Exploring ocean biogeochemistry by single-cell microprobe analysis of protist elemental composition.
The biogeochemical cycles of many elements in the ocean are linked by their simultaneous incorporation into protists. In order to understand these elemental interactions and their implications for global biogeochemical cycles, accurate measures of cellular element stoichiometries are needed. Bulk analysis of size-fractionated particulate material obscures the unique biogeochemical roles of different functional groups such as diatoms, calcifying protists, and diazotrophs. Elemental analysis of individual protist cells can be performed using electron, proton, and synchrotron X-ray microprobes. Here we review the capabilities and limitations of each approach and the application of these advanced techniques to cells collected from natural communities. Particular attention is paid to recent studies of plankton biogeochemistry in low-iron waters of the Southern Ocean. Single-cell analyses have revealed significant inter-taxa differences in phosphorus, iron, and nickel quotas. Differences in the response of autotrophs and heterotrophs to iron fertilization were also observed. Two-dimensional sub-cellular mapping indicates the importance of iron to photosynthetic machinery and of zinc to nuclear organelles. Observed changes in diatom silicification and cytoplasm content following iron fertilization modify our understanding of the relationship between iron availability and silicification. These examples demonstrate the advantages of studying ocean biogeochemistry at the level of individual cells.


Seasonal variations in pancreatic surgery outcome A retrospective time-trend analysis of 2748 Whipple procedures.
Observing cyclic patterns in surgical outcome is a common experience. We aimed to measure this phenomenon and to hypothesize possible causes using the experience of a high-volume pancreatic surgery department. Outcomes of 2748 patients who underwent a Whipple procedure at a single high-volume center from January 2000 to December 2018 were retrospectively analyzed. Three different hypotheses were tested: the effect of climate changes, the "July effect" and the effect of vacations. Clavien-Dindo ≥ 3 morbidity was similar during warm vs. cold months (22.5% vs. 19.8%, p = 0.104) and at the beginning of activity of new trainees vs. the rest of the year (23.5 vs. 22.5%, p = 0.757). Patients operated when a high percentage of staff is on vacation showed an increased Clavien-Dindo ≥ 3 morbidity (22.3 vs. 18.5%, p = 0.022), but similar mortality (2.3 vs. 1.8%, p = 0.553). The surgical waiting list was also significantly longer during these periods (37 vs. 27 days, p = 0.037). Being operated in such a period of the year was an independent predictor of severe morbidity (OR 1.271, CI 95% 1.086-1.638, p = 0.031). Being operated when more staff is on vacation significantly affects severe morbidity rate. Future healthcare system policies should prevent the relative shortage of resources during these periods.


Visual outcomes in patients with zonular dialysis following cataract surgery.
PurposeTo evaluate the visual outcomes of patients with zonular dialysis following cataract surgery.Patients and methodsMedical records of all patients with documented zonular dialysis, either pre- or peri-operatively, undergoing cataract surgery between 2004-2010 at Queen Alexandra Hospital, Portsmouth, were retrospectively reviewed. Baseline demographics and biometry were analysed, and ocular co-morbidities documented. Intraoperative complications and the use of a capsular tension ring (CTR) were identified. Early and late best-corrected visual acuity (BCVA) post surgery were determined using LogMar values. Univariate and multivariate linear regression analysis was performed to determine associations with BCVA post surgery, and further subgroup analysis performed in groups determined by CTR use.ResultsThe records of 22 312 consecutive eyes undergoing cataract surgery were reviewed. The incidence of zonular dialysis was 0.50% (111 eyes). A CTR was inserted in 46 eyes. Using a multivariate linear regression model, better initial pre-operative BCVA (P=0.019), the use of a CTR (P=0.014), and the absence of vitreous loss during surgery (P=0.008, β 0.45) were associated with improved early postoperative BCVA (mean follow-up 6.6 weeks). Better medium-term postoperative BCVA was significantly associated with preoperative BCVA (P=0.002) and the use of a CTR during surgery (P=0.004, β -0.41).ConclusionsThe overall incidence of zonular dialysis is low. CTR use intra-operatively suggests improved early and medium-term BCVA and should be considered in all cases of zonular dialysis.


Combined application of targeted and untargeted proteomics identifies distinct metabolic alterations in the tetraacetylphytosphingosine (TAPS) producing yeast Wickerhamomyces ciferrii.
The Wickerhamomyces ciferrii strain NRRL Y-1031 F-60-10A is a well-known producer of tetraacetylphytosphingosine (TAPS) and used for the biotechnological production of sphingolipids and ceramides. It was our aim to gain new biological insights into the sphingolipid metabolism by employing a dual platform mass spectrometry strategy. The first step comprised metabolic (15)N-labeling in combination with label-free proteomics using high resolution LTQ Orbitrap mass spectrometry. Then selected reaction monitoring tandem mass spectrometry served for the targeted quantification of sphingoid base biosynthesis enzymes. The non-producer strain NRRL Y-1031-27 served as a reference strain. In total, 1697 proteins were identified, and 123 enzymes of main metabolic pathways were observed as differentially expressed. Major findings were: 1) the likely presence of higher carbon flux in NRRL Y-1031 F-60-10A, resulting in higher availability of pyruvate and acetyl-CoA; 2) indications of oleaginous yeast-like behavior in NRRL Y-1031 F-60-10A; and 3) approx. 2-fold higher abundance of eight sphingolipid biosynthesis enzymes in NRRL Y-1031 F-60-10A. Taken together, in NRRL Y-1031 F-60-10A, the levels of glycolytic enzymes apparently gear carbon flux towards higher acetyl-CoA synthesis rates, while simultaneously reducing protein biosynthesis in the process. Thereby, C2 building blocks for acyl-moieties and downstream sphingoid base acetylation are provided to maintain TAPS production. First quantitative proteome study for a phytosphingosine-producing yeast.


Signaling pathways regulating the expression of Prx1 and Prx2 in the chick mandibular mesenchyme.
Prx1 and Prx2 are members of the aristaless-related homeobox genes shown to play redundant but essential roles in morphogenesis of the mandibular processes. To gain insight into the signaling pathways that regulate expression of Prx genes in the mandibular mesenchyme, we used the chick as a model system. We examined the patterns of gene expression in the face and the roles of signals derived from the epithelium on the expression of Prx genes in the mandibular mesenchyme. Our results demonstrated stage-dependent roles of mandibular epithelium on the expression of Prx in the mandibular mesenchyme and provide evidence for positive roles of members of the fibroblast and hedgehog families derived from mandibular epithelium on the expression of Prx genes in the mandibular mesenchyme. Our studies suggest that endothelin-1 signaling derived from the mesenchyme is involved in restricting the expression of Prx2 to the medial mandibular mesenchyme.


Correction to: AUGS/IUGA scientific meeting : Abstracts of poster presentations, IUGA 44th annual meeting - Nashville, TN, USA, September 24-28, 2019.
In the originally published version of the Abstracts article, the figure presented in Poster 204 entitled "Exosomes derived from fibroblasts in anterior vaginal wall associated with the pathogenesis of female stress urinary incontinence by promoting fibroblasts migration".


Benefit versus risk for oral, inhaled, and nasal glucocorticosteroids.
Many factors affect the benefit-to-risk analysis of the use of the glucocorticosteroids in the treatment of allergic inflammation. For most patients the benefit-to-risk ratio favors topical over oral administration, but in severe disease oral administration may be required. It is clear from pharmacokinetic and pharmacodynamic studies that inhaled glucocorticosteroids in particular are absorbed into the systemic circulation and that they can have clinical adverse effects when given in high doses. Therefore, with inhaled and nasal glucocorticosteroids, the dose that will achieve the optimal benefit/risk ratio is the lowest dose that controls symptoms and prevents exacerbations requiring treatment with oral glucocorticosteroids. This optimal dose needs to be determined on an individual basis and is likely to vary as disease severity changes over time.


Mapping of the pressure response element of the c-fos gene by direct DNA injection into beating hearts.
In vivo gene transfer into various organs of postnatal animals has recently been demonstrated. We applied this technique in order to map the promoter element of the protooncogene c-fos in the myocardium that responds to left ventricular pressure overload in vivo. Beating rat hearts were directly injected with a recombinant plasmid containing a 356-base pair upstream regulatory element of the mouse c-fos gene fused to a reporter gene. Two days after the injection, the hearts were excised and buffer-perfused. Acute pressure overload was applied for 2 h by inflating an intraventricular balloon. Pressure overload increased the myocardial reporter gene activity by 3-8-fold (p < 0.01). Deletion and point mutations in the serum response element (SRE) of the c-fos promoter resulted in loss of pressure-induced reporter gene expression, indicating that the SRE is necessary for pressure response. The SRE alone was sufficient to confer pressure responsiveness to the minimal c-fos promoter, confirming that the pressure response element coincides with the SRE. A construct containing a point mutation at the p62TCF binding site of the SRE did not respond to pressure overload or to a protein kinase C activator but retained responsiveness to a calcium ionophore. This suggests that pressure overload may activate the c-fos promoter by a protein kinase C-dependent pathway. Thus, a directly injected gene can be regulated in the host cells by a physical stimulus to the intact organ. The in vivo DNA injection technique is a useful method to study complex organ physiology at the molecular level.


Retraction Note: Silencing of Glutathione S-Transferase Pi Inhibits Cancer Cell Growth via Oxidative Stress Induced by Mitochondria Dysfunction.
Editor's Note: this Article has been retracted; the Retraction Note is available at https://www.nature.com/articles/s41598-020-77205-9.


NMR study of self-paired parallel duplex of d(AAAAACCCCC) in solution.
The oligonucleotide d(A5C5) in solution forms a parallel self-duplex at neutral and low pH values. H2O NMR spectra at pH 5.1 indicate the presence of five imino resonances at lower temperatures; and the structure is stable up to 60 degrees C. These signals can arise only from the hemiprotonated C+.C pairs [Westhof, E. and Sundaralingham, M. (1980) Biochemistry 77, 1852-1856; Westhof, E. and Sundaralingham, M. (1980) J. Mol. Biol. 142, 331-361] and constitute the first direct observation of C+.C hemiprotonated pairs in solution. The cross peaks from H1's and more than five distinct AH8's in 500 MHz 1H 2D-NOESY spectra indicate that there are two conformationally different and energetically similar A-tracts. There is good qualitative agreement between NOESY data and two theoretically derived structures in which A-tracts are reverse Watson-Crick and reverse Hoogsteen base-paired, respectively.


