Measurements of Intra- and Extra-Cellular 5-Methyltetrahydrofolate Indicate that Bifidobacterium Adolescentis DSM 20083T and Bifidobacterium Pseudocatenulatum DSM 20438T Do Not Actively Excrete 5-Methyltetrahydrofolate In vitro.
Certain intestinal bifidobacteria have the ability to synthesize folates. In vitro experiments revealed a high production, cellular accumulation, and release of reduced folate vitamers like 5-methyltetrahydrofolate and tetrahydrofolate in folate-free medium (FFM). However, it is still unclear to which extent synthesized folates are polyglutamylated and probably not available for transport, and if they are actively released by excretion. To address these questions, we characterized intra- and extra-cellular pteroylmonoglutamates and polyglutamylated 5-methyltetrahydrofolate (5-CH3-H4PteGlu2-4) in Bifidobacterium adolescentis DSM 20083T and Bifidobacterium pseudocatenulatum DSM 20438Tin vitro. Folates were measured by means of stable isotope dilution assays (SIDA) coupled with LC-MS/MS analysis using [2H4]-5-methyltetrahydrofolic acid, [2H4]-tetrahydrofolic acid, and [2H4]-5-formyltetrahydrofolic acid as internal standards. Cell viability was examined by fluorescence microscopy. Quantitation of folate production by B. adolescentis during the stationary phase revealed a linear increase of dead cells paralleled by increasing concentration of 5-formyltetrahydrofolate and 5-methyltetrahydrofolate (100% 5-CH3-H4PteGlu4) in FFM, whereas the intracellular concentrations of these vitamers remained constant. After 24 h, B. adolescentis (125 mg cells, wet weight) produced a total amount of 0.846 nmol 5-CH3-H4folate: 0.385 ± 0.059 nmol (46 ± 7%) and 0.461 ± 0.095 nmol (54 ± 11%) measured in the intracellular (viable cells; 52 ± 3% measured by fluorescence microscopy) and extracellular (lysed cells; 48 ± 3%) fraction, respectively. For B. pseudocatenulatum (124 mg cells, wet weight), 1.135 nmol 5-CH3-H4folate was produced after 24 h, and a similar proportionality between intra- and extra-cellular folate concentrations and viable/lysed cells was observed. These results indicate that the strains tested produce and accumulate 5-CH3-H4PteGlu4 for cellular metabolism, and that extracellular concentrations of the vitamer arise from cell lysis.


Identification of two novel frameshift mutations in the KCNJ11 gene in two Italian patients affected by Congenital Hyperinsulinism of Infancy.
Congenital Hyperinsulinism of Infancy (CHI) is a genetically heterogeneous disorder characterized by profound hypoglycemia related to inappropriate insulin secretion. Two histopathologically and genetically distinct groups are recognized among patients with CHI due to ATP-sensitive potassium channel (KATP) defects: a diffuse type (Di-CHI), which involves the whole pancreas, and a focal form (Fo-CHI), which shows adenomatous islet-cell hyperplasia of a particular area within the normal pancreas. The beta-cell KATP channel consists of two essential subunits: Kir6.2 encoded by the KCNJ11 gene which is the pore-forming unit and belongs to the inwardly rectifying potassium channel family, and SUR1 (sulfonylurea receptor 1) encoded by the ABCC8 gene, which belongs to the ATP-binding cassette (ABC) transporter family. The KATP channel is an octameric complex of four Kir6.2 and four SUR1 subunits. More than one hundred mutations have been found in KATP channel genes ABCC8 and KCNJ11, but to date only twenty mutations have been identified in KCNJ11, most of them are missense mutations and only one is a single base deletion. The Fo-CHI has been demonstrated to arise in individuals who have a germline mutation in the paternal allele of ABCC8 or KCNJ11 in addition to a somatic loss of the maternally derived chromosome region 11p15 in adenomatous pancreatic beta-cells, while Di-CHI predominantly arises from the autosomal recessive inheritance of KATP channel gene mutations. Here we describe the molecular findings in nine children who presented, in the neonatal period, with signs and symptoms of hypoglycemia and diagnosed affected by CHI according to international diagnostic criteria. Direct sequencing of the complete coding exon and promoter region of KCNJ11 gene showed, in two Italian patients, two new heterozygous mutations which result in the appearance of premature translation termination codons resulting in the premature end of Kir6.2. Interestingly most of the CHI mutations detected in other population studies are situated in the ABCC8 gene.


Failure of sentinel lymph node mapping in a patient with endometrial cancer with chronic lymphocytic leukemia: A case report.
We aimed to report a detection failure of sentinel lymph node (SLN) mapping via indocyanine green in a patient with endometrial cancer with a previous history of chronic lymphocytic leukemia (CLL), which is a potential risk factor to obstruct lymphatic channels. A 64-year-old woman with a 12-year history of CLL presented to the clinic with grade 2 endometrioid carcinoma. The patient underwent laparoscopic surgical staging. Indocyanine green was used intraoperatively to locate the SLN. No lymph node or lymphatic vessels were identified during SLN mapping. At the final pathology, the morphological findings of CLL were detected in the lymph nodes without metastasis of endometrial cancer. Sentinel lymph node mapping failure due to obstruction of lymphatic channels in a patient with CLL was demonstrated in this study. This is the first report to the best of our knowledge showing SLN mapping failure in the presence of lymphoproliferative diseases.


Chemical composition and antiprolifrative activity of Artemisia persica Boiss. and Artemisia turcomanica Gand. essential oils.
Essential oils obtained from aerial parts of Artemisia persica and Artemisia turcomanica were analyzed by GC/MS. While 28 components representing 91.01 % of A. persica were identified, the identity of 50 components, constituting 81.93 % of the total oil, was confirmed in A. turcomanica. β-thujone was the main compound (75.23%) in A. persica while the major identified phytochemicals in A. turcomanica were 1,8-cineol (19.23%), camphor (15.55%) and filifolone (15.53%). Both of the essential oils were predominantly made up of monoterpenes. Time- and dose-dependent cytotoxic effects of A. persica and A. turcomanica on MCF-7 cell line evaluated by MTT assay at 24, 48 and 72 h, showed that the highest cytotoxic effect of A. persica and A. turcomanica were appeared at 72 h incubation. At that incubation period, CI50 of A. persica was found to be 0.15 μg/ml, while that of A. turcomanica was 0.1 μg/ml. Thus, cytotoxicity of A. turcomanica was slightly higher than A. persica which could be attributed to the higher content of sesquiterpene present in A. turcomanica. As a conclusion, these volatile oils could have chemotherapeutic potentials.


