Three-dimensional histochemistry and imaging of human gingiva.
In the present study, 3D histochemistry and imaging methodology is described for human gingiva to analyze its vascular network. Fifteen human gingiva samples without signs of inflammation were cleared using a mixture of 2-parts benzyl benzoate and 1-part benzyl alcohol (BABB), after being immunofluorescently stained for CD31, marker of endothelial cells to visualize blood vessels in combination with fluorescent DNA dyes. Samples were imaged in 3D with the use of confocal microscopy and light-sheet microscopy and image processing. BABB clearing caused limited tissue shrinkage 13 ± 7% as surface area and 24 ± 1% as volume. Fluorescence remained intact in BABB-cleared gingiva samples and light-sheet microscopy was an excellent tool to image gingivae whereas confocal microscopy was not. Histochemistry on cryostat sections of gingiva samples after 3D imaging validated structures visualized in 3D. Three-dimensional images showed the vascular network in the stroma of gingiva with one capillary loop in each stromal papilla invading into the epithelium. The capillary loops were tortuous with structural irregularities that were not apparent in 2D images. It is concluded that 3D histochemistry and imaging methodology described here is a promising novel approach to study structural aspects of human gingiva in health and disease.


[Transhiatal esophagectomy for cardia and esophagus cancer].
To present the result of transhiatal esophagectomies with simultaneous repair. The study included 67 procedures. In 35 cases surgery was carried out for adenocarcinoma of distal esophagus or cardia with high transition to esophagus, in 32 cases - for epidermal carcinoma of the esophagus. Gastric graft and left half of the colon were used in 60 and 7 cases respectively for simultaneous repair. 29 patients underwent transhiatal instrumental esophagectomy using author's original technique.


Effects of tumour necrosis factor-alpha (TNF-alpha), IL-1 beta and monocytes on lymphokine-activated killer (LAK) induction from natural killer (NK) cells and T lymphocytes.
Roles of monocytes and cytokines were investigated on LAK induction from T and NK cells. Monocytes augmented more T-LAK induction than did NK-LAK. Expression of IL-1 beta, TNF-alpha and interferon-gamma (IFN-gamma)-mRNA and their cytokine production were superior in NK cells compared with T cells in parallel with their LAK activities. An increase of TNF-alpha, IL-1 beta and IFN-gamma production was induced by co-culturing NK or T cells with autologous monocytes. The augmentation of T cell cytokine production and T-LAK activity by monocytes was more prominent than that of NK cells. TNF-alpha and IL-1 beta were generated 24 h after IL-2 stimulation, and these cytokines were able to almost substitute for monocytes in LAK induction. Conversely, LAK induction was almost completely suppressed by both anti-IL-1 beta and anti-TNF-alpha antibodies, if they were added within 24 h after the start of the LAK induction. IFN-gamma, which was produced at a later stage, scarcely affected LAK induction in spite of the cooperation with TNF-alpha. The results obtained indicate conclusively that the superiority of NK-LAK depends on their superior productivity of both IL-1 beta and TNF-alpha, and that the up-regulation of LAK induction by monocytes is largely due to the enhanced generation of both cytokines.


[Pivotal role of cell hyperactivation in the pathogenesis of different diseases and its mechanism].
Inflammasomes are key inflammatory signaling platforms that detect several stimulus derived from microbial substances and sterile environmental insults, eliciting pyroptosis and the release of cytokines. Recent studies have found that inflammasomes could also elicit cell hyperactivation which is defined as living cells that release interleukin-1 (IL-1). Hyperactive cells promote long-term IL-1 release and then activate adaptive immune response, which has a pivotal role in the infection of Salmonella and Staphylococcus aureus, as well as in non-lethal inflammatory sepsis. So it is of great significance to explore the pathogenesis of cell hyperactivation in several inflammatory diseases. Here, we summarized the possible pathogenesis of cell hyperactivation in different inflammatory diseases, such as infection, sepsis and acute respiratory distress syndrome (ARDS), providing a theoretical basis for clinical treatment of these diseases.


Referrals for suspected ventriculo-peritoneal shunt dysfunction and necessity for further imaging.
Shunt series (SS) X-rays are often performed as additional routine initial investigations for suspected ventriculo-peritoneal shunt dysfunction, despite low diagnostic utility and radiological implications. We reviewed such referrals at our neurosurgical unit and found an 8.3% sensitivity rate for SS; consistent with current evidence. SS should not be performed routinely.


Measured muscle sodium content in biopsy specimens is a reflection of true intracellular content.
Determined on the basis of small skeletal muscle biopsies, muscle sodium content has a very high coefficient of variation. Furthermore, at least some of the measured sodium must originate from the extracellular space. In order to assess the applicability of the measurement of intracellular sodium on small muscle biopsy specimens, the measured sodium content was related to the content of dry solids in 25 muscle biopsy samples, and compared with the theoretical content of sodium with varying extra- and intracellular water content in biopsy samples. Four of the 25 measurements were clearly outliers. Disregarding these outliers, it was found that muscle sodium content varied with intracellular water content, whereas the theoretical effect of addition of extracellular water could not account for the observed values. The difference depended upon the specified conditions, but the slope of the theoretical regression line (-25.92 mmol x (kg dry weight)(-1) x %(-1), which was closest to the observed slope, -8.55 mmol x (kg dry weight)(-1) x %(-1), differed substantially (p < 0.0001). No association was found between the primarily intracellular ions muscle potassium and muscle magnesium on the one hand and either muscle sodium or muscle water content on the other. The measured sodium content in muscle biopsy specimens, which are freeze-dried and dissected, seems to reflect the true intracellular sodium content to some extent. The total content of sodium seems to be closely related to the content of water within the skeletal muscle cells.


Optimizations for identifying reference genes in bone and cartilage bioengineering.
Reverse transcription quantitative real-time polymerase chain reaction (RT-qPCR) remains one of the best-established techniques to assess gene expression patterns. However, appropriate reference gene(s) selection remains a critical and challenging subject in which inappropriate reference gene selction can distort results leading to false interpretations. To date, mixed opinions still exist in how to choose the most optimal reference gene sets in accodrance to the Minimum Information for Publication of Quantitative Real-Time PCR Experiments (MIQE) guideline. Therefore, the purpose of this study was to investigate which schemes were the most feasible for the identification of reference genes in a bone and cartilage bioengineering experimental setting. In this study, rat bone mesenchymal stem cells (rBMSCs), skeletal muscle tissue and adipose tissue were utilized, undergoing either chondrogenic or osteogenic induction, to investigate the optimal reference gene set identification scheme that would subsequently ensure stable and accurate interpretation of gene expression in bone and cartilage bioengineering. The stability and pairwise variance of eight candidate reference genes were analyzed using geNorm. The V0.15- vs. Vmin-based normalization scheme in rBMSCs had no significant effect on the eventual normalization of target genes. In terms of the muscle tissue, the results of the correlation of NF values between the V0.15 and Vmin schemes and the variance of target genes expression levels generated by these two schemes showed that different schemes do indeed have a significant effect on the eventual normalization of target genes. Three selection schemes were adopted in terms of the adipose tissue, including the three optimal reference genes (Opt3), V0.20 and Vmin schemes, and the analysis of NF values with eventual normalization of target genes showed that the different selection schemes also have a significant effect on the eventual normalization of target genes. Based on these results, the proposed cut-off value of Vn/n + 1 under 0.15, according to the geNorm algorithm, should be considered with caution. For cell only experiments, at least rBMSCs, a Vn/n + 1 under 0.15 is sufficient in RT-qPCR studies. However, when using certain tissue types such as skeletal muscle and adipose tissue the minimum Vn/n + 1 should be used instead as this provides a far superior mode of generating accurate gene expression results. We thus recommended that when the stability and variation of a candidate reference genes in a specific study is unclear the minimum Vn/n + 1 should always be used as this ensures the best and most accurate gene expression value is achieved during RT-qPCR assays.


Susceptibility to oral squamous cell carcinoma: correlation with variants of CYP1A1-MspI, GSTT1, GSTM1, ALDH2, EC-SOD and Lifestyle factors.
In order to investigate the association between polymorphisms in genes encoding metabolizing enzymes (CYP1A1-MspI, EC-SOD (extracellular superoxide dismutase), GSTT1, GSTM1, ALDH2), cigarette and alcohol consumption, and the risk of oral squamous cell carcinoma, we conducted a prospective case-control study comprised of 750 individuals with oral squamous cell carcinoma (OSCC) and 750 healthy individuals. Data about smoking and drinking habits were collected along with other demographic and clinical information. Peripheral blood samples were collected for DNA extraction, and polymerase chain reaction (PCR) and PCR-RFLP (restriction fragment length polymorphism) were used to determine genotypes of CYP1A1, EC-SOD, GSTT1, GSTM1, ALDH2. The results showed that smoking and alcohol consumption were significantly more common among patients than controls (p <0.05). There were significant differences in the genotype distribution for each locus between groups, with the CYP1A1 (m2/ m2), EC-SOD (C/G), GSTT1 [-], GSTM1 [-] and ALDH2 (non G/G) genotypes being more common among patients (p <0.05). Furthermore, the majority of patients had at least two or more variant genotypes, while controls had one or no variant genotype (p <0.05). Finally, multiple variant genotypes combined with smoking, drinking, or both smoking and drinking significantly increased the risk of OSCC, with greater increase for heavier smoking/drinking. In brief, genetic polymorphism of CYP1A1, EC-SOD, GSTT1, GSTM1, and ALDH2 and smoking and drinking history are closely associated with susceptibility to OSCC.


A somatostatin analogue decreases embryonic loss following superovulation in rats by normalizing insulin-like growth factor-I action in the uterus.
This study was designed to determine whether the somatostatin analogue, octreotide, could prevent embryonic loss by normalizing increased uterine insulin-like growth factor-I (IGF-I) action related to hyperoestrogenaemia following superovulation. Superovulated immature and oestradiol-17beta-treated adult rats were infused with 100 or 300 microg/ml of octreotide respectively, or injected daily with 1 or 10 microg of octreotide from day 1 to day 3 of pregnancy. On day 3, embryos were collected from the oviducts and uteri. Uterine luminal fluid was subjected to embryo culture. The amounts of uterine IGF-I and IGF binding proteins (IGFBP) were determined by radioimmunoassay and ligand binding assay respectively. Octreotide infusion normalized uterine IGF-I action following superovulatory and oestradiol-17beta treatment, by reducing IGF-I concentrations and increasing IGFBP concentrations. Octreotide infusion increased the number of normal embryos by 2.7-fold and 1.7-fold in superovulated and oestradiol-17beta-treated rats respectively, and reversed the detrimental effects of uterine luminal fluid on embryonic development caused by superovulatory and oestradiol-17beta treatment. Daily injections with octreotide had similar but reduced effects in all parameters examined in both treatment groups. In conclusion, octreotide may reduce embryonic loss, at least in part, by normalizing IGF-I action following superovulation.