The thymus atrophy-inducing organotin compound DBTC inhibits the binding of thymocytes to thymic epithelial cells.
In this study, it is examined whether the organotin compound di-n-butyltindichloride (DBTC), which has been shown to inhibit immature thymocyte proliferation, is able to disturb the binding between thymocytes and thymic epithelial cells (TEC). To that end, an enzyme-linked binding assay was developed in which the amount of binding of Thy-1+ (mAb ER4)-thymocytes to the rat-derived TEC-line IT45R1 (IT45R1-TEC) could be detected. It was found that preincubation of thymocytes with 3-5 microM DBTC for 30 min inhibited the binding by 50-60% during a 1 h adhesion period. By extending the preincubation period to 1 h and the adhesion period to 22 h, 0.1 microM DBTC was already sufficient to reduce the binding with 60-80%. Further characterization of the binding revealed that splenic lymphocytes were unable to bind to the MHC class II-negative IT45R1-TEC. Since dextran sulfate inhibited the binding as well, sulfated polysaccharide-binding molecules such as Thy-1 and CD2 are likely to be involved in the binding. Electron microscopy showed filament-containing microvilli at the site of interaction. The results are discussed in relation to the mechanism of DBTC-induced thymus atrophy.


Kidney fibrosis: Emerging diagnostic and therapeutic strategies.
An increasing number of patients worldwide suffers from chronic kidney disease (CKD). CKD is accompanied by kidney fibrosis, which affects all compartments of the kidney, i.e., the glomeruli, tubulointerstitium, and vasculature. Fibrosis is the best predictor of progression of kidney diseases. Currently, there is no specific anti-fibrotic therapy for kidney patients and invasive renal biopsy remains the only option for specific detection and quantification of kidney fibrosis. Here we review emerging diagnostic approaches and potential therapeutic options for fibrosis. We discuss how translational research could help to establish fibrosis-specific endpoints for clinical trials, leading to improved patient stratification and potentially companion diagnostics, and facilitating and optimizing development of novel anti-fibrotic therapies for kidney patients.


Engaging and supporting fathers to promote breast feeding: a concept analysis.
Empirical evidence demonstrates that fathers have a strong influence on a mother's decision to initiate and continue breast feeding. However, no clear delineation of what behaviours and attributes constitute father support or differentiate it from other kinds of support is provided in the current literature. The purpose of this study was to analyse the concept of 'father support' in relation to maternity services and broader health settings, thereby clarifying meaning to enable comprehension and application in practice, education, and research. A concept analysis combining the evolutionary model of concept development with the inter-related theoretical, fieldwork and analytical phases of the hybrid model of concept development. Children's Centres in East and West Sussex in Southern England. Repeated qualitative research over two phases with 16 parents of breast fed infants through seven focus groups and five telephone interviews. CINAHL, PsycINFO, AMED, MEDLINE, OVID and EMBASE databases were searched for articles published in English between 1999 and 2013 using the keywords breast feeding, father, and support. Seven same-sex focus groups and five individual interviews were also conducted over two research phases with the parents of breast fed infants to expand and exemplify, and then validate the analysis of the literature search. Five main attributes of father support in relation to breast feeding were identified: (1) knowledge about breast feeding; (2) positive attitude to breast feeding; (3) involvement in the decision-making process; (4) practical support; and (5) emotional support. Multiple antecedents and consequences to these attributes were also identified. This study has contributed to clarifying the meaning of father support in relation to breast feeding and provides an important starting point for the development of a theoretical and practical model of optimal breast feeding that takes into account father support. Identification of attributes, antecedents, and consequences of father support may assist practitioners to reflect on current working practices and service delivery models, and offer important educational opportunities for the training of student midwives and other health professionals.


[The calculation and regulation of dust exposure doses to decrease the level of occupational diseases of dust etiology].
One of the real ways to reduce occupational diseases caused by dust is to cut the duration of contact with dust. The studies helped to suggest a method calculating occupational risk correlated to dust content of the air. The authors evaluated the duration of contact with dust so as to reduce occupational risk to the minimum. Those measures appeared to lower but not eliminate the occupational risk.


Insight into the nonlinear effect of COVID-19 on well-being in China: Commuting, a vital ingredient.
COVID-19 had a devastating impact on people's work, travel, and well-being worldwide. As one of the first countries to be affected by the virus and develop relatively well-executed pandemic control, China has witnessed a significant shift in people's well-being and habits, related to both commuting and social interaction. In this context, what factors and the extent to which they contribute to well-being are worth exploring. Through a questionnaire survey within mainland China, 688 valid sheets were collected, capturing various aspects of individuals' life, including travel, and social status. Focusing on commuting and other factors, a Gradient Boosting Decision Tree (GBDT) model was developed based on 300 sheets reporting working trips, to analyze the effects on well-being. Two indicators, i.e., the Relative Importance (RI) and Partial Dependency Plot (PDP), were used to quantify and visualize the effects of the explanatory factors and the synergy among them. Commuting characteristics are the most critical ingredients, followed by social interactions to explain subjective well-being. Commuting stress poses the most substantial effect. Less stressful commuting trips can solidly improve overall well-being. Better life satisfaction is linked with shorter confinement periods and increased restriction levels. Meanwhile, the switch from in-person to online social interactions had less impact on young people's life satisfaction. Older people were unsatisfied with this change, which had a significant negative impact on their life satisfaction. From the synergy of commuting stress and commuting time on well-being, the effect of commuting time on well-being is mediated by commuting stress in the case of China. Even if one is satisfied with online communication, the extent of enhancement on well-being is minimal, for it still cannot replace face-to-face interaction. The findings can be beneficial in improving the overall well-being of society during the pandemic and after the virus has been eradicated.


Predictive factor of recurrence after curative resection for stage I-II colon cancer.
Colon cancer has become a common malignant neoplasm in Tunisia. Patients with negative lymph node have a 5 years recurrence rate of 21.1%. Studies reporting the prognostic factors of recurrence for patients with stage I-II colon cancer are limited. This study aimed to determine factors predicting recurrence for patients with stage I-II colon cancer after curative resection. This was a retrospective cohort study. Were included patients who underwent curative surgery for stage I or II colon cancer. Enrolled variables were subdivided into: Pre-operative, Intraoperative and Post-operative variables. Main outcome measures were local recurrence and distant metastasis detected during follow-up. Eighteen men and 17 women with median age of 61 years, ranging from 33 to 89, were enrolled in this study. Twenty-eight patients out of 35 were classified T3 and T4 colon cancer. The mean number of lymph nodes harvested was 16.23 (median= 17; range: 4-44). Ten patients (28%) had colloid component in the tumor. At a median follow-up of 23 months (range: 6-56 months), recurrence was observed in five cases (14%). Variables associated to recurrence were Carcinoembryonic antigen level (p= 0.03), serum albumin level (p=0.029) and the presence of colloid component (0.02). Multivariate logistic regression retained colloid component as the only predictive factor of recurrence (OR=1.2, 95%CI [1.019-1.412], p=0.028). This study showed that the percentage of mucinous component equal or greater than 25% was the only predictive factor of recurrence for curatively resected, stages I and II, colon cancer.


Hypotension produced by intravenous apomorphine in the anaesthetized dog is not centrally mediated.
1 Intravenous administration of apomorphine (1.25 to 20 microgram/kg) in the anaesthetized dog produced a dose-dependent decrease in blood pressure which was antagonized by haloperidol but not influenced by propranolol or atropine. 2 Intracarotid administration of apomorphine produced a systemic hypotension which was significantly smaller than that seen with intravenous injection. 3 Doses of apomorphine that caused a decrease in blood pressure on intravenous injection, had no effect on blood pressure or caused retching accompanied by an increase in blood pressure on intravertebral or intracisternal administration. The animals showed a marked hypotension on intravertebral or intracisternal injection of clonidine. 4 From these results it is concluded that the hypotension seen with intravenous apomorphine cannot be explained by a central site of action.


Putative TRP channel antagonists, SKF 96365, flufenamic acid and 2-APB, are non-competitive antagonists at recombinant human α1β2γ2 GABA(A) receptors.
Although transient receptor potential (TRP) channel biology research has expanded rapidly in recent years, the field is hampered by the widely held, but relatively poorly investigated, belief that most of the pharmacological tools used to investigate TRP channel function may not be particularly selective for their intended targets. The objective of this study was therefore to determine if this was indeed the case by systematically evaluating the effects of three routinely used putative TRP channel antagonists, SKF 96365, flufenamic acid (FF) and 2-aminoethoxydiphenyl borate (2-APB) against one of the most widely expressed CNS receptor subtypes CNS, the human α1β2γ2 GABA(A) receptor. Using whole cell patch-clamp recording to record responses to rapidly applied GABA in the absence and presence of the three putative antagonists in turn we found that SKF 96365 (1-100 μM) and FF (1-100 μM) significantly inhibited GABA responses of recombinant human α1β2γ2 GABA(A) receptor stably expressed in HEK293 cells with IC(50) values of 13.4 ± 5.1 and 1.9 ± 1.4 μM, respectively, suppressing the maximal response to GABA at all concentrations used in a manner consistent with a non-competitive mode of action. SKF 96365 and FF also both significantly reduced desensitisation and prolonged the deactivation kinetics of the receptors to GABA (1mM; P<0.05). 2-APB (10-1000 μM) also inhibited responses to GABA at all concentrations used with an IC(50) value of 16.7 ± 5.4 μM (n=3-5) but had no significant effect on the activation, desensitisation or deactivation kinetics of the GABA responses. Taken together this investigation revealed that these widely utilised TRP channel antagonists display significant 'off-target' effects at concentrations that are routinely used for the study of TRP channel function in numerous biological systems and as such, data which is obtained utilising these compounds should be interpreted with caution.


Rb protein is essential to the senescence-associated heterochromatic foci formation induced by HMGA2 in primary WI38 cells.
Cellular senescence is an irreversible form of cell cycle arrest that provides a barrier to neoplastic transformation. The integrity of the Rb (Retinoblastoma) pathway is necessary for the formation of the senescence-associated heterochromatin foci (SAHF) that offers a molecular basis for the stability of the senescent state. Surprisingly, although high mobility group A2 protein (HMGA2) can promote tumorigenesis and inhibit Rb function in tumor cells, high-level expression of HMGA2 is sufficient to induce SAHF formation in primary cells. It therefore becomes significant to determine whether Rb protein is necessary in HMGA2-induced SAHF formation. In this study, we established the cellular senescence and SAHF assembly WI38 cell model by ectopic expression of HMGA2, in which typical senescent markers were seen, including notable upregulation of p53, p21 and p16, and elevated SA-β-galactosidase staining together with downregulation of E2F target genes. We then showed that the Rb pathway inhibitor E7 protein was able to partly abolish the ability of SAHF formation after HMGA2 expression in WI38 cells, indicating that Rb is a crucial factor for HMGA2-induced SAHF formation. However, Rb depletion did not completely rescue the cell growth arrest induced by HMGA2, suggesting that Rb is not an exclusive pathway for HMGA2-induced senescence in WI38 cells.