Intraparenchymal Renal Artery Pseudoaneurysm and Arteriovenous Fistula on a Solitary Kidney Occurring 38 Years after Blunt Trauma.
Pseudoaneurysm and arteriovenous fistulae of the renal artery are rare complications of kidney trauma. They commonly result from open traumas and occur within days after the injury. Common symptoms include acute haematuria, pain, or hypertension. We report the case of a fifty-three-year-old man presenting with symptomatic complex chronic high flow kidney arteriovenous fistula with interposition of a pseudoaneurysmal pouch and arterial aneurysmal dilatation in a solitary left kidney 38 years after a blunt trauma. Those conditions were successfully treated by endovascular embolization followed by regular radiologic, biological, and clinical follow-up. To the best of our knowledge, few similar cases were reported more than 20 years after trauma. However, no case combining an arteriovenous fistula and a pseudoaneurysm revealing as late as 38 years after trauma was found. In addition, management of those conditions on a solitary kidney and outcomes has not been described. We believe that our case depicts the clinical presentation and management of this rare entity that should not be unrecognized due to its potential lethal implications.


[Complications of cerebrospinal fluid shunt].
The objective of this study was to know the incidence, mode of presentation, clinical and microbiological aspects, as well as the management of ventriculo-peritoneal shunt malfunction. A retrospective study was carried out where thirty-seven children with malfunctioning shunts were studied. The mechanical causes of shunt malfunction included disconnection, fracture and obstruction, while infection was considered as either a clinical or analytical change (blood and/or CSF). The mean age of the patients was 5.5 years. The most common indication for the insertion of the ventriculoperitoneal shunt was congenital hydrocephalus. The average stay in the ICU was 8.2 days. Mechanical dysfunction was detected in 25 patients (68%) and infectious etiology in 12. The most common clinical manifestations were vomiting, fever and headache. CSF biochemical alteration was found in 8 of the infected group. Staphylococcus was isolated in six of the 8 cases. The ventriculoperitoneal shunt was changed during the first 24 hours in 17 of 25 patients (68%) with mechanical malfunction, while 11 cases with an infective cause (92%) were managed with an external shunt. The interval between the insertion of the ventriculoperitoneal shunt and the malfunction was over 6 months in 20 cases (54%). Ventriculoperitoneal shunts are the main method for treating hydrocephaly, although they are not without complications which may require surgical procedures. Appropriate surgical technique, asepsis, as well as prophylactic antibiotics are essential to decrease the incidence of complications.


Strand specificity and absence of hot spots for p53 mutations in ultraviolet B-induced skin tumors of XPA-deficient mice.
We examined the spectrum of p53 mutations found in 40 UV-induced skin tumors of xeroderma pigmentosum group A gene (XPA)-deficient mice. p53 mutations were detected in 48% of the tumors. Nearly all of the mutations were induced at dipyrimidine sites. Ninety-three % of the mutations were G.C-->A.T transitions at dipyrimidine sites, including tandem transitions (CC-->TT), which are the hallmark of the UVB-induced mutation. Seventy-two % of the mutations at dipyrimidine sites could be ascribed to damage on the transcribed strand. In addition, no evident mutational hot spots were detected. This is in contrast to the UVB-induced skin tumors of normal mice, in which 92% of p53 mutations occurred as a result of DNA damage on the nontranscribed strand, and clear hot spots were observed. Thus, XPA-deficient mice showed significant mutation features that might be characteristic of the absence of nucleotide excision repair and may provide a good animal model for the analysis of the high incidence of skin cancer in xeroderma pigmentosum group A patients.


Is an antiemetic prophylactic treatment needed for patients submitted to consecutive days of 5-fluorouracil? An observational study.
The necessity of an antiemetic prophylaxis in patients treated with chemotherapy of low emetogenic potential, such as 5-fluorouracil +/- folinic acid fractionated over several consecutive days, is controversial. The aim of the study was to evaluate the therapeutic behavior of oncologists on this issue. All consecutive in and out patients who started chemotherapy in 33 Italian oncological departments from June 24 to July 6,1996, were studied. The antiemetic prescription pattern and its effectiveness, in patients submitted to 5-fluorouracil +/- folinic acid were evaluated. Of the 1956 patients submitted to cancer chemotherapy, 259 patients received 5-fluorouracil +/- folinic acid. Of these, 186 patients were treated for 5 consecutive days, 47 for 4 days, 20 for 3 days and 6 for 2 days. A total of 219 (84.5%) received an antiemetic prophylaxis: 43.4% a 5-HT3 antagonist +/- steroids, 37.5% an antidopaminergic drug, 10.9% a steroid +/- antidopaminergic drug, and 8.2% other drugs. Only 40 patients (15.5%) did not receive an antiemetic prophylaxis. Overall complete protection from vomiting/nausea was 225/259 (86.9%)/163/259 (62.9%). The complete protection from vomiting/nausea during the 5 days in the 186 patients was not significantly different among patients receiving or not an antiemetic prophylaxis (88.1%/64.9% vs 88.9%/55.6%). At unifactorial analysis, the previous experience of vomiting/nausea caused by chemotherapy was found to be a significant prognostic factor. In fact, overall complete protection from vomiting/nausea was significantly inferior in patients who had previous experience of vomiting/nausea (65.1%/35.0%) with respect to those who did not (91.2%/75.4%, P < 0.001/ > 0.001, respectively). The study showed that in clinical practice patients submitted to 5-fluorouracil +/- folinic acid obtained a similar high protection from vomiting and nausea regardless of whether or not antiemetic prophylaxis was given. It would be therefore reasonable not to treat patients undergoing such chemotherapy, whereas patients with previous experience of vomiting/nausea caused by chemotherapy should be given an antiemetic prophylaxis.


A Case Report of Neovascular Glaucoma Due to Carotid Artery Occlusive Disease. Diagnosis of Neovascular Glaucoma and Histological Characteristics.
Purpose: The methods of diagnosing neovascular glaucoma were evaluated in a case with carotid artery occlusive disease. A trabeculectomy specimen taken from this patient was examined histologically to find the characteristic picture of this slowly progressive neovascular glaucoma.Materials and Methods: The left eye of a 48-year old man with neovascular glaucoma due to carotid artery occlusive disease had been previously diagnosed as uveitis with ocular hypertension. His left eye was examined by fluorescein fundus and gonioangiography. Trabeculectomy was performed because of uncontrolled intraocular pressure and the trabeculectomy specimen was examined histologically including immunohistochemistry with antibodies against von Willebrand factor antigen.Results: The characteristic picture of fluorescein fundus angiography was filling delay in the choroid and stained walls of the retinal arteries and veins. Fluorescein gonioangiography using a fundus camera clearly demonstrated new vessels in the pupil and angle with intense fluorescein leak in spite of faint neovascularization in those areas when observed by slit lamp with gonioscopy. Histological examination of the trabeculectomy specimen revealed proliferation of new vessels in the spaces of the trabecular meshwork and focal occlusion of Schlemm's canal.Conclusion: Fluorescein fundus angiography and gonioangiography by fundus camera is very useful for diagnosis of neovascular glaucoma due to carotid artery occlusive disease. Neovascular proliferation in the trabecular meshwork with slight of peripheral anterior synechia and focal occlusion of Schlemm's canal may be peculiar to such slowly progressive neovascular glaucoma.