Percutaneous radiofrequency ablation of small renal cell carcinoma: technique, complications, and outcomes.
Imaging-guided radiofrequency ablation (RFA) is an option for treatment in patients with early-stage small renal cell carcinomas (RCCs). RFA has been introduced to treat focal renal tumors, particularly incidental lesions <3 cm in elderly patients and those with comorbid conditions. Other uses have included treatment in patients with von Hippel-Lindau syndrome and other diseases that predispose patients to multiple renal carcinomas, where renal parenchymal preservation is desired. It appears that this technique has a low complication rate, preserves renal function, is well tolerated by the patients, and, in a high percentage of patients, can eradicate small renal tumors. Techniques, patient selection, complications, and results are discussed.


The detection and phylogenetic analysis of porcine deltacoronavirus from Guangdong Province in Southern China.
Porcine deltacoronavirus (PDCoV) is a newly discovered coronavirus that causes diarrhoea, vomiting and dehydration in sucking and nursing piglets. It was first reported in Hong Kong in 2012 and has since been discovered in the United States, Canada, South Korea, mainland China, Thailand and Laos. PDCoV has been experimentally proved to lead to diarrhoea in swine and it was detected positive in pigs in Guangdong, southern China. In our study, 252 faecal and intestinal samples from sucking piglets and sows with diarrhoea were surveyed for common enteric viruses. We found a prevalence of PDCoV (21.8%), porcine epidemic diarrhoea virus (65.5%), transmissible gastroenteritis virus (0%), rotavirus group A (25.0%) and porcine kobuvirus (68.7%). We isolated 13 PDCoV strains and discovered that PDCoV infections were often co-infections with kobuvirus rather than the commonly linked porcine epidemic diarrhoea virus. Phylogenetic analysis of S gene and N gene revealed that 11 of 13 PDCoV strains belonged to Chinese lineage. As for the left two strains, one single strain (CHN-GD16-05) belonged to American and Korean lineages while another strain (CHN-GD16-03) was similar to a Thai strain, but only in the S gene. This suggested a possible recombination event between the Thai and the newly described Chinese strain.


Unintentional child poisonings treated in United States hospital emergency departments: national estimates of incident cases, population-based poisoning rates, and product involvement.
The goals were to develop national estimates of unintentional child poisoning cases treated in US hospital emergency departments, to determine population-based poisoning rates, and to evaluate characteristics of the victims and the products involved. Cases reported through the US Consumer Product Safety Commission National Electronic Injury Surveillance System, involving a national probability sample of US hospital emergency departments, were used as a basis for developing national estimates of product-related poisonings involving children<5 years of age treated in US hospital emergency departments in 2004. There were an estimated 86194 child poisoning incidents treated in US hospital emergency departments in 2004, amounting to 429.4 poisonings per 100000 children. Approximately 70% of the poisonings involved children 1 or 2 years of age, slightly more than one half involved boys, and 13.3% resulted in hospital admission. Approximately 59.5% of the poisonings involved oral prescription drugs, oral nonprescription drugs, or supplements. Other major product categories resulting in poisonings included cleaning products (13.2%), drugs and ointment preparations intended for external use (4.9%), and personal care products (4.7%). Approximately 54.7% of the poisonings involved products already subject to child-resistant packaging requirements under the Poison Prevention Packaging Act. Despite advances in recent years, unintentional child poisonings remain an important public health concern. The circumstances surrounding poisonings need to be evaluated further, and intervention strategies need to be developed.


On the Possible Trade-Off between Shoot and Root Biomass in Wheat.
Numerous studies have shown that under a limited water supply, a larger root biomass is associated with an increased above-ground biomass. Root biomass, while genetically controlled, is also greatly affected by the environment with varying plasticity levels. In this context, understanding the relationship between the biomass of shoots and roots appears prudent. In this study, we analyze this relationship in a large dataset collected from multiple experiments conducted up to different growth stages in bread wheat (Triticum aestivum L.) and its wild relatives. Four bread wheat mapping populations as well as wild and domesticated members of the Triticeae tribe were evaluated for the root and shoot biomass allocation patterns. In the analyzed dataset the root and shoot biomasses were directly related to each other, and to the heading date, and the correlation values increased in proportion to the length of an experiment. On average, 84.1% of the observed variation was explained by a positive correlation between shoot and root biomass. Scatter plots generated from 6353 data points from numerous experiments with different wheats suggest that at some point, further increases in root biomass negatively impact the shoot biomass. Based on these results, a preliminary study with different water availability scenarios and growth conditions was designed with two cultivars, Pavon 76 and Yecora Rojo. The duration of drought and water level significantly affected the root/shoot biomass allocation patterns. However, the responses of the two cultivars were quite different, suggesting that the point of diminishing returns in increasing root biomass may be different for different wheats, reinforcing the need to breed wheats for specific environmental challenges.


Florence Nightingale: The Making of a Hospital Reformer.
The first of two articles is to show how Florence Nightingale became a leading, effective hospital reformer. The aim of the first paper is to relate how Nightingale was influenced by the great defects in the war hospitals of the Crimean War (1854-1856) and how she learned the lessons from those defects to set a different course. The article shows how her famous Notes on Nursing is a positive treatment of the lessons learned, turning the sanitary defects, notably in ventilation, into chapters of the book. The importance of the pavilion model of hospital design is highlighted. There is coverage of the advances made by Semmelweis at the Vienna General Hospital. This is a purely historical study drawing on the extensive publications by Nightingale, augmented by her (massive) surviving correspondence and notes. The search for archival materials was done for the publication of the 16-volume Collected Works of Florence Nightingale, written by the author of this article. The collected works was peer reviewed, and the research process succeeded in locating material in more than 200 archives worldwide.


Preoperative chemoradiotherapy in locally advanced cervical cancer: long-term outcome and complications.
To demonstrate the efficacy and feasibility of preoperative chemoradiotherapy in a consecutive series of 100 locally advanced cervical cancer (LACC) patients. Between October 1997 and December 2004, 100 LACC patients were consecutively staged and treated at the Catholic University of the Sacred Heart of Rome. Radiotherapy was administered to the whole pelvic region (1.8 Gy/day, totaling 39.6 Gy) in combination with cisplatin (20 mg/m(2)) and 5-FU (1000 mg/m(2)) (both on days 1-4 and 27-30). Radical surgery was performed 5-6 weeks after the end of the treatment. A clinical complete or partial response was observed in 96 patients (56 and 40, respectively). Radical surgery was performed in 95 patients and an overall complication rate of 12.6% was observed in the early postoperative time. At pathological examination, 43 of 95 patients (45.2%) undergoing radical surgery showed complete response to treatment, 28 patients (29.5%) only had a microscopic disease, 18 patients (19%) had a partial response and 6 (6.3%) had no change of disease. With a median follow-up time of 25 months, the 5-year disease-free survival was 76% and the 5-year overall survival was 78%. These data confirm the possibility of achieving encouraging rates of local control and OS in LACC patients submitted to chemoradiation plus surgery, with a low rate of toxicity and complications.


The effect of a cationic porphyrin on Pseudomonas aeruginosa biofilms.
Current studies have indicated the utility of photodynamic therapy using porphyrins in the treatment of bacterial infections. Photoactivation of porphyrins results in the production of singlet oxygen ((1)O(2)) that damages biomolecules associated with cells and biofilms, e.g., proteins, polysaccharides, and DNA. The effect of a cationic porphryin on P. aeruginosa PAO1 biofilms was assessed by exposing static biofilms to 5,10,15,20-tetrakis(1-methyl-pyridino)-21H,23H-porphine, tetra-p-tosylate salt (TMP) followed by irradiation. Biofilms were visualized using confocal laser scanning microscopy (CLSM) and cell viability determined using the LIVE/DEAD BacLight viability assay and standard plate counts. At a concentration of 100 μM TMP, there was substantial killing of P. aeruginosa PAO1 wild-type and pqsA mutant biofilms with little disruption of the biofilm matrix or structure. Exposure to 225 μM TMP resulted in almost complete killing as well as the detachment of wild-type PAO1 biofilms. In contrast, pqsA mutant biofilms that contain less extracellular DNA remained intact. Standard plate counts of cells recovered from attached biofilms revealed a 4.1-log(10) and a 3.9-log(10) reduction in viable cells of wild-type PAO1 and pqsA mutant strains, respectively. Our results suggest that the action of photoactivated TMP on P. aeruginosa biofilms is two-fold: direct killing of individual cells within biofilms and detachment of the biofilm from the substratum. There was no evidence of porphyrin toxicity in the absence of light; however, biofilms pretreated with TMP without photoactivation were substantially more sensitive to tobramycin than untreated biofilms.


[A pharmacological analysis of the anxiolytic activity of uridine].
The effects of prazosin (0.6 and 3 mg/kg), propranolol (5 mg/kg), haloperidol (0.1 and 0.5 mg/kg) and ciproheptaline (0.3 and 0.6 mg/kg) on the anxiolytic (anticonflict) action of a pyrimidine ribonucleoside uridine, a hypothetic endogenic regulator of anxiety states were studied in the experiments on male CBWA mice. It was found that the degree of the anxiolytic effect of uridine decreases at the blockade of alpha 1, beta 2, D2 and H1-receptors and significantly increases at the blockade of 5-HT2-receptors. This suggests the involvement of the mentioned receptors in the processes of realization of uridine anxiolytic activity as well as the presence of the central serotonin-negative component in the mechanisms of action of uridine.


[Unusual haemostasis disorder in a seriously burned patient].
We report the case of a woman severely burned during a gas explosion with an unusual disorder of haemostasis. The prothrombin time was very elevated, essentially by a drop in factor II. This abnormality persisted for a long time after the correction of the dilutional and consumption coagulopathy which followed immediately the burn. Blood analysis found fluindione and benzodiazepine derivatives. Such a disorder during a potentially suicidal burn must evoke a toxic aetiology.