L-glutamate and gamma-aminobutyric acid uptake in synaptosomes from the cerebral cortex of dogs with congenital chronic hepatic encephalopathy.
High affinity [3H]gamma-aminobutyric acid (GABA) and [3H]L-glutamate uptake were determined in synaptosomes prepared from the cerebral cortex of dogs with congenital hepatic encephalopathy and control dogs. The Km value for GABA uptake was increased by 35% but there was a concomitant 34% increase in Vmax suggesting that GABA uptake capacity was not changed in HE dogs. In contrast, mean Vmax for glutamate uptake in HE dogs was 85% greater than mean Vmax in control dogs; mean Km was increased by 25% in HE dogs. Therefore, overall synaptosomal high affinity glutamate uptake capacity was increased in HE dogs compared to controls.


Headache in Adolescents.
The most common headache disorders in adolescents are tension-type headache, migraine, and posttraumatic headache. These disorders in adolescents may have different characteristics than in adults but can be similarly disabling. This review highlights the emerging abortive and preventive treatment options for the adolescent population. Although future high-quality headache studies in this age group are still needed, current evidence for the safety and efficacy of various treatment modalities is also discussed.


Same environment, stratified impacts? Air pollution, extreme temperatures, and birth weight in South China.
This paper investigates whether associations between birth weights and prenatal ambient environmental conditions-pollution and extreme temperatures-differ by 1) maternal education; 2) children's innate health; and 3) interactions between these two. We link birth records from Guangzhou, China, during a period of high pollution, to ambient air pollution (PM10 and a composite measure) and extreme temperature data. We first use mean regressions to test whether, overall, maternal education is an "effect modifier" in the relationships between ambient air pollution, extreme temperature, and birth weight. We then use conditional quantile regressions to test for effect heterogeneity according to the unobserved innate vulnerability of babies after conditioning on other confounders. Results show that 1) the negative association between ambient exposures and birth weight is twice as large at lower conditional quantiles of birth weights as at the median; 2) the protection associated with college-educated mothers with respect to pollution and extreme heat is heterogeneous and potentially substantial: between 0.02 and 0.34 standard deviations of birth weights, depending on the conditional quantiles; 3) this protection is amplified under more extreme ambient conditions and for infants with greater unobserved innate vulnerabilities.


Polyalkylphenyl-sulphonic acid with acid groups of variable strength from compost.
The insoluble organic fraction (humin-like material, HLM) from rice hull-dairy cattle compost is well converted into water soluble HLM-sulphonate by reaction in liquid SO3. Microanalytical, potentiometric, molecular weight, and NMR data are consistent with a highly homogeneous polymeric arylsulphonate having 4000 Da MW, 1.3 sulphonic groups per aromatic ring and significant content of carboxylic and phenolic groups. By comparison with structure-property relationships for commercial lignosulphonates derived from the pulp and paper industry, the above arylsulphonate is likely to be a candidate at a variety of applications in the chemical industry and in agriculture. Therefore, sulphonation is a means for upgrading composts HLM to the same uses as for commercial lignosulphonates.


Increased anticardiolipin antibodies associated with the development of anetoderma in HIV-1 disease. Military Medical Consortium for the Advancement of Retroviral research (MMCARR).
Anetoderma has been reported in patients with HIV-1 disease. In patients with autoimmune disease, anetoderma has been associated with increased levels of antiphospholipid antibodies (APL) that include anticardiolipin antibodies (ACA) and lupus anticoagulant (LA). This has led to speculation that the autoimmune phenomena seen in HIV-1 disease and the immune dysregulation induced by HIV-1 disease may play a role in the development of these lesions. We have seen both primary and secondary lesions of anetoderma in patients followed for HIV-1 disease. In this study, we wanted to determine whether there was an association in the development of anetoderma and elevated anticardiolipin antibodies (ACA) in HIV-1 patients. Quantitative ACA levels were measured in eight HIV-1-infected patients with anetoderma and four HIV-1-infected patients without anetoderma. Anticardiolipin antibodies were moderately elevated in seven of eight patients with lesions and were borderline in the four HIV-1-positive patients without lesions of anetoderma. There appears to be a correlation between increased ACA and the development of cutaneous lesions of anetoderma in HIV-1 disease. Patterns of immune dysregulation, including APL, may predispose to the development of lesions of anetoderma in HIV-1-positive patients. Although some of the lesions appear to represent primary anetoderma, the majority of our patients develop lesions in areas secondary to well characterized eruptions.


Congenital malformations as a cause of neonatal and postnatal death in Massachusetts (1970-1980).
Massachusetts birth and death certificate tapes for the years 1970-1980 were linked and analyzed to determine causes of death in the neonatal and postneonatal periods and to identify any related sociodemographic factors. Our analysis suggests that, although the neonatal mortality rate declined by about 43 percent, the postneonatal mortality rate remained relatively unchanged. Perinatal problems remained the principal cause of death during the neonatal period, throughout the decade. In the postneonatal period, congenital malformations became a leading cause of death toward the end of the decade because of a reduction in mortality from infectious diseases and perinatal problems. Infants born to mothers under 18 and over 34 years of age had the highest death rates from congenital birth defects. Higher mortality rates caused by congenital malformations were found in the more industrialized areas of Massachusetts. Further declines in infant mortality rates in Massachusetts will depend on preventive measures to reduce the incidence of congenital malformations.


Omega-3 PUFA attenuate mice myocardial infarction injury by emerging a protective eicosanoid pattern.
Omega-3 polyunsaturated fatty acid (n-3 PUFA) supplementation is a recommended preventive approach against cardiovascular diseases, but its mechanism of protection against myocardial infarction (MI) injury is not fully understood. Eicosanoid metabolomics demonstrated an abnormal eicosanoid profile was in the plasma of mice receiving MI surgery. 19,20-EDP, 17,18-EEQ, 14,15-EET and 9,10-EpOME were decreased, and PGE2 was increased by the surgery. N-3 PUFA-rich diets feeding or transgene of Fat-1 shifted the eicosanoid profile to an n-3 PUFA dominant style and attenuated the myocardial infarction injury. Multiple logistic regression analysis suggested the degree of MI injury was related with an eicosanoid pattern, composed by eicosanoids derived from both n-3 and n-6 PUFA in the three enzymatic pathways. These results suggested the benefits of n-3 PUFA on MI was achieved synergistically.


Nosocomial pneumonia. A multivariate analysis of risk and prognosis.
One hundred and twenty consecutive episodes of nosocomial pneumonia (NP) in 118 nonneutropenic adults admitted to a 1,000-bed teaching hospital were studied in order to investigate the prognosis and risk factors. The overall fatality rate was 36.6 percent. The identification of a "high-risk" microorganism (Pseudomonas aeruginosa, Enterobacteriaceae, and other Gram-negative bacilli, Streptococcus faecalis, Staphylococcus aureus, Candida sp, Aspergillus sp, and episodes of polymicrobial pneumonia), bilateral involvement on chest x-ray examination, the presence of respiratory failure, inappropriate antibiotic therapy, and age older than 60 years or an underlying condition ultimately or rapidly fatal were those factors selected by a stepforward logistic regression analysis as independently worsening the prognosis. A series of variables frequently quoted as predisposing to NP was determined to be either present or absent in the same 120 cases of NP and in an equal number of randomly selected control subjects. After adjusting for confounding, factors significantly predisposing to NP were tracheal intubation, depressed level of consciousness, underlying chronic lung disease, thoracic or upper abdominal surgery, prior episode of a large volume aspiration, and age older than 70 years. Since some of the factors influencing the risk or the prognosis of NP are amenable to medical intervention, a percentage of NP might be prevented and its prognosis can be improved.


Intrathoracic rhabdoid carcinoma: a clinicopathological, immunohistochemical, and ultrastructural study of 6 cases.
We evaluated the clinicopathological spectrum of intrathoracic rhabdoid carcinoma, including its immunophenotype and ultrastructural features. Our series included 6 cases arising from the lung (4 cases) and the anterior mediastinum (2 cases). The patients were 4 men and 1 woman aged between 40 and 63 years (median, 53 years). Microscopically, all of the lesions were composed of loosely cohesive, large, atypical polygonal cells, with glassy cytoplasms and eccentric nuclei. Chromatin texture was finely granular or open. On immunohistochemical stain, the neoplastic cells were positive for vimentin in all cases, positive for vimentin, keratins and/or epithelial membrane antigen in all cases, and negative for other antigens. In 1 case, neoplastic cells were also positive for CD34. Electron microscopic study showed prominent perinuclear whorls of densely packed intermediate filaments or mitochondria. All patients were treated with combined chemotherapy and radiotherapy. One patient died 8 months after the diagnosis was made. Of the remaining patients, 3 are alive with evidence of disease progression, including brain metastases. We concluded that rhabdoid neoplasms arising in the thoracic cavity are aggressive tumors of epithelial lineage and should be categorized as true "rhabdoid carcinomas" instead of being ambiguously designated as "intrathoracic rhabdoid tumors" or "carcinomas with rhabdoid phenotype."


Salmon fibrin glue in rats: antibody studies.
Fibrin sealants and topical thrombin preparations are often used for haemostatic and sealing applications in clinical practice. Some of these preparations contain coagulation factors from bovine sources. To minimize the risk of infection and immunogenicity connected with mammalian blood products, proteins derived from the plasma of farmed Atlantic salmon have been considered as an alternative to these mammalian sources. The purpose of this study is to characterize the immunogenicity of salmon fibrin glue in an animal model focusing on crossreactivity of IgG antibodies to host endogenous counterparts. After two immunizations with salmon fibrin glue, rats developed antibodies of IgG and IgM type to both fibrin glue components. Weak crossreactivity to endogenous fibrinogen and thrombin was seen in a subset of rats after the second application of salmon proteins. Coagulation tests showed that salmon fibrin application has no effect on coagulation profiles in mammalian hosts, consistent with previous reports that found no evidence of significant crossreactivity with host proteins. These studies support the potential suitability of salmon fibrin glue for the development of preparations with clinical impact. Before human use can be considered, however, additional data about safety of this preparation in other animal models, including large animal studies, should be obtained.