Lung contusion after extracorporeal shock wave lithotripsy for pancreatic stones: A case report.
Pancreatic extracorporeal shock wave lithotripsy (P-ESWL) is recommended as the first-line treatment for large pancreatic stones. While complications such as post-P-ESWL pancreatitis, bleeding, infection, steinstrasse, and perforation have been reported in the past 30 years, lung contusion has never been reported. The present case demonstrates lung contusion as a complication after P-ESWL. A 48-year-old man was admitted to our department due to painful chronic pancreatitis with pancreatic duct stones. Computed tomography revealed normal lungs. P-ESWL was performed. The shock wave head contacted with right upper quadrant and the path of shock wave was at a 45° angle to the ventral midline. After P-ESWL, multiple patchy high-density shadows in the lower lobe of right lung were found, which was normal before P-ESWL. The patient had no symptoms of lung injury. Laboratory studies revealed elevated D-dimer from 0.33 to 0.74 ug/mL, which was consistent with abnormal clotting of lung contusion. Chest computed tomography showed slight pleural effusion. Considering the interval between 2 X-rays was only 3 hours, we inferred that lung contusion was related to P-ESWL. The patient displayed stable vital signs, therefore, no specific interventions were conducted. Three days after P-ESWL, endoscopic retrograde cholangiopancreatography was performed and the lung shadows were partially absorbed. Considering the location of shock wave head, it was possible to cause lung contusion in lower lobe of right lung. More than 10,000 P-ESWL therapeutic sessions had been performed in our center since 2010, and it is the first case about lung contusion as a complication. It is also the first report to describe lung contusion after P-ESWL. Although the patient was asymptomatic, it should raise awareness of clinicians.


Plasmid pKpQIL encoding KPC-3 and TEM-1 confers carbapenem resistance in an extremely drug-resistant epidemic Klebsiella pneumoniae strain.
An extremely drug-resistant (XDR) clone of KPC-3-producing Klebsiella pneumoniae emerged in Israel in 2006, causing a nationwide outbreak. We aimed to characterize the local KPC-3-encoding plasmid carried by these isolates and study its contribution to antibiotic resistance. Mechanisms of carbapenem resistance were investigated in seven selected isolates (isolated between 2006 and 2008) belonging to the epidemic clone. Isolates underwent MIC testing, and were examined for the presence of KPC, Tn4401, class I integron elements and additional antibiotic resistance genes. Plasmids were analysed by transformation, transconjugation, restriction mapping, curing and complementation experiments. Outer membrane protein (OMP) analysis was performed. OMP analysis did not reveal loss of porins. KPC-3-producing K. pneumoniae isolates possessed various plasmids but all harboured a common self-transmissible 105 kb plasmid, termed pKpQIL, encoding bla(TEM-1) and bla(KPC-3). Curing of pKpQIL led to a complete loss of resistance to cephalosporins and carbapenems, proving its crucial role in carbapenem resistance. Transformation of plasmid pKpQIL into the cured Klebsiella strain resulted in full reconstitution of carbapenem resistance. The presence of all Tn4401 transposon elements located upstream of the KPC-3 gene was detected by PCR and sequencing. pKpQIL lacked additional antibiotic resistance genes. Our findings demonstrate the presence of pKpQIL, a 105 kb KPC-3- and TEM-1-encoding plasmid, in the XDR K. pneumoniae epidemic strain in Israel. pKpQIL is unique and appears consistently in all isolates of this clone over the years. The extensive beta-lactam resistance phenotype of this clone is primarily mediated by this single self-transmissible plasmid.


Significance of post-chemoradiation biopsy in predicting residual esophageal carcinoma in the surgical specimen.
Pathologic complete response in the resected esophagus can be achieved in approximately 30% of patients with locally advanced esophageal or gastroesophageal junction carcinoma after preoperative chemoradiation therapy. These patients tend to have a longer survival than those who have less than pathologic complete response. Post-chemoradiation esophageal biopsy (PCEB) is used to check for the presence of residual tumor before a definitive resection is performed, but the clinical significance of PCEB findings is not clear due to the possibility of sampling bias and the superficial nature of the specimen obtained. We evaluated the use of PCEB (defined as biopsy taken within 30 days before esophagectomy) in predicting residual cancer in post-treatment esophagectomy specimens. PCEB was performed in 65 of 183 (36%) patients with locally advanced esophageal or gastroesophageal junction carcinoma, who received preoperative chemoradiation therapy. The cancer status in PCEB was correlated with the residual cancer in the esophagectomy specimens. PCEB had no cancer in 80% (52 of 65) of patients (Bx-negative) and cancer in 20% (13 of 65) of patients (Bx-positive). There was no difference in the presence of residual cancer (either in esophagus or lymph node) in esophagectomy specimens between Bx-negative patients (77%, 40 of 52) or Bx-positive patients (92%, 12 of 13), P = 0.44. The positive predictive value of biopsy was 92% (12 of 13), negative predictive value 23% (12 of 52), sensitivity 23% (12 of 52) and specificity 92% (12 of 13). There was no difference in the residual cancer staging in the esophagectomy specimen between Bx-positive and Bx-negative patients. In contrast, residual metastatic carcinoma in lymph nodes was more frequent in Bx-positive patients (69.2%, 9 of 13) than in Bx-negative patients (28.8%, 15 of 52), P = 0.01. Our data suggest that PCEB is a specific but not a sensitive predictor of residual cancer following esophagectomy. Bx-positive patients tend to have more frequent residual tumor in lymph nodes. The utility of PCEB in predicting residual cancer in the lymph nodes needs to be explored further along with molecular predictors of response to preoperative therapy.