MobiDis: Toward a Patient Centric Healthcare Information System.
The paper presents some results of the MobiDis project. MobiDis is an information system that includes healthcare consumers and providers in a unique, virtual organisation aimed at promoting a patient centric paradigm in healthcare. It allows logons from desktop or laptop computers, as well as wireless PDAs or tablet PCs connected to Internet. In MobiDis the clinical data of each consumer are stored in the consumer's virtual healthcare record (VHR), a highly structured entity that exists on the network and is simultaneously updated with information from multiple locations. The MobiDis architecture creates an environment for VHRs by providing them with a large variety of services. In order to prove that our proposed architectural solution meets the project goals a prototype was developed. The paper describes the MobiDis architecture and the VHR services, and briefly presents the prototype.


The addition of oat fiber and natural alternative sweeteners in the manufacture of plain yogurt.
Calorie-reduced yogurts that were fortified with 1.32% oat fiber were prepared from lactose-hydrolyzed milk, alone and supplemented with 2 and 4% sucrose or with 1.6, 3.6, and 5.5% fructose. Treated samples were compared with unsweetened yogurt and with yogurts sweetened with 2, 4, and 6% sucrose. Addition of 5.5% fructose increased fermentation time by 60%, slowing down the production of lactic, pyruvic, acetic, and propionic acids and the consumption of hippuric and orotic acids. Lactose hydrolysis had an inhibitory effect on starter activity at the beginning of fermentation and a stimulatory effect at the end of fermentation. Fiber addition led to increases in concentrations of acetic and propionic acid. Lactobacilli counts were lower in samples treated with fructose. The use of hydrolyzed milk had a stimulatory effect on total bacteria and lactobacilli counts throughout the cold storage period. After 28 d of storage, lactobacilli counts were consistently higher in fiber-fortified yogurts, but total bacteria counts were lower. Apparent viscosity increased with the addition of sweetener and fiber. Lactose-hydrolyzed and fructose yogurts had the highest viscosity values. Samples sweetened with sucrose received the highest scores for flavor. Fiber addition decreased overall flavor quality. The lactose-hydrolyzed yogurts received the highest flavor scores, independent of fiber fortification. Fiber addition improved the body and texture of unsweetened yogurts but lowered overall scores for body and texture in yogurts sweetened with sucrose.


Perinatal protein deprivation facilitates morphine cross-sensitization to cocaine and enhances ΔFosB expression in adult rats.
Previous studies have indicated that neural changes induced by early nutritional insult cause an altered response to pharmacological treatments, including addictive drugs. This study evaluates the influence of perinatal protein malnutrition in developing cross-sensitization to cocaine-induced rewarding effects in animals pre-exposed to morphine. Different groups of well-nourished (C-rats) and protein-deprived animals (D-rats) were treated twice a day for three days with increasing doses of morphine or with saline. After 3days, the incentive motivational effects of cocaine were assessed in a Conditioned Place Preference paradigm in both groups. In saline pre-treated animals, dose-response curves to cocaine revealed a conditioning effect in D-rats at doses of 5, 7.5 and 10mg/kg, while this effect was observed in C-rats only with 10 and 15mg/kg. Furthermore, when animals of both groups were pre-treated with escalating doses of morphine, cross-sensitization to the conditioning effect of cocaine was elicited only in D-rats with low doses of cocaine (5 and 7.5mg/kg). In contrast, under the same experimental conditions, C-rats show no cross-sensitization. To correlate this differential rewarding response with a molecular substrate linked to the behavioral changes observed after repeated drug exposure, ΔFosB expression was assessed in different brain regions. D-rats showed a significant increase in this transcription factor in the nucleus accumbens, amygdala and medial prefrontal cortex. These results demonstrated that perinatal protein deprivation facilitates rewarding effects and the development of cross-sensitization to cocaine, which correlates with an upregulation of ΔFosB in brain areas related to the reward circuitry.


[Hormone tolerance mechanism of prostate cancer].
One of the delicate problems in a cancer treatment is the acquisition of a tolerance for a medicine. Antiandrogen treatment is valid in treatment of prostate cancer, but the disease develops into hormone-refractory prostate cancer after a uniformity period. The molecular mechanisms of recurrence and endocrine therapy failure in prostate cancer have remained unclear. In this paper we discuss the hormone-refractory prostate cancer that has been reported by many investigators.


Development of loop-mediated isothermal amplification with Plasmodium falciparum unique genes for molecular diagnosis of human malaria.
In order to achieve better outcomes for treatment and in the prophylaxis of malaria, it is imperative to develop a sensitive, specific, and accurate assay for early diagnosis of Plasmodium falciparum infection, which is the major cause of malaria. In this study, we aimed to develop a loop-mediated isothermal amplification (LAMP) assay with P. falciparum unique genes for sensitive, specific, and accurate detection of P. falciparum infection. The unique genes of P. falciparum were randomly selected from PlasmoDB. The LAMP primers of the unique genes were designed using PrimerExplorer V4. LAMP assays with primers from unique genes of P. falciparum and conserved 18S rRNA gene were developed and their sensitivity was assessed. The specificity of the most sensitive LAMP assay was further examined using genomic DNA from Plasmodium vivax, Plasmodium yoelii and Toxoplasma gondii. Finally, the unique gene-based LAMP assay was validated using clinical samples of P. falciparum infection cases. A total of 31 sets of top-scored LAMP primers from nine unique genes were selected from the pools of designed primers. The LAMP assay with PF3D7_1253300-5 was the most sensitive with the detection limit 5 parasites/μl, and it displayed negative LAMP assay with the genomic DNA samples of P. vivax, P. yoelii, and T. gondii. The LAMP assay with PF3D7_0112300 (18S rRNA) was less sensitive with the detection limit 50 parasites/μl, and it displayed negative LAMP assay with the genomic DNA samples of P. yoelii and T. gondii, but displayed positive LAMP detection with P. vivax. The positive detection rate of the LAMP assay with PF3D7_1253300-5 was 90% (27/30), higher than that (80%, 24/30) of the positive rate of PF3D7_0112300 (18S rRNA) in examining clinical samples of P. falciparum infection cases. The LAMP assay with the primer set PF3D7_1253300-5 was more sensitive, specific, and accurate than those with PF3D7_0112300 (18S rRNA) in examining P. falciparum infection, and therefore it is a promising tool for diagnosis of P. falciparum infection.


Gut sterilization to prevent nosocomial infection.
Nosocomial infections, particularly respiratory tract infections, are common complications that not only lengthen ICU stay but also increase mortality. Since it has been recognized that colonization of the oropharynx and proximal gastrointestinal tract by Gram-negative bacteria is predictive of subsequent pneumonia, attempts have been directed toward sterilizing the upper gastrointestinal tract of pathogenic organisms. Selective decontamination of the digestive tract (SDD) utilizes the administration of topical, nonabsorbable antimicrobials to the gastrointestinal tract and parenteral antibiotics to eradicate the carriage of pathogenic bacteria and, hopefully, improve outcome in critically ill patients. This review of 19 recently published studies indicates that SDD significantly reduces colonization with pathogenic bacteria and nosocomial respiratory tract infections. Reduction in mortality, however, is not a proven effect of SDD.


n-alkanes on Pt(111) and on C(0001)Pt(111): chain length dependence of kinetic desorption parameters.
We have measured the desorption of seven small n-alkanes (C(N)H(2N+2), N=1-4,6,8,10) from the Pt(111) and C(0001) surfaces by temperature programed desorption. We compare these results to our recent study of the desorption kinetics of these molecules on MgO(100) [J. Chem. Phys. 122, 164708 (2005)]. There we showed an increase in the desorption preexponential factor by several orders of magnitude with increasing n-alkane chain length and a linear desorption energy scaling with a small y-intercept value. We suggest that the significant increase in desorption prefactor with chain length is not particular to the MgO(100) surface, but is a general effect for desorption of the small n-alkanes. This argument is supported by statistical mechanical arguments for the increase in the entropy gain of the molecules upon desorption. In this work, we demonstrate that this hypothesis holds true on both a metal surface and a graphite surface. We observe an increase in prefactor by five orders of magnitude over the range of n-alkane chain lengths studied here. On each surface, the desorption energies of the n-alkanes are found to increase linearly with the molecule chain length and have a small y-intercept value. Prior results of other groups have yielded a linear desorption energy scaling with chain length that has unphysically large y-intercept values. We demonstrate that by allowing the prefactor to increase according to our model, a reanalysis of their data resolves this y-intercept problem to some degree.


Characteristics of Computed Tomography Images for Patients with Acute Liver Injury Caused by Sepsis under Deep Learning Algorithm.
This study was aimed at exploring the application of image segmentation based on full convolutional neural network (FCN) in liver computed tomography (CT) image segmentation and analyzing the clinical features of acute liver injury caused by sepsis. The Sigmoid function, encoder-decoder, and weighted cross entropy loss function were introduced and optimized based on FCN. The Dice value, precision, recall rate, volume overlap error (VOE), relative volume difference (RVD), and root mean square error (RMSE) values of the optimized algorithms were compared and analyzed. 92 patients with sepsis were selected as the research objects, and they were divided into a nonacute liver injury group (50 cases) and acute liver injury group (42 cases) based on whether they had acute liver injury. The differences in the proportion of patients with different disease histories, the proportion of patients with different infection sites, the number of organ failure, and the time of admission to intensive care unit (ICU) were compared between the two groups. It was found that the optimized window CT image Dice value after preprocessing (0.704 ± 0.06) was significantly higher than the other two methods (P < 0.05). The Dice value, precision, and recall rate of the optimized-FCN algorithm were (0.826 ± 0.06), (0.91 ± 0.08), and (0.88 ± 0.09), respectively, which were significantly higher than other algorithms (P < 0.05). The VOE, RVD, and RMSE values were (21.19 ± 1.97), (10.45 ± 1.02), and (0.25 ± 0.02), respectively, which were significantly lower than other algorithms (P < 0.05). The proportion of patients with a history of drinking in the nonacute liver injury group was lower than that in the acute liver injury group (P < 0.05), and the proportion of patients with a history of hypotension was greatly higher than that in the nonacute liver injury group (P < 0.01). CT images of sepsis patients with acute liver injury showed that large areas of liver parenchyma mixed with high-density hematoma, the number of organ failures, and the length of stay in ICU were significantly higher than those in the nonacute liver injury group (P < 0.05). It showed that the optimization algorithm based on FCN greatly improved the performance of CT image segmentation. Long-term drinking, low blood pressure, number of organ failures, and length of stay in ICU were all related to sepsis and acute liver injury. Conclusion in this study could provide a reference basis for the diagnosis and prognosis of acute liver injury caused by sepsis.