Dissociation of actions of BRL 34915 in the rat portal vein.
In rat portal vein, 0.5 and 5.0 microM BRL 34915 [(+/-)-6-cyano-3,4-dihydro-2,2-dimethyl-trans-4-(2-oxo-1-pyrrolidyl++ +)-2H- benzo[b]pyran-3-ol] abolished spontaneous rhythmic movements and norepinephrine (NE)-induced tension responses, respectively. Only the higher (5 microM) concentration of BRL 34915 increased 42K efflux and inhibited the NE-induced increase in 42K efflux. These results suggest that BRL 34915 inhibits spontaneous rhythmic movements and NE-induced tension responses by differing mechanisms of action and that only the block of the NE-induced tension response is related to K+ permeability or conductance changes.


Coherence and horizontal movements of pathces of Holopedium gibberum (Cladocera).
The horizontal spatial dynamics of Holopedium gibberum in a small lake was followed during two summers. H. gibberum individuals aggregated into a single patch measured in tens of meters. Patch movement was extensive throughout the limnetic region, bore no apparent relationship to wind action, and patch structure remained cohesive regardless of weather conditions. Grazing activity by H. gibberum within the patch was intensive, reducing phytoplankton density to half that outside the patch.


Feasibility of aortic valve assessment with low dose prospectively triggered adaptive systolic (PTAS) cardiac computed tomography angiography.
Cardiac computed tomography angiography (CTA) is feasible for aortic valve evaluation, but retrospective gated protocols required high radiation doses for aortic valve assessment. A prospectively triggered adaptive systolic (PTAS) cardiac CT protocol was recently described in arrhythmia using second-generation dual-source CT. In this study, we sought to evaluate the feasibility of PTAS CTA to assess the aortic valve at a low radiation dose. A retrospective cohort of 29 consecutive patients whom underwent PTAS protocols for clinical indications other than aortic valve assessment and whom also received echocardiography within 2 months of CT, was identified. Images were reviewed for aortic valve morphology (tricuspid/bicuspid/prosthetic) and stenosis (AS) by experienced blinded readers. Accuracy versus echocardiography and radiation doses were assessed. PTAS CTA protocols using second-generation dual-source CT for aortic valve evaluation are feasible at low doses. This protocol should be investigated further in larger cohorts.


Intracranial recurrence and distant metastasis of scalp dermatofibrosarcoma protuberans.
Only few cases of scalp dermatofibrosarcoma protuberans with intracranial and distant metastasis have been reported. Here we report a case of scalp dermatofibrosarcoma protuberans with frequent local recurrence, intracranial invasion and with distant lung metastasis during 6 years of treatment. We would like to emphasize difficulties in surgical treatment of such invasive and locally recurrent tumors of scalp, and necessity to understand new molecular pathogenesis of dermatofibrosarcoma protuberans and potential treatment strategy with imatinib for patients with surgically untreatable disease. Close surveillance of patients with scalp dermatofibrosarcoma is necessary due recurrence nature of tumor.


The Association Between Common Labor Drugs and Suckling When Skin-to-Skin During the First Hour After Birth.
Intrapartum drugs, including fentanyl administered via epidural and synthetic oxytocin, have been previously studied in relation to neonatal outcomes, especially breastfeeding, with conflicting results. We examined the normal neonatal behavior of suckling within the first hour after a vaginal birth while in skin-to-skin contact with mother in relation to these commonly used drugs. Suckling in the first hour after birth has been shown in other studies to increase desirable breastfeeding outcomes. Prospective comparative design. Sixty-three low-risk mothers self-selected to labor with intrapartum analgesia/anesthesia or not. Video recordings of infants during the first hour after birth while being held skin-to-skin with their mother were coded and analyzed to ascertain whether or not they achieved Stage 8 (suckling) of Widström's 9 Stages of newborn behavior during the first hour after birth. A strong inverse correlation was found between the amount and duration of exposure to epidural fentanyl and the amount of synthetic oxytocin against the likelihood of achieving suckling during the first hour after a vaginal birth. Results suggest that intrapartum exposure to the drugs fentanyl and synthetic oxytocin significantly decreased the likelihood of the baby suckling while skin-to-skin with its mother during the first hour after birth.


Some thermodynamic effects of varying nonpolar surfaces in protein-ligand interactions.
Understanding how making structural changes in small molecules affects their binding affinities for targeted proteins is central to improving strategies for rational drug design. To assess the effects of varying the nature of nonpolar groups upon binding entropies and enthalpies, we designed and prepared a set of Grb2-SH2 domain ligands, Ac-pTyr-Ac6c-Asn-(CH2)n-R, in which the size and electrostatic nature of R groups at the pTyr+3 site were varied. The complexes of these ligands with the Grb2-SH2 domain were evaluated in a series of studies in which the binding thermodynamics were determined using isothermal titration calorimetry, and binding interactions were examined in crystallographic studies of two different complexes. Notably, adding nonpolar groups to the pTyr+3 site leads to higher binding affinities, but the magnitude and energetic origins of these effects vary with the nature of the R substituent. For example, enhancements to binding affinities using aliphatic R groups are driven by more favorable changes in binding entropies, whereas aryl R groups improve binding free energies through a combination of more favorable changes in binding enthalpies and entropies. However, enthalpy/entropy compensation plays a significant role in these associations and mitigates against any significant variation in binding free energies, which vary by only 0.8 kcal•mol-1, with changes in the electrostatic nature and size of the R group. Crystallographic studies show that differences in ΔG° or ΔH° correlate with buried nonpolar surface area, but they do not correlate with the total number of polar or van der Waals contacts. The relative number of ordered water molecules and relative order in the side chains at pTyr+3 correlate with differences in -TΔS°. Overall, these studies show that burial of nonpolar surface can lead to enhanced binding affinities arising from dominating entropy- or enthalpy-driven hydrophobic effects, depending upon the electrostatic nature of the apolar R group.


Membranes nephropathy associated with renal cell carcinoma. Evidence against a role of renal tubular or tumor antibodies in pathogenesis.
A patient with the nephrotic syndrome due to membranous nephropathy was found to have renal cell carcinoma. Since membranous nephropathy in patients with malignancies has been attributed to a tumor antigen-antibody complex form of glomerulonephritis, an attempt was made to implicate tumor antigens and/or renal tubular epithelial antigens in the pathogenesis of membranous nephropathy in our patient with renal cell carcinoma. Antibodies directed against tumor antigens and renal tubular antigens and renal tubular eipthelial antigens were sought in his serum and in eluates of his glomeruli; no such antibodies were found. The concurrence of the two renal lesions may have been fortuitous in this patient. However, their association temporally suggests that they were related, and our immunologic studies demonstrate that tumor antigen-antibody complexes are not invariably involved in the pathogenesis of malignancy-associated membranous nephropathy.


Differences between males and females with spinal cord injury in the experience of subliminal and explicit sexual pictures.
Observational cross-sectional study. To determine the differences between persons with spinal cord injury (SCI) and control individuals in terms of conscious and unconscious sexual responses to subliminally presented visual sexual stimuli. Spinal cord injury rehabilitation center in northern Italy. A two-part behavioral experiment was conducted on 40 participants (27 individuals with SCI; 13 controls). In first part, all participants were subliminally exposed to a prime picture (neutral or sexual) and asked to rate the extent to which they were emotionally aroused, while watching a set of explicit target pictures (neutral or sexual). In the second part, choice reaction time task was employed, wherein participants were shown a subliminal prime picture (neutral or sexual) followed by an explicit target picture (neutral or sexual) superimposed by a black dot and were asked to locate the dot as fast as possible. In the first part, men with SCI reported higher levels of emotional arousal to explicit sexual target pictures compared to other groups. In the second part, slower choice reaction times were found in the SCI group, particularly with sexual prime picture. Moreover, females with SCI spent more time during implicit motor learning tasks with sexual target pictures than other groups. We found differences in the experience of subliminal and explicit sexual pictures not only between the two groups, but also between females and males with SCI. Attention should thus be paid when considering sexual experience at subliminal and conscious level in SCI population for future research and rehabilitative protocols.


Retinol bound to cellular retinol-binding protein is a substrate for cytosolic retinoic acid synthesis.
Retinol bound to cellular retinol-binding protein (CRBP) was found to be oxidized to retinoic acid by a soluble activity from calf liver. Cytosolic retinoic acid synthesis from retinol-CRBP was strictly dependent on the exogenous supply of either NAD or NADP. NAD-supported reactions carried out in the presence or in the absence of dimethyl sulfoxide yielded apparent Km and Vmax values for the retinol-CRBP complex of 3.5 +/- 0.6 microM, 611 +/- 49 pmol h-1 (mg of protein)-1, and 0.84 +/- 0.12 microM, 601 +/- 38 pmol h-1 (mg of protein)-1, respectively. The corresponding values for the oxidation of free retinol, dissolved in dimethyl sulfoxide, were 7.1 +/- 0.3 microM and 948 +/- 47 pmol h-1 (mg of protein)-1. Since the dissociation constant of the bovine retinol-CRBP complex is less than 10(-8) M, whereas the Km for retinol-CRBP is of the same order as the Km for free retinol, synthesis of retinoic acid from retinol-CRBP does not rely on prior dissociation of retinol. ApoCRBP proved to be a specific inhibitor of retinoic acid synthesis from CRBP-bound retinol. Its inhibitory effect was indistinguishable from the dilution of the radioactive retinol-CRBP substrate that was obtained by the addition of unlabeled holoCRBP. In contrast, the oxidation of CRBP-bound retinol was not inhibited by the addition of other retinoid binding proteins nor by the addition of either free retinol or retinol complexed with proteins distinct from CRBP. These results indicate that the protein moiety of holoCRBP is specifically recognized by the cytosolic enzyme system that catalyzes retinoic acid synthesis from CRBP-bound retinol.


Comparison of heart rate variability between surgical and interventional closure of atrial septal defect in children.
This study was designed to compare reduced heart rate variability (HRV) in children with atrial septal defect (ASD) after surgical ASD closure with interventional (Amplatzer device) closure. Reduced HRV was observed in all children with ASD before treatment. HRV was further impaired 1 month after surgical ASD closure but not after catheter device closure. Three months after treatment, HRV indexes tended to normalize in both groups.