Tissue expression of cytokines (IL-1alpha, IL-2, IL-6, IL-12, TNF-alpha) in B-cell lymphomas in children.
In most malignant tumours disturbed production of cytokines and of their receptors can be noted. The main immune system cells engaged in tumour regression with the involvement of cytokines include helper lymphocytes (Th1), cytotoxic lymphocytes (Tc) and dendritic cells. It has been noted that cytokines are also produced by the cells of a tumour. The present study aimed at immunocytochemical localisation of cytokines (IL-1alpha, IL-2, IL-6, IL-12, TNF-alpha) in B-cell lymphomas in children (n = 10). The studies were performed on surgical material following formalin fixation and embedding in paraffin. The control material involved human reactive lymph nodes (n = 5). IL-2, IL-6, IL-12 were detected in 4/10 examined cases. Expression of IL-1alpha was highly pronounced, as in the control, and was detected in 6/9 examined patients. No TNF-alpha could be demonstrated in tumours even if lymph node expression of the cytokine was demonstrated in the control. The studies pointed to a decreased production of antitumour cytokines in B-cell lymphomas in children, which might correspond to the progression of the tumour.


A Social-Ecological Approach to Identify Facilitators and Barriers of Home Modifications.
Aging individuals may face difficulty with independently navigating and interacting with their home environment. Evidence-based interventions promoting home modifications are needed to support aging-in-place across the lifespan. This study identified the facilitators and barriers to implementing home modifications from the perspectives of residents and professionals (N = 16). Guided by a social-ecological model, researchers utilized directed content analysis of focus group interviews. While participants discussed facilitators and barriers mainly on the individual level, factors were presented at the relationship, community, and societal level of the model. Overall, the findings suggest a potential for targeted interventions on all levels of the model to promote adoption of home modifications.


N,N,N',N'-Tetra-methyl-ethylene-diammonium tetra-chloridocobaltate(II).
The asymmetric unit of the title compound, [(CH(3))(2)NH(CH(2))(2)NH(CH(3))(2)][CoCl(4)], contains a tetra-chlorido-cobalt-ate(II) dianion and two halves of two centrosymmetric, crystallographically-independent, dications. One independent dication is disordered between two conformations in a 0.784 (13):0.216 (13) ratio. In the crystal, inter-molecular N-H⋯Cl hydrogen bonds link cations and anions into chains propagated in [01]. These hydrogen bonds contribute to the distorted tetra-hedral geometry at the Co(II) atom.


Prediction and modeling of pre-analytical sampling errors as a strategy to improve plasma NMR metabolomics data.
Biobanks are important infrastructures for life science research. Optimal sample handling regarding e.g. collection and processing of biological samples is highly complex, with many variables that could alter sample integrity and even more complex when considering multiple study centers or using legacy samples with limited documentation on sample management. Novel means to understand and take into account such variability would enable high-quality research on archived samples. This study investigated whether pre-analytical sample variability could be predicted and reduced by modeling alterations in the plasma metabolome, measured by NMR, as a function of pre-centrifugation conditions (1-36 h pre-centrifugation delay time at 4 °C and 22 °C) in 16 individuals. Pre-centrifugation temperature and delay times were predicted using random forest modeling and performance was validated on independent samples. Alterations in the metabolome were modeled at each temperature using a cluster-based approach, revealing reproducible effects of delay time on energy metabolism intermediates at both temperatures, but more pronounced at 22 °C. Moreover, pre-centrifugation delay at 4 °C resulted in large, specific variability at 3 h, predominantly of lipids. Pre-analytical sample handling error correction resulted in significant improvement of data quality, particularly at 22 °C. This approach offers the possibility to predict pre-centrifugation delay temperature and time in biobanked samples before use in costly downstream applications. Moreover, the results suggest potential to decrease the impact of undesired, delay-induced variability. However, these findings need to be validated in multiple, large sample sets and with analytical techniques covering a wider range of the metabolome, such as LC-MS. The sampleDrift R package is available at https://gitlab.com/CarlBrunius/sampleDrift. firstname.lastname@example.com. Supplementary data are available at Bioinformatics online.


Epidemiology of the pseudo-exfoliation syndrome.
1) Pseudo-exfoliation syndrome (PE syndrome) is a condition with worldwide distribution. 2) Marked geographical variations have been demonstrated. 3) Environmental factors may be of etiological significance for the development of PE syndrome. 4) The PE syndrome's ability to promote glaucoma may vary from one area to another. 5) There seems to be a trend towards lower debut age of PE syndrome at lower latitudes. 6) More prevalence studies from different parts of the world are needed. A standardized set-up of such works would facilitate comparison of the results, and so some guidelines have been proposed.


Reinforcement of eye movement with concurrent schedules.
Human macrosaccadic eye movements to two areas of a four-dial display were conditioned by concurrent variable-interval schedules of signals. Reinforcers (signals) were delivered to the two right-hand dials on one schedule and to the two left-hand dials on another, independent schedule. The use of a changeover delay between crossover eye movements and reinforcement had the effect of changing the pattern of scanning from fixating four dials in succession or in a Z-shaped pattern to scanning vertically the dials on either side with fewer crossovers. In the presence of a changeover delay, subjects matched relative eye-movement rates and relative reinforcement rates on each schedule. Rate of crossover eye movements, with a changeover delay in effect, was also inversely related to the difference in reinforcements arranged by the concurrent schedules. The results suggest that for stimuli whose critical components are arranged spatially, conditioned eye movements play an important part in selective stimulus control.


Thiomicrorhabdus indica sp. nov., an obligately chemolithoautotrophic, sulfur-oxidizing bacterium isolated from a deep-sea hydrothermal vent environment.
A new mesophilic bacterium, designated strain 13-15AT, was isolated from the deep-sea water from the Carlsberg Ridge, northwestern Indian Ocean. Cells were short rods and motile with a single polar flagellum. Growth was observed in the presence of 85-1700 mM NaCl (optimum 680 mM), at 10-45 °C (optimum, 28 °C) and pH 5.0-9.0 (optimum, pH 7.0). The isolate was an obligate chemolithoautotroph capable of growth using thiosulfate, sulfide, elemental sulfur or tetrathionate as the sole energy source, carbon dioxide as the sole carbon source, and molecular oxygen as the sole electron acceptor. Molecular hydrogen did not support growth. The major fatty acids were C16 : 1 (45.0 %), C18 : 1 (22.5 %) and C16 : 0 (20.1 %). The G+C content of the genomic DNA was 41.6 mol%. The results of phylogenetic analysis based on 16S rRNA gene sequences showed that the novel isolate belonged to the genus Thiomicrorhabdus and was most closely related to Thiomicrorhabdus hydrogeniphila MAS2T (94.8 % sequence similarity). On the basis of the taxonomic data obtained in this study, strain 13-15AT represents a novel species of the genus Thiomicrorhabdus, for which the name Thiomicrorhabdus indica sp. nov. is proposed, with the type strain 13-15AT (=MCCC 1A13986T=KCTC 15750T).