[Case of intravascular lymphoma with a longitudinal spinal lesion diagnosed by multiple biopsies].
A 45-year-old man was admitted to our hospital with flaccid paraplegia. Neurological examination at a local hospital, 2 months before admission to our hospital, showed sensory impairment of the right posterior surface of the thigh and a decreased Achilles tendon reflex. Spinal magnetic resonance imaging (MRI) showed a T₂ weighted high-intensity area at the Th10-11 level that was more pronounced in the gray matter. The patient developed flaccid paraparesis and urinary retention. No improvement was observed after 2 rounds of methylprednisolone (mPSL) pulse therapy. Spinal cord biopsy showed demyelinated axons and myelinophagia without any tumorous lesion. Myelopathy exacerbated, and hence, plasma exchange was performed. However, this was ineffective. We suspected that myelopathy was caused by intravascular lymphoma (IVL) because of the presence of a low-grade fever, weight loss, and elevated serum soluble IL-2 receptor titers. Random biopsies, including skin, rectal, bone marrow, muscle, and renal biopsies, and splenectomy were performed to make a definite diagnosis of IVL myelopathy. Among these biopsies, the diagnosis of IVL myelopathy was confirmed from the renal specimen. The patient underwent chemotherapy at our hospital, and the IVL remitted. The results of this study confirm that sufficient systemic investigation by using tissue biopsy specimens should be performed in order to confirm the diagnosis of IVL myelopathy.


Anton's syndrome as a presentation of decompression illness.
We present a case of a patient with Anton's syndrome due to decompression illness (DCI) after recreational scuba diving. Visual anosognosia, or denial of loss of vision, which is associated with lack of awareness regarding visual loss in the setting of cortical blindness, is known as Anton's syndrome (also termed Anton-Babinski syndrome). Our patient presented with progressive neurological DCI treated with repeated recompression. The anosogosia resolved after 48 h. Subsequent echocardiography revealed a persistent (patent) foramen ovale.


Edwardsiella tarda bacteremia and septic arthritis in a patient with diabetes mellitus.
Edwardsiella tarda is an uncommon pathogen in the family Enterobacteriaceae which usually infects patients with underlying diseases. Its habitats include fresh water, a variety of animals and human feces. We report a case of E. tarda bacteremia and septic arthritis with underlying diabetes mellitus, the first found in Thailand.


Phylogenetic and immunoinformatic analysis of VP4, VP7, and NSP4 genes of rotavirus strains circulating in children with acute gastroenteritis in Indonesia.
Rotavirus is a major cause of diarrhea in Indonesian children. However, rotavirus vaccines have not been introduced in the national immunization program of Indonesia. Understanding the genetic diversity and conserved antigenic regions of circulating strains are therefore essential to assess the potential efficacy of rotavirus vaccines. We collected fecal samples from hospitalized children less than 5 years of age with acute diarrhea. Rotavirus genotyping was performed by reverse transcriptase polymerase chain reaction, followed by sequencing of the VP4, VP7, and NSP4 genes of representative strains. Phylogenetic analysis was performed to investigate their relationship with globally circulating strains. Conservational analysis, immunoinformatics, and epitope mapping in comparison to vaccine strains were also performed. The sequence analyses showed that differences of multiple amino acid residues existed between the VP4, VP7, and NSP4 antigenic regions of the vaccine strains and the Indonesian isolates. However, many predicted conserved epitopes with higher antigenicity were observed in the vaccine and Indonesian strains, conferring the importance of these epitopes. The identified epitopes showed a higher potential of rotavirus vaccine to be employed in Indonesia. It could also be helpful to inform the design of a peptide vaccine based on the conserved regions and epitopes in the viral proteins.


Introduction by molecular cloning of artifactual inverted sequences at the 5' terminus of the sense strand of bovine parathyroid hormone cDNA.
To study the structure and function of the gene for parathyroid hormone, we obtained recombinant plasmids containing bovine parathyroid hormone cDNA. The nucleotide sequence at the 5' terminus (relative to the sense strand) of the cDNA insert in a recombinant plasmid, pPTHi4, was different from that previously reported for the bovine parathyroid hormone cDNA insert of another recombinant plasmid, pPTHm1 [Kronenberg, H. M., McDevitt, B. F., Majzoub, J. A., Nathans, J., Sharp, P. A., Potts, J. T., Jr. & Rich, A. (1979) Proc. Natl. Acad. Sci. USA 76, 4981-4985]. The first 50 nucleotides of the pPTHm1 insert were an inverted complement of nucleotides 2-51 of the pPTHi4 insert. the cDNA insert of another plasmid, pPTHi8, contained a sequence identical to nucleotides 2-51 of the pPTHi4 insert but also contained an additional 42 bases at the 5' terminus. The first 41 bases of the pPTHi8 insert were an inverted repeat of an internal sequence of the pPTHi4 insert corresponding to nucleotides 184-224. Restriction endonuclease analysis of pPTHi8 indicated that the internal sequence corresponding to this region was retained. The nucleotide sequence of a restriction fragment hybridized to parathyroid hormone mRNA and extended toward the 5' terminus of the mRNA with reverse transcriptase confirmed that the sequence at the 5' terminus of the pPTHi4 insert was an accurate copy of the parathyroid hormone mRNA sequence. These data suggest that two types of sequence rearrangements may occur at the 5' terminus, as occurred in pPTHm1, and (ii) an inverted repeat of an internal sequence, as occurred in pPTHi8.


Helmet-Induced Occipital Neuralgia in a Military Aviator.
Headaches among military personnel are very common and headgear wear is a frequently identified culprit. Helmet wear may cause migrainous headaches, external compression headache, other primary cranial neuralgias, and occipital neuralgia. The clinical features and the response to treatment allow distinction between the different types of headaches. Headaches among aviators are particularly concerning as they may act as distractors while flying and the treatment options are often incompatible with flying status. A 24-yr-old door gunner presented with suboccipital pain associated with the wear of his helmet. He described the pain as a paroxysmal stabbing sensation coming in waves. The physical exam and history supported the diagnosis of primary occipital neuralgia. Systemic pharmacological options were discussed with the soldier, but rejected due to his need to remain in flying status. An occipital nerve block was performed with good clinical results, supporting the diagnosis of occipital neuralgia and allowing him to continue as mission qualified. Occipital neuralgia can be induced by helmet wear in military personnel. Occipital nerve block can be performed in the deployed setting, allowing the service member to remain mission capable and sparing him/her from systemic side effects.Chalela JA. Helmet-induced occipital neuralgia in a military aviator. Aerosp Med Hum Perform. 2018; 89(4):409-410.


Preventing chronic disease among the aged: a call for evidence-based design research.
The quality of the built environment and the public health and well-being of its inhabitants are profoundly interwoven. Among all age groups, the aged are particularly susceptible to disengagement or avoidance of the built environment and the physical exercise options it affords; this can have a deleterious influence on personal health. In this discussion, concepts drawn from the fields of architecture, landscape research, urban and regional planning, and environmental gerontology are drawn together in the context of a hybrid conceptual and operative model. This model is put forth to assist in the acquisition of knowledge in the field to further understanding of how chronic diseases among the aged can be reduced through the provision and utilization of sufficiently supportive outdoor physical activity options in the everyday environment. This hybrid model, the Prospect-Refuge Competency Index (PRCI), combines key elements of prospect-refuge theory and environmental press-competency theory. It can be applied to diverse settings and user constituencies. The discussion concludes with the presentation of a set of hypotheses for the neighborhood/community level and the residential/exterior environs level of inquiry.


The cornerstone K-RAS mutation in pancreatic adenocarcinoma: From cell signaling network, target genes, biological processes to therapeutic targeting.
RAS belongs to the super family of small G proteins and plays crucial roles in signal transduction from membrane receptors in the cell. Mutations of K-RAS oncogene lead to an accumulation of GTP-bound proteins that maintains an active conformation. In the pancreatic ductal adenocarcinoma (PDAC), one of the most deadly cancers in occidental countries, mutations of the K-RAS oncogene are nearly systematic (>90%). Moreover, K-RAS mutation is the earliest genetic alteration occurring during pancreatic carcinogenetic sequence. In this review, we discuss the central role of K-RAS mutations and their tremendous diversity of biological properties by the interconnected regulation of signaling pathways (MAPKs, NF-κB, PI3K, Ral…). In pancreatic ductal adenocarcinoma, transcriptome analysis and preclinical animal models showed that K-RAS mutation alters biological behavior of PDAC cells (promoting proliferation, migration and invasion, evading growth suppressors, regulating mucin pattern, and miRNA expression). K-RAS also impacts tumor microenvironment and PDAC metabolism reprogramming. Finally we discuss therapeutic targeting strategies of K-RAS that have been developed without significant clinical success so far. As K-RAS is considered as the undruggable target, targeting its multiple effectors and target genes should be considered as potential alternatives.


Yarrowia lipolytica cells mutant for the PEX24 gene encoding a peroxisomal membrane peroxin mislocalize peroxisomal proteins and accumulate membrane structures containing both peroxisomal matrix and membrane proteins.
Peroxins are proteins required for peroxisome assembly and are encoded by the PEX genes. Functional complementation of the oleic acid-nonutilizing strain mut1-1 of the yeast Yarrowia lipolytica has identified the novel gene, PEX24. PEX24 encodes Pex24p, a protein of 550 amino acids (61,100 Da). Pex24p is an integral membrane protein of peroxisomes that exhibits high sequence homology to two hypothetical proteins encoded by the open reading frames YHR150W and YDR479C of the Saccharomyces cerevisiae genome. Pex24p is detectable in wild-type cells grown in glucose-containing medium, and its levels are significantly increased by incubation of cells in oleic acid-containing medium, the metabolism of which requires intact peroxisomes. pex24 mutants are compromised in the targeting of both matrix and membrane proteins to peroxisomes. Although pex24 mutants fail to assemble functional peroxisomes, they do harbor membrane structures that contain subsets of peroxisomal proteins.


Roles of the Aspergillus nidulans homologues of Tup1 and Ssn6 in chromatin structure and cell viability.
For three different carbon catabolite repressible promoters, alcA, alcR and the bidirectional promoter prnD-prnB, a deletion of rcoA, the Aspergillus nidulans homologue of TUP1, does not result in carbon catabolite derepression. Surprisingly, it results in disruption of the chromatin default structure of alcR and prnD-prnB promoters. In these promoters, and at variance with the wild type, repression occurs in the absence of nucleosome positioning. For alcR, repression occurs together with a nucleosome pattern identical to that found under conditions of full expression, and for prnD-prnB it occurs with a novel pattern that does not correspond to the pattern seen under conditions of repression in a wild-type strain. Deletion of the putative RcoA partner, SsnF, is lethal in A. nidulans.