Doxylamine and diphenhydramine pharmacokinetics in women on low-dose estrogen oral contraceptives.
Thirteen women chronically using low-dose estrogen-containing oral contraceptives (50 micrograms or less of ethinyl estradiol or its equivalent for a minimum of 3 months) and 12 age-matched drug-free control women received a single 25 mg oral dose of doxylamine succinate in the fasting state. Ten women taking oral contraceptives and ten controls received a single 50 mg oral dose of diphenhydramine hydrochloride. Multiple plasma samples drawn during 30 hours following the dose of doxylamine, and 12 hours after diphenhydramine dosage, were analyzed by gas chromatography using nitrogen-phosphorus detection. Mean pharmacokinetic variables for doxylamine in control and oral contraceptive groups were: peak plasma concentration, 103 vs 100 ng/ml; time of peak, 2.40 vs 1.87 hours after dosage, elimination half-life, 10.1 vs 10.2 hours; and total clearance, 3.70 vs 3.88 ml/min/kg. Mean pharmacokinetic variables for diphenhydramine in control and oral contraceptive groups were: peak plasma concentration, 63.7 vs 73.8 ng/ml; time of peak, 2.7 vs 2.2 hours after dosage; elimination half-life, 6.0 vs 5.1 hours; and total clearance, 21.8 vs 25.5 ml/min/kg. None of these differences were statistically significant. Thus, low-dose estrogen-containing oral contraceptives do not significantly influence the pharmacokinetics of the antihistamines doxylamine or diphenhydramine.


Inhibitor for FcεRI expression on mast cell from Verbasucum thapsus L.
We found out 2',3'-dihydroxypuberulin from South American medicinal plant, V. thapsus L., as a candidate of an anti-allergic lead which inhibits the expression of high-affinity receptor of IgE (FcεRI) on the surface of mast cells. Furthermore, the analysis of structure-activity relationship by using synthesized 2',3'-dihydroxypuberulin analogs revealed that both hydroxy groups in the side chain and both of methyl moieties on phenolic hydroxy groups were crucial for potent activity, but absolute configuration of C-3' position wasn't. The active principle, 2',3'-dihydroxypuberulin, was disclosed to down-regulate the mRNA level of β-chain of FcεRI, different from previous reported active natural product reducing γ-chain level.


Heterologous expression of oxalate decarboxylase in Lactobacillus plantarum NC8.
Lactic acid bacteria (LABs) are being used as a probiotic very often for various enteric problems. Many genetically modified LABs are created by different workers for various novel applications. In this study we examine the expression of heterologous oxalate decarboxylase (oxdc) in Lactobacillus plantarum NC8. Generally, this enzyme is not present in Lactobacillus spp. Oxdc gene from Bacillus subtilis was polymerase chain reaction-amplified and cloned in a shuttle vector pSIP400 series, downstream of the inducible promoter, P(orfx). In the presence of an inducing peptide, Sakacin-P, the expression of OxdC was observed in sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The cell-free extract and the purified protein from the recombinant LABs showed the presence of OxdC activity. The above recombinant LABs, with desired modifications, can be used as a possible probiotic for the degradation of intestinal dietary oxalate for preventing enteric hyperoxaluria.


Empathetic Concern in Emerging Adolescents: The Role of Theory of Mind and Gender Roles.
The current study examined the roles of gender, and gender-role orientation in young adolescents' empathetic concern. In addition, this study aimed to explore the contribution of Theory of Mind in participants' empathetic concern. Finally, this study examined whether gender and gender-role orientation were implicated in emerging adolescents' Theory of Mind understanding. One-hundred-fifty 11- to 12-year-olds (79 self-identified females) completed questionnaires measuring their empathetic concern, Theory of Mind, and their perceived gender-role orientation. Results showed that gender-role orientation, specifically, femininity and masculinity predicted empathetic concern above and beyond gender. In addition, the effects of cognitive and affective Theory of Mind are explored and discussed in relation to empathetic concern. Finally, neither gender nor gender-role orientation was found to contribute to participants' Theory of Mind understanding. These findings suggest that emerging adolescents' perceived gender roles, as well as their ability to consider another's beliefs, play a role in their expression of empathetic concern.


Segregation of genetic determinants for murine glucuronidase synthesis and loss in CXB recombinant-inbred strains.
A set of recombinant-inbred strains developed from mouse strains BALB/c and C57BL/6 includes two beta-glucuronidase phenotypes that are not seen in either of the progenitor strains. These new recombinant phenotypes indicate that glucuronidase levels are regulated by genes additional to the Gur locus, which is closely linked to the glucuronidase structural gene (Gus) and is known to regulate the rate of glucuronidase synthesis. In this study, induced rates of glucuronidase synthesis were determined for these recombinant-inbred strains, and rate constants for enzyme loss were calculated. The rate of synthesis was found to segregate with the Gus gene in all of the strains, and only the determinants for rate of enzyme loss recombined to give new phenotypes. It was concluded that at least two genes affect the rate of enzyme loss, that these genes are not closely linked to each other or to the Gur-Gus region on chromosome 5, and that no major determinants of glucuronidase synthesis segregate independently of Gur.


Comparison of a row-column speller vs. a novel lateral single-character speller: assessment of BCI for severe motor disabled patients.
Non-invasive brain-computer interface (BCI) based on electroencephalography (EEG) offers a new communication channel for people suffering from severe motor disorders. This paper presents a novel P300-based speller called lateral single-character (LSC). The LSC performance is compared to that of the standard row-column (RC) speller. We developed LSC, a single-character paradigm comprising all letters of the alphabet following an event strategy that significantly reduces the time for symbol selection, and explores the intrinsic hemispheric asymmetries in visual perception to improve the performance of the BCI. RC and LSC paradigms were tested by 10 able-bodied participants, seven participants with amyotrophic lateral sclerosis (ALS), five participants with cerebral palsy (CP), one participant with Duchenne muscular dystrophy (DMD), and one participant with spinal cord injury (SCI). The averaged results, taking into account all participants who were able to control the BCI online, were significantly higher for LSC, 26.11 bit/min and 89.90% accuracy, than for RC, 21.91 bit/min and 88.36% accuracy. The two paradigms produced different waveforms and the signal-to-noise ratio was significantly higher for LSC. Finally, the novel LSC also showed new discriminative features. The results suggest that LSC is an effective alternative to RC, and that LSC still has a margin for potential improvement in bit rate and accuracy. The high bit rates and accuracy of LSC are a step forward for the effective use of BCI in clinical applications.


Molecular probes for general testicular and specific spermatogenic function.
Northern analysis of human testis poly(A+) RNA with a mixture of oligonucleotide primer extended cDNA probes revealed several similar RNAs. These RNAs were subsequently cloned into a VPCS (vector-primer-cloner-sequencer) plasmid. One of these clones, NDHu1, was represented within the library a number of times and hybridized strongly to a poly(A+) RNA of congruent to 1.2 kb. Sequence analysis identified this clone as the URF 1 subunit of the mitochondrial NADH dehydrogenase (NDHu1). Comparison of the relative levels of the NDHu1 and human protamine 1 (HP1) transcripts revealed that HP1 was less abundant than NDHu1. This was unexpected, since it is known that within differentiating mammalian spermatid cells, protamine (HP1) is an abundant transcript. This suggested that the ratio of the relative levels of these two very different mRNAs was indicative of the relationship between specific spermatogenic function (germ cell transcription, determined by the level of the HP1 transcript) and general testicular cell function (determined by the level of the mitochondrial mRNAs, i.e. NDHu1). This correlation was maintained when several individuals expressing various degrees of testicular dysfunction were examined. This study suggests that these probes may be useful markers for general testicular and specific spermatogenic function.


Modernizing residential treatment centers for children and youth--an informed approach to improve long-term outcomes: the Damar pilot.
Much controversy exists regarding the effectiveness of residential treatment. Recently, emerging research has demonstrated that community-based residential treatment has more positive long-term outcomes for youth. This article describes a community-based program that was implemented at a residential treatment agency serving youth. Targeted recidivism variables that were used to guide the study are described. Results demonstrated significant behavioral improvements, as well as improved post-discharge status. Conclusions and recommendations are also provided.


State of the art and new developments in molecular diagnostics for hemoglobinopathies in multiethnic societies.
For detecting carriers of thalassemia traits, the basic part of diagnostics consists of measurement of the hematological indices followed by mostly automatic separation and measurement of the Hb fractions, while direct Hb separation either on high pressure liquid chromatography or capillary electrophoresis is sufficient to putatively identify carriers of the common Hb variants like HbS, C, E, D, and O-Arab. A putative positive result is reported together with an advice for parents, partner, or family analysis. For couples, presumed at-risk confirmation at the DNA level is essential. In general, this part of diagnostics is done in specialized centers provided with sufficient experience and the technical tools needed to combine hematological and biochemical interpretation with identification of the mutations at the molecular level. State-of-the-art tools are usually available in centers that also provide prenatal diagnosis and should consist of gap-PCR for the common deletions, direct DNA sequencing for all kind of point-mutations and the capacity to uncover novel or rare mutations or disease mechanisms. New developments are MLPA for large and eventually unknown deletion defects and microarray technology for fine mapping and primer design for breakpoint analysis. Gap-PCR primers designed in the region flanking the deletion breakpoints can subsequently be used to facilitate carrier detection of uncommon deletions in family members or isolated populations in laboratories where no microarray technology or MLPA is available.


Postoperative Complications in Medicare Beneficiaries Following Endothelial Keratoplasty Surgery.
To determine national-level incidence rates of major postoperative complications following endothelial keratoplasty (EK) procedures and to stratify these rates based on EK indications over an 8-year period using Medicare claims data. Retrospective, cohort study. Setting: population-based; study population: Medicare beneficiaries aged ≥65 years who underwent EK procedures; main outcome measurements: 1) occurrence of major postoperative complications (i.e., endophthalmitis, choroidal hemorrhage, infectious keratitis, cystoid macular edema [CME], retinal detachment [RD], or RD surgery) following EK surgery; 2) time-to-event analysis for glaucoma surgery; and 3) occurrence of graft complications. A total of 94,829 EK procedures (n = 71,040 unique patients) were included in the analysis. Of the total, 29% of patients had pre-existing glaucoma. The overall 90-day cumulative incidence of postoperative endophthalmitis and choroidal hemorrhage following EK was 0.03% and 0.05%, respectively. The overall 1-year cumulative rates of RD or RD surgery, infectious keratitis, and CME were 1.0%, 0.8%, and 4.1%, respectively. Approximately 7.6%, 12.2%, and 13.8% of all eyes in this study needed glaucoma surgery at 1-, 5-, and 8-years of follow-up, respectively. The probability of glaucoma surgery among patients with pre-existing glaucoma was 29% vs. 8% among those without pre-existing glaucoma at 8 years. The cumulative probabilities of developing any graft complications were 13%, 23.2%, and 27.1% at 1, 5, and 8 years, respectively, of follow-up. On average, patients undergoing EK procedures for a prior failed graft had the highest rate of complications, whereas those with Fuchs' corneal endothelial dystrophy had the lowest. The incidence of major postoperative complications including endophthalmitis, retinal detachment, and choroidal hemorrhage following EK procedures is low. A high proportion of eyes undergoing EK eventually require glaucoma surgery and experience graft-related complications. Postoperative outcomes are typically worse for patients undergoing EK for prior failed grafts than for those undergoing EK for Fuchs' corneal endothelial dystrophy.


Myocardial involvement in children with post-COVID multisystem inflammatory syndrome: a cardiovascular magnetic resonance based multicenter international study-the CARDOVID registry.
Recent evidence shows an association between coronavirus disease 2019 (COVID-19) infection and a severe inflammatory syndrome in children. Cardiovascular magnetic resonance (CMR) data about myocardial injury in children are limited to small cohorts. The aim of this multicenter, international registry is to describe clinical and cardiac characteristics of multisystem inflammatory syndrome in children (MIS-C) associated with COVID-19 using CMR so as to better understand the real extent of myocardial damage in this vulnerable cohort. Hundred-eleven patients meeting the World Health Organization criteria for MIS-C associated with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), having clinical cardiac involvement and having received CMR imaging scan were included from 17 centers. Median age at disease onset was 10.0 years (IQR 7.0-13.8). The majority of children had COVID-19 serology positive (98%) with 27% of children still having both, positive serology and polymerase chain reaction (PCR). CMR was performed at a median of 28 days (19-47) after onset of symptoms. Twenty out of 111 (18%) patients had CMR criteria for acute myocarditis (as defined by the Lake Louise Criteria) with 18/20 showing subepicardial late gadolinium enhancement (LGE). CMR myocarditis was significantly associated with New York Heart Association class IV (p = 0.005, OR 6.56 (95%-CI 1.87-23.00)) and the need for mechanical support (p = 0.039, OR 4.98 (95%-CI 1.18-21.02)). At discharge, 11/111 (10%) patients still had left ventricular systolic dysfunction. No CMR evidence of myocardial damage was found in most of our MIS-C cohort. Nevertheless, acute myocarditis is a possible manifestation of MIS-C associated with SARS-CoV-2 with CMR evidence of myocardial necrosis in 18% of our cohort. CMR may be an important diagnostic tool to identify a subset of patients at risk for cardiac sequelae and more prone to myocardial damage. The study has been registered on ClinicalTrials.gov, Identifier NCT04455347, registered on 01/07/2020, retrospectively registered.


Inhibition of cytokine-induced prostanoid biogenesis by phytochemicals in human colonic fibroblasts.
Many of the inflammatory pathways regulating the production of prostanoids are implicated in the development of colon cancer. Diets rich in fruits and vegetables are associated with decreased rates of colon cancer and this may reflect anti-inflammatory properties of some phytochemicals in plant-based foods. In order to ascertain which of the many dietary compounds may be protective, a cell-based screening method was established to determine their effects on the production of prostanoids. By up-regulating prostaglandin H synthase-2 in human colonic fibroblast cells with cytokines, we have investigated the potential protective effect of a structurally related group of phytochemicals on prostanoid biogenesis. Several of the compounds significantly inhibited prostanoid biogenesis, by up to 81% and others enhanced prostanoid production. All of the compounds that enhanced prostanoid production belonged to the hydroxylated benzoic acid family and good correlation was observed with their redox activity and the ability to enhance prostanoid production. Common structural features of the inhibitors were the presence of 4-hydroxyl and 3-methoxyl substituents on the aromatic ring and/or the presence of a three-carbon side-chain on C1.


High molecular weight vimentin complex is formed after proteolytic digestion of vimentin by caspase-3: detection by sera of patients with interstitial pneumonia.
In a previous study, we demonstrated anti-vimentin antibodies in sera of patients with interstitial pneumonia. We hypothesized that antibodies in sera might detect vimentin fragments formed during the process of apoptosis. To prove this, recombinant human vimentin was digested by recombinant human caspase-3 or caspase-8. Then, Western blotting using several commercially available antibodies against human vimentin or patients' sera which had anti-vimentin autoantibodies, was performed. As a result, after recombinant human vimentin was digested by caspase-3 or caspase-8, several vimentin fragments were formed and detected by 2 kinds of monoclonal anti-vimentin antibodies (clone 3B4 and clone V9) as well as by polyclonal sheep anti-human vimentin antibody. It was demonstrated that high molecular weight vimentin was formed after the digestion of vimentin by caspase-3, which was only detected by patients' sera. The high molecular weight vimentin was not formed after digestion of vimentin by caspase-8. Our present results show that high molecular weight vimentin was formed after the digestion of vimentin by caspase-3. In addition, it is suggested that this high molecular weight vimentin acted as an autoantigen to form anti-vimentin autoantibody in vivo.


Self-organization of organoids from endoderm-derived cells.
Organoids constitute biological systems which are used to model organ development, homeostasis, regeneration, and disease in vitro and hold promise for use in therapy. Reflecting in vivo development, organoids form from tissue cells or pluripotent stem cells. Cues provided from the media and individual cells promote self-organization of these uniform starting cells into a structure, with emergent differentiated cells, morphology, and often functionality that resemble the tissue of origin. Therefore, organoids provide a complement to two-dimensional in vitro culture and in vivo animal models of development, providing the experimental control and flexibility of in vitro methods with the three-dimensional context of in vivo models, with fewer ethical restraints than human or animal work. However, using organoids, we are only just beginning to understand on the cellular level how the external conditions and signaling between individual cells promote the emergence of cells and structures. In this review, we focus specifically on organoids derived from endodermal tissues: the starting conditions of the cells, signaling mechanisms, and external media that allow the emergence of higher order self-organization.


Potential antifertility agents from plants: a comprehensive review.
Traditional medicines are practiced worldwide for regulation fertility since ancient times. This review provides a comprehensive summary of medicinal flora inhabitating throughout the world regarding their traditional usage by various tribes/ethnic groups for fertility regulation in females. Bibliographic investigation was carried out by analyzing classical text books and peer reviewed papers, consulting worldwide accepted scientific databases from the last six decades. Plants/their parts/extracts traditionally used for abortion, contraception, emmenagogue and sterilization purposes have been considered as antifertility agents. Research status of selected potential plant species has been discussed. Further, compounds isolated from plants with attributed fertility regulating potentials are also classified into three categories: (a) phytoconstituents with anti-implantation activity, (b) phytoconstituents with abortifacient activity and (c) phytoconstituents with contraceptive activity. 577 plant species belonging to 122 families, traditionally used in fertility regulation in females, have been recorded, of which 298 plants have been mentioned as abortifacients (42%), 188 as contraceptives (31%), 149 as emmenagogues (24%), and 17 as sterilizers. Among 122 plant families, fabaceae constitutes 49.2%, asteraceae 40.98%, euphorbiaceae 19.7%, apiaceae 16.4%, poaceae 12.3%, labiateae 11.5%, and others in lesser proportion. Various plant parts used in fertility regulation include leaves (25%), roots (22%), fruits (15%), seeds (12%), stem/stem bark (37%), and flowers (4%). Some active compounds, isolated from about various plant species, have been reported to possess significant antifertility potential. This review clearly indicates that it is time to increase the number of experimental studies to find out novel potential chemical entities from such a vast array of unexploited plants having traditional role in fertility regulation. Also, the mechanisms of action by which plant extracts and their active compounds exert antifertility effects remain to be studied.


Beclin1 haploinsufficiency compromises mesenchymal stem cell-offered cardioprotection against myocardial infarction.
Mesenchymal stem cells (MSCs)-based therapy has displayed some promises in ischemia heart diseases although its efficacy may be affected by changes in surrounding environments. This study evaluated the role of autophagy insufficiency using Beclin1 haploinsufficiency (BECN+/-) on intra-myocardial MSC transplantation-evoked effect against myocardial infarction. Donor MSCs from C57BL/6 mice were labelled with cell-tracker CM Dil and were delivered into LV free wall adjacent to infarct region in wild-type (WT) and BECN+/- recipient mice following ligation of left main coronary artery (MI-MSCs). Ten days following MI, myocardial function was assessed using echocardiography. Cardiomyocyte contractility and intracellular Ca2+ were monitored using cardiomyocytes from the area-at-risk adjacent to infarct. CM-Dil labeled cells were tracked in MSCs recipient mice using fluorescence microscopy. Lectin, Masson trichrome staining and Western blot analysis were employed to determine cardiomyocyte area, scar fibrosis, apoptosis and inflammation. MI insult triggered scar fibrosis, LV chamber dilation, decreased fractional shortening, ejection fraction, cardiomyocyte shortening, maximal velocity of shortening and relengthening as well as prolonged relengthening, which were abrogated or attenuated by MSCs therapy in WT but not BECN+/- mice. MI decreased intracellular Ca2+ rise and decay in response to electrical stimuli without affecting resting intracellular Ca2+, which were reconciled by MSCs in WT but not BECN+/- mice. MSCs further attenuated MI-induced mitochondrial ultrastructural injury, apoptosis, inflammation and autophagy defects in peri-infarct area in WT but not BECN+/- mice. Collectively, our results suggested that autophagy insufficiency dampened in MSCs-elicited cardioprotection associated with dampened apoptosis and inflammation.


Factors associated with low birth weight at Debre Markos Referral Hospital, Northwest Ethiopia: a hospital based cross-sectional study.
To assess the prevalence and associated factors of low birth weight among newborns delivered at Debre Markos Referral Hospital, Northwest Ethiopia. From the total of 368 newborn baby/mother pairs planned to be participated, 338 agreed and involved in the study giving a response rate of 91.2%. In this study, the prevalence of low birth weight was 21.6 (95% CI 17.5, 26%). Being rural residence (AOR 2.0, 95% CI 1.0, 4.1), duration of pregnancy (AOR = 7.6, 95% CI 3.3, 17.4), and having complications during pregnancy (AOR 2.6, 95% CI 1.2, 5.7) were found to be factors significantly associated with low birth weight.


Micromechanical oscillators as rapid biosensor for the detection of active growth of Escherichia coli.
A rapid biosensor for the detection of bacterial growth was developed using micromechanical oscillators coated by common nutritive layers. The change in resonance frequency as a function of the increasing mass on a cantilever array forms the basis of the detection scheme. The sensor is able to detect active growth of Escherichia coli cells within 1 h which is significantly faster than any conventional plating method which requires at least 24 h. The growth of E. coli was confirmed by scanning electron microscopy. This new sensing method for the detection of active bacterial growth allows future applications in, e.g., rapid antibiotic susceptibility testing by adding antibiotics to the nutritive layer.