Exploration of digestion-resistant immunodominant epitopes in shrimp (Penaeus vannamei) allergens.
The digestion products of Penaeus vannamei still had sensitizing and eliciting capacity; however, the underlying mechanism has not been identified. This study analyzed the structural changes of shrimp proteins during digestion, predicted the linearmimotopepeptides and first validated the allergenicity of immunodominantepitopes with binding ability. The results showed that the shrimp proteins were gradually degraded into small peptides during digestion, which might lead to the destruction of linear epitopes. However, these peptides carried IgE epitopes that still trigger allergic reactions. Eighteen digestion-resistant epitopes were predicted by multiple immunoinformatics tools and digestomics. Five epitopes contained more critical amino acids and had strong molecular docking (P1: DSGVGIYAPDAEA, P2: EGELKGTYYPLTGM, P3: GRQGDPHGKFDLPPGV, P4: IFAWPHKDNNGIE, P5: KSTESSVTVPDVPSIHD), and these epitopes were identified as novel IgE binding immunodominantepitopes in Penaeus vannamei. These findings provide novel insight into allergenic epitopes, which might serve as key targets for reducing the allergenicity in shrimp.


[The Darmstadt Johannes Alexandrinus fragment].
Three latin fragments (about 1050), included in the envelope of a manuscript, written about 1530 in Cologne, could be identified as parts of the latin version of an Alexandrian commentary (probably written by Johannes Alexandrinus) on Galen's book 'De sectis'. C. D. Pritchet edited this text in 1982, but he thought that he was printing a translation of this commentary on Galen's book, written by Burgundio de Pisa in the year 1185. By means of an exact comparative analysis of the fragments with Galen's original Greek text, we were able to show that these fragments offer almost the same text as Pritchet. But it is remarkable that there are some differences between our version and the text of Pritchet. It seems that our version proves the existence of an until now unknown medieval revision of the Galen-commentary by Johannes Alexandrinus.


Systemic Manifestations of Immunoglobulin G4-Related Disease: A Pictorial Essay.
Immunoglobulin G4 (IgG4)-related disease is a systemic fibro-inflammatory disease characterized by pathologic findings in various organs. Imaging is critical for the diagnosis and treatment assessment of patients with IgG4-related disease. In this pictorial essay, we review the key features of multiple imaging modalities, typical pathologic findings, and differential diagnosis of IgG4-related disease. This systematic pictorial review can further our understanding of the broad-spectrum manifestations of this disease.


The Regulatory Review Process in South Africa: Challenges and Opportunities for a New Improved System.
The aims of this study were to assess the regulatory review process in South Africa from 2015 to 2017, identify the key milestones and timelines; evaluate the effectiveness of measures to ensure consistency, transparency, timeliness, and predictability in the review process; and to provide recommendations for enhanced regulatory practices. A questionnaire was completed by the Medicines Control Council (MCC) to describe the organization of the authority, record key milestones and timelines in the review process and to identify good review practices (GRevPs). Currently, the MCC conducts a full assessment of quality, efficacy, and safety data in the review of all applications. The overall regulatory median approval time decreased by 14% in 2017 (1411 calendar days) compared with that of 2016, despite the 27% increase in the number of applications. However, the MCC has no target for overall approval time of new active substance applications and no targets for key review milestones. Guidelines, standard operating procedures, and review templates are in place, while the formal implementation of GRevPs and the application of an electronic document management system are planned for the near future. As the MCC transitions to the newly established South Africa Health Products Regulatory Authority, it would be crucial for the authority to recognize the opportunities for an enhanced regulatory review and should consider models such as abridged assessment, which encompass elements of risk stratification and reliance. It is hoped that resource constraints may then be alleviated and capacity developed to meet target timelines.


[The losses of nutrients by fire in a savanna vegetation of Trachypogon].
The losses of nutrients by fire were assessed in a Trachypogon savanna located in Calabozo, Venezuela. About 95% of the biomass, 97% of N, 61% of P, 76% of K and 65% of Ca and Mg were transferred to the atmosphere. Ash deposition returned between 21-34% of Mg, Ca, K and P and 0.2% of N. Previous data indicated that precipitation and atmospheric fixation (in the case of N) could replace the rest of N, Ca, K and Mg transferred to the atmosphere. As a consequence of frequent burning (annual or biannual), the soil of the savanna showed lower organic matter and available P and K content when it is compared with a 32 year protected savanna.


Rheumatic diseases in an MRL strain of mice with a deficit in the functional Fas ligand.
To characterize Fas antigen expression on the cell surface, and to determine the effect of this expression in rheumatic diseases using a newly established gld-congenic MRL strain of mice (MRL/gld), which is defective in its functional Fas ligand (Fas-L). Flow cytometric analyses of lymphoid cells and macrophages were performed using anti-Fas and other cell surface markers. Histopathologic manifestations were examined using immunochemistry and light and electron microscopy. Serum levels of IgG and anti-DNA antibodies were measured by single radial immunodiffusion and enzyme-linked immunosorbent assay, respectively. MRL/gld mice developed systemic lymphadenopathy with an accumulation of Thy1.2+, B220+ and CD4-, CD8- T cells, which both express the Fas antigen. Splenic B cells positive for surface IgM and/or surface IgD, and resident peritoneal macrophages exhibited up-regulated expression of the Fas antigen, at much higher levels than those observed in MRL/MpJ-+/+ (MRL/+) mice. Forms of rheumatic disease were observed in these mice, although not in C3H/HeJ-gld/gld mice. These forms included diffuse glomerulonephritis, granulomatous arteritis, and arthritis, and were associated with the infiltration of mononuclear cells expressing the Fas antigen. Serum levels of IgG and anti-DNA antibodies were significantly increased in MRL/gld mice compared with MRL/+ mice. Rheumatic disease was generated by the gld gene in mice with an MRL background, as it is by the lpr gene, which is a Fas deletion mutant, associated with autoimmune traits. Rheumatic disease in this MRL strain was initiated by an incapacity for Fas/Fas-L-induced apoptosis, resulting in the development of autoimmunity and allowing for a persistent immune response in the affected lesions.


Towards practice development--a vision in reality or a reality without vision?
This paper describes the development of a conceptual framework for practice development. Drawing on the authors' combined experiences of facilitating developments in practice, a conceptual framework is proposed. It is argued that much practice development in health care today lacks a systematic approach and is often undertaken by individual practitioners who are poorly prepared for their roles. A short history of practice development is outlined to contextualize current development activities. The proposed framework is located in a critical social science philosophy and it is suggested that such a philosophy enables individual growth and development, empowerment of practitioners and the generation of cultural change that sustains continuous growth and innovation in practice. An example of the framework in use is described and recommendations proposed to enable organizations to embrace a systematic approach to practice development.


