Bacterial Communities Associated with Four Cyanobacterial Genera Display Structural and Functional Differences: Evidence from an Experimental Approach.
To overcome the limitations associated with studying the interactions between bacterial communities (BCs) and cyanobacteria in natural environments, we compared the structural and functional diversities of the BCs associated with 15 non-axenic cyanobacterial strains in culture and two natural BCs sampled during cyanobacterial blooms. No significant differences in richness and diversity were found between the natural and cultivated BCs, although some of the cyanobacterial strains had been isolated 11 years earlier. Moreover, these BCs shared some similar characteristics, such as a very low abundance of Actinobacteria, but they display significant differences at the operational taxonomic unit (OTU) level. Overall, our findings suggest that BCs associated with cyanobacteria in culture are good models to better understand the interactions between heterotrophic bacteria and cyanobacteria. Additionally, BCs associated with heterocystous cyanobacterial strains cultivated in Z8X culture medium without nitrate (Aphanizomenon-Dolichospermum) demonstrated significant differences compared to BCs associated with non-heterocystous strains cultivated in Z8 culture medium (Planktothrix-Microcystis) in terms of their composition and their ability to utilize different carbon sources, suggesting the potential influence of cyanobacterial metabolism and/or culture media on associated BCs. Finally, half of the dominant OTUs in these BCs were specifically associated with cyanobacteria or other phytoplankton, whereas the remaining OTUs were generally associated with ecosystems containing high organic matter content, such as sludge or intestines.


Gender difference in long-term prognosis among patients with cardiovascular disease.
Differences in prognosis between women and men with atherosclerosis are currently under attention. Previous studies describe contradictory results and are restricted to one cardiovascular bed, while atherosclerosis is a systemic disease. We therefore studied the prognosis of women versus men in the SMART study, a large cohort of patients with clinically manifest atherosclerosis with extensive baseline and follow-up information. 5349 patients (1347 women, 4002 men) with at least one type of atherosclerotic vascular disease (coronary artery disease, cerebrovascular disease, peripheral arterial disease or abdominal aortic aneurysm) were included in the SMART study, an ongoing long-term follow-up study. They all received a standardized cardiovascular work-up with a personalized therapy advice. All future cardiovascular events were collected prospectively. All-cause mortality, cardiovascular mortality and cardiovascular outcome (composite of myocardial infarction, stroke and cardiovascular death) were evaluated using Cox regression and expressed as hazard ratios (HR) with 95% confidence intervals (95%CI). Men served as the reference category. Different models were used to adjust for differences in baseline characteristics. Women and men had a mean age of 60 years and their median follow-up (range) was five years (13.5). The hazard ratios of all-cause mortality, cardiovascular death and cardiovascular outcome were 0.62 (95%CI: 0.51-0.75), 0.59 (95%CI: 0.46-0.75) and 0.73 (95%CI: 0.60-0.87). Neither differences in risk-factor profile nor the different vascular beds involved could explain this advantage. Women with cardiovascular disease who received a similar standardized cardiovascular work-up and personalized therapy advice as men had a favourable long-term outcome.


The Impact of Protective Gloves on Manual Dexterity in Cold Environments-A Pilot Study.
Our research aimed to determine the impact of two types of protective gloves. The research tested the glove performance on men exposed to a range of temperatures reflecting the working conditions in fruit and vegetable processing. The gloves were assessed for performance within the time required to complete a specific manual task and for performance relative to the subjective thermal sensations in the male subjects. Six males participated in a total of 3 study variants: at +5 °C (with double gloves and single glove), at -1 °C (with double gloves and single glove) and in reference conditions +20 °C (without gloves), in which they performed manual tasks. The measurement of manual task performance time was used to assess manual dexterity. Subjective thermal sensations were determined. Differences in the time required to complete specific tasks were observed between the variants with gloves (both at a temperature of +5 °C and -1 °C), and without gloves (p < 0.05). The type of protective gloves had an impact on the time needed to complete manual tasks and therefore may affect manual dexterity.


Atmospheric polycyclic aromatic hydrocarbons in remote European and Atlantic sites located above the boundary mixing layer.
Ambient air concentrations of polycyclic aromatic hydrocarbons (PAH) were determined at five elevated mountain sites on the European continent and the Atlantic Ocean. All sites can be considered remote background areas since they are situated above the timberline and they lack local emission sources of these compounds. Average gas phase concentrations of SigmaPAH were 165, 1,475, 1,553, 1,822 and 4,443 pg m(-3) for Tenerife, Pyrenees, Central Norway, Tyrolean Alps and High Tatras, respectively. Particulate phase concentrations were 55, 70, 383, 196 and 708 pg m(-3), respectively. The PAH profiles of samples from the different sites are very similar, being typical of PAH mixtures after long-range atmospheric transport. Part of the fluctuations in PAH concentrations are explained by the influence of temperature on the particulate/gas phase partitioning. The differences in PAH levels between sites, with the lowest concentrations found in Tenerife and the highest in the High Tatras, suggest the geographical influence of regional emissions on the sites, especially in the cold periods and for the sites in the eastern sector of the European continent. This is supported by air mass back-trajectories analysis for the samples on the different sites. The influence of the continent is not detectable in the case of the elevated site of Tenerife where the free troposphere has been sampled. The results in this study are consistent with the PAH levels found in soils and/or high mountain lake sediments from these areas.


Penile prosthesis implantation: A single center 25 years of experience.
We analyzed the long-term results of malleable and inflatable penile prosthesis: the rates of complications, revision surgery and patient satisfaction. Between January 1996 and December 2020, a total of 138 patients underwent implantation of a malleable or inflatable types penile prosthesis. The data of the patients were derived from the patient files and digital recording system. The patients were contacted face-to-face interaction if possible. If it is not possible, phone connection was made. The median age of patients was 56 (21-83) years. Fourteen (10.2%) 1-piece, 46 (33.3%) 2-piece and 78 (56.5%) 3-piece penile prosthesis were implanted. The mostly used perioperative antibiotic was teicoplanin+amikacin in 107 patients. Total complication rate was 30% (n=42). Prosthesis infection constituted 9% (n=13) of these complications. The total satisfaction rate was 89.1%. There was no correlation between the complications and antibiotic regimen or prosthesis type (P=0.488, P=0.454, respectively). The rate of infection showed a slight increase in 3-piece penile prosthesis without any statistically significance (P=0.633). The patients with complications expressed more dissatisfaction compared to those without complications (P=0.001). In our series, penile prosthesis implantation seems to be a recommended treatment method with high patient satisfaction in the treatment of refractory erectile dysfunction. 4.


Expression of root glutamate dehydrogenase genes in tobacco plants subjected to boron deprivation.
Recently it has been reported that boron (B) deficiency increases the expression of Nicotiana tabacum asparagine synthetase (AS) gene in roots, and that AS might play a main role as a detoxifying mechanism to convert ammonium into asparagine. Interestingly, glutamate dehydrogenase (GDH) genes, Ntgdh-NAD;A1 and Ntgdh-NAD;B2, were up-regulated when tobacco roots were subjected to B deprivation for 8 and 24 h. In addition, aminating and deaminating GDH (EC 184.108.40.206) activities were higher in B-deficient than in B-sufficient plants after 24 h of B deficiency. Ammonium concentrations were kept sufficiently low and with similar values in B-deficient roots when compared to control. Glucose and fructose contents decreased after 24 h of B deprivation. This drop in hexoses, which was corroborated by metabolomic analysis, correlated with higher GDH gene expression. Furthermore, metabolomic profiling showed that concentrations of several organic acids, phenolics, and amino acids increased after 24 h of B deficiency. Our results suggest that GDH enzyme plays an important role in metabolic acclimation of tobacco roots to B deprivation. A putative model to explain these results is proposed and discussed.


Micropapillary ductal carcinoma in situ of the breast: an inter-institutional study.
The clinical significance of micropapillary growth pattern in ductal carcinoma in situ is controversial and the impact of nuclear grading in terms of recurrence of this lesion is yet to be clarified. Our aim was to evaluate, on a series of micropapillary in situ carcinomas, the histological features correlated with recurrence and whether the micropapillary subtype had a different behavior from other non-micropapillary ductal carcinoma in situ. We collected 55 cases of micropapillary in situ carcinomas from four institutions. All cases were reviewed for nuclear grade, extent, necrosis, microinvasion and tested for estrogen and progesterone receptors, Ki67, HER2, EGFR and p53 expression. Clinical data, type of surgery and follow up were obtained for all patients. Our results showed that the nuclear grade is crucial in determining the biology of micropapillary carcinoma in situ, so that the high nuclear grade micropapillary ductal carcinoma in situ more frequently overexpressed HER2, showed higher proliferation index, displayed necrosis and microinvasion and was more extensive than low/intermediate nuclear grade. Logistic regression analysis confirmed the high nuclear grade (Odds ratio: 6.86; CI: 1.40-33.57) as the only parameter associated with elevated risk of local recurrence after breast-conserving surgery. However, the recurrence rate of 19 micropapillary carcinoma in situ, which were part of a cohort of 338 consecutive ductal carcinoma in situ, was significantly higher (log-rank test, P-value=0.019) than that of non-micropapillary, independently of the nuclear grade. In conclusion, although nuclear grade may significantly influence the biological behavior of micropapillary ductal carcinoma in situ, micropapillary growth pattern per se represents a risk factor for local recurrence after breast-conserving surgery.


Increased urinary N-telopeptide cross-linked type 1 collagen predicts bone loss in patients with inflammatory bowel disease.
Reduced bone mineral density (BMD) is common in patients with inflammatory bowel disease (IBD), but the factors associated with its longitudinal rate of change have not been established. We prospectively assessed the rate of change in BMD, and its association with biochemical markers of bone turnover. Twenty-two patients with Crohn's disease and 14 ulcerative colitis patients age 37.1 +/- 11.6 yr were followed for 2 yr. Lumbar spine (L2-L4) and femoral neck BMD were measured by dual x-ray absorptiometry at baseline and 24 months. Bone-specific alkaline phosphatase, osteocalcin, urinary N-telopeptide crosslinked type 1 collagen (NTx), parathyroid hormone, and 25-hydroxyvitamin-D were determined at baseline. At baseline, 59% of Crohn's patients and 43% of ulcerative colitis patients were osteoporotic, with spine or femoral neck BMD T-score < -2.5. Spine BMD, and spine and femoral neck T-scores were lower and disease duration was longer in nine patients with ileal resection compared with nonoperated patients (0.84 +/- 0.15 g/cm2 vs 0.96 +/- 0.11 g/cm2, -3.0 +/- 1.5 vs -1.7 +/- 1.3, -3.2 +/- 1.5 vs -2.2 +/- 1.0, respectively; all p < 0.05). At 24 months, 13/36 (36%) and 14/36 (39%) patients experienced spinal and femoral neck bone loss, respectively, with mean annual percent BMD changes of -2.0% and -1.5%, respectively. NTx, a bone resorption marker, inversely correlated with spinal BMD rate of change (r = -0.4, p < 0.05). Using quartiles analysis, patients with the highest NTx (Q4) experienced the greatest decrease in spine BMD compared with patients with the lowest NTx (Q1). Spine and femoral neck bone loss continues over time in more than one-third of IBD patients. Increased NTx level predicts spinal bone loss in IBD patients.


Electrochemical detection of heavy metal ions in water.
Heavy metal ions are one of the main sources of water pollution. Most heavy metal ions are carcinogens that pose a threat to both ecological balance and human health. With the increasing demand for heavy metal detection, electrochemical detection is favorable due to its high sensitivity and efficiency. Here, after discussing the pollution sources and toxicities of Hg(ii), Cd(ii), As(iii), Pb(ii), UO2(ii), Tl(i), Cr(vi), Ag(i), and Cu(ii), we review a variety of recent electrochemical methods for detecting heavy metal ions. Compared with traditional methods, electrochemical methods are portable, fast, and cost-effective, and they can be adapted to various on-site inspection sites. Our review shows that the electrochemical detection of heavy metal ions is a very promising strategy that has attracted widespread attention and can be applied in agriculture, life science, clinical diagnosis, and analysis.


Detection of angiotensin-converting enzyme gene insertion/deletion polymorphism from paraffin-embedded tissues: the Hisayama study.
Previous studies have suggested that archival materials from formalin-fixed paraffin-embedded blocks are unsuitable for most molecular techniques because the extracted DNA can be severely degraded. Therefore, the present study was designed to investigate the accuracy of genotyping for the insertion (I)/deletion (D) polymorphism of the angiotensin-converting enzyme (ACE) gene from paraffin-embedded tissues of autopsy cases from Hisayama Town, Japan. The genotype was determined using the double polymerase chain reaction method and to test the accuracy of the method, the polymorphism was investigated using paraffin-embedded tissues from 18 cases whose ACE genotypes (6 cases for each genotype) were known in advance from analysis of fresh-frozen tissue samples. Genotyping using paraffin-embedded tissues was then determined for 968 autopsy subjects. The genotype could be determined in 16 of the 18 test samples (88.9%) and there was no discrepancy with the results obtained from the fresh-frozen tissues. Of the 968 autopsy cases, the frequency of the DD, ID, and II genotypes was 12.4%, 47.3%, and 40.3%, respectively, a distribution that did not deviate from the Hardy-Weinberg equilibrium (chi(22df )= 0.67, p=0.72). These findings suggest the accuracy of the present method of ACE genotyping from paraffin-embedded tissues.


A systematic approach to the assessment of erythropoiesis.
The pathogenesis of anaemia may be simple or complex and the differential diagnosis can be difficult. An appreciation of the erythropoietic processes is required, together with regular review of investigations, to ensure that appropriate protocols are adopted. The application of tests, which define different facets of erythropoiesis, should be appropriate to the clinical circumstances. In some situations, such as the anaemia of chronic disorders, pregnancy and chronic renal failure, a detailed analysis of erythropoiesis is often required. Guidelines for investigating anaemia due to megaloblastosis or haemoglobinopathy are well established, whereas disturbances of iron metabolism are often difficult to classify. These require a clear distinction between storage and functional iron to differentiate whether the defect is due to readily treatable simple iron deficiency or more complex mechanisms, which do not respond to iron supplementation. Determination of red cell haemoglobin content, reticulocyte analysis and the assay of serum transferrin receptors are new generation parameters developed to address this. Practice pressures and new treatment options have contributed to investigations becoming more complex, especially those of the secondary anaemias, as new tests have become more readily available and often automated. This has resulted in reduced turnaround times and clinical demand has driven request patterns. Initiatives to develop evidence-based anaemia management protocols are welcomed but, wherever possible, should be developed through collaboration between the haematology department and the user unit, and based on available guidelines.


Dyslexics' eye fixations may accommodate to hemispheric desynchronization.
We argue that the vertical division of the fovea and the hemispheric division of the brain condition reading, and that hemispheric desynchronization is a proximal cause of dyslexia. We predict that dyslexics' fixation behaviour in reading accommodates to problematic hemispheric transfer/coordination, with fixations projecting more letter-information directly to the left hemisphere to facilitate processing. We analysed eye movements of 24 dyslexics and 24 normal readers. Dyslexics fixated closer to word beginning than did normal readers, projecting more of the word directly to the left hemisphere. Both groups produced comparable fixation durations at the beginning of the word; further into the word the dyslexics produced longer fixation durations. The results support a model of dyslexia based on hemispheric desynchronization.


Implications of the mitochondrial interactome of mammalian thioredoxin 2 for normal cellular function and disease.
Multiple thioredoxin isoforms exist in all living cells. To explore the possible functions of mammalian mitochondrial thioredoxin 2 (Trx2), an interactome of mouse Trx2 was initially created using (i) a monothiol mouse Trx2 species for capturing protein partners from different organs and (ii) yeast two hybrid screens on human liver and rat brain cDNA libraries. The resulting interactome consisted of 195 proteins (Trx2 included) plus the mitochondrial 16S RNA. 48 of these proteins were classified as mitochondrial (MitoCarta2.0 human inventory). In a second step, the mouse interactome was combined with the current four-membered mitochondrial sub-network of human Trx2 (BioGRID) to give a 53-membered human Trx2 mitochondrial interactome (52 interactor proteins plus the mitochondrial 16S RNA). Although thioredoxins are thiol-employing disulfide oxidoreductases, approximately half of the detected interactions were not due to covalent disulfide bonds. This finding reinstates the extended role of thioredoxins as moderators of protein function by specific non-covalent, protein-protein interactions. Analysis of the mitochondrial interactome suggested that human Trx2 was involved potentially in mitochondrial integrity, formation of iron sulfur clusters, detoxification of aldehydes, mitoribosome assembly and protein synthesis, protein folding, ADP ribosylation, amino acid and lipid metabolism, glycolysis, the TCA cycle and the electron transport chain. The oxidoreductase functions of Trx2 were verified by its detected interactions with mitochondrial peroxiredoxins and methionine sulfoxide reductase. Parkinson's disease, triosephosphate isomerase deficiency, combined oxidative phosphorylation deficiency, and lactate dehydrogenase b deficiency are some of the diseases where the proposed mitochondrial network of Trx2 may be implicated.


Responding to, and learning from, peer review feedback.
What is peer review in research? Peer review is the process of assessing the scientific quality of a research proposal, research report and/or paper by an independent expert, usually an academic or clinical expert.


Trends in human immunodeficiency virus infection among drug users in a detoxification unit.
In a cross-sectional study of 1111 injection drug users (IDUs) admitted to a hospital detoxification unit between 1987 and 2001, characteristics of substance abuse and blood samples were obtained at study entry. The median age at admission was 27 years, the median age at first injection of drugs was 19 years, and the median duration of injection drug use was 84 months. Overall prevalence of human immunodeficiency virus (HIV) infection was 60% (58% in men, 66% in women; P=.026). Age at start of and duration of injection drug use were associated with HIV infection (P<.001). The rate of HIV infection in persons with short duration of drug use (<24 months) was higher among women (45%) than among men (21%) (P=.002). The prevalence of HIV infection among IDUs <21 years old at first use of injection drugs and among patients reporting <48 months of injection drug use was significantly lower in those who started injection drug use after 1992 (19% vs. 53% of IDUs; odds ratio, 4.42; 95% confidence interval, 1.80-10.85). Awareness of acquired immunodeficiency syndrome, preventive measures, and substance abuse treatment have contributed to the stabilization of the HIV epidemic. The higher prevalence of HIV infection among women IDUs and among young IDUs who recently began injecting drugs emphasizes the need to target HIV-transmission risk reduction interventions early.


Progress report on new antiepileptic drugs: A summary of the Sixteenth Eilat Conference on New Antiepileptic Drugs and Devices (EILAT XVI): I. Drugs in preclinical and early clinical development.
The Eilat Conferences have provided a forum for discussion of novel treatments of epilepsy among basic and clinical scientists, clinicians, and representatives from regulatory agencies as well as from the pharmaceutical industry for 3 decades. Initially with a focus on pharmacological treatments, the Eilat Conferences now also include sessions dedicated to devices for treatment and monitoring. The Sixteenth Eilat Conference on New Antiepileptic Drugs and Devices (EILAT XVI) was held in Madrid, Spain, on May 22-25, 2022 and was attended by 157 delegates from 26 countries. As in previous Eilat Conferences, the core of EILAT XVI consisted of a sequence of sessions where compounds under development were presented and discussed. This progress report summarizes preclinical and, when available, phase 1 clinical data on five different investigational compounds in preclinical or early clinical development, namely GAO-3-02, GRT-X, NBI-921352 (formerly XEN901), OV329, and XEN496 (a pediatric granular formulation of retigabine/ezogabine). Overall, the data presented in this report illustrate novel strategies for developing antiseizure medications, including an interest in novel molecular targets, and a trend to pursue potential new treatments for rare and previously neglected severe epilepsy syndromes.


Molar crown and root size relationship in anthropoid primates.
Mandibular corpus form is thought to reflect masticatory function and the size of the dentition, but there is no universal association between crown dimensions and corpus size across anthropoids. Previous research was based on the assumption that crown size is an appropriate proxy for overall tooth size, but this hypothesis remains largely untested. This study assesses the relationship between the volume and surface area of molar crowns and roots by examining two main hypotheses: (1) crown size correlates significantly with root size, and (2) the proportion of root-to-crown surface area is related to dietary proclivity. Permanent M2s (n=58) representing 19 anthropoid species were CT scanned and the volume and surface area of the crown and root were measured. Interspecific correlation and regression analyses reveal significant isometric relationships between crown and root volume and a positive allometric relationship between root and crown surface area (i.e. as crown surface area increases, root surface area becomes disproportionately greater). Intraspecifically, crown and root surface area correlate significantly in some species where such analyses were possible. In general, hard object feeders exhibit relatively larger root surface area per unit crown surface area compared to soft and tough object feeders. The results also show that despite differences in food specialization closely related species have similar root-to-crown surface area proportions, thus indicating a strong phylogenetic influence. Since it is possible that, at least in some species, crown and root size vary independently, future studies should elucidate the relationship between tooth root size and mandible form.


Hsp70 chaperone ligands control domain association via an allosteric mechanism mediated by the interdomain linker.
Hsp70 chaperones assist in protein folding, disaggregation, and membrane translocation by binding to substrate proteins with an ATP-regulated affinity that relies on allosteric coupling between ATP-binding and substrate-binding domains. We have studied single- and two-domain versions of the E. coli Hsp70, DnaK, to explore the mechanism of interdomain communication. We show that the interdomain linker controls ATPase activity by binding to a hydrophobic cleft between subdomains IA and IIA. Furthermore, the domains of DnaK dock only when ATP binds and behave independently when ADP is bound. Major conformational changes in both domains accompany ATP-induced docking: of particular importance, some regions of the substrate-binding domain are stabilized, while those near the substrate-binding site become destabilized. Thus, the energy of ATP binding is used to form a stable interface between the nucleotide- and substrate-binding domains, which results in destabilization of regions of the latter domain and consequent weaker substrate binding.


A metastasizing composite carcinoma of the thyroid with distinct medullary and papillary components.
Thyroid tumors with differentiation of both parafollicular and follicular epithelial cells are rare. The majority of those reported have been composed of a single cell population with combined features of the two cell types. We describe a 48-year-old man with a thyroid tumor composed of two discrete cell populations: thyroglobulin-positive papillary carcinoma intermixed with calcitonin-containing medullary carcinoma. The tumor metastasized to regional lymph nodes and maintained this composite differentiation. We propose that such tumors with two discrete intermingled cell populations be called composite thyroid carcinomas, as opposed to mixed thyroid tumors, which describes tumors with a single cell population that exhibits features of two cell types. Dual differentiation in thyroid neoplasms has been interpreted as indicative of a common stem cell origin. In this case, the presence of two mature, highly differentiated elements may suggest a proliferative response of two distinct cell populations to a common tumorigenic stimulus.


Aberrant Expression of HOXA5 and HOXA9 in AML.
Aberrant expression of HOX gene expression has been observed in cancer. The purpose of this study was to investigate the alteration of HOXA5 and HOXA9 expression and their clinical significance in acute meloid leukemia (AML). The expression of HOXA5 and HOXA9 genes of bone marrow samples from 75 newly diagnosed AML patients and 22 healthy controls for comparison were examined by Real- time quantitative PCR (RQ-PCR) assay. Statistical analysis was conducted to evaluate HOXA5 and HOXA9 expression as possible biomarkers for AML. The results showed that the complete remission rate (52.6%) of the patients who highly expressed HOXA5 and HOXA9 was significantly lower than that (88.9%) in patients who lowly express the genes (P=0.015). Spearmann correlation coefficients indicated that the expression levels for HOXA5 and HOXA9 genes were highly interrelated (r=0.657, P<0.001). Meanwhile, we detected significant correlations between HOXA9 expression and age in this limited set of patients (P=0.009). The results suggest a prognostic impact of increased expression of HOXA5 and HOXA9 in AML patients.


[Coronary artery blood flow velocity non-invasively measured using a vessel-tracking pulsed Doppler system].
A newly-developed noninvasive method was used to measure left coronary blood flow during phantom experiments. Two techniques were used in which: (1) the sample position can always be set in a fluctuating vessel using a wall echo-tracking method with a phase-locked-loop, and (2) the Doppler reference signal was generated separately synchronous with the wall echo signal. These techniques were combined, using a commercially available pulsed Doppler apparatus (SSH-40B: Toshiba). Basic experiments were performed using a blood vessel phantom to verify the validity of these systems. Blood flow velocity in the fluctuating tube could be measured clearly using a vessel-tracking method. The blood flow velocity of the left anterior descending artery was measured in three normal subjects and in seven patients from the third intercostal space along the left sternal border. The velocity pattern was characterized by a crescendo-decrescendo shape in diastole. The peak velocity which appeared in diastole ranged from 19 to 69 cm/sec, with no difference by disease entity. However, in all cases, the blood flow velocity signals were marred by extraneous signals, making it impossible to measure blood flow velocity during systole. Further improvement of the system is mandatory in order to use this flowmeter clinically.


The spider procedure: a new Z-plasty-based local flap procedure.
Using neighboring similar skin, local flap procedures provide optimal aesthetic and functional results in reconstruction of skin defects. Therefore, a number of local flap procedures have been described. However, as skin defects present limitless variations in size and shape, new flap procedures are still needed. Here, we describe a new Z-plasty-based local flap technique, "the spider procedure," for closure of the skin defects. In this procedure, first, the existing defect is surgically converted to a triangle in shape. Then, using a modified 5-flap Z-plasty pattern, the flaps are outlined. By transposing the elevated flaps in a Z-plasty manner, a tension-free closure is achieved by means of tissue relaxation provided by opposing Z-plasties. Over 9 years, this technique has been used for closure of various skin defects in 42 patients (19 men and 23 women) aged 17 to 57 years. This procedure was used for facial defects in 20 patients, abdominal defects in 6, upper extremity defects in 7, and lower extremity defects in 9 other patients. The defect size ranged between 1.5 and 17 cm in diameter. A tension-free defect closure was obtained in all patients. Except 2 patients with minor flap tip necrosis who underwent secondary healing, all patients healed uneventfully and no patient required revision surgery in our series. There was no patient with dog-ear formation. A mean follow-up of 2 years (6 months-9 years) revealed an aesthetically acceptable scar formation in all patients. On the basis of our clinical results, the spider procedure seems to be a useful alternative for the closure of various skin defects in all regions of the body. Using the advantage of extra tissue relaxation provided by opposing Z-plasties, it enables a surgeon to obtain tension-free closure of considerably large skin defects without dog-ear formation.


Omental and subcutaneous adipose tissue steroid levels in obese men.
We examined plasma and fat tissue sex steroid levels in a sample of 28 men aged 24.8-62.2 years (average BMI value of 46.3 +/- 12.7 kg/m(2)). Abdominal adipose tissue biopsies were obtained during general or obesity surgery. Omental and subcutaneous adipose tissue steroid levels were measured by gas chromatography and chemical ionization mass spectrometry after appropriate extraction procedures. BMI and waist circumference were negatively correlated with plasma testosterone (r = -0.49 and -0.50, respectively, p < 0.01) and dihydrotestosterone (r = -0.58 and -0.56, respectively, p < 0.01), and positively associated with estrone levels (r = 0.64 and 0.62, respectively, p < 0.001). Regional differences in adipose tissue steroid levels were observed for dihydrotestosterone (p < 0.005), androstenedione (p < 0.0001) and dehydroepiandrosterone levels (p < 0.05), which were all significantly more concentrated in omental versus subcutaneous fat. Positive significant associations were found between circulating level of a steroid and its concentration in omental and subcutaneous adipose tissue, for estrone (r = 0.72 and 0.57, respectively, p < 0.01), testosterone (r = 0.66 and 0.58, respectively, p < 0.01) and dihydrotestosterone (r = 0.58 and 0.45, respectively, p < 0.05). Positive correlations were observed between plasma dehydroepiandrosterone-sulfate and omental (r = 0.56, p < 0.01) as well as subcutaneous adipose tissue dehydroepiandrosterone level (r = 0.38, p = 0.05). Positive significant associations were found between omental adipocyte responsiveness to positive lipolytic stimuli (isoproterenol, dibutyryl cyclic AMP and forskolin) and plasma or omental fat tissue androgen levels. In conclusion, although plasma androgen or estrogen levels are strong correlates of adipose tissue steroid content both in the omental and subcutaneous fat depots, regional differences may be observed. Androgen concentration differences in omental versus subcutaneous adipose tissue suggest a depot-specific impact of these hormones on adipocyte function and metabolism.


Real-time in vivo bioluminescence imaging of lentiviral vector-mediated gene transfer in mouse testis.
Although much research has focused on transferring exogenous genes into living mouse testis to investigate specific gene functions in spermatogenic, Sertoli, and Leydig cells, relatively little is known regarding real-time gene expression in vivo. In this study, we constructed a bicistronic lentiviral vector (LV) encoding firefly luciferase and enhanced green fluorescence protein (EGFP); this was a highly efficient in vivo gene transfer tool. After microinjecting LV into the seminiferous tubules the ICR mouse testis, we detected luciferase and EGFP expression in vivo and ex vivo in the injected tubules using bioluminescence imaging (BLI) with the IVIS-200 system and fibered confocal fluorescence microscopy (CellViZio), respectively. In addition, with an in vivo BLI system, luciferase expression in the testis was detected for approximately 3 mo. Furthermore, EGFP expression in seminiferous tubules was confirmed in excised testes via three-dimensional fluorescent imaging with a confocal laser-scanning microscope. With immunostaining, EGFP expression was confirmed in several male germ cell types in the seminiferous tubules, as well as in Sertoli and Leydig cells. In conclusion, we demonstrated that real-time in vivo BLI analysis can be used to noninvasively (in vivo) monitor long-term luciferase expression in mouse testis, and we verified that EGFP expression is localized in seminiferous tubules after bicistronic LV-mediated gene transfer into mouse testes. Furthermore, we anticipate the future use of in vivo BLI technology for real-time study of specific genes involved in spermatogenesis.


Glycans from avian influenza virus are recognized by chicken dendritic cells and are targets for the humoral immune response in chicken.
To increase our understanding of the interaction between avian influenza virus and its chicken host, we identified receptors for putative avian influenza virus (AIV) glycan determinants on chicken dendritic cells. Chicken dendritic cells (DCs) were found to recognize glycan determinants containing terminal αGalNAc, Galα1-3Gal, GlcNAcβ1-4GlcNAcβ1-4GlcNAcβ (chitotriose) and Galα1-2Gal. Infection of chicken dendritic cells with either low pathogenic (LP) or highly pathogenic (HP) AIV results in elevated mRNA expression of homologs of the mouse C-type lectins DEC205 and macrophage mannose receptor (MMR), whereas expression levels of the human dendritic cell-specific intercellular adhesion molecule-3-grabbing non-integrin (DC-SIGN) homolog remained unchanged. Following uptake and subsequent presentation of avian influenza virus by DCs, adaptive immunity, including humoral immune responses are induced. We have investigated the antibody responses against virus glycan epitopes after avian influenza virus infection. Using glycan micro-array analysis we showed that chicken contained antibodies that predominantly recognize terminal Galα1-3Gal-R, chitotriose and Fucα1-2Galβ1-4GlcNAc-R (H-type 2). After influenza-infection, glycan array analysis showed that both levels and repertoire of glycan-recognizing antibodies decreased. However, analysis of the sera by ELISA indicated that the levels of different isotypes of anti-glycan Abs against specific glycan antigens was increased after influenza-infection, suggesting that the presentation of the glycan antigens and iso-type of the Abs are critical parameters to take into account when measuring anti-glycan Abs. This novel approach in avian influenza research may contribute to the development of a broad spectrum vaccine and improves our mechanistic understanding of innate and adaptive responses to glycans.


Patterning of the third pharyngeal pouch into thymus/parathyroid by Six and Eya1.
Previous studies have suggested a role of the homeodomain Six family proteins in patterning the developing vertebrate head that involves appropriate segmentation of three tissue layers, the endoderm, the paraxial mesoderm and the neural crest cells; however, the developmental programs and mechanisms by which the Six genes act in the pharyngeal endoderm remain largely unknown. Here, we examined their roles in pharyngeal pouch development. Six1-/- mice lack thymus and parathyroid and analysis of Six1-/- third pouch endoderm demonstrated that the patterning of the third pouch into thymus/parathyroid primordia is initiated. However, the endodermal cells of the thymus/parathyroid rudiments fail to maintain the expression of the parathyroid-specific gene Gcm2 and the thymus-specific gene Foxn1 and subsequently undergo abnormal apoptosis, leading to a complete disappearance of organ primordia by E12.5. This thus defines the thymus/parathyroid defects present in the Six1 mutant. Analyses of the thymus/parathyroid development in Six1-/-;Six4-/- double mutant show that both Six1 and Six4 act synergistically to control morphogenetic movements of early thymus/parathyroid tissues, and the threshold of Six1/Six4 appears to be crucial for the regulation of the organ primordia-specific gene expression. Previous studies in flies and mice suggested that Eya and Six genes may function downstream of Pax genes. Our data clearly show that Eya1 and Six1 expression in the pouches does not require Pax1/Pax9 function, suggesting that they may function independently from Pax1/Pax9. In contrast, Pax1 expression in all pharyngeal pouches requires both Eya1 and Six1 function. Moreover, we show that the expression of Tbx1, Fgf8 and Wnt5b in the pouch endoderm was normal in Six1-/- embryos and slightly reduced in Six1-/-;Six4-/- double mutant, but was largely reduced in Eya1-/- embryos. These results indicate that Eya1 appears to be upstream of very early events in the initiation of thymus/parathyroid organogenesis, while Six genes appear to act in an early differentiation step during thymus/parathyroid morphogenesis. Together, these analyses establish an essential role for Eya1 and Six genes in patterning the third pouch into organ-specific primordia.


Phosphorus metabolism disorders in erythrocytes and lymphocytes among patients with inflammatory breast cancer, infiltrative stomach and colorectal cancer.
Energy and plastic potential dysfunction of erythrocytes and lymphocytes among people with inflammatory breast cancer, infiltrative stomach cancer, and infiltrative colon cancer is characterized by a more aggressive clinical course and poor prognosis. We explored the features of energy metabolism and phosphorus metabolism disorders in the erythrocytes and lymphocytes of patients with inflammatory breast cancer, infiltrative stomach cancer, and infiltrative colon cancer as a predicting factor in the course of the disease. 49 people were examined; the 1st group had infiltrative stomach cancer (n=17); the 2nd group had infiltrative colon cancer (n=11); the 3rd group had inflammatory breast cancer (n=21). Glycerol-3-phosphate dehydrogenase activity was 1.8 times reduced (p≤0.005), and the activity of glyceraldehyde-3-phosphate dehydrogenase in erythrocytes of patients with cancer at the main localization increased 2.5 times, compared with normal. Inflammatory breast cancer patients had a statistically significant decrease (p<0.005) in erythrocytes adenosine triphosphate content by an average of 56.5% compared with the normal ratio, and in cases of patients with gastric and colorectal cancer, a decrease of 67%. Excessive use of phosphorus for energy metabolism and adenosine triphosphate production destroys the balance of energetic and plastic potentials of erythrocytes and lymphocytes in inflammatory breast cancer, infiltrative stomach, and infiltrative colorectal cancers patients.


Gastric tumor from metastasis of breast cancer.
Metastatic tumours of the stomach have been reported to result from various types of cancer. Among them, gastric metastasis from breast cancer has been recognised in 0.3-18% patients (1-4). Here, a rare case of metastatic gastric tumour derived from breast carcinoma is reported. Gastric endoscopy confirmed a large, friable mass (approximately 5 cm in diameter) in the upper part of the gastric body. The mass within the stomach was difficult to distinguish from primary gastric cancer, although biopsies of this lesion revealed the characteristics of adenocarcinoma. In addition, immunohistochemistry showed the positive expression of mammaglobin. Taken together, the evidence pointed to metastasis of breast cancer to the stomach. The patient was treated with hormonal therapy (letrozole), and the size of the metastasis in the stomach was markedly reduced. Therefore, a gastric metastasis from breast cancer was diagnosed successfully using immunohistochemistry and unnecessary surgery was avoided. In conclusion, although gastric metastatic tumours derived from breast carcinoma are rare, their accurate pre-operative diagnosis and appropriate systemic treatment is essential.


The Use of Virtual Reality in the Study of People's Responses to Violent Incidents.
This paper reviews experimental methods for the study of the responses of people to violence in digital media, and in particular considers the issues of internal validity and ecological validity or generalisability of results to events in the real world. Experimental methods typically involve a significant level of abstraction from reality, with participants required to carry out tasks that are far removed from violence in real life, and hence their ecological validity is questionable. On the other hand studies based on field data, while having ecological validity, cannot control multiple confounding variables that may have an impact on observed results, so that their internal validity is questionable. It is argued that immersive virtual reality may provide a unification of these two approaches. Since people tend to respond realistically to situations and events that occur in virtual reality, and since virtual reality simulations can be completely controlled for experimental purposes, studies of responses to violence within virtual reality are likely to have both ecological and internal validity. This depends on a property that we call 'plausibility' - including the fidelity of the depicted situation with prior knowledge and expectations. We illustrate this with data from a previously published experiment, a virtual reprise of Stanley Milgram's 1960s obedience experiment, and also with pilot data from a new study being developed that looks at bystander responses to violent incidents.


Antigen-specific B cells efficiently present low doses of antigen for induction of T cell proliferation.
Previous experiments suggested a role for specific B cells in the induction of antigen (SRBC)-specific T cell proliferation. Two models were proposed: in the first, B cells directly presented antigen to T cells; alternatively, B cells secreted antibody, which opsonized antigen for presentation by macrophages. Experiments to distinguish between these possibilities are presented here. Three lines of evidence support the conclusion that antigen is presented directly by specific B cells. First, nonimmune splenic adherent cells (SAC), which efficiently induced proliferation of appropriately primed T cells to antigens such as OVA and GAT, were unable to induce SRBC-specific proliferation. Secondly, a slope analysis of the logarithmic plot of T cell proliferation vs the number of irradiated B cells suggested that two cells were limiting within the presenting population. The addition of IL 1 or SAC reduced the slope to 1 (although in serum-free conditions, the addition of IL 1, but not SAC, reduced the slope of the line). Specificity of the B cells for the antigen continued to be required in the presence of exogenous IL 1 or SAC. These results suggested that presentation by specific B cells and the amount of IL 1 were the limiting requirements for the induction of SRBC-specific T cell proliferation. The third line of evidence was the demonstration of a restricted interaction between T cells and B cells. The addition of irradiated, allogeneic SRBC-specific B cells to T cell lines and syngeneic SAC failed to support proliferative responses. We further show that a GAT-specific T cell clone was triggered to proliferate by either SAC or B cells, but that antigen-specific B cells were necessary at low doses of antigen. This finding is important in two respects. First, the T cell clone previously has been shown to act as a helper; secondly, when low doses of antigen are used, the requirement for priming of the B cells to the specific antigen is true for a soluble, as well as a particulate, antigen. We propose that at low (physiologic) doses of antigen, presentation to secondary T cells takes place mainly at the surface of antigen-specific B cells. At high doses of antigen,h presentation can also be accomplished by nonspecific cells such as other B cells, macrophages, or dendritic cells.


Immediate Neutrophil-Variable-T Cell Receptor Host Response in Bacterial Meningitis.
Bacterial meningitis is a life-threatening disease that evokes an intense neutrophil-dominated host response to microbes invading the subarachnoid space. Recent evidence indicates the existence of combinatorial V(D)J immune receptors in neutrophils that are based on the T cell receptor (TCR). Here, we investigated expression of the novel neutrophil TCRαβ-based V(D)J receptors in cerebrospinal fluid (CSF) from human patients with acute-phase bacterial meningitis using immunocytochemical, genetic immunoprofiling, cell biological, and mass spectrometric techniques. We find that the human neutrophil combinatorial V(D)J receptors are rapidly induced in CSF neutrophils during the first hours of bacterial meningitis. Immune receptor repertoire diversity is consistently increased in CSF neutrophils relative to circulating neutrophils and phagocytosis of baits directed to the variable immunoreceptor is enhanced in CSF neutrophils during acute-phase meningitis. Our results reveal that a flexible immune response involving neutrophil V(D)J receptors which enhance phagocytosis is immediately initiated at the site of acute bacterial infection.


Evaluating cognitive impairment in the clinical setting: practical screening and assessment tools.
HIV-associated neurocognitive disorders (HAND) remain a substantial problem in the era of combination antiretroviral therapy. Neither the Mini Mental State Exam nor the HIV Dementia Scale is sufficiently sensitive for HAND. The Montreal Cognitive Assessment shows promise, but current data suggest that adding an additional test will be needed to improve sensitivity for the clinical setting. Patient reporting of symptoms is insensitive as most cases of HAND are asymptomatic. Examination of cerebrospinal fluid (CSF) is sometimes warranted in select patients to evaluate for CSF HIV RNA detectability. CSF escape of virus, when CSF HIV RNA is detectable but plasma HIV RNA is not, appears to be a relatively uncommon event in the clinical setting where the level of detectability for typical clinical assays is around 50 copies/mL. In cases of CSF escape, cognitive improvement has been linked to changes in antiretroviral regimens that are aimed at either overcoming antiretroviral resistance or improving central nervous system (CNS) penetration-effectiveness. Currently, for most patients with HAND in the absence of unusual features, there are insufficient data for a recommendation to routinely intensify therapy with a neurointensive antiretroviral regimen; however, there is considerable uncertainty given emerging data and variability in approach among experts in the field. This article summarizes a case-based presentation by Victor G. Valcour, MD, at the 14th Annual Clinical Conference for the Ryan White HIV/AIDS Program held in Tampa, Florida, in June 2011. The Clinical Conference is sponsored by the IAS-USA under the Health Resources and Services Administration (HRSA) contract number HHSH250200900010C.


Peptides as tight junction modulators.
The first step in drug absorption is the passage of drug molecules across epithelial cell sheets. Epithelial cell sheets are pivotal for the maintenance of homeostasis in the body by acting as a biological barrier that separates the inside of the body from the outside environment. Intercellular space between the adjacent epithelial cells is tightly sealed by tight junctions (TJs), which prevent solutes from freely moving across the epithelial cell sheets. Modulation of the TJ barrier has been a potent strategy for drug absorption. Absorption enhancers have been investigated since the 1980s, and sodium caprate is clinically used as an absorption enhancer. However, the biochemical constituents and structures of TJs were not elucidated until 1993. Occludin, a tetra-transmembrane protein, was identified to be a structural component of TJs in 1993. Claudin, another tetra-transmembrane protein, was identified as a structural and functional component of TJs in 1998. Modulation of occludin- or claudin-barrier is novel methods to enhance drug absorption. Recently, synthetic TJ-binding peptides, a kinase of claudin and peptide fragments of toxins have been developed. In the present review, we summarize the recent progress in TJ-modulating peptides and discuss their potencies.


Chronic Alcohol Exposure Enhances Lipopolysaccharide-Induced Lung Injury in Mice: Potential Role of Systemic Tumor Necrosis Factor-Alpha.
It is well known that liver and lung injury can occur simultaneously during severe inflammation (e.g., multiple organ failure). However, whether these are parallel or interdependent (i.e., liver-lung axis) mechanisms is unclear. Previous studies have shown that chronic ethanol (EtOH) consumption greatly increases mortality in the setting of sepsis-induced acute lung injury (ALI). The potential contribution of subclinical liver disease in driving this effect of EtOH on the lung remains unknown. Therefore, the purpose of this study was to characterize the impact of chronic EtOH exposure on concomitant liver and lung injury. Male mice were exposed to EtOH-containing Lieber-DeCarli diet or pair-fed control diet for 6 weeks. Some animals were administered lipopolysaccharide (LPS) 4 or 24 hours prior to sacrifice to mimic sepsis-induced ALI. Some animals received the tumor necrosis factor-alpha (TNF-α)-blocking drug, etanercept, for the duration of alcohol exposure. The expression of cytokine mRNA in lung and liver tissue was determined by quantitative PCR. Cytokine levels in the bronchoalveolar lavage fluid and plasma were determined by Luminex assay. As expected, the combination of EtOH and LPS caused liver injury, as indicated by significantly increased levels of the transaminases alanine aminotransferase/aspartate aminotransferase in the plasma and by changes in liver histology. In the lung, EtOH preexposure enhanced pulmonary inflammation and alveolar hemorrhage caused by LPS. These changes corresponded with unique alterations in the expression of pro-inflammatory cytokines in the liver (i.e., TNF-α) and lung (i.e., macrophage inflammatory protein-2 [MIP-2], keratinocyte chemoattractant [KC]). Systemic depletion of TNF-α (etanercept) blunted injury and the increase in MIP-2 and KC caused by the combination of EtOH and LPS in the lung. Chronic EtOH preexposure enhanced both liver and lung injury caused by LPS. Enhanced organ injury corresponded with unique changes in the pro-inflammatory cytokine expression profiles in the liver and the lung.


Cognitive Reserve, Early Cognitive Screening, and Relationship to Long-Term Outcome after Severe Traumatic Brain Injury.
The objective was to investigate the relationship between early global cognitive functioning using the Barrow Neurological Institute Screen for Higher Cerebral Functions (BNIS) and cognitive flexibility (Trail Making Test (TMT), TMT B-A), with long-term outcome assessed by the Mayo-Portland Adaptability Index (MPAI-4) in severe traumatic brain injury (sTBI) controlling for the influence of cognitive reserve, age, and injury severity. Of 114 patients aged 18-65 with acute Glasgow Coma Scale 3-8, 41 patients were able to complete (BNIS) at 3 months after injury and MPAI-4 5-8 years after injury. Of these, 33 patients also completed TMT at 3 months. Global cognition and cognitive flexibility correlated significantly with long-term outcome measured with MPAI-4 total score (rBNIS = 0.315; rTMT = 0.355). Global cognition correlated significantly with the participation subscale (r = 0.388), while cognitive flexibility correlated with the adjustment (r = 0.364) and ability (r = 0.364) subscales. Adjusting for cognitive reserve and acute injury severity did not alter these relationships. The effect size for education on BNIS and TMT scores was large (d ≈ 0.85). Early screenings with BNIS and TMT are related to long-term outcome after sTBI and seem to measure complementary aspects of outcome. As early as 3 months after sTBI, educational level influences the scores on neuropsychological screening instruments.


Successful allogeneic transplantation of patients with suspected prior invasive mold infection.
Prior invasive fungal infection (IFI) has historically limited the use of allogeneic stem cell transplantation for patients with hematologic malignancies. Transplantation of such patients frequently resulted in recurrent infection and high mortality rates. Several new antifungal agents have been introduced over the past 5 years with broader spectra of activity against molds such as Aspergillus and a favorable toxicity profile. In this study, we present a series of 16 consecutive patients with hematologic malignancy and prior invasive fungal infection who underwent allogeneic transplantation. Of these patients, the majority of whom were treated with voriconazole and/or caspofungin, only four experienced recurrent fungal infection and recurrent fungal infection was the primary cause of death in only one patient. The estimated 45% 2-year survival in this series is similar to that for other patients with high risk hematologic malignancy undergoing stem cell transplantation. We conclude that suspected prior invasive fungal infection should not preclude the use of allogeneic stem cell transplantation.


Studies on microbial transformation of meloxicam by fungi.
Screening-scale studies were performed with 26 fungal cultures for their ability to transform the anti-inflammatory drug meloxicam. Among the different fungi screened, a filamentous fungus, Cunninghamella blakesleeana NCIM 687, transformed meloxicam to three metabolites in significant quantities. The transformation of meloxicam was confirmed by high-performance liquid chromatography (HPLC). Based on the liquid chromatography-tandem mass spectrometry (LC-MS/MS) data, two metabolites were predicted to be 5-hydroxymethyl meloxicam and 5-carboxy meloxicam, the major mammalian metabolites reported previously. A new metabolite was produced, which is not detected in mammalian systems. Glucose medium, pH of 6.0, temperature of 27 degrees , 5-day incubation period, dimethylformamide as solvent, and glucose concentration of 2.0%were found to be suitable for maximum transformation of meloxicam when studied separately. It is concluded that C. blakesleeana can be employed for biotransformation of drugs for production of novel metabolites.


Smart Thixotropic Hydrogels by Disulfide-Linked Short Peptides for Effective Three-Dimensional Cell Proliferation.
Supramolecular assembly of short peptides is a crucial process and has shown numerous potential applications as biomaterials. In the present work, the hydrogelation process of short peptides containing C-terminal "Lys-Cys" (KC) residues have been studied in detail. The N-terminal capping is found to be essential for effective gelation. Out of 12 peptides we studied, two of them could form hydrogels efficiently: Ac-VVKC-NH2 and Ac-FFKC-NH2. In both cases, the monomer-to-dimer formation through disulfide linkages by Cys residues controls the aggregation process. Interestingly, the presence of H2O2 facilitated the dimerization and thereby reduced the gelation time but could not impart much effect on the mechanical properties of the gels. Detailed rheological study revealed that both hydrogels are thixotropic in nature. Moreover, they are responsive to glutathione (GSH) due to the presence of disulfide linkages. However, the hydrogel of Ac-FFKC-NH2 is found to be stronger and more effective for biological applications. The thixotropic nature as well as a model drug release study in response to varying GSH concentration indicates the possible use of the hydrogel as an injectable local drug delivery vehicle. The hydrogel of Ac-FFKC-NH2 is noncytotoxic in nature. Three-dimensional cell proliferation has been found to be more effective than 2D, as it mimics the in vivo situation more closely if not exactly. In the present study, we have shown that both differentiated RAW macrophages and undifferentiated THP-1 monocytes could proliferate significantly within the 3D matrix of the hydrogel, without depicting any apparent cytotoxicity. Thus, the hydrogel of Ac-FFKC-NH2 has potential for application in localized drug administration and as a supporting biomaterial to study basic phenomena involving cell behavior.


[Compliance and resistance to child vaccination: a study among Swiss mothers].
This study examines mothers' practices and attitudes in relation to their child's immunization. In this area, like in others, gaps are being observed between the public health model of risk management and the population expectations and behaviors. Data were collected using a standardized questionnaire from 1295 women having, in 1999, one child between 7 and 8 years of age of Swiss nationality and attending the public school system in Geneva. Four immunization types were established based on (1) the degree of satisfaction (in relation to past choices), (2) the perception of adequate knowledge about immunization and (3) practices. The mothers were divided into the following categories: compliant (57%), compliant ambivalent (19%), moderately resistant (17%) and resistant (7%). Mothers having reached an intermediate education level are more likely to be resistant. Furthermore, women having negative attitudes towards biomedical institutions, having consulted alternative practitioners and those considering that they have a certain individual control over the health of the family are more often resistant to immunization. Our results confirm the existence of a resistance to child immunization among a section of the population. It should not be attributed to mothers' ignorance, but rather could reflect their perplexity towards the choices they are expected to make. In that regards, the importance some parents give to individual control over health can be in contradiction with community objectives of public health. Actions for the promotion of immunization should primarily target families who are uncertain (compliant ambivalent and moderately resistant). At the same time, the difficulties entailed in the - individual and collective - management of risks require that new forms of dialogue be developed between experts and the public.


Exciton-Related Raman Scattering, Interband Absorption and Photoluminescence in Colloidal CdSe/CdS Core/Shell Quantum Dots Ensemble.
By using the numerical discretization method within the effective-mass approximation, we have theoretically investigated the exciton-related Raman scattering, interband absorption and photoluminescence in colloidal CdSe/CdS core/shell quantum dots ensemble. The interband optical absorption and photoluminescence spectra have been revealed for CdSe/CdS quantum dots, taking into account the size dispersion of the ensemble. Numerical calculation of the differential cross section has been presented for the exciton-related Stokes-Raman scattering in CdSe/CdS quantum dots ensemble with different mean sizes.


Patient record access--the time has come.
There are now a number of systems across the world that enables patients to view their electronic health records. These include kiosks that have fingerprint authentication and also net-based access. The paper outlines the approach taken by the UK NHS explains the legal underpinning of access. Starting form the premise that record access is here to stay the paper outlines the research on benefits and risks of record access, concluding that, with simple precautions, record access is safe and affords many benefits to both patients and clinicians. It goes on to consider possible impacts of record access on the way records might be written as a co-produced document and emphasizes that national standards for record sharing need to be written.


Twin mole and viable fetus: The case for misdiagnosis.
To report the Sheffield Trophoblast Centre experience of twin molar gestations and review this in the light of international experience. Thirty patients with possible twin molar gestations were registered from 1986 to 2004 (during which period 7,200 cases of mole were seen). The accuracy of suspected clinical and histologic diagnoses was investigated. In 10 cases twin mole/fetus had been suspected clinically but not confirmed when products of conception were examined. In 3 of these cases the pregnancy had been therapeutically terminated because of clinical (ultrasound) suspicion of coexisting molar pregnancy. In the 19 cases where twin mole/fetus was suspected, central histopathology review was possible in 14 cases. Only 7 were confirmed. In 2 further cases twin molar gestation was diagnosed on specimens referred for central review as partial mole singleton pregnancies. For confirmed cases the pregnancy outcome was term delivery in 5 cases and miscarriage in 4. Clinical and histopathologic diagnosis of twin molar pregnancies is inaccurate in many suspected cases; therefore, a second (expert) opinion should be sought. When the diagnosis is accurate, maternal and fetal complications are common. However, in suspected cases the pregnancy may be allowed to proceed, with caution, if the mother wishes.


Regional variation in tooth size and pathology in fossil hominids.
Tooth size and dental pathology in fossil hominids were studied to test for regional differences in these parameters. The results showed little regional variation in tooth size for the Middle and Upper Pleistocene sites compared (except for Krapina) but considerable differences in the severity of attrition and dental pathology. These differences were considered indicative of regional differences in the functional load borne by the teeth, and in view of the similar technological status of the groups studied, were attributed to environmental differences in the diet. Since, in all regions, reduction in tooth size appeared to continue at the same rate for the periods investigated, no association can be established between presumed selective pressures related to differences in functional demands made on the dentition, and tooth reduction.


Folliculosebaceous Cystic Hamartoma With Prominent Adipose Tissue Resembling Spindle Cell Lipoma.
Folliculosebaceous cystic hamartomas (FSCH) are a rare form of cutaneous hamartoma composed of follicular, sebaceous, and mesenchymal elements. These lesions are most often seen in the central face and scalp and rarely exceed 2 cm in size. Here, we report a case of a folliculosebaceous cystic hamartoma with a prominent adipose tissue component resembling a spindle cell lipoma. The patient is a 36-year-old man with a slowly enlarging, flesh-colored, lobulated lesion on his right ala that had been previously biopsied. The clinical differential included a hypertrophic scar or recurrent hamartoma. A full-thickness excisional biopsy was performed, which revealed prominent, large, irregularly shaped, folliculosebaceous structures, including multiple dilated follicles associated with prominent hyperplastic-appearing sebaceous glands, findings consistent with FSCH. The mesenchymal component contained a proliferation of spindle-shaped cells associated with mucin and thickened, ropey-appearing bundles of collagen, features mimicking a spindle cell lipoma. This combination of a folliculosebaceous cystic hamartoma with prominent adipose tissue resembling spindle cell lipoma is unusual and has only been previously reported once in the literature, and therefore, our case contributes to the expanding knowledge of this rare variant of FSCH.


Activating photoactivatable proteins with laser light to visualize membrane systems and membrane traffic in living cells.
Eukaryotic cells are composed of an intricate system of internal membranes that are organized into different compartments--including the endoplasmic reticulum (ER), the nuclear envelope, the Golgi complex (GC), lysosomes, endosomes, caveolae, mitochondria, and peroxisomes--that perform specialized tasks within the cell. The localization and dynamics of intracellular compartments are now being studied in living cells because of the availability of green fluorescent protein (GFP)-fusion proteins and recent advances in fluorescent microscope imaging systems, such as the confocal laser-scanning microscope (CLSM). This protocol describes the steps for activating one of the first photoactivatable proteins, PA-GFP.


Structure-activity relationship for substance P inhibition of carbamylcholine-stimulated 22Na+ flux in neuronal (PC12) and non-neuronal (BC3H1) cell lines.
The inhibition of carbamylcholine-stimulated 22Na+ flux by substance P and various peptide analogs was examined in PC12 cells, a line which contains a neuronal-type nicotinic receptor, and BC3H1 cells, a line which contains a muscle-type nicotinic receptor. Substance P produces a noncompetitive inhibition of carbamylcholine-stimulated 22Na+ influx in both cell lines (IC50 = 1.2 microM on PC12 cells and 8.2 microM on BC3H1 cells). The structure-activity relation for substance P analogs was qualitatively similar in both cell lines; however, there were quantitative differences. Substance P was the most potent peptide tested. Analogs of substance P with amino acids removed from the N-terminus resulted in significant decreases in potency, whereas removal of amino acids from the C-terminus resulted in analogs virtually devoid of activity. Compounds purported to be substance P antagonists had actions similar to substance P in reducing carbamylcholine-stimulated 22Na+ flux. The related tachykinins physalaemin and eledoisin had low potencies on both cell lines. These results indicate that the site through which substance P exerts its inhibitory effects on activation of nicotinic receptors is different from the receptors described previously for substance P in more classical systems. In addition, our results indicate that substance P has an effect on both the neuronal-type nicotinic receptor (alpha-bungarotoxin insensitive) expressed on PC12 cells and the muscle-type nicotinic receptor (alpha-bungarotoxin sensitive) expressed on BC3H1 cells.


The effects of a prolonged undernutrition on serum lipids and fatty acid composition of reindeer calves during winter and spring.
We examined the effects of undernutrition on lipid metabolism in reindeer (<1 year) during mid-winter and spring, with particular focus on the proportions of polyunsaturated fatty acids (PUFAs) in major serum lipids. The reindeer (n=8) were fed their winter feed, lichen, ad libitum for 5 weeks, followed by 40% restriction of energy for 8 weeks and refeeding to normal for 6 weeks. The concentrations of major serum lipids, cholesterol and phospholipids decreased significantly during the ad libitum period (by 50 and 44%, respectively). The proportion of major PUFA, linoleic acid in serum cholesteryl esters, decreased from 48.2 to 38.4% during the ad libitum period (P < 0.01), and to 29.2% during the restriction period (P < 0.001). The proportion of linoleic acid in phospholipids decreased from 27.9 to 15.6% during the ad libitum period (P < 0. 001), and to 13.0% during the restriction (P < 0.01). Also alpha-linolenic acid in the major lipids decreased significantly during the ad libitum and restriction periods. The decreases in the major lipids and linoleic acid were reversed during the refeeding. The control group (n=8) which was fed high-quality concentrates ad libitum gained weight most of the spring but showed similar although slower decreases in the major serum lipids and PUFAs than the lichen group. Our results indicate that feeding reindeer on lichen during winter leads to the retardation of growth and reductions in major serum lipids and their principal C18-PUFA proportions. The decreased proportions of the principal PUFAs most probably reflect their low dietary intake but may have been modified also by seasonal factors.


Low-dose thalidomide plus monthly high-dose oral dexamethasone (Thali-Dexa): results, prognostic factors and side effects in eight patients previously treated with multiple myeloma.
Thalidomide is active both as single agent and in combination-therapy against refractory or relapsing multiple myeloma. Eigth patients previously treated were given Thalidomide 100mg/daily plus Dexametasone 40mg/daily for four days each month (Thali-Dexa) and followed for response, prognostic factors and side effects. Two patients had early death (one from massive cerebral ischemic stroke, the other from dementia and progressive renal failure), one patient progressed during Thali-Dexa (thalidomide 200mg) and was rescued with chemotherapy, two patients required increasing thalidomide dosage (to 200 and 400mg, respectively) because of progressive disease, three patients had stable disease remission lasting from 4m+ to 16m+. Thali-Dexa is a useful agent but age and vascular/metabolic diseases may increase the risk of severe side effects. Early decrease in erythrocyte sedimentation rate seems to correlate with better disease control.


Occupational risks related to vibrations using a brush cutter for green area management.
Weed control is one of the most important issues in the maintenance sectors of both agriculture and green areas. Small tools are employed for controlling grass and other growths on steep verges and river banks. This leads the operators being exposed to many risks among which vibration is one. The purpose of this study is to measure and evaluate hand-arm vibration and to verify the daily exposure to which workers are often subjected while weeding. Two cutting heads, a brush knife and a mowing head were compared. Both were mounted on the same cow-horn brush cutter. The vibration total value was expressed as the root-mean-square (rms) of three component values according to the axes X, Y and Z. The signal was frequency weighted using the weighting curve Wh, as described in the ISO 5349-1 (2001) standard. In addition, the daily vibration exposure was calculated and compared with the thresholds set by EU Directive 2002/44/EC (2005). The obtained results showed that the exposure action value (EAV) of 2.5 ms-2 was exceeded while using both cutting heads. The exposure limit value (ELV) using the brush knife also exceeded 5 ms-2. The results highlighted important aspects in terms of exposure values that should be considered with the view of preventing the risk of Hand-Arm Vibration Syndrome (HAVS) to which the operators who frequently use these tools are exposed. Specific measures should therefore be taken to protect the exposed workers.


Neoadjuvant and Adjuvant Therapy for HER2 Positive Disease.
Since the initial description of the HER2 proto-oncogene as a poor prognostic factor in breast cancer in 1987, to the first randomized trial of a monoclonal antibody directed against HER2 in combination with chemotherapy for the treatment of metastatic HER2-positive breast cancer published in 2001, to the American Society of Clinical Oncology (ASCO) 2005 Annual Meeting in which we saw the unprecedented collective presentations demonstrating the dramatic benefit of trastuzumab in the adjuvant setting-the clinical landscape of HER2-overexpressing breast cancer has forever changed. More recently, there has been increasing use of preoperative chemotherapy and anti-HER2 targeted therapies in primary operable HER2 disease in the research domain and in clinical practice. In the next few years, we will see if dual adjuvant anti-HER2 antibody inhibition produces clinically significant improvements in outcome; understand if there is a role of small molecule inhibitors of the HER family of receptors either in combination or sequential to trastuzumab; further refine the relationship between pathologic complete response (pCR) and long-term clinical outcomes; and find predictive biomarkers to identify cohorts of patients that may need differential combinations and/or durations of anti-HER2 therapies.


Structural analogues of diosgenyl saponins: synthesis and anticancer activity.
Saponins display various biological activities including anti-tumor activity. Recently intensive research has been focused on developing saponins for tumor therapies. The diosgenyl saponin dioscin is one of the most common steroidal saponins and exhibits potent anticancer activity in several human cancer cells through apoptosis-inducing pathways. In this paper, we describe the synthesis of several diosgenyl saponin analogues containing either a 2-amino-2-deoxy-beta-d-glucopyranosyl residue or an alpha-l-rhamnopyranosyl-(1-->4)-2-amino-2-deoxy-beta-d-glucopyranosyl residue with different acyl substituents on the amino group. The cytotoxic activity of these compounds was evaluated in MCF-7 breast cancer cells and HeLa cervical cancer cells. Structure-activity relationship studies show that the disaccharide saponin analogues are in general less active than their corresponding monosaccharide analogues. The incorporation of an aromatic nitro functionality into these saponin analogues does not exhibit significant effect on their cytotoxic activity.


[Recent advances in strawberry transgenic research].
Strawberry (Fragaria ananassa Duch.) is one of most important fruit crops cultivated widely in world. Genetic transformation has launched a new era in strawberry breeding and germplasm creativity. It offers a direct method of creating varieties that selectively targets gene or a few heterologous traits for introduction into the strawberry plant. Great advances have been made in strawberry genetic transformation in the past years. This paper reviews the recent progress in genetic transformation of strawberry on promoting resistance to viruses and fungi, insects, herbicides, stress and quality improvement. Problems and the prospects for application of genetic transformation in strawberry were discussed.


[Giant cell tumor--osteoclastoma: part II].
Authors present current views on the comprehensive diagnostics, treatment and prognosis of giant cell tumours. They include an exact algorithm of diagnostics necessary for a timely treatment. They describe an often unexpected behaviour of these tumours which is frequently active and aggressive, to a small extent these tumours show the tendency to become malignant. The treatment prefers curretage, antibiotic irrigation and filling in with bone cement. In case of recurrence and an extensive process, block resection and replacement by an individual implant, in hardly accessible locations (spine, sacrum, pelvis) or radiotherapy is recommended. The authors present in detail results of a 10-year systematic follow-up of 56 patients, of which 61.6% were in active stage, 36.8% in aggressive stage. The most frequently used method (in 73.2%) was curretage, irrigation and filling in with bone grafts, in 12.5% of cases block resection and replacement by bone grafts. Total results after all these types of operations evaluated in the interval of 3 years after the operation were very good, i.e. 85.7% remissions, there were 14.3% of recurrence. Of these setbacks 5.4% were in remission for more than 3 years and thus conduced to the increase of the number of remissions to 94.6% which can be evaluated as an excellent result. Local recurrence occurs in 10-15% with the application and 15-25% without the application of local adjuvant therapy. In the third stage in case of extensive destruction we perform resection en bloc, radiotherapy in hardly accessible locations (spine, sacrum, pelvis). In case of extensive destruction in the region of joints the method of choice is the application of both individual and total replacement.


The SAMHD1-mediated block of LINE-1 retroelements is regulated by phosphorylation.
The restriction factor SAMHD1 regulates intracellular nucleotide level by degrading dNTPs and blocks the replication of retroviruses and DNA viruses in non-cycling cells, like macrophages or dendritic cells. In patients, inactivating mutations in samhd1 are associated with the autoimmune disease Aicardi-Goutières Syndrome (AGS). The accumulation of intracellular nucleic acids derived from endogenous retroelements thriving in the absence of SAMHD1 has been discussed as potential trigger of the autoimmune reaction. In vitro, SAMHD1 has been found to restrict endogenous retroelements, like LINE-1 elements (L1). The mechanism, however, by which SAMHD1 blocks endogenous retroelements, is still unclear. Here, we show that SAMHD1 inhibits the replication of L1 and other endogenous retroelements in cycling cells. By applying GFP- and neomycin-based reporter assays we found that the anti-L1 activity of SAMHD1 is regulated by phosphorylation at threonine 592 (T592). Similar to the block of HIV, the cofactor binding site and the enzymatic active HD domain of SAMHD1 proofed to be essential for restriction of L1 elements. However, phosphorylation at T592 did not correlate with the dNTP hydrolase activity of SAMHD1 in cycling 293T cells suggesting an alternative mechanism of regulation. Interestingly, we found that SAMHD1 binds to ORF2 protein of L1 and that this interaction is regulated by T592 phosphorylation. Together with the finding that the block is also active in cycling cells, our results suggest that the SAMHD1-mediated inhibition of L1 is similar but not identical to HIV restriction. Our findings show conclusively that SAMHD1 restricts the replication of endogenous retroelements in vitro. The results suggest that SAMHD1 is important for maintaining genome integrity and support the idea of an enhanced replication of endogenous retroelements in the absence of SAMHD1 in vivo, potentially triggering autoimmune diseases like AGS. Our analysis also contributes to the better understanding of the activities of SAMHD1 in antiviral defense and nucleotide metabolism. The finding that the phosphorylation of SAMHD1 at T592 regulates its activity against retroelements but not necessarily intracellular dNTP level suggests that the dNTP hydrolase activity might not be the only function of SAMHD1 important for its antiviral activity and for controlling autoimmunity.


Localized fluid collections after liver transplantation.
Liver transplantation is an accepted effective therapy for patients with diseases that lead to hepatic failure. In addition to vascular, rejection-related, biliary, and lymphoproliferative complications, posttransplantation fluid collections, such as hematomas, seromas, bilomas, localized ascites, and abscesses can affect graft survival. In this study, we have presented, ultrasonography and computed tomography images as well as reviewed the literature. Between December 1988 and March 2005, 138 patients (94 men [68.11%], 44 women [31.89%] of mean age, 26.77 years (range, 1 to 64) underwent liver transplantation in our institution from living-related donors in 85 and deceased donors in 53 recipients. This retrospective study analyzed the fluid collections diagnosed after liver transplantation. Among 138 liver transplants we identified 56 localized intra-abdominal fluid collections, including 46 hematomas, 4 bilomas, and 6 abscesses. Fluid collections were noticed between 0 and 1095 days (mean = 13.50 +/- 152.82 days). Thirty-three collections were identified in living-related, and 23 in deceased donor recipients. The collections showed an average volume of 375 cm3, with a range of 1 to 4368 cm3. Two cases of bilomas were related to hepatic arterial insufficiency, one from a hepatic artery occlusion and the other from a significant stenosis. Localized intra-abdominal fluid collections were commonly seen after liver transplantations, diminished in size without treatment, but required aspiration or drainage in some patients.


Release of soluble peptidoglycan from growing conococci: demonstration of anhydro-muramyl-containing fragments.
Previous analysis of soluble peptidoglycan (PG) fragments released by exponentially growing gonococci implicated the combined action of both hexosaminidase and amidase activities in PG turnover. Current studies further characterized PG fragments which were labeled in the glycan with D-glucosamine and in the peptide moiety with meso-diaminopimelic acid of L- and D-alanine. Labeled PG fragments were isolated by gel filtration and characterized on the bases of (i) KD values, (ii) free amino group analysis using fluorodinitrobenzene, (iii) borohydride reduction, (iv) alkali-catalyzed beta-elimination, (v) paper chromatography in various solvents, (vi) electrophoretic mobility at various pH values, (vii) digestibility by Charonia lampas glycosidases, and (viii) content of labeled D- and L-alanine. A set of well-characterized PG fragments was used as standards. The monomer fraction (the major extracellular product) was found to contain two components. Most (about 80%) appeared to be N-acetylglucosaminyl-beta-1 leads to 4-1,6-anhydro-N-acetylmuramyl-L-ala-D-glu-meso-diaminopimelic acid; the remainder was the corresponding disaccharide tetrapeptide containing a C-terminal D-alanine. An unusual feature of these products was the presence of the anhydro-muramyl (non-reducing) ends, reflecting the activity of a gonococcal transglycosylase, and the near absence of products containing detectable reducing ends. Otherwise, the structures of the monomer fragments were typical of those expected for a gram-negative bacterium (chemotype I). The corresponding peptide-cross-linked dimer and the free disaccharide also contained nonreducing ends, exclusively. Free peptides (products of amidase activity) consisted of both tripeptide and tetrapeptide. In summary, all gonococci examined appear to possess an unusual transglycosylase activity which contributes to the release of soluble PG fragments containing nonreducing, anhydro-muramyl ends. The release of these fragments in vivo might be a unique aspect of gonococci-host interactions.


Isolated mitral valve replacement. With Carpentier-Edwards bioprosthesis :independent risk factors for long-term survival and prosthesis failure.
From 1976 through 1986, 188 patients (female/male ratio: 2/1, age 20-77 years, mean 58 years) with isolated mitral valve disease underwent valve replacement using the Carpentier-Edwards porcine bioprothesis (CEPB). Nine hospital deaths (4.8%) were excluded from further analysis. Follow-up was 0.2-11.3 years (mean 5.2 years); preoperatively, 74% had atrial flutter/fibrillation, and 75% were in NYHA-classes III-IV. All patients were put on life-long coumadin treatment. Preoperative predictability of long-term survival and prosthesis-related complications was examined using Cox regression analysis. Five preoperative variables were found to have independent predictive value as regards long-term survival: myxomatous degeneration of the valve (p = 0.002), chronic regurgitation (p = 0.003), age (p = 0.004), NYHA-class III-IV (p = 0.05), and atrial flutter/fibrillation (p = 0.05). A prognostic index calculated form the final Cox model identified six risk groups (I-VI) having cumulative 10-year survivals +/- SE of: I (n = 9) 100%, II (n = 10) 90 +/- 9%, III (n = 30) 73 +/- 10%, IV (n = 70) 51 +/- 9%, V (n = 43) 17 +/- 10%, and VI (n = 17, 7-year survival) 16 +/- 13% (p less than 0.0001). The incidence of late valve-related complications (%/patient-years) were: hemorrhage, 1.2; thromboembolism, 0.5; Endocarditis, 1.0; paravalvular leak, 0.2; and primary tissue failure, 1.5. Previous closed comissurotomy adversely influenced the occurrence of hemorrhage, while calcified mitral annulus were predictive of endocarditis. Younger age (less than or equal to 45 years) had a strong predictive influence of primary tissue failure.(ABSTRACT TRUNCATED AT 250 WORDS)


[The double control system of heart relaxation].
The systolic performance of the heart is regulated by two separate mechanisms: the first involves an adaptation to load (Frank-Starling), and the second, variations in contractility. Similarly, recent experimental studies have shown a double mechanism of control of the relaxation phase. Cardiac relaxation, as regards the muscle and muscle-pump system, is governed by the constant interrelation of the sensitivity of the contractile system to conditions of load (load dependence) and by the sensitivity to decreasing activation (inactivation dependence). The load dependence of cardiac muscle requires an important and effective intracellular system of calcium sequestration. For given load conditions, relaxation may be modulated by subtle changes in the dissipation of activation. Such variations of dissipation of activation may result from metabolic (coronary circulation), neuro humoral (sympathetic tone), and pharmacological factors (catecholamine, vasodilator therapy, digitalis) affecting the heart as a whole. Also, for a given load dependence, relaxation will be affected by changes in the conditions of load, which are complex in the whole heart (return force, end systolic deformation, arterial impedence, the law of Laplace, increased intra myocardial pressure due to rapid filling of the coronary reservoir). This double system of control of relaxation, with load dependence predominating is very valuable during the rapid filling phase of the heart. In this phase, relaxation is mainly under load and not in activation dependence. This allows perfect return to the presystolic configuration despite the low filling pressures. The continuous interaction of the cardiac load complex during relaxation and its metabolic and neuro humoral control makes it difficult to assess the relative importance of each factor of this double control. The interaction is even more complex in diseased hearts where regional functional abnormalities are often observed. Similarly, any pharmacological intervention may simultaneously affect load and inactivation dependence.


Quantification of Hypotony Maculopathy Using Spectral-Domain Optical Coherence Tomography.
We provide a free-to-use, open-source algorithm to quantify macular hypotony based on optical coherence tomography (OCT) images. This numerical approach calculates a metric that measures the deviations of Bruch's membrane from a smooth ideal retinal layer. Hypotony maculopathy is a recurrent complication of glaucoma surgical interventions in which extremely low intraocular pressure triggers changes in the shape of retinal layers. Abnormal folds can often be observed in the retina using standard fundoscopy, but OCT is particularly important to appreciate the severity of symptoms at different depths. Despite the need for metrics that could be used for the informed clinical decision to evaluate the progression and resolution of macular hypotony, algorithms that quantify the retinal folds are not available in the literature or included in clinical imaging equipment. The purpose of this work is to introduce a simple algorithm that can be used to assess hypotony maculopathy from OCT B-Scans and volumes and a free, open-source implementation. The pipeline we present is based on a straightforward segmentation of Bruch's membrane complex. The principal idea of quantification is to compute a smoothed version of this complex and analyze the deviations from an ideal interface. Such deviations are then measured and added to create a metric that characterizes each OCT B-Scan. A full OCT volume reconstruction is thus characterized by the average metric obtained from all planes. We tested the metric we proposed against the assessment of 3 experts and obtained a very good correspondence, with Pearson correlation coefficients higher than 0.8. Furthermore, agreement with automatic analysis seemed better than between experts. We describe the pipeline in detail and illustrate the results with a group of patients, comparing baseline images, severe hypotony maculopathy, and a variety of outcomes. The tool we introduce and openly provide fills a clinical gap to quantitatively grade hypotony maculopathy. It offers a metric of relatively simple interpretation that can be used to help clinicians in cases where the regression of symptoms is not obvious to the naked eye. Our pilot study demonstrates reliable results, and an open-source implementation facilitates easy improvements to our algorithm.


Treatment of Bartter's syndrome in early childhood with prostaglandin synthetase inhibitors.
The diagnosis of Bartter's syndrome was made in a 9-month-old boy investigated for poor weight and height gain. Initial treatment with oral potassium supplements and later spironolactone had little or no effect on his growth, although plasma potassium rose to normal after spironolactone. At 33 months indomethacin therapy was started with dramatic results. His symptoms went and his height and weight accelerated into the normal range. In view of the toxicity of indomethacin it was replaced after 12 months by another prostaglandin synthetase inhibitor, ketoprofen, with a satisfactory result. During the change-over period from indomethacin to ketoprofen the expected deterioration in clinical well-being was observed, accompanied by a rise in urinary prostaglandins and plasma renin activity. Prostaglandin synthetase inhibitors provide the best available treatment for Bartter's syndrome.


Use of a self-report constipation questionnaire with older adults in long-term care.
The Patient Assessment of Constipation (PAC), a symptom and quality-of-life inventory, was developed among community-dwelling adults under age 65. The purpose of this study was to evaluate the feasibility and validity of an interview format of the PAC for older adults residing in long-term care. The PAC was administered to 24 nursing home residents aged 65 and older. Cognitive interviewing methods were used to evaluate subjects' understanding and interpretation of PAC items. The PAC interview required an average of 11 min, 15 s, to complete. Administration modifications for this population included rereading Likert anchor points and the stem question for each item. Subject item interpretation was generally accurate, as was interpretation of Likert anchors. The PAC is easily administered and well understood by older adult residents in long-term care and provides a brief method for obtaining constipation-specific symptom and quality-of-life data from this population.


Levels of heparin-releasable TFPI are increased in first-ever lacunar stroke patients.
New insights in the pathophysiology of lacunar stroke (LS) suggest that it is caused by increased permeability of the blood-brain barrier due to endothelial activation. Because endothelial cells are the major production and storage site of tissue factor pathway inhibitor (TFPI), this protein can be used as marker of endothelial activation. In this observational study we measured the different pools of TFPI, as a marker of endothelial function, in first-ever lacunar stroke patients. We determined antigen levels of total and free full-length (FL) TFPI using ELISA in 149 patients and 42 controls. Heparin-releasable free FL TFPI was determined in a random subset of 17 patients and 15 controls. By brain MRI, we classified LS patients as having isolated lacunar infarct (ILA) or silent ischemic lesions (SILs). Plasma levels of total TFPI were highest in patients with SILs compared with those with ILA, but this association disappeared after correction for age and levels of low-density lipoprotein cholesterol. However, levels of heparin-releasable free FL TFPI were higher in patients than in controls. Although ambient plasma levels of total TFPI were not different in subtypes of LS, the increased levels of heparin-releasable TFPI in patients suggest a role of endothelial activation in the pathogenesis of LS.


[Spontaneous occlusion of the circle of Willis (moyamoya disease). Diagnosis and therapy].
Spontaneous occlusion of the circle of Willis (moyamoya disease), a rare cerebrovascular disease, may lead to repeated hemodynamic stroke due to unremitting changes of the basal cranial vessels. Moyamoya can be diagnosed on its specific epidemiological, clinical, and radiological features. In patients with distinctive Moyamoya disease, improvement of cerebral blood supply by means of neurosurgical revascularization enables definitive treatment. This review summarizes current knowledge of the clinical and diagnostic features of this disease and illustrates its pathological and pathophysiological characteristics. Additionally, indications and techniques of current neurosurgical revascularization procedures are illustrated.


Cloacibacterium rupense sp. nov., isolated from freshwater lake sediment.
A Gram-negative, yellow-pigmented bacterium, designated strain R2A-16(T), was isolated from sediment of Rupa Lake in Nepal and analysed using a polyphasic taxonomic approach. Phylogenetic analyses based on 16S rRNA gene sequences showed that strain R2A-16(T) is affiliated to the genus Cloacibacterium of the family Flavobacteriaceae; 16S rRNA gene sequence similarity between strain R2A-16(T) and Cloacibacterium normanense CCUG 46293(T) was 98.07 %. The isolate contained iso-C(15 : 0) (35.6 %) as the major fatty acid and menaquinone MK-6 as the predominant respiratory quinone. The G+C content of the genomic DNA was 33.3 mol%. On the basis of its phenotypic properties and phylogenetic distinctiveness, strain R2A-16(T) represents a novel species of the genus Cloacibacterium, for which the name Cloacibacterium rupense sp. nov. is proposed; the type strain is R2A-16(T) (=CGMCC 1.7656(T) =NBRC 104931(T)).


OSAS surgery and postoperative discomfort: phase I surgery versus phase II surgery.
This study aims to investigate the reasons that discourage the patients affected by OSAS to undergo orthognathic surgery and compares the postoperative discomfort of phase I (soft tissue surgery) and phase II (orthognathic surgery) procedures for treatment of OSAS. A pool of 46 patients affected by OSAS was divided into two groups: "surgery patients" who accepted surgical treatments of their condition and "no surgery patients" who refused surgical procedures. The "surgery patients" group was further subdivided into two arms: patients who accepted phase I procedures (IP) and those who accepted phase II (IIP). To better understand the motivations behind the refusal of II phase procedures, we asked the patients belonging to both the IP group and "no surgery" group to indicate the main reason that influenced their decision to avoid II phase procedures. We also monitored and compared five parameters of postoperative discomfort: pain, painkiller assumption, length of hospitalization, foreign body sensation, and diet assumption following IP and IIP procedures. The main reason to avoid IIP procedures was the concern of a more severe postoperative discomfort. Comparison of the postoperative discomfort following IP versus IIP procedures showed that the former scored worse in 4 out of 5 parameters analyzed. IIP procedures produce less postoperative discomfort. IIP procedures, namely, orthognathic surgery, should be the first choice intervention in patients affected by OSAS and dentoskeletal malformation.


Conceptual design and performance predictions for 2D beam emission spectroscopy turbulence measurements at Wendelstein 7-X.
A conceptual design for a 2D beam emission spectroscopy diagnostic system to measure ion gyro-scale plasma turbulence at Wendeslstein 7-X is described. The conceptual design identifies field-aligned viewing geometries and ports for cross-field turbulence measurements in the neutral beam volume. A 2D sightline grid covers the outer plasma region, and the grid configuration provides sufficient k-space coverage in radial and poloidal directions for ion temperature gradient and trapped-electron mode turbulence measurements. Emission intensity estimates, optical transmission losses, and detector noise levels indicate that the measurements will be sensitive to plasma density fluctuations as small as δn/n ≈ 0.5% with a bandwidth of 1 MHz. Implementation challenges include a small beam emission Doppler shift due to nearly radial heating beams and reduced optical throughput due to collection aperture limitations.