Non-Invasive Local Acoustic Therapy Ameliorates Diabetic Heart Fibrosis by Suppressing ACE-Mediated Oxidative Stress and Inflammation in Cardiac Fibroblasts.
The extent of myocardial fibrosis is closely related to the prognosis of diabetic cardiomyopathy (DCM). Low-intensity pulsed ultrasound (LIPUS) has been reported to have multiple biological effects. However, the effect of LIPUS on diabetic heart fibrosis remains unclear. The present study aimed to investigate the effect of LIPUS on diabetic heart fibrosis and explore its underlying mechanisms. High glucose (HG) was applied to cultured neonatal rat cardiac fibroblasts (NRCFs) to mimic the in vivo hyperglycemia microenvironment. LIPUS (19.30 mW/cm2 to 77.20 mW/cm2) dose-dependently inhibited HG-induced fibrotic response in NRCFs. Also, LIPUS downregulated NADPH oxidase 4 (NOX4)-associated oxidative stress and nod-like receptor protein-3 (NLRP3) inflammasome activation in NRCFs. In vivo, diabetes in mice was induced with streptozotocin (STZ). Mice in the LIPUS group and STZ + LIPUS group were treated with LIPUS (77.20 mW/cm2) twice a week for 12 weeks and then euthanized at 12 weeks or 24 weeks post-diabetes. Treatment with LIPUS significantly ameliorated the progression of cardiac fibrosis (Masson staining 6.5 ± 2.3% vs. 2.8 ± 1.5%, P < 0.001) and dysfunction (E/A ratio 1.35 ± 0.14 vs. 1.59 ± 0.11, P < 0.05), as well as NOX4-associated oxidative stress (relative expression fold of NOX4 1.43 ± 0.12 vs. 1.07 ± 0.10, P < 0.01; relative DHE fluorescence 1.51 ± 0.13 vs. 1.28 ± 0.06, P < 0.05) and NLRP3 inflammasome activation (relative expression fold of NLRP3 1.57 ± 0.12 vs. 1.05 ± 0.16, P < 0.01), at 12 weeks post-diabetes. At 24 weeks post-diabetes, the heart function in diabetic mice treated with LIPUS was still significantly better than untreated diabetic mice (E/A ratio 1.08 ± 0.12 vs. 1.49 ± 0.14, P < 0.001). Further exploration revealed that LIPUS significantly attenuated the upregulated angiotensin-converting enzyme (ACE) and angiotensin II (AngII), in both HG-induced NRCFs and diabetic hearts (relative expression of ACE in myocardium 3.77 ± 0.55 vs. 1.07 ± 0.13, P < 0.001; AngII in myocardium 115.5 ± 21.77 ng/ml vs. 84.28 ± 9.03 ng/ml, P < 0.01). Captopril, an ACE inhibitor, inhibited NOX4-associated oxidative stress and NLRP3 inflammasome activation in both HG-induced NRCFs and diabetic hearts. Our results indicate that non-invasive local LIPUS therapy attenuated heart fibrosis and dysfunction in diabetic mice and the effect could be largely preserved at least 12 weeks after suspending LIPUS stimulation. LIPUS ameliorated diabetic heart fibrosis by inhibiting ACE-mediated NOX4-associated oxidative stress and NLRP3 inflammasome activation in cardiac fibroblasts. Our study may provide a novel therapeutic approach to hamper the progression of diabetic heart fibrosis.


Adriamycin effects on hydroperoxide metabolism and growth of human breast tumor cells.
Human breast tumor cells MCF-7 were grown during 5 days in the presence of Adriamycin and the IC50 was 50 nM with the highest sublethal concentration 0.1 microM. At this latter concentration Adriamycin produced a complete inhibition of cell division and a partial reversion to a normal breast epithelial appearance. Similar effects of Adriamycin were observed in cells cultured in the presence of 10% FBS and in a chemically defined medium, with Se-glutathione peroxidase activities of 3.8 and 1.3 U/mg of protein, respectively. Cell size and cell oxygen uptake were increased by 41% and by 50%, respectively, in Adriamycin-treated cells. The spontaneous chemiluminescence of monolayers of intact MCF-7 cells (81 +/- 9 cps/mg protein) was increased by 48% in the Adriamycin-treated cultures (120 +/- 11 cps/mg of protein) in agreement with a 91% higher concentration of malondialdehyde in the same cultures. Adriamycin treatment produced a 71% increase in the steady state concentration of H2O2, which was estimated assuming diffusion equilibrium with the external medium, from 1.38 microM in the control cells to 2.38 microM in the treated cells. Cyanide-insensitive respiration was also higher in the cells exposed to the drug than in the control cells. Adriamycin did not affect the activity of the antioxidant enzymes, Cu-Zn and Mn-superoxide dismutase, Se and non-Se-glutathione peroxidase, and catalase. These results contribute to the current hypothesis that oxygen free radicals produced by Adriamycin redox cycling are responsible for at least part of the cytotoxic effects due to this drug.


Impurity profiling quality control testing of synthetic peptides using liquid chromatography-photodiode array-fluorescence and liquid chromatography-electrospray ionization-mass spectrometry: the obestatin case.
Following several conflicting publications, the inability to reproduce the original findings on in vitro obestatin binding and activation of GPR39 receptors was recently reported by its discoverers, and several hypotheses to rationalize these findings were proposed. Based on one of these postulations (i.e., presence of impurities), peptide identity and impurity profiles were thoroughly evaluated on obestatin peptides obtained from five different manufacturers, as used by the different research groups. We found that one of the products examined was in reality a totally different peptide and that the quality of two-thirds of the other peptides was insufficient for in vitro and in vivo experiments (i.e., peptide purity less than 95% and/or individual impurities exceeding 1%). These observations question the divergent conclusions reported in the literature about the activity of obestatin. Therefore, we strongly recommend appropriate quality control testing before using any peptides for biomedical research purposes.


Acute and repeated administration of cocaine differentially regulates expression of PSA-NCAM-positive neurons in the rat hippocampus.
Recent data indicating that addictive substances are able to alter brain plasticity and its morphology inclined us to determine whether acute and chronic cocaine administration could modify the expression of a polysialylated form of the neuronal cell adhesion molecule (PSA-NCAM) in the dentate gyrus of the rat hippocampus. Alterations in the PSA-NCAM expression are known to effect a variety of neuroanatomical rearrangements in the brain structure. Cocaine was administered acutely (15 mg/kg, i.p.) or repeatedly (15 mg/kg, i.p. once a day for five consecutive days). The number of PSA-NCAM immunopositive cells was determined at six time points after cocaine treatment: 6 h and 1, 2, 4, 6, and 10 days (both in acute and repeated treatment). It was found that a single injection of cocaine induced a time-dependent decrease in the number of PSA-NCAM cells in the dentate gyrus. The decrease was observed on day 1 after cocaine treatment and lasted for at least 6 days. In contrast, an increase in the number of PSA-NCAM-positive cells in the dentate gyrus was observed 2 and 4 days after the last dose of repeated cocaine. It is concluded that cocaine can evoke long-lasting changes in the PSA-NCAM protein expression in the dentate gyrus and that the direction of cocaine-induced PSA-NCAM changes depends on the regimen of cocaine administration. It is postulated that cocaine may have impact on hippocampal plasticity and subsequent processes that are controlled by plastic changes in the hippocampal structure.


Developing and delivering high quality surgery into community in LMIC: Experience in Kerala, India.
The road to the delivery of high-quality cleft surgery into communities in the LMIC is beset with numerous hurdles. These include the availability of trained health care personnel, the availability of infrastructure in health care, parent awareness of the need for timely interventions in these children, presence of affordable treatment centres with adequate means of transport to and from these facilities for patients coming from the interior rural locations, but most importantly, the financial considerations that are the underlying limitations in most of these above mentioned considerations. We would like to approach this problem by describing the evolution of our own centre over several decades. Our centre, The Charles Pinto Centre for Cleft lip and palate at the Jubilee Mission Hospital is located in Thrissur, in the beautiful state of Kerala. In the late 1950s, Dr HS Adenwalla who had trained under experts in the art of Cleft Lip and Palate Surgery in Mumbai joined the hospital and transformed it through sheer dedication and meticulous clinical expertise into a major hub for treatment and also training in this speciality. In this endeavour he was greatly helped by non governmental organizations of which Smile Train deserves special mention in view of its model of functioning, empowering local partners by adequate funding for treatment, training and establishment of infrastructure, while ensuring strict adherence to safety standards. We believe that it is essential that there should be trained and dedicated health care personnel who are willing to treat such patients without an eye on the monetary benefits accrued, and also enough funding either from the government or from the NGOs to provide equipment and infrastructure for safe, high quality and accessible treatment and the establishment of multidisciplinary management. Lastly, the patients should also be made aware of the need for timely treatment and long-term follow- up.


Air quality impacts in the vicinity of a chemical herder mediated in-situ burn for Arctic oil spill response.
Chemical herder augmented in-situ burning (ISB) is one of the most viable oil spill response techniques in partially ice-covered waters. Herein we report on the impacts of herder-mediated ISB tests on ambient air quality by making atmospheric measurements during ISB field tests in partially ice-covered waters in Fairbanks, Alaska. Concentrations of PM2.5, six combustion gases (CO, CO2, NO, NO2, NOx, SO2), volatile organic compounds (VOCs), and herding agent (OP-40) in the airborne plume (6-12 m downwind) were measured during three ISB events. PM2.5 concentrations significantly (p-value = 0.8014) exceeded NAAQS (24 h) exposure limits, while the remaining pollutants were found significantly (p-value <0.05) below the established exposure limits. OP-40 herder was not detected in the collected aerosol samples. To our knowledge, this is the first study on atmospheric emissions in the vicinity of a field-scale herder-augmented oil-spill ISB study in a high-latitude Arctic environment and provides information that is helpful to ensure the safety and well-being of on-site response personnel.