[Treatment of facial lesion in infantile and childhood atopic dermatitis].
Treatment of infantile and childhood atopic dermatitis needs special consideration compared to adult's counterpart. Repeated exposure to irritation by slaver or rubbing makes facial lesions difficult to treat in infants and children. We considered the deterioration factor and the treatment that we performed. We considered 6 infantile and childhood atopic dermatitis who had severe condition on their face. All the patients needed enough application of topical steroids. Exacerbation of atopic dermatitis masked concomitant skin infection such as impetigo or Kaposi's varicelliform eruption. Guardian usually hesitated periorbital and perioral application of topical steroids and tacrolimus. In order to obtain the guardian's understanding and cooperation, it was very important to establish a good doctor/guardian relationship by educational approach including an enough explanation about treatment policy. It was important for the treatment of infantile and childhood atopic dermatitis to identify and eliminate triggering factors. And we need to obtain the guardian's understanding and cooperation for our treatment policy.


The NORPLANT system of contraception.
The NORPLANT system is the first new methodology to be introduced in the past 30 years in the field of contraception. It is a subdermal, reversible system of Silastic capsules containing the synthetic progestin levonorgestrel. Advantages and disadvantages are discussed. Compliance-free use and high effectiveness are its main characteristics.


Expression of chemically synthesized alpha-neo-endorphin gene fused to E. coli alkaline phosphatase.
An alpha-neo-endorphin (alpha NE) gene, which we previously synthesized chemically and inserted into E. coli beta-galactosidase gene of pK013 plasmid, has been excised and fused to E. coli alkaline phosphatase (APase) gene. One of the transformants was named E15/pA alpha NE1. Under the APase gene regulation, APase-alpha NE chimeric protein was expressed at 1.3 X 10(6) molecules per cell, and accounted for about 60% of total cellular proteins. The HPLC pattern of CNBr treated E15/pA alpha NE1 was very simple reflecting the high content of the chimeric protein and low numbers of methionine residues in it. A series of genes encoding APase-alpha NE chimeric proteins in which 30 to 94 C-terminal amino acid residues were replaced by (met)-alpha NE, was cloned in E. coli. Transportation of the chimeric proteins to periplasmic space was studied. All chimeric proteins were apparently processed by signal peptidase but few, if any, was transported to the periplasmic space.


Factors associated with the level of prolactin in patients under remission from Alcohol Use Disorder: A gender perspective.
Prolactin mirrors the dopaminergic activity in the brain which is key to understanding alcohol use disorders (AUD). Still, patients with AUD are a heterogenous group and there seem to be gender differences in the relationship between alcohol use and the level of prolactin. In this study, we examined gender-wise relationship of alcohol use trait- and state-related factors with the level of prolactin among AUD inpatients in remission. This cross-sectional study examined the level of prolactin along with general patient characteristics and alcohol use trait- and state-related factors that could influence the level of prolactin in 112 AUD inpatients at three rehabilitation clinics in Norway. Logistic regression was performed to identify the gender-specific predictors of level of prolactin. Male and female AUD patients had similar level of prolactin. Among females, younger age, early alcohol debut, and absence of parental drinking problem predicted higher level of prolactin. In males, presence of other substance dependence predicted a lower level of prolactin. There were gender differences in the factors associated with the level of prolactin among the AUD patients. Especially in the female AUD patients under remission, alcohol use trait-related factors were better predictors of the level of prolactin than the alcohol use state-related factors, indicating that individuals might characteristically have varying degree of dopamine reactivity.


Hepatitis C virus-associated hepatocellular carcinoma after sustained virologic response.
The introduction of a direct-acting antiviral (DAA) for patients with hepatitis C virus (HCV) infection, could lead to higher sustained virologic response (SVR) rates with fewer adverse events, and it could shorten the treatment duration relative to the interferon era. Although most recent clinical studies have demonstrated that the occurrence rates of hepatocellular carcinoma (HCC) are decreased by SVR with both interferon-based and interferon-free-regimens, there are several reports about the unexpected observation of high rates of early tumor occurrence and recurrence in patients with HCV-related HCC undergoing interferon-free therapy despite SVR. Several mechanisms of HCC occurrence and rapid immunological changes, including cytokines and chemokines during and after DAA treatment, have also been reported. We focused on the possibilities that HCC occurs or recurs during and after DAA treatment, based on the reported clinical and basic studies. Further studies and observations will be needed to determine the short-term and long-term effects on hepatocarcinogenesis caused by the eradication of HCV with DAAs. New serum biomarkers and a follow-up system for HCV-patients with SVR should be established.


Diagnosis and classification of personality disorders: novel approaches.
To provide an update of the recent studies, which have evaluated the radical changes in personality disorder classification in DSM-5 and ICD-11. Although the DSM-5 Committee rejected the personality disorders Work Group proposal for personality disorder classification, the model was published in DSM-5 Section III. This Alternative Model of Personality Disorders (AMPD) has been widely adopted by the research community resulting in multiple studies evaluating its reliability and clinical utility. The ICD-11 Personality Classification has recently been accepted by the WHO and is also receiving increasing study. Both models emphasize personality disorder severity, which most studies report is consistently linked to impairment and outcome. Both models propose five descriptive domains, which appear to capture most of the current personality disorder diagnoses, and can also be linked to disease extremes of normal personality such as the Five Factor Model. The changes in DSM-5 AMPD and ICD-11 represent a significant paradigm shift in the diagnosis of personality disorders. Early research suggests that the changes may be beneficial for clinicians and researchers. The models more closely align with the large body of literature supporting dimensional models of normal personality. The severity dimensions are consistent with the large body of evidence that personality disorder severity is a strong determinant of impairment and outcome. It remains to be seen if clinicians will use the classification to plan and predict treatment for a wide range of mental disorders.


[Interaction of tetrahydroisoquinolines and 3-aminotetralines with opioid mu-receptors].
The interaction of the tetrahydroisoquinoline (THIQ) and 3-aminotetraline (3-AT derivatives with opioid mu-receptors has been studied. It is shown that THIQ and 3-AT derivatives bind to a site on the mu-receptor which these compounds are likely to share with "classical" opiates, whose structure also includes the 3-AT group. The binding site for nonpeptide substances is in a strong allosteric interaction with the binding site for enkephalins. Some biological effects of THIQ and 3-AT derivatives can be explained in terms of their interaction with opioid receptors. One may speculate that the evolution of the endogenous opioid receptor ligands proceeded from simple 3-AT derivatives towards morphinans and, probably, benzomorphans.