Caryolan-1-ol, an antifungal volatile produced by Streptomyces spp., inhibits the endomembrane system of fungi.
Streptomyces spp. have the ability to produce a wide variety of secondary metabolites that interact with the environment. This study aimed to discover antifungal volatiles from the genus Streptomyces and to determine the mechanisms of inhibition. Volatiles identified from Streptomyces spp. included three major terpenes, geosmin, caryolan-1-ol and an unknown sesquiterpene. antiSMASH and KEGG predicted that the volatile terpene synthase gene clusters occur in the Streptomyces genome. Growth inhibition was observed when fungi were exposed to the volatiles. Biological activity of caryolan-1-ol has previously not been investigated. Fungal growth was inhibited in a dose-dependent manner by a mixture of the main volatiles, caryolan-1-ol and the unknown sesquiterpene, from Streptomyces sp. S4-7. Furthermore, synthesized caryolan-1-ol showed similar antifungal activity. Results of chemical-genomics profiling assays showed that caryolan-1-ol affected the endomembrane system by disrupting sphingolipid synthesis and normal vesicle trafficking in the fungi.


Increased alanine uptake and lipid synthesis from alanine in isolated hepatocytes of Wistar-Kyoto fatty rats: an inhibitory effect of biguanides.
To examine the pathophysiological characteristics of non-insulin-dependent diabetes mellitus, alanine metabolism in isolated hepatocytes of male Wistar-Kyoto (WKY) fatty rats (genetically obese and hyperglycemic) and their lean littermates was investigated. The effects of glucagon and the biguanides, metformin and buformin, on alanine metabolism were also studied by measuring alanine uptake and lipid synthesis from alanine. WKY fatty rats showed higher plasma insulin and lipid concentrations than lean rats at 5 as well as at 12 weeks of age. Alanine uptake into hepatocytes was increased in fatty rats only at 12 weeks of age compared with lean rats. Lipid synthesis from alanine in hepatocytes was increased in fatty rats at 5 and 12 weeks of age compared with lean rats. Glucagon increased alanine uptake into hepatocytes but did not affect lipid synthesis from alanine in both fatty and lean rats. Low concentrations (0.1 mM) of biguanides decreased lipid synthesis from alanine only in fatty rats without inhibiting alanine uptake into hepatocytes. These observations suggest that lipid synthesis from alanine in hepatocytes of WKY fatty rats is accelerated prior to the onset of diabetes mellitus, which might be associated with the development of diabetes, and that an inhibitory effect on increased lipid synthesis is one of the pharmacodynamic actions of biguanides.


Effect of water on exenatide acylation in poly(lactide-co-glycolide) microspheres.
Peptide or protein degradation often occurs when water flows into the dosage form. The aim of this study was to investigate the effect of water on exenatide acylation in poly(lactide-co-glycolide) (PLGA) microspheres. Exenatide-loaded PLGA microspheres were incubated at different relative humidities (RH) as well as in solutions of different pH for 20 days. The stability of exenatide was monitored using HPLC and HPLC-MS analysis. The alteration of exenatide conformation caused by water was investigated by FT-IR spectroscopy. Exenatide and glycolide were incubated in DMSO-water solutions to verify the effect of exenatide conformation state on the peptide acylation. Exenatide was relatively stable in microspheres at lower RH, and the absorbed water could act as a plasticizer and thus promote the peptide acylation by PLGA. However, when the microspheres were incubated at 100% RH, the excessively absorbed water could cause conformation recovery of exenatide and play an inhibitory effect on acylation. The formation of acylated exenatide incubated in acetate buffer saline of pH 6.0 was more than that of pH 4.5 and 3.0. Stability studies of exenatide in glycolide solutions showed that exenatide in nonnative monomer state was easier to be acylated by eletrophiles than that in aggregation state.


Studies on the preparation and characterization of monodisperse polystyrene latices. VI. Preparation of zwitterionic latices.
The preparation of zwitterionic latex particles is reported by using a mixed anionic and cationic initiator system without requiring surface-active agents. Isoelectric points were found from microelectrophoresis experiments and were in the pH range of 3.5-5. Close to the isoelectric point, the latices coagulated as expected, and good stability was achieved outside this narrow range. This range of stability was in good agreement with predictions from current theory. Redispersion after coagulation was found to be difficult as was expected for a hydrophobic colloid. The electrokinetic behavior did not result in the maximum in zeta potential at an electrolyte concentration of 1 mM unlike the situation for other hydrophobic polystyrene latex particles, and hence these systems may be even better models for other colloidal studies.


Comparison of kinetic Monte Carlo and molecular dynamics simulations of diffusion in a model glass former.
We present results from kinetic Monte Carlo (KMC) simulations of diffusion in a model glass former. We find that the diffusion constants obtained from KMC simulations have Arrhenius temperature dependence, while the correct behavior, obtained from molecular dynamics simulations, can be super-Arrhenius. We conclude that the discrepancy is due to undersampling of higher-lying local minima in the KMC runs. We suggest that the relevant connectivity of minima on the potential energy surface is proportional to the energy density of the local minima, which determines the "inherent structure entropy." The changing connectivity with potential energy may produce a correlation between dynamics and thermodynamics.


A homologous gene of OsREL2/ASP1, ASP-LSL regulates pleiotropic phenotype including long sterile lemma in rice.
Panicle is a harvesting organ of rice, and its morphology and development are closely associated with grain yield. The current study was carried on a mutant screened through an EMS (ethyl-methane sulphonate) mutagenized population of a Japonica cultivar Kitaake (WT). A mutant, named as asp-lsl (aberrant spikelet-long sterile lemma), showed a significant decrease in plant height, number of tillers, thousand-grains weight, seed setting rate, spikelet length, kernel length and effective number of grains per panicle as compared to WT. Asp-lsl showed a pleiotropic phenotype coupled with the obvious presence of a long sterile lemma. Cross-sections of lemma showed an increase in the cell volume rather than the number of cells. Genetic segregation analysis revealed its phenotypic trait is controlled by a single recessive nuclear gene. Primary and fine mapping indicated that candidate gene controlling the phenotype of asp-lsl was located in an interval of 212 kb on the short arm of chromosome 8 between RM22445 and RM22453. Further sequencing and indels markers analysis revealed LOC_Os08g06480 harbors a single base substitution (G→A), resulting in a change of 521st amino acid(Gly→Glu. The homology comparison and phylogenetic tree analysis revealed mutation was occurred in a highly conserved domain and had a high degree of similarity in Arabidopsis, corn, and sorghum. The CRISPR/Cas9 mutant line of ASP-LSL produced a similar phenotype as that of asp-lsl. Subcellular localization of ASP-LSL revealed that its protein is localized in the nucleus. Relative expression analysis revealed ASP-LSL was preferentially expressed in panicle, stem, and leaves. The endogenous contents of GA, CTK, and IAA were found significantly decreased in asp-lsl as compared to WT. Current study presents the novel phenotype of asp-lsl and also validate the previously reported function of OsREL2 (ROMOSA ENHANCER LOCI2), / ASP1(ABERRANT SPIKELET AND PANICLE 1).


Insights into the Effect of Sulfur Incorporation into Tungsten Diphosphide for Improved Hydrogen Evolution Reaction.
Exploring the highly active and stable nonprecious metal electrocatalysts is particularly important for the advancement of water electrolysis, whereas it remains a challenge to efficiently improve the intrinsic electrocatalytic activity. Herein, we reasonably constructed a self-supporting nanosheet array material with sulfur incorporated into WP2. Because of the tunability of electronic configuration and the formation of partial metal phase sulfides, the optimized catalyst exhibits a low overpotential of 115 mV at 10 mA cm-2, along with superb durability over 24 h in acidic media. Furthermore, theoretical calculations reveal that sulfur substitution effectively manipulates the local electronic configuration of WP2, which reduces the interaction between the catalyst surface and hydrogen atoms, thus improving the intrinsic activity of the hydrogen evolution reaction. This work provides valuable insight into the rational fabrication of highly efficient flexible electrode materials based on resourceful electrocatalysts for electrochemical water splitting.


[Medical history education in Finland - some reflections].
Since the nineteen twenties in Finland there has to a varied extent been lectured in "Medical History". It was not a compulsory subject but from the nineteen thirties and until the end of the nineteen fifties it was included in the programme for the University of Helsinki. After a pause of 15 years lecturing in Medical History began again in 1975, with an offer of a course of 20 hours. Two years later this course was extended to 36 hours and was now compulsory for everyone, but in 1980 the hours were reduced to 16. In 1985 the course was changed again, now it became a voluntary with 15 hours lecturing.


Membrane damage in dehydrated bacteria and its repair.
The dehydration of bacteria by vacuum exposure results in damage to the cell membrane. This membrane damage does not necessarily lead to cell death. A part of the dehydrated bacteria is capable of eliminating the membrane damage by repair processes. Repair can proceed rapidly under conditions that permit synthesizing activities. The kinetics of this repair process were studied by means of the membrane-mediated biosynthesis of the cell wall as well as by the recovery of resistance to small concentrations of lysozyme. Repair is a precondition for cell proliferation. At low temperature cells can conserve their membrane damage and the repair process can be initiated when conditions become favourable.


Official dosimetry with personal electronic dosemeters--the framework in Germany.
In Germany, personal electronic dosemeters (AEPDs) are presently applied mainly for operational radiation protection monitoring particularly in nuclear power engineering companies, large hospitals and research centres. This is done in addition to the official dosimetry of record. Therefore, frequently, double monitoring occurs-officially and operationally. A crucial advantage of AEPDs compared with passive dosemeters is the ability to adapt the monitoring period to the working time in controlled areas and to allow an immediate readout of the dose after leaving the controlled area, e.g. a job-related monitoring is possible by correlating the readout dose with the job performed. Germany started a general research project, consisting of two parts, for an optimised implementation of personal electronic dosemeters into official dosimetry. The use of AEPDs as official dosemeters depends on an approval by Federal and Federal State ('Länder') authorities as an official dosimetry system, which has to comply with special requirements ensuring that the legal requirements are fulfilled. The formulation of these special requirements is in the focus of part one of the research project, supervised by the Federal Office for Radiation Protection (BfS) and performed by the Gesellschaft für Anlagen- und Reaktorsicherheit (GRS) mbH. As a result of part one, a framework was developed which is the basis for a future technical implementation project. Part one is described in the paper, while part two is still to be initiated and will deal with the implementation and testing phase of the introduction of personal electronic dosemeters as official dosemeters.


[Pseudo-Conn's syndrome with hypokalemic paralysis due to diuretics and licorice abuse].
Dyskalemic paralyses are characterised by single or periodic episodes with muscle weakness that affect mostly the proximal skeletal muscles. Symptoms may last for a few hours or persist for several days, spontaneous recovery is common. Familial cases can be distinguished from secondary, non-familial forms which are based on other diseases, for example, of the thyroid gland, kidneys or gastrointestinal tract. Familial cases are mostly inherited in an autosomal-dominant pattern and belong to the channelopathies. Both groups are characterised by changed potassium levels in the blood during an episode. A detailed and accurate medical history (plus family history, use of medication and eating habits) often easily leads to the diagnosis. Provoking tests or instrumental and histological investigations can help to solve difficult cases. Treatment focuses on relieving acute symptoms and attacks can be managed by correcting the blood potassium to a normal level. Changing eating and/or exercise habits and also permanent medical treatment helps to prevent further attacks.


Micro-Raman detection of nuclear membrane lipid fluctuations in senescent epithelial breast cancer cells.
Originally identified in cultured cells, oncogenic cellular senescence is a growth-arrest mechanism which may inhibit tumor development by limiting the ability of cells to divide. However, literature shows that these cells secrete tumor-inducing and tumor-suppressing proteins leading to poor prognosis. Understanding the progression of oncogenic cellular senescence and associated mechanisms provides important implications for improving tumorigenesis therapeutic treatments. Micro-Raman spectroscopic imaging has grown in popularity as an imaging technique compared to the standard imaging predecessors and can be attributed to its numerous benefits such as no sample perturbation and the provision of direct chemical information. Through the use of label-free micro-Raman spectroscopy, control and senescent cells were noninvasively imaged. Resulting spectral images were processed using chemometric techniques, and average nuclei spectra from each sample set were compared. In turn, changes in the -cis and -trans unsaturated lipid isomer content were found to differ among proliferating and senescent cells. This may lead to increased nuclear fluidity and may contribute to the inability of senescent cells to complete the cell cycle. In the tumor environment, this detected increase in nuclear envelope fluidity could lead to downstream gene expression modifications and increased nucleo-cytoplasmic RNA translocation. Understanding nuclear envelope fluidity could provide insight into secretory profiles of senescent cells and their role in carcinogenesis, meriting further investigation into novel therapeutic technique development for oncogenic cellular senescence.


Landmark clinical trials influencing surgical management of non-invasive and invasive breast cancer.
The surgical management of breast cancer has changed considerably since the use of the Halstedian radical mastectomy early in the 20th century. Over the last 50 years, several landmark clinical trials from the USA and Europe have resulted in a paradigm shift in the management of breast cancer toward less radical forms of surgery with the combined use of multi-modality treatments including systemic chemotherapy, endocrine therapy, and radiotherapy. Advances in such research have established a new worldwide standard of care for breast cancer surgical management and treatment, which has become more patient centric and which places a higher emphasis on cosmesis and improved patient quality of life. In this chapter, we review the landmark clinical trials that have influenced surgical management for non-invasive and invasive breast cancer and that serve to guide current clinical practices to date.


Effect of office-based brief high-impact exercise on bone mineral density in healthy premenopausal women: the Sendai Bone Health Concept Study.
Although there is ample evidence supporting the effectiveness of physical activity in the prevention and treatment of osteoporosis, there are no previous studies to examine the effect of office-based brief high-impact exercise (HIE) on bone mineral density (BMD) in healthy premenopausal women. This study evaluated the effects of office-based HIE on BMD in healthy premenopausal Japanese women. Ninety-one healthy premenopausal women were randomized to receive stretching exercise (SE) or HIE (stretching, along with up to 5 × 10 vertical and versatile jumps) for 12 months. The BMD of the lumbar spine and proximal femur was measured using dual-energy X-ray absorptiometry. Several cardiovascular risk factors and leg strength also were assessed. An accelerometer-based recorder was used to measure daily impact loading in four 1-week samples. The progression of the HIE program was ensured by the accelerometer. Thirty-three women (71.7%) in the SE group and 34 (75.6%) in the HIE group completed the study. There was a significant difference in the change in the femoral neck BMD between the groups in favor of the HIE group [0.6% (95% CI: -0.4, 1.7) vs. -1.0% (95% CI: -2.2, 0.2)]. Adiponectin, LDL, HDL, and the leg strength of participants in both the groups improved during the intervention. These finding suggested that office-based brief HIE can be recommended for premenopausal women for preventing bone mineral loss.


Nerve lesions in proximal humeral fractures.
In the literature nerve injury is not frequently considered a problem in proximal humeral fractures. Only a few studies exist concerning traction injury of nerves in fractures of the proximal humerus after low-velocity trauma. Almost all of them are retrospective and did not use electromyography. Patients with identical fractures can show quite different outcomes, which vary between complete recovery and severely limited shoulder function. On the assumption that nerve lesions can play a role in the recovery of conservatively and operatively treated proximal humeral fractures, we started a prospective follow-up study with electromyographic investigation. For this study, 143 consecutive proximal humeral fractures due to low-velocity trauma were included. According to the Neer classification, 93 were nondisplaced and 50 were displaced fractures. Denervation on the electromyogram was found in 96 patients (67%). The nerves most frequently involved were the axillary nerve (83 [58%]) and the suprascapular nerve (69 [48%]). Frequently a combination of nerve lesions was seen. Nerve lesions were much more frequent in displaced fractures (82% [41/50]) than in nondisplaced fractures (59% [55/93]). Complicating nerve lesions in patients older than 20 years of age were seen in about the same percentage of patients per decade. Nerve injury and the corresponding loss of muscle strength recovered well in all patients; however, the duration of the recovery was prolonged in cases with nerve lesions. Restoration of the function of the shoulder was less favorable. It is important to realize that, in both conservative and operative treatment of proximal humeral fractures, a paresis due to nerve injury can affect the restoration of shoulder motions. An electromyogram can be useful in the investigation of nerve lesions, because detection only by clinical examination proved to be very difficult. Because of the favorable electrophysiological recovery, no indication for exploration of nerve lesions was present in this series of fractures due to low-velocity trauma.


The potential of localized heating as a adjunct to radiation therapy.
Experimental studies have shown that (a) tumor cells may be more sensitive to heat than normal cells; (b) hyperthermia inactivates cellular repair mechanisms for radiation damage; and (c) heat may lower the OER for ionizing radiation (anoxic cells are at least as sensitive to hyperthermia as oxygenated cells). Localized hyperthemia produced by localized current fields in the range of 100 kHz-10 MHz by direct contact electrodes offers two major advantages: the eletrode configurations may be manipulated to obtain desired thermal dose distributions, and, since the mode of heating is essentially instantaneous, accurate temperature control can be maintained during treatment.


Novel harmonic regularization approach for variable selection in Cox's proportional hazards model.
Variable selection is an important issue in regression and a number of variable selection methods have been proposed involving nonconvex penalty functions. In this paper, we investigate a novel harmonic regularization method, which can approximate nonconvex Lq (1/2 < q < 1) regularizations, to select key risk factors in the Cox's proportional hazards model using microarray gene expression data. The harmonic regularization method can be efficiently solved using our proposed direct path seeking approach, which can produce solutions that closely approximate those for the convex loss function and the nonconvex regularization. Simulation results based on the artificial datasets and four real microarray gene expression datasets, such as real diffuse large B-cell lymphoma (DCBCL), the lung cancer, and the AML datasets, show that the harmonic regularization method can be more accurate for variable selection than existing Lasso series methods.


Performance of planar-waveguide external cavity laser for precision measurements.
A 1542-nm planar-waveguide external cavity laser (PW-ECL) is shown to have a sufficiently low level of noise to be suitable for precision measurement applications. Its frequency noise and intensity noise was comparable or better than the non-planar ring oscillator (NPRO) and fiber laser between 0.1 mHz to 100 kHz. Controllability of the PW-ECL was demonstrated by stabilizing its frequency to acetylene ((13)C(2)H(2)) at 10(-13) level of Allan deviation. The PW-ECL also has the advantage of the compactness of a standard butterfly package, low cost, and a simple design consisting of a semiconductor gain media coupled to a planar-waveguide Bragg reflector.


MiRNA 126 as a New Predictor Biomarker in Venous Thromboembolism of Persistent Residual Vein Obstruction: A Review of the Literature Plus a Pilot Study.
Venous thromboembolism (VTE) is the third most common cardiovascular disease. Interleukins (ILs) and micro-ribonucleic acids (miRNAs) have been proposed as molecules able to modulate endothelial inflammation and platelet hyperactivity. At present, no early biomarkers are available to predict the outcome of VTE. We investigated in a pilot study a selected number of miRNAs and ILs as prognostic VTE biomarkers and reviewed literature in this setting. Twenty-three patients (aged 18-65) with a new diagnosis of non-oncological VTE and free from chronic inflammatory diseases were enrolled. Twenty-three age- and sex-matched healthy blood donors were evaluated as control subjects. Serum miRNAs (MiRNA 126, 155, 17.92, 195), inflammatory cytokines (IL-6, tumor necrosis factor-α, IL-8), and lymphocyte subsets were evaluated in patients at enrolment (T0) and in controls. In VTE patients, clinical and instrumental follow-up were performed assessing residual vein obstruction, miRNA and ILs evaluation at 3 months' follow-up (T1). At T0, IL-8, activated T lymphocytes, Treg lymphocytes, and monocytes were higher in patients compared with healthy controls, as were miRNA 126 levels. Moreover, miRNA 126 and IL-6 were significantly increased at T0 compared with T1 evaluation in VTE patients. Higher levels of MiR126 at T0 correlated with a significant overall thrombotic residual at follow-up. In recent years an increasing number of studies (case-control studies, in vivo studies in animal models, in vitro studies) have suggested the potential role of miRNAs in modulating the cellular and biohumoral responses involved in VTE. In the frame of epidemiological evidence, this pilot study with a novel observational approach supports the notion that miRNA can be diagnostic biomarkers of VTE and first identifies miRNA 126 as a predictor of outcome, being associated with poor early recanalization.


Flavonoids from Potentilla parvifolia Fisch. and Their Neuroprotective Effects in Human Neuroblastoma SH-SY5Y Cells in vitro.
Potentilla parvifolia Fisch. (Rosaceae) is a traditional medicinal plant in P. R. China. In this study, seven flavonoids, ayanin (1), tricin (2), quercetin (3), tiliroside (4), miquelianin (5), isoquercitrin (6), and astragalin (7), were separated and purified from ethyl acetate extractive fractions from ethanol extracts of P. parvifolia using a combination of sevaral chromatographic methods. The human neuroblastoma SH-SY5Y cells were differentiated with all trans-retinoic acid and treated with okadaic acid to induce tau protein phosphorylation and synaptic atrophy, which could establish an Alzheimer's disease cell model. The neuroprotective effects of these flavonoids in cellular were evaluated in vitro by this cell model. Results from the Western blot and morphology analysis suggested that compounds 3 and 4 had the better neuroprotective effects.


Gender Differences in Sexual Attraction and Moral Judgment: Research With Artificial Face Models.
Sexual attraction in humans is influenced by cultural or moral factors, and some gender differences can emerge in this complex interaction. A previous study found that men dissociate sexual attraction from moral judgment more than women do. Two experiments consisting of giving attractiveness ratings to photos of real opposite-sex individuals showed that men, compared to women, were significantly less influenced by the moral valence of a description about the person shown in each photo. There is evidence of some processing differences between real and artificial computer-generated faces. The present study tests the robustness of González-Álvarez's findings and extends the research to an experimental design using artificial face models as stimuli. A sample of 88 young adults (61 females and 27 males, average age 19.32, SD = 2.38) rated the attractiveness of 80 3D artificial face models generated with the FaceGen Modeller 3.5 software. Each face model was paired with a "good" and a "bad" (from a moral point of view) sentence depicting a quality or activity of the person represented in the model (e.g., she/he is an altruistic nurse in Africa vs. she/he is a prominent drug dealer). Results were in line with the previous findings and showed that, with artificial faces as well, sexual attraction is less influenced by morality in men than in women. This gender difference is consistent with an evolutionary perspective on human sexuality.


[How to raise awareness about antibiotics and vaccination among French teenagers?]
Introduction: Bacterial resistance to antibiotics is a growing concern in Europe, vaccination coverage is inadequate, and young adults, who are the highest antibiotic consumers, have the lowest level of knowledge. Extension of the e-Bug project, a European educational school programme concerning microbes, antibiotics and vaccination, to 15-18-year-olds implies the development of educational resources based on social marketing to promote sustainable behavioural changes. The objectives of the present study, which is part of the needs assessment in France, are to explore the attitudes, knowledge and behaviour of 15-18-year-olds concerning antibiotics and vaccination and their health education needs. Methods: Individual semi-structured interviews were conducted using purposeful sampling of 15- to 18-year-olds, until data saturation, and were transcribed and analysed. Results: The results confirm that teenagers are high antibiotic consumers with poor knowledge. They are interested in their health, but less so in antibiotics and vaccination. Health information is mainly obtained from the Internet, but tends to be criticised. General practitioners play an important role in terms of health information and are also influenced by peers and family. Teenagers believe school should play an important role in health education, which they would prefer to be more interactive: debates, case-stories, sharing experience with peers, parents and health professionals, based on relevant concrete examples, via new technologies. Conclusion: Knowledge gaps revealed among young people during this survey confirm that this age-group should be targeted by a specifically tailored approach. Responders are receptive to information provided by reliable and easily accessible sources. High schools, as well as general practitioners, play an important role in health education in general for this age-group and particularly concerning antibiotics and vaccinations. The Internet is a relevant medium for disseminating information. Teaching should be interactive and take social influences (peers, family) into account.


Association Between Tacrolimus Pharmacokinetics and Cytochrome P450 3A5 and Multidrug Resistance Protein 1 Exon 21 Polymorphisms.
Individual differences in the pharmacokinetics (PK) of tacrolimus (TAC), an immunosuppressive drug, are reportedly associated with single-nucleotide polymorphisms (SNPs) of cytochrome P450 (CYP) 3A5 and multidrug resistance protein 1 (MDR1). We determined the effect of SNPs in CYP3A5 and MDR1 exons 21 and 26 on TAC PK parameters. Thirty-eight Japanese patients who underwent renal transplantation were genotyped for CYP3A5 and exons 21 and 26 of MDR1 with the use of polymerase chain reaction-restriction fragment length polymorphism analysis. TAC concentrations were determined 3 weeks after renal transplantation and PK parameters calculated. The area under the blood concentration-time curve (AUC) in CYP3A5 expressers was significantly higher than that in CYP3A5 nonexpressers (CYP3A5*3/*3). Patients with the MDR1 exon 21 A allele (G2677A) showed higher dose-adjusted AUC (AUC/D) and lower doses of TAC than those who did not possess that allele. Furthermore, patients with both CYP3A5*3/*3 and MDR1 G2677A showed significantly lower TAC doses and higher dose-adjusted trough levels (C/D) and AUC/D than those without those genotypes. There was no significant association between MDR1 exon 26 polymorphism and the PK of TAC. Patients with both CYP3A5*3/*3 and MDR1 G2677A had higher blood TAC concentrations than those without those genotypes. Japanese patients should be carefully monitored for consideration of lower TAC doses, because 24% of Japanese patients have double mutations.


Change in Blood and Benign Breast Biomarkers in Women Undergoing a Weight-Loss Intervention Randomized to High-Dose ω-3 Fatty Acids versus Placebo.
The inflammation-resolving and insulin-sensitizing properties of eicosapentaenoic (EPA) and docosahexaenoic (DHA) fatty acids have potential to augment effects of weight loss on breast cancer risk. In a feasibility study, 46 peri/postmenopausal women at increased risk for breast cancer with a body mass index (BMI) of 28 kg/m2 or greater were randomized to 3.25 g/day combined EPA and DHA (ω-3-FA) or placebo concomitantly with initiation of a weight-loss intervention. Forty-five women started the intervention. Study discontinuation for women randomized to ω-3-FA and initiating the weight-loss intervention was 9% at 6 months and thus satisfied our main endpoint, which was feasibility. Between baseline and 6 months significant change (P < 0.05) was observed in 12 of 25 serum metabolic markers associated with breast cancer risk for women randomized to ω-3-FA, but only four for those randomized to placebo. Weight loss (median of 10% for trial initiators and 12% for the 42 completing 6 months) had a significant impact on biomarker modulation. Median loss was similar for placebo (-11%) and ω-3-FA (-13%). No significant change between ω-3-FA and placebo was observed for individual biomarkers, likely due to sample size and effect of weight loss. Women randomized to ω-3-FA exhibiting more than 10% weight loss at 6 months showed greatest biomarker improvement including 6- and 12-month serum adiponectin, insulin, omentin, and C-reactive protein (CRP), and 12-month tissue adiponectin. Given the importance of a favorable adipokine profile in countering the prooncogenic effects of obesity, further evaluation of high-dose ω-3-FA during a weight-loss intervention in obese high-risk women should be considered. PREVENTION RELEVANCE: This study examines biomarkers of response that may be modulated by omega-3 fatty acids when combined with a weight-loss intervention. While focused on obese, postmenopausal women at high risk for development of breast cancer, the findings are applicable to other cancers studied in clinical prevention trials.


Daughter cells as an important factor in determining the physiological state of yeast populations.
Cells of Candida utilis grown in a single-stage chemostat at D = 0.05, 0.1, 0.25, and 0.35 hr-1 were separated into a fraction of scar-bearing mother cells and a fraction of scar-free daughter cells. The scar-free cells were transferred into small batch cultures where the length of the maturation phase, changes in length and width of cells, specific growth rate, and specific rate of RNA and protein synthesis were examined for 5 hr. The daughter cells grown at D = 0.05 hr-1 were very small at the moment of separation from the mother cells (about one-third of the mother cell). Their maturation phase (in a batch culture), at the beginning of which they attain the specific growth rate approaching the mumax of the strain used, lasts for 3 hr. On the other hand, daughter cells grown at D = 0.35 hr-1 are almost the same size as the mother cells at the moment of separation. After transfer to a batch culture they begin to bud almost immediately. Similarly, in their other morphological and physiological parameters they differ strikingly from immature daughter cells which are formed at low specific growth rates. The importance of these differences from the point of view of mathematical modeling of growth processes is discussed.


Numerical and experimental studies of long-range magnetic dipolar interactions.
We describe several numerical methods developed to analyze the behavior of spin polarized liquids in the presence of long-range magnetic dipolar interactions and external field gradients. Two of the methods use a discrete lattice of spins. In the first we calculate the magnetic field from the lattice of spins directly, either in the rotating frame, or in the lab frame. In the second method we include the dipolar fields from linear magnetization gradients analytically and calculate the dipolar fields from higher order gradients in Fourier space, where they are a local function of the magnetization. In the third method the magnetization is expanded in a Taylor series and the dipolar fields are calculated analytically for each term. The results of these calculations are compared to experimental data, in which we use two superconducting quantum interference device magnetometers adjacent to a spherical sample of hyperpolarized liquid 129Xe to detect the evolution of magnetization gradients. In particular, we observe an increase by a factor of 100 of the spin dephasing time in a longitudinal magnetic field gradient due to dipolar interactions of the spins. While each of the numerical techniques has certain limitations, they are generally in agreement with each other and with experimental data.


Cloning of HSP90, expression and localization of HSP70/90 in different tissues including lactating/non-lactating yak (Bos grunniens) breast tissue.
The aim of this study is to investigate the expression and localization of HSP70/90 in different tissues and explore the regulation effects of HSP70/90 at lactation period of female yaks. HSP90 mRNA was cloned from the heart samples of female yaks, Quantitative real-time (qRT-PCR), Western blotting (WB), immunohistochemistry and immunofluorescence assays were utilized to analyze the expressions of HSP70/90 mRNA and protein in different tissues. Sequence analysis showed that HSP90 is a conserved molecular chaperone of female yaks. The qRT-PCR, WB results showed that the expressions of HSP70/90 mRNA and protein were significantly different in different tissues, and 3-fold higher expression during the lactation period than the non-lactation period of breast tissue (P < 0.01). Immunohistochemistry and immunofluorescence assays results showed that HSP70/90 were located in the cardiac muscle cells, cerebellar medulla, theca cells lining at the reproductive system, and the mammary epithelia of the breasts. In addition, the expression level of HSP70 was higher than those of HSP90 in all examined tissues. Therefore, our results strongly suggest that the expression and localization of HSP70/90 could provide significant evidence to further research in tissue specific expression, and lactation function of female yaks.