Laparoscopic right donor nephrectomy: technique and comparison with left nephrectomy.
Laparoscopic donor nephrectomy (LDN) preferentially involves the left kidney to optimize vessel length, but occasionally, right nephrectomy is preferred. Right LDN differs markedly in anatomic relations and the need for a fourth port. This retrospective study compares donor outcomes and graft function of right and left LDN and describes the technique. Consecutive patients undergoing right LDN from March 26, 1996 to December 31, 2000 were compared with those undergoing left LDN. Age, height, weight, body mass index, creatinine, creatinine clearance, operative time, warm ischemia time, analgesic requirements, serial postoperative creatinine, time to diet resumption, and hospital stay were compared. A second cohort matched for age, gender, race, and temporal left LDN also were compared with the group undergoing right LDN. No significant differences were found for any of the parameters measured. This study demonstrates that despite substantial differences in the procedures, donor outcome and graft survival are similar for right and left LDN.


Population pharmacokinetics of ciclosporin in Chinese children with aplastic anemia: effects of weight, renal function and stanozolol administration.
To develop a population pharmacokinetic model for the immunosuppressant ciclosporin in Chinese children with aplastic anemia and to identify covariates influencing ciclosporin pharmacokinetics. A total of 102 children with either acquired or congenital aplastic anemia aged 8.8±3.6 years (range 0.9-17.6 years) were included. Therapeutic drug monitoring (TDM) data for ciclosporin were collected. The population pharmacokinetic model of ciclosporin was described using the nonlinear mixed-effects modeling (NONMEM) VI software. The final model was validated using bootstrap and normalized prediction distribution errors. A one-compartment model with first-order absorption and elimination was developed. The estimated CL/F was 15.1, which was lower than those of children receiving stem cell or kidney transplant reported in the West (16.9-29.3). The weight normalized CL/F was 0.45 (range: 0.27-0.70) Lh(-1)·kg(-1). The covariate analysis identified body weight, serum creatinine and concomitant administration of the anabolic steroid stanozolol as individual factors influencing the CL/F of ciclosporin. Our model could be used to optimize the ciclosporin dosing regimen in Chinese children with aplastic anemia.


Flagellar rotational features of an optically confined bacterium at high frequency and temporal resolution reveal the microorganism's response to changes in the fluid environment.
Rotations of the flagella control the movement of a peritrichous (multiflagellar) bacterium in fluids, the run and tumble events being caused through modulations in the flagella's collective rotation speed and pattern. Observing such modulations is a challenge in free swimming bacteria. In this work, we present a setup to measure the collective flagellar rotational features of an optically confined Bacillus subtilis bacterium. We adopt a Continuous Wavelet Technique (CWT) while monitoring the rotational patterns in frequency and time, thus achieving optimal resolution in both the domains. This enables in marking the events wherein subtle changes in the flagellar rotational pattern occur. These studies unravel a fact, hitherto unknown, that variations in swimming speed that are seen in pure run sequences are also caused by modulations in the rotating flagella. Further, we have monitored the flagellar rotation for durations over a minute and observe a gradual slowing down of the rotation before ceasing completely due to the trapping laser induced photodamage. We have observed a significant alteration in the rate of rotational fall off in real time with changes in pH or the nutrient concentration in the fluid. This work serves to demonstrate the advantage of optical confinement of a bacterium in its pristine form for carrying out such studies and can serve as a marker for work that assesses membrane photodamage in active matter. Details on the role of flagella in propulsion and on other factors influencing the rotations, can be of significance in the design of artificial microswimmers.


Risk factors for venous thrombosis in Swedish children and adolescents.
To identify prothrombotic risk profiles in children and adolescents referred to a regional coagulation centre in southern Sweden for a first thrombotic event. One hundred and twenty-eight consecutive children and adolescents (newborn to 20 y) referred for evaluations of a first episode of venous thrombosis were investigated. Clinical data were collected retrospectively, and the following variables were investigated: protein C, protein S, antithrombin; resistance to activated protein C; the genotypes FV-G1691A, F II-G20210A, MTHFR-C677T, MTHFR- A1298C; coagulation factors VIII and XI. 104/128 subjects (81%) had identifiable acquired risk factors, most often indwelling catheters and hormone therapy. Predisposing genetic factors related to thromboembolic events were revealed in 53/83 (64%) of subjects who agreed to follow-up blood sampling, and 17/83 (20%) had two or more inherited risk factors. Combinations of genetic and acquired risk factors were identified in 45/83 (54%) of the subjects, and 77/83 (93%) had at least one such risk factor. Both sexes had one peak in frequency at less than 1 y of age and then an increase during adolescence, more in females than in males. Plasma values for coagulation factors VIII and XI were age appropriate and showed a normal Gaussian distribution. This study identified prothrombotic risk profiles in almost all children and adolescents with venous thrombosis, which underlines the importance of careful evaluation of genetic and acquired risk factors.


Role of radiotherapy in the management of bladder cancer: Recommendations of the French society for radiation oncology.
We present the recommendations of the French society of oncological radiotherapy on the indications and techniques for external beam radiotherapy for bladder cancer.


[A study of fluoride and dexamethasone affecting the expression of growth factors in osteoblast].
Investigate the expression of growth factors or receptors in osteoblast, to understand the possible mechanism of low dose fluoride and dexamethasone on the development and differentiation of osteoblast. Investigate the expression of platelet derived growth factors (PDGF-A) and insulin-like growth factors (IGF-I-Ralpha) in osteoblast cultured in vitro by immunohistochemistry, furthermore, to deal with the results by image analysis. There were no significant effects of PDGF-A both of low doses fluoride and dexamethasone treated on the MC3T3-E1 osteoblast during the various stages of the development. Otherwise, the expression of IGF-I-Ralpha might increase treated with them at the early stage of the cell culture, and dexamethasone did more significantly. Low doses of fluoride or dexamethasone may promote the expression of IGF-I-Ralpha in osteoblast precursors, which maybe contribute to the effects of fluoride and dexamethasone on the proliferation and differentiation of osteoblast.


Seasonal and spatial trends in particle number concentrations and size distributions at the children's health study sites in Southern California.
Continuous measurements of particle number (PN), particle mass (PM(10)) and gaseous copollutants (NO(x), CO and O3) were obtained at eight sites (urban, suburban and remote) in Southern California during years 2002 and 2003 in support of University of Southern California Children's Health Study. We report the spatial and temporal variation of PNs and size distributions within these sites. Higher average total PN concentrations are found in winter (November to February), compared to summer (July to September) and spring (March to June) in all urban sites. Contribution of local vehicular emissions is most evident in cooler months, whereas effects of long-range transport of particles are enhanced during warmer periods. The particle size profile is most represented by a combination of the spatial effects, for example, sources, atmospheric processes and meteorological conditions prevalent at each location. Afternoon periods in the warmer months are characterized by elevated number concentrations that either coincide or follow a peak in ozone concentrations, suggesting the formation of new particles by photochemistry. Results show no meaningful correlation between PN and mass, indicating that mass based standards may not be effective in controlling ultrafine particles. The study of the impact of the Union worker's strike at port of Long Beach in October 2002 revealed statistically significant increase in PN concentrations in the 60-200 nm range (P<0.001), which are indicative of contributions of emissions from the idling ships at the port.


Immuno-pharmacological characterization of group II metabotropic glutamate receptors controlling glutamate exocytosis in mouse cortex and spinal cord.
We recently proposed the existence of mGlu3 -preferring autoreceptors in spinal cord terminals and of mGlu2 -preferring autoreceptors in cortical terminals. This study aims to verify our previous conclusions and to extend their pharmacological characterization. We studied the effect of LY566332, an mGlu2 receptor positive allosteric modulator (PAM), and of LY2389575, a selective mGlu3 receptor negative allosteric (NAM) modulator, on the mGlu2/3 agonist LY379268-mediated inhibition of glutamate exocytosis [measured as KCl-evoked release of preloaded [3 H]-D-aspartate]. The mGlu2 PAM BINA and the mGlu3 NAM ML337, as well as selective antibodies recognizing the N-terminal of the receptor proteins, were used to confirm the pharmacological characterization of the native receptors. Cortical synaptosomes possess LY566332-sensitive autoreceptors that are slightly, although significantly, susceptible to LY2389575. In contrast, LY566332-insensitive and LY2389575-sensitive autoreceptors are present in spinal cord terminals. BINA and ML337 mimicked LY566332 and LY2389575, respectively, in controlling LY379268-mediated inhibition of glutamate exocytosis from both cortical and spinal cord synaptosomes. Incubation of cortical synaptosomes with anti-mGlu2 antibody prevented the LY379268-induced inhibition of glutamate exocytosis, and this response was partially reduced by the anti-mGlu3 antibody. Incubation of spinal cord synaptosomes with the anti-mGlu3 antibody abolished LY379268-mediated reduction of glutamate exocytosis from these terminals, while the anti-mGlu2 antibody was inactive. Western blot analysis and confocal microscopy data were largely consistent with these functional observations. We confirmed that mGlu3 -preferring autoreceptors exist in spinal cord terminals. Differently, cortical glutamatergic terminals possess mGlu2 /mGlu3 heterodimers, whose inhibitory effect is largely mediated by mGlu2 receptors.


FcγRIIB: a modulator of cell activation and humoral tolerance.
An immune response needs to be tightly regulated to prevent excessive inflammation, which may result in the destruction of healthy tissues. At the molecular level, the strength of an immune response is determined by the integration of a multitude of positive and negative signals. This review will focus on IgG-dependent immune responses and discuss how the inhibitory receptor FcγRIIB may be involved in regulating both the afferent and efferent phases of such a response. Furthermore, we will discuss recent evidence suggesting that FcγRIIB may have important functions beyond the negative regulation of signals transduced by the B-cell receptor or activating FcγRs and could be responsible for the activity of agonistic antibodies in vivo.


Organization of the terminal two enzymes of the heme biosynthetic pathway. Orientation of protoporphyrinogen oxidase and evidence for a membrane complex.
Protoporhyrinogen oxidase (EC 126.96.36.199), the penultimate enzyme of the heme biosynthetic pathway, catalyzes the removal of six hydrogens from protoporphyrinogen IX to form protoporphyrin IX. The enzyme in eukaryotes is associated with the inner mitochondrial membrane. In the present study we have examined requirements for solubilization of this enzyme and find that it behaves as an intrinsic membrane protein that is solubilized only with detergents such as sodium cholate. The in situ orientation of the enzyme with respect to the inner mitochondrial membrane places the active site on the cytosolic face of this membrane rather than the matrix side where the active site of ferrochelatase, the terminal pathway enzyme, is located. Examination of the kinetics of the two terminal enzymes in mitochondrial membranes demonstrates that substrate channeling occurs between these terminal two-pathway enzymes. However, examination of solubilized and membrane-reconstituted enzymes shows no evidence for a stable complex. Based upon these and previous data a model for the terminal three-pathway enzymes is presented.