The Role of IL-22 in Wound Healing. Potential Implications in Clinical Practice.
Wound healing is a complex process that is mediated and influenced by several cytokines, chemokines, and growth factors. Interleukin-22 (IL-22) is a cytokine that plays a critical role in tissue regeneration. Our study is a systematic review that addressed the implications of IL-22 in the healing of wounds caused by external factors. Thirteen studies were included in our review, most of them being experimental studies. Three clinical studies underlined the potential role of IL-22 in day-to-day clinical practice. IL-22 plays a central role in wound healing, stimulating the proliferation, migration, and differentiation of the cells involved in tissue repair. However, overexpression of IL-22 can cause negative effects, such as keloid scars or peritoneal adhesions. The results of the presented studies are promising, but further research that validates the roles of IL-22 in clinical practice and analyzes its potential implication in surgical healing is welcomed.


Non-histone chromosomal proteins: their role in the regulation of histone-gene expression.
Histone-gene expression was studied during the cell cycle of continuously dividing HeLa S3 cells and after stimulation of confluent monolayers of WI-38 human diploid fibroblasts to proliferate. The presence of histone-mRNA sequences was assayed by hybridization to a 3H-labelled single-stranded DNA complementary to histone-mRNA molecules. In HeLa S3 cells histone mRNA sequences were found in the nucleus and associated with polyribosomes during S-phase, but not during G1-phase. Transcripts of S-phase chromatin contained histone-mRNA sequences, but those of G1-phase chromatin did not. Similarly, in WI-38 cells association of histone-mRNA sequences with polyribosomes and transcription of histone-mRNA sequences from chromatin parallel DNA replication. Taken together these results suggest that the regulation of histone-gene expression resides, at least in part, at the transcriptional level. Chromatin-reconstitution studies provide evidence that non-histone chromosomal proteins play a key role in activation of histone-gene transcription during the period of the cell cycle when DNA is replicated. Phosphate groups associated with the S-phase non-histone chromosomal proteins appear to be functionally involved in the control of histone-gene readout.


Use of market research data by state chronic disease programs, Illinois, 2012-2014.
Market research data complement traditional epidemiologic data by allowing users to examine health behavior and patterns by census block or census tract. Market research data can identify products and behaviors that align or do not align with public health program goals. Illinois is a recipient of an award from the Directors of Health Promotion and Education to use industry market research data collected by The Nielsen Company for public health purposes. Illinois creates customized community profiles using market research data on tobacco use characteristics to describe the demographics, habits, and media preferences of smokers in certain locations. Local agencies use profiles to plan and target marketing initiatives, reach disparate groups within overall community populations, and restructure program objectives and policy initiatives. Local market research data provide detailed information on the characteristics of smokers, allowing Illinois communities to design public health programs without having to collect data on their own.


The effects of a group visit program on outcomes of diabetes care in an urban family practice.
Diabetes mellitus is a prevalent chronic health condition associated with significant morbidity and mortality. Those with diabetes must acquire self-efficacy in the tasks necessary for them to successfully manage their disease. In this study, a controlled pre- and post-design was used to determine the effect of an adult support and education group visit program embedded in an urban academic family medicine practice on weight and the achievement of treatment goals for hemoglobin A1C, low-density lipoprotein (LDL) blood concentration, and blood pressure (BP) several months after it was implemented. Participants in the program were matched to a comparison group based on age, gender, race/ethnicity, and zip code group, a surrogate marker for socioeconomic status. The distribution of demographic characteristics and co-morbidities was similar between the groups. Significant increases occurred in the proportion of participants achieving both an A1C concentration <7% (CMH=4.6613, p = 0.0309) while controlling for baseline AIC concentration, and a BP<140/90 mm Hg (CMH=5.61, p = 0.018) controlling for baseline BP compared to the comparison group. The hemoglobin A1C concentration declined in 76.9% of the participants in the group visit program compared to 54.3% in the comparison group (CMH=8.9911, p = 0.0027). The increase in the proportion of group visit participants achieving the target LDL concentrations did not achieve statistical significance. The proportion of participants who lost weight was similar to that in the comparison group. Early experience with the program was encouraging and suggested it may improve patients' management of their diabetes mellitus in an urban, predominantly African American population.


Acidity and proton affinity of hypoxanthine in the gas phase versus in solution: intrinsic reactivity and biological implications.
Hypoxanthine is a mutagenic purine base that most commonly arises from the oxidative deamination of adenine. Damaged bases such as hypoxanthine are associated with carcinogenesis and cell death. This inevitable damage is counteracted by glycosylase enzymes, which cleave damaged bases from DNA. Alkyladenine DNA glycosylase (AAG) is the enzyme responsible for excising hypoxanthine from DNA in humans. In an effort to understand the intrinsic properties of hypoxanthine, we examined the gas-phase acidity and proton affinity using quantum mechanical calculations and gas-phase mass spectrometric experimental methods. In this work, we establish that the most acidic site of hypoxanthine has a gas-phase acidity of 332 +/- 2 kcal mol-1, which is more acidic than hydrochloric acid. We also bracket a less acidic site of hypoxanthine at 368 +/- 3 kcal mol-1. We measure the proton affinity of the most basic site of hypoxanthine to be 222 +/- 3 kcal mol-1. DFT calculations of these values are consistent with the experimental data. We also use calculations to compare the acidic and basic properties of hypoxanthine with those of the normal bases adenine and guanine. We find that the N9-H of hypoxanthine is more acidic than that of adenine and guanine, pointing to a way that AAG could discriminate damaged bases from normal bases. We hypothesize that AAG may cleave certain damaged nucleobases as anions and that the active site may take advantage of a nonpolar environment to favor deprotonated hypoxanthine as a leaving group versus deprotonated adenine or guanine. We also show that an alternate mechanism involving preprotonation of hypoxanthine is energetically less attractive, because the proton affinity of hypoxanthine is less than that of adenine and guanine. Last, we compare the acidity in the gas phase versus that in solution and find that a nonpolar environment enhances the differences in acidity among hypoxanthine, adenine, and guanine.