Nitrogen dioxide exposure and airway responsiveness in individuals with asthma.
Controlled human exposure studies evaluating the effect of inhaled nitrogen dioxide (NO2) on the inherent responsiveness of the airways to challenge by broncho-constricting agents have had mixed results. In general, existing meta-analyses show statistically significant effects of NO2 on the airway responsiveness of individuals with asthma. However, no meta-analysis has provided a comprehensive assessment of the clinical relevance of changes in airway responsiveness, the potential for methodological biases in the original papers, and the distribution of responses. This paper provides analyses showing that a statistically significant fraction (i.e. 70% of individuals with asthma exposed to NO2 at rest) experience increases in airway responsiveness following 30-min exposures to NO2 in the range of 200 to 300 ppb and following 60-min exposures to 100 ppb. The distribution of changes in airway responsiveness is log-normally distributed with a median change of 0.75 (provocative dose following NO2 divided by provocative dose following filtered air exposure) and geometric standard deviation of 1.88. About a quarter of the exposed individuals experience a clinically relevant reduction in their provocative dose due to NO2 relative to air exposure. The fraction experiencing an increase in responsiveness was statistically significant and robust to exclusion of individual studies. Results showed minimal change in airway responsiveness for individuals exposed to NO2 during exercise.


Ab initio potential energy surface, infrared spectra, and dynamics of the ion-molecule complexes between Br- and H2, D2, and HD.
A three-dimensional potential energy surface (PES) for the Br(-)-H(2) complex is computed using the ab initio CCSD(T) method and an extended basis set. The PES has two equivalent minima at the linear geometries (equilibrium interfragment distance R(e)=3.34 A and interaction energy D(e)=670 cm(-1)) separated by the barrier at the T-shaped configuration (interfragment distance R(e)=4.03 A and barrier height of 610 cm(-1)). Ab initio points are fitted to a flexible analytical form and used in the variational rovibrational energy level calculations. Simulated infrared spectra of the Br(-)-H(2) and Br(-)-D(2) complexes in the monomer stretching excitation region are in good agreement with the measured ones. Nonstatistical intensity ratios of the complexes of para- and ortho-monomers are qualitatively explained by monomer ligand exchange reactions. Predissociation of the complexes containing vibrationally excited monomers is analyzed and shown to proceed through the near-resonant vibration-to-rotation energy transfer. For complexes involving Br(-) and the HD monomer, two energetically low-lying states are predicted, corresponding predominately to the Br(-)-DH and Br(-)-HD isomeric forms. The results demonstrate the close similarity of the bromide containing complexes to their analogs containing the chloride anion.


Antenatally detected Klinefelter's syndrome in twins.
An increased incidence of twinning has been reported among patients with Klinefelter's syndrome (XXY) and their relatives, but no data have been reported about the incidence of XXY among twins. Results from our institution's antenatal testing clinic provide data about twins and XXY. In 1842 pregnancies tested, 21 were noted to be twin gestations. Five fetuses with XXY were detected in all pregnancies, of which there was a pair of MZ twins concordant for XXY and a pair of DZ twins discordant for XXY. The incidence of XXY in twins was thus 7,1%. The fetal phenotype of XXY was examined in aborted MZ twins and found to be mild. Notably, the testicular histology was normal. These cases confirm the previously reported association of twinning and XXY. The association of twinning and XXY implies a commonality of causation; however, a maternal age effect cannot be excluded. Our data suggest that there is an increased risk of XXY among twins.


TOMBU and COMBU as Novel Uronium-type peptide coupling reagents derived from Oxyma-B.
Here we describe two novel uronium salts, TOMBU and COMBU, derived from the recently described Oxyma-B for use in peptide bond synthesis. These coupling reagents are more stable than COMU in DMF. Furthermore, using various peptide synthetic models in solution and solid-phase synthesis, we reveal that they show better performance than HBTU in terms of preserving chiral integrity and coupling yields, but slightly worse performance than COMU.


Transient translocation of hemidesmosomal bullous pemphigoid antigen 1 from cytosol to membrane fractions by 12-O-tetradecanoylphorbol-13-acetate treatment and Ca2+-switch in a human carcinoma cell line.
We previously showed that 12-O-tetradecanoylphorbol-13-acetate (TPA) and Ca2+-switch from low (0.07 mM) to normal (1.87 mM) concentration in culture medium, which were also linked to activation of protein kinase C (PKC), lead to phosphorylation of 180 kDa-bullous pemphigoid antigen (BPAG) 2, but not of 230 kDa-BPAG1, and possibly to its disassembly from hemidesmosomes in a human squamous cell carcinoma cell line (DJM-1). In this study, we examined the effects of TPA and Ca2+-switch on intracellular localization of BPAG1 by immuno-blotting and immuno-fluorescence microscopy with monoclonal antibodies to the antigen after sub-cellular fractionation. In DJM-1 cells cultured in low Ca2+ medium, BPAG1 was detected as phosphate buffered saline-soluble (cytosolic), Triton X-100 soluble (roughly membrane-associated) and Triton X-100 insoluble (cytoskeleton-bound) forms, whereas in normal Ca2+-grown cells only as cytosolic and cytoskeleton-bound forms. In normal Ca2+-cultured cells, TPA (50 nM) caused a complete translocation of BPAG1 from cytosol to membrane fractions within 10 min, that was inhibited by pretreatment with H7 (a selective PKC inhibitor) at 40 microM. After 30 min and 4 h of TPA-treatment, BPAG1 was exclusively detected in cytoskeleton fractions. Morphologically, immuno-fluorescence microscopy showed that treatment caused a marked reduction of BPAG1 from the cytoplasm and generated a linear pattern at cell-cell contacts, suggesting translocation of BPAG1 from the cytosol to the plasma membrane. In contrast, the Ca2+-switch from low to normal caused a prominent increase of BPAG1, both in cytosolic and membrane-associated forms after 4 h, that was inhibited both with H7 and cycloheximide (an inhibitor of protein synthesis) at 70 microM, suggesting a role for PKC and BPAG1 synthesis in these Ca2+-induced effects. These results suggest that TPA and Ca2+-switch induced BPAG1 translocation to membrane fractions possibly mediated by PKC-activation. Furthermore, whereas TPA affects the redistribution of BPAG1 among their pools without inducing their synthesis, Ca2+-switch induces both membrane translocation and synthesis of BPAG1, suggesting involvement of signaling other than PKC pathways in control of BPAG1 synthesis.


Implementation and evaluation of a training program to improve resident teaching skills.
This study was undertaken to develop, implement, and evaluate a program designed to improve residents' teaching skills. A training program to improve the teaching skills of obstetrics and gynecology residents was developed and implemented. All residents completed a survey of their teaching skills, participated in the program, and then completed a follow-up survey. The surveys were compared using the Wilcoxon signed rank test. Medical students also completed surveys about participating residents' teaching skills either before or after the intervention. The training program was well received by the residents. Resident self-rating scores significantly improved (P<.05) in a number of areas, such as ability to teach physical examination skills, and explaining topics in a clear and concise manner. Interestingly, student ratings of resident teaching skills did not improve. Although the training program improved resident self-ratings of teaching skills, this was not reflected in student ratings of resident teaching skills. Possible explanations for this difference will be explored.


Fate of Staphylococcus aureus on vacuum-packaged ready-to-eat meat products stored at 21 degrees C.
The U.S. Department of Agriculture has established standards for the composition and shelf stability of various ready-to-eat meat products. These standards may include product pH, moisture:protein ratio, and water activity (aw) values. It is unclear how closely these standards are based on the potential for pathogen growth or toxin production. Because the vacuum packaging used on most ready-to-eat meat products inhibits mold, Staphylococcus aureus is the pathogen most likely to grow on products with reduced aw and increased percentage of water-phase salt. In this study, 34 samples of various ready-to-eat meat products were inoculated with a three-strain mixture of S. aureus, vacuum packaged, and stored at 21 degrees C for 4 weeks. S. aureus numbers decreased by 1.1 to 5.6 log CFU on fermented products (pH < or = 5.1) with a wide range of salt concentrations and moisture content. Similarly, S. aureus numbers decreased by 3.2 to 4.5 log CFU on dried nonacidified jerky (aw < or = 0.82; moisture:protein ratio of < or =0.8). Products that were not fermented or dried clearly supported S. aureus growth and cannot be considered shelf stable. The product pH and moisture:protein ratio were the two compositional factors most highly correlated (R2 = 0.84) with S. aureus survival and growth for the types of products tested, but pH and aw or pH and percentage of water-phase salt also may provide useful predictive guidance (R2 = 0.81 and 0.77, respectively).


Role of sex, age, previous valve lesion, and pregnancy in the clinical expression and outcome of Q fever after a large outbreak.
Q fever is a zoonosis caused by Coxiella burnetii. After a large outbreak occurred in the Chamonix Valley in the French Alps in 2002, an extensive surveillance was conducted, to describe the variations in the clinical expression of acute Q fever according to host factors, as well as to monitor the risk of evolution of acute Q fever to chronic Q fever in patients at risk. Three groups of patients with risk factors for evolution of acute Q fever to chronic Q fever were considered: 376 pregnant women, 19 immunocompromised patients, and 91 patients with valvular or vascular abnormalities. A group of 578 people without risk factors for evolution of acute Q fever to chronic Q fever was also tested. Diagnosis of Q fever was based on serologic testing by immunofluorescence assay. Between 30 August 2002 and 31 July 2003, a total of 1946 serum samples obtained from 1064 persons were tested. A total of 101 patients (9.3%) had acute Q fever diagnosed, and 5 patients (0.5%) had chronic Q fever diagnosed. A diagnosis of acute Q fever was established for 11 pregnant women (2.6% of 379 pregnancies), 5 patients with valvular disease (5.5%), and 85 people without risk factors (14.7%) (71 [27.9%] of 254 symptomatic patients and 14 [4.3%] of 324 asymptomatic patients). A new pregnancy in a woman with negative results of serologic tests for Q fever exposes the woman to a new risk for acute Q fever able to evolve to chronic Q fever. The rates of clinical expression were 90.6% in adult men, 75% in adult women, and 33.3% in children, and they were significantly lower (9.1%) in pregnant women. Evolution to chronic Q fever was observed in 5 patients. This study emphasizes the importance of active surveillance in postepidemic conditions, especially among patients at risk, as well as the importance of systematic serologic testing during pregnancy.