 21 patients completed courses with propranolol and 17 completed courses with acebutolol. The mean number of PVCs per hour during placebo, propranolol and acebutolol treatment were 267, 87 and 119, respectively. Using paired t test statistics on observation differences, both propranolol and acebutolol significantly reduced the number of PVCs per hour compared with placebo, whereas similar analysis revealed no significant difference in the antiarrhythmic effect. However, with the current sample size the power of the test is too low for the latter conclusion to be stated with confidence. Side effects were mild and infrequent, requiring discontinuation of acebutolol in 2 patients and discontinuation of propranolol in 1. Thus, acebutolol is a safe and effective antiarrhythmic agent and compares favorably with propranolol.


Natriuretic peptide receptor: structure and signaling.
The ANP receptor is a single-transmembrane sequence receptor coupled to guanylate cyclase (GCase). It belongs to a family of GCase-coupled receptors that share a common overall molecular configuration. Collectively, theses GCase-coupled receptors belong to a larger family of single-transmembrane sequence receptors that include growth hormone and cytokine receptors. The signal transduction mechanism of these receptors has not been thoroughly understood. Receptor dimerization (or oligomerization) has been suggested as the mechanism. However, at least for the ANP receptor, dimerization has been seen to occur in the absence of the ligand, suggesting that an additional, as yet unknown effect of hormone binding is responsible for receptor activation. To understand the signaling mechanism, some of the functions and subsites of the ANP receptor critical for signaling have been identified, including the binding stoichiometry, receptor self-association, the juxtamembrane hinge structure containing a signature motif critical for GCase signaling, ANP-binding site residues, chloride-dependence of ANP binding, disulfide linkages, and glycosylation structures. These structures and the functional sites have been identified in the crystal structure of dimerized recombinant extracellular domain of the ANP receptor. The intracellular domain contains a kinase-homologous domain that regulates the activity of the GCase domain responding to ANP binding and also to binding of the allosteric effector ATP. Moreover, this regulatory role of the kinase-homologous domain is modulated by its own phosphorylated state. Although considerable data have been accumulated, the mechanism of ANP receptor signaling has not been well defined. Further studies are necessary to understand how ANP binds to the receptor, what conformational effect is caused by ANP binding, how this effect is transduced across the cell membrane, and how this transmembrane effect leads to stimulation of the GCase catalytic activity.


Red blood cell transfusion during septic shock in the ICU.
Transfusion of red blood cells (RBCs) remains controversial in patients with septic shock, but current practice is unknown. Our aim was to evaluate RBC transfusion practice in septic shock in the intensive care unit (ICU), and patient characteristics and outcome associated with RBC transfusion. Prospective cohort study of all adult patients with septic shock (n = 164) in six general ICUs during a 3-month period. Characteristics, other treatments, monitoring and outcome were compared in RBC-transfused and -non-transfused patients. Ninety-nine patients (95% confidence interval 87-111) received a median 900 ml (interquartile range 490-1405) of RBC during septic shock in ICU. Among transfused patients, there were more females [49/99 (49%) vs. 22/65 (34%), P = 0.048] and surgical patients [39/99 (39%) vs. 14/65 (22%), P = 0.02] than among patients not transfused. Also, admission simplified acute physiology score II was higher and minimal haemoglobin levels (days 1-3) were lower in transfused patients compared with those not transfused. In contrast, age, markers of shock and severity organ failure assessment score on day 1 and 90-day mortality did not differ between RBC-transfused and -non-transfused patients. Most patients with septic shock received RBCs during shock, and these patients had higher disease severity and lower haemoglobin levels than those not transfused. In spite of this, mortality did not differ between groups neither in the unadjusted or adjusted analyses. However, neither the design nor the sample size allows us to make inferences about treatment effects, which underlines the need for large randomised, clinical trials on transfusion in septic shock.


Cytotoxic activity of anti-mucin 1 chimeric antigen receptor T cells expressing PD-1-CD28 switch receptor against cholangiocarcinoma cells.
Cholangiocarcinoma (CCA) is a lethal bile-duct cancer that is difficult to treat by current standard procedures. This drawback has prompted us to develop adoptive T-cell therapy for CCA, which requires an appropriate target antigen for binding of chimeric antigen receptor (CAR) T cells. Mucin 1 (MUC1), an overexpressed protein in CCA cells, is a potential target antigen for the CAR T-cell development. However, MUC1 overexpression also is associated with the upregulation of programmed death-ligand 1 (PD-L1), an immune checkpoint protein that prohibits anti-tumor functions of T cells, probably causing poor overall survival of patients with CCA. To overcome this problem, we developed anti-MUC1-CAR T cells containing PD-1-CD28 switch receptor (SR), namely αM.CAR/SR T cells, to target MUC1 and switch on the inhibitory signal of PD-1/PD-L1 interaction to activate CD28 signaling. Our lentiviral construct contains the sequences that encode anti-MUC1-single chain variable fragment, CD137 and CD3ζ, linked with P2A, PD-1 and CD28. Initially, the upregulations of MUC1 and PD-L1 proteins were confirmed in CCA cell lines. αM.CAR and SR were co-expressed in 53.53 ± 13.89% of transduced T cells, mainly CD8+ T cells (85.7 ± 0.75%, P<0.0001) with the effector memory phenotype (59.22 ± 16.31%, P < 0.01). αM.CAR/SR T cells produced high levels of intracellular tumor necrosis factor-α and interferon-γ in response to the activation by CCA cells expressing MUC1, including KKU-055 (27.18 ± 4.38% and 27.33 ± 5.55%, respectively, P < 0.05) and KKU-213A (47.37 ± 12.67% and 54.55 ± 8.66%, respectively, P < 0.01). Remarkably, the cytotoxic function of αM.CAR/SR T cells against KKU-213A cells expressing PD-L1 was significantly enhanced compared with the αM.CAR T cells (70.69 ± 14.38% versus 47.15 ± 8.413%, respectively; P = 0.0301), correlated with increased granzyme B production (60.6 ± 9.89% versus 43.2 ± 8.95%, respectively; P = 0.0402). Moreover, the significantly enhanced disruption of KKU-213A spheroids by αM.CAR/SR T cells (P = 0.0027), compared with αM.CAR T cells, was also observed. Taken together, the cytotoxic function of αM.CAR/SR T cells was enhanced over the αM.CAR T cells, which are potential to be further tested for CCA treatment.


Delayed exposed skin grafting: a 10 year experience of the technique.
We describe the routine use of the technique of delayed exposed skin grafting in 2062 patients over a 10 year period. Points of technique have been reviewed and tested in a prospective trial. Delayed graft application requires some 20 minutes (range 5-90 minutes) of nursing time, and daily care of 10 to 20 minutes. Wound healing was achieved in all patients in this series even when a second application of stored skin was applied, without the need for a further operation. Take was achieved in 75% of patients by 8 days after application. The technique offers a saving in theatre time for the patient and the surgeon, although hospital in-patient time is increased.


Ganglioneuroma : primary tumor or maturation of a suspected neuroblastoma?
Ganglioneuroma is a benign neurogenic tumor. These tumors are originating from neuroepithelium along sympathetic ganglia. Main localization is the mediastinum in children older than 10 years. An association with malignant neuroblastoma is rarely observed and it still remains a topic under current discussion. We describe the clinical course of a 17 year-old female patient with a large presacral mass causing amenorrhoea and weight loss. Eleven years before presentation, an incidental urine test showed an elevation of vanillylmandelic acid (VMA) and homovanillic acid (HMA) and a neuroblastoma was suspected. However, further investigations showed no tumor and the test results turned out to be normal within 1 year. Now, a malignant neurogenic tumor was again suspected, but a CT-guided biopsy revealed a benign tumor. The mass was originating from the left sacral nerve roots. A tumor resection via a dorsal approach was performed. Final histology showed a differentiated ganglioneuroma. This is, to our knowledge, the first report describing a patient with elevated VMA/HMA and suspected neuroblastoma who developed a ganglioneuroma 11 years later. The association of ganglioneuroma and neuroblastoma and the abnormal urine tests pointing toward a neuroblastoma 11 years ago remains unclear and the possible answers are discussed in our report.


Endocrine studies of normal and polycystic ovarian tissues in vitro.
The purpose of this study was to compare the steroidogenic potential of the granulosa, theca, and medullary tissues from polycystic and normal ovaries. These ovarian endocrine compartments were isolated from appropriate ovaries and were cultured in vitro for three days in the absence (control) and presence of follicle-stimulating hormone (FSH)/luteinizing hormone (LH) (1 lU/ml), N6,O2-dibutyryladenosine-3':5''-cyclic monophosphoric acid (Bu2cAMP) (10(-2)M), and adrenocorticotropic hormone (ACTH) (1.3 U/ml). After the incubation, steroids in the media were measured by radioimmunoassay. Granulosa cells (10(5) cells per dish) from 4 to 7 mm follicles of normal and polycystic ovaries secreted progesterone spontaneously during the culture period and the production of progesterone was markedly stimulated (between tenfold and thirtyfold) by gonadotropins and Bu2cAMP but not by ACTH. Little, if any, androgen (androstenedione, dehydroepiandrosterone, and testosterone) or estrogen (estrone and estradiol) accumulated in the media of any granulosa cell culture. The control cultures of theca tissue from normal and polycystic ovaries secreted large amounts of androstenedione and progesterone and the production of these steroids by normal and polycystic ovary theca was stimulated in most cases by LH/FSH and Bu2cAMP but not by ACTH. Both normal and polycystic ovary theca secreted some testosterone and dehydroepiandrosterone but little, if any, estrone or estradiol accumulated in any theca culture. The medullary tissue of normal and polycystic ovaries produced only trace amounts of steroids in vitro except for the results from one polycystic ovary with hyperthecosis in which case significant quantities of C19 and C18 steroids were secreted. These experiments have demonstrated that isolated granulosa and theca cells from midantral follicles of normal and polycystic ovaries have a similar capacity to secrete C21 and C19 steroids in the absence and presence of trophic agents. Therefore, it seems probable that chronic anovulation in patients with polycystic ovaries is not caused by an obvious deficiency in the de novo steroidogenic potential of the multiple midantral follicles of the polycystic ovaries or by the absence of gonadotropin receptors on the polycystic ovary follicular cells.


Graphical representation of multidimensional EEG data and classificatory aspects.
EEG data are intrinsically multidimensional and this applies also to a set of broad band spectral parameters (6 parameters per derivation). Non-metric multidimensional scaling (MDS) and principal component analysis (PCA) are compared with respect to their capacity to achieve an adequate few-dimensional representation for the graphical comparison of groups and the identification of subgroups. In an application to neurophysiological aspects of mental retardation and learning disability, MDS proved to be somewhat superior to PCA. Both methods profited from the introduction of (1-p)-convex hulls to define a normative region.


Injection practices of healthcare professionals in a Tertiary Care Hospital.
Unsafe injection practices are prevalent worldwide and may result in spread of infection. Thus the present study was planned to observe the injection practices of healthcare professionals (HCP), including aseptic precautions and disposal of used syringes/needle. Injection practices were observed in the outpatients and inpatients departments. Questionnaire was designed, tested and administered for this purpose. 130 patients receiving injections were observed. Overall injection practices of the HCP were satisfactory. However, unsafe practices with respect to not washing hands (95.4%), not wearing/changing gloves (61.6%), recapping of needles (12.2%), wiping of needle with swab (15.4%) and breaking of ampoule with solid object (44.4%) were observed. The problem of unsafe injections can be successfully addressed by organizing continuing medical education/symposium/workshops for improving the knowledge, attitude and practices of the HCP. Periodic monitoring and such interventions may also further improve safe injection practices.


Blood pressure control in type II diabetics with diabetic retinopathy.
Large clinical trials have emphasized that blood pressure control provides a major clinical benefit in reducing the risk of blindness in patients with diabetic retinopathy. This audit was carried out to assess the quality of care for hypertension in 100 consecutive type II diabetics with diabetic retinopathy. The target blood pressure of 140/80 mmHg was achieved only in 38% of the patients. We also observed that 65% of the patients requiring diabetic macular laser treatment in this cohort had suboptimal control. The factors associated with suboptimal blood pressure control are identified and discussed. Despite the unequivocal fact that lowering BP significantly reduces morbidity and mortality in diabetics, the majority of patients are not treated to a goal BP.


Three-dimensional relation of skin markers to lumbar vertebrae of healthy subjects in different postures measured by open MRI.
The debate is to which extent external skin markers represent true underlying vertebral position and motion. Skin markers and lumbar vertebrae L3 and L4 were examined by vertically open magnetic resonance imaging (MRI) within different postures to investigate whether, and to which extent the position and orientation of skin markers represent the corresponding information of assigned underlying vertebra. Nine healthy volunteers sat within an open MRI scanner in five different seating postures: upright, low flexion, heavy flexion, upright left turn and upright right turn. Skin markers were fixed at lumbar levels L3 and L4. A set of landmarks defines corresponding positions on the vertebrae. Translation-vectors quantify the change of co-ordinates while changing position. Orientation (Cardan-angles) of each level in space was calculated from co-ordinates of three skin-markers and the corresponding vertebral landmarks respectively. The close relation between the position of the individual skin marker and its corresponding landmark on the vertebrae is conserved through all postures (regression coefficients: 0.720<b<0.972, 0.916<r<0.993, p<0.0001). Despite the high sensitivity to measurement errors there is a strong relation between the translation-vectors of external markers and the corresponding translation of the landmark on the vertebrae (0.68<r<0.99, p<0.0001). Within the resolution of the MR-imaging the orientation of the vertebrae can be predicted from the external markers (regression coefficients: 0.936<b<1.189, 0.769<r<0.959, p<0.0001). Positions and motions of skin markers can be taken as a first estimate to calculate the position and spatial orientation of underlying vertebrae. The use of skin markers may help to investigate spinal positions in daily life situations where internal imaging is not possible.


Climbing the e-waste mountain.
The benefits of the information revolution are clear for all to see. Devices such as PCs, faxes, mobile phones, music players and a host of others open up exciting possibilities for individuals and businesses alike. Yet there is a downside to this digital era: the growing mountain of electronic waste. How we tackle this dilemma will have major implications for sustainability.


A glimpse of vitamin D status in Mainland China.
As an essential dietary micronutrient, vitamin D plays a pivotal role in promoting calcium absorption in the intestine and maintaining a healthy skeletal system throughout life. Beyond bone health, an emerging volume of scientific studies shows that vitamin D also may provide cardiovascular, metabolic, and immunologic benefits and reduce mortality. To our knowledge, in mainland China no national surveys have been conducted to date to depict the overall vitamin D status in the population. Therefore, the purpose of this contribution was to provide the best possible evaluation of vitamin D deficiency/insufficiency in China by reviewing publications that measured plasma/serum 25-hydroxyvitamin-D (25[OH]D) levels in various age groups and in different areas of China from January 2000 to June 2012. From these investigations conducted throughout the country and from newborns to adults to the elderly, it has been found that vitamin D deficiency/insufficiency is prevalent in the Chinese population in almost all age groups and areas if individuals are not taking vitamin D-fortified products/supplements or are lacking sufficient sunshine exposure. Some studies showed severe deficiency (25[OH]D <25 nmol/L) in Nanjing (north latitude 31) during the winter months and in Beijing (north latitude 40) in the fall. This unoptimistic situation represents a significant but modifiable public health risk that deserves greater attention and more efficient and timely management.


[Possibilities of quantitative criteria of the effectiveness of treatment of glial brain tumors].
The authors suggest a quantitative criterion for evaluating the efficacy of radiotherapy in patients with glial tumors of the brain. Three types of course followed by the disease are distinguished: regressive, progressive, and undulating. The use of the quantitative criterion made it possible to detect patients with individual increased sensitivity to macrofractional radiotherapy and to come to the decision on the expediency of conducting the next series of irradiation in each concrete case.


Effect of contaminants on the adhesion of light-bodied silicones to putty silicones in putty-wash impression technique.
This study was concerned with the adhesion of a light-bodied silicone to a putty silicone in a putty-wash impression technique when the preliminary putty impression was contaminated with human saliva or residues from acrylic resins used in the fabrication of provisional restorations by direct technique. Data indicated that condensation and addition silicones differed in their susceptibility toward the tested contaminants. Salivary contamination and chemical residues from the autopolymerizing acrylic resins weakened the bond strength and caused adhesive failure dependent upon the type of silicone impression material used.


Oxaliplatin immunohybrid nanoparticles in vitro synergistic suppression evaluation in treatment of colorectal cancer.
In the present study, we have investigated the enhanced synergistic and apoptotic activity of immunohybrid nanoparticles encapsulating oxaliplatin and covalently conjugated with TRAIL (Apo-2L/CD-253). Time-dependent cytotoxicity activity of nanoparticles was determined by MTT assay in HT-29 cells. Nuclear morphological changes and assessment of apoptotic ratio was analyzed by DAPI (4'6-diamidino-2-phenylindole) staining and annexin-propidium iodide (PI) assay. Cell-cycle analysis of oxaliplatin in HT-29 cell was analyzed by flow cytometry at 72 h. Furthermore, molecular mechanisms related to oxaliplatin-induced anticancer activity was explored by western blot analysis. Our study revealed appreciable time-dependent cytotoxicity, apoptotic, and synergistic activity of oxaliplatin immunohybrid nanoparticles.


Physiology of Leymus chinensis under seasonal grazing: Implications for the development of sustainable grazing in a temperate grassland of Inner Mongolia.
Plants have different physiological characteristics as the season changes, grazing management in compliance with plant growth and development characteristics may provide new ideas for sustainable livestock development. However, there has been little research on seasonal grazing and plants physiological responses under it. Here, we studied a typical steppe ecosystem of Inner Mongolia, with Leymus chinensis as the dominant species, in five grazing treatments: continuous grazing, seasonal grazing (which started in spring or in early and late summer), and no grazing (the control). We analyzed growth and resistance of L. chinensis in the five treatments by measuring annual primary productivity, morphological traits and various physiological processes. Compared with continuous grazing, seasonal grazing significantly alleviated grassland degradation. The plants were less affected by stress under spring grazing, with net photosynthesis and non-photochemical quenching closer to the control values and with a lower malondialdehyde content. The annual primary production of plants under grazing started in the early and late summer were 3-4 times the value under continuous grazing. Regrowth under early-summer grazing was greatly improved, and stress resistance was stronger with a higher proline content and high antioxidant enzyme activity. And nutrient accumulation at the end of the growing season such as abundant soluble sugars were transferred from aboveground tissue to the roots in September under late-summer grazing, which benefited regrowth the next year. All these physiological processes were regulated by hormonal changes. Our results highlight how plants response grazing stress in different growing seasons and suggest that seasonal grazing can improve the stress resistance and regrowth capacity of forage vegetation, and applying this knowledge can promote more sustainable grazing practices.