Dysexecutive syndrome in Parkinson's disease: the GREFEX study.
The objectives of this study were to characterize the frequencies and profiles of behavioral and cognitive dysexecutive syndromes in PD (based on validated battery and diagnostic criteria) and to develop a shortened diagnostic battery. Eighty-eight non-demented patients with a diagnosis of PD were examined with an executive validated battery. Using a validated framework, the patients' test results were interpreted with respect to normative data from 780 controls. A dysexecutive syndrome was observed in 80.6% of the patients [95% confidence interval: 71.1-90.1]. The dysexecutive profile was characterized by prominent impairments in deduction, flexibility, inhibition and initiation in the cognitive domain, and by global hypoactivity with apathy and hyperactivity in the behavioral domain. This finding implies that patients with PD should be assessed with cognitive tests and a validated inventory for behavioral dysexecutive syndromes. A shortened battery (based on three cognitive tests and three behavioral domains) provided high diagnostic accuracy.


Age-related changes in serum undercarboxylated osteocalcin and its relationships with bone density, bone quality, and hip fracture.
The effect of the degree of carboxylation of osteocalcin (OC) on the properties of bone is unclear. The aim of this study was to relate serum concentrations of total OC (tOC) and undercarboxylated OC (ucOC), measured with a two-site immunoassay, to bone mineral density (BMD) at the femoral neck and ultrasonic transmitted velocity (UTV) at the os calcis in 257 women aged 60-99 years, 22 of whom had sustained a hip fracture. There was an increase in tOC (r = 0.19, P = 0.003) and ucOC (r = 0.20, P = 0.002) with age. No significant difference in tOC or ucOC between subjects with and without hip fracture was found. Serum tOC was negatively correlated with femoral neck BMD (r = -0.23, P = 0.0001) and os calcis UTV (r = -0.29, P = 0.0001) and partial correlations indicated that these relationships were independent of age. Serum ucOC also correlated negatively with os calcis UTV (r = -0.21, P = 0.001) and less strongly with femoral neck BMD (r = -0.13, P = 0.052). After adjusting for age, only the relationship between ucOC and os calcis UTV remained significant (r = -0.16, P = 0.017). It is concluded that in women over 60 years, the increase in tOC reflects an age-related rise in bone remodeling, whereas the increase in ucOC reflects an age-related fall in vitamin K status. The stronger relationship of ucOC with UTV than BMD suggests that the rise in ucOC may perhaps relate more to changes in bone quality than mineral content. Higher serum ucOC concentrations in subjects with a history of hip fracture could not be confirmed.


Beyond Woodward-Fieser Rules: Design Principles of Property-Oriented Chromophores Based on Explainable Deep Learning Optical Spectroscopy.
An adequate understanding of molecular structure-property relationships is important for developing new molecules with desired properties. Although deep learning optical spectroscopy (DLOS) has been successfully applied to predict the optical and photophysical properties of organic chromophores, how specific functional groups and solvents affect the optical properties is not clearly understood. Here, we employed an explainable DLOS method by applying the integrated gradients method to DLOS. The integrated gradients method allows us to obtain attributions, indicating how much the functional group contributes to the optical properties including the absorption wavelength and bandwidth, extinction coefficients, emission wavelength and bandwidth, photoluminescence quantum yield, and lifetime. The attributions of 54 functional groups and 9 solvent molecules to seven optical properties are quantified and can be used to estimate the optical properties of chromophores as in the Woodward-Fieser rule. Unlike the Woodward-Fieser rule for only the absorption wavelength, the attributions obtained in this work can be applied to estimate all seven optical properties, which makes a significant extension of the Woodward-Fieser rules. In addition, we demonstrated a strategy for utilizing the attributions in the design of molecules and in tuning the optical properties of the molecules. The design of molecular structures using attributions can revolutionize the development of optimal molecules.


Ultra-broadband optical parametric generation in periodically poled stoichiometric LiTaO3.
We report on broadband gain in an optical parametric generator based on periodically poled 1 mol% magnesium-doped stoichiometric LiTaO3 (PPMg:SLT). More than an octave-spanning parametric gain, stretching from near to mid-infrared, is generated by pumping the crystals close to the point where, at parametric degeneracy, the waves experience zero group-velocity dispersion. Using a picosecond Ti:sapphire source, we measured the broadest parametric gain bandwidths, 180 THz at 10 dB, in PPMg:SLT gratings with a period of 25 µm pumped at 860 nm.


Quantitative detection of amplification of proto-oncogenes in breast cancer.
In the present study, dot-blot hybridization, serial dilution analysis and densitometric scanning were used to detect amplification of proto-oncogenes including c-erbB2, c-myc, int-2 and c-Ha-ras in 101 paraffin-embedded breast cancers. Expression of c-erbB2 was also examined by immunohistochemistry. Amplification of c-erbB2, c-myc and int-2 genes was found in 34.7%, 17.8% and 11.9% of breast cancers respectively. However amplification of c-Ha-ras was not detected in all cases. In 11.9% of cases co-amplification of two or more oncogenes was observed. Positive immunostaining of c-erbB2 was seen in 23.8% of the cases and it was significantly associated, but not always corresponding to the amplification of the gene. There was no difference between primary and metastatic breast cancer in the alterations of proto-oncogenes examined in this study, which suggested that the amplification and overexpression of these proto-oncogenes occurred prior to and maintained in the process of metastasis of breast cancer. Statistical analysis showed that high-scale of immunopositive staining of c-erbB2 and high-fold co-amplification of proto-oncogenes were significantly correlated with large size of the tumour and the number of involved lymph nodes. Our results indicate that the alterations of multiple oncogenes are involved in the development of breast cancer and some of them may have prognostic importance for breast cancer patients.


EZH2-mediated repression of GSK-3β and TP53 promotes Wnt/β-catenin signaling-dependent cell expansion in cervical carcinoma.
Enhancer of zeste homolog 2 (EZH2), a catalytic core component of the Polycomb repressive complex 2 (PRC2), stimulates the silencing of target genes through histone H3 lysine 27 trimethylation (H3K27me3). Recent findings have indicated EZH2 is involved in the development and progression of various human cancers. However, the exact mechanism of EZH2 in the promotion of cervical cancer is largely unknown. Here, we show that EZH2 expression gradually increases during the progression of cervical cancer. We identified a significant positive correlation between EZH2 expression and cell proliferation in vitro and tumor formation in vivo by the up-regulation or down-regulation of EZH2 using CRISPR-Cas9-mediated gene editing technology and shRNA in HeLa and SiHa cells. Further investigation indicated that EZH2 protein significantly accelerated the cell cycle transition from the G0/G1 to S phase. TOP/FOP-Flash reporter assay revealed that EZH2 significantly activated Wnt/β-catenin signaling and the target genes of Wnt/β-catenin pathway were up-regulated, including β-catenin, cyclin D1, and c-myc. Moreover, dual-luciferase reporter and chromatin immunoprecipitation (ChIP) assays confirmed that EZH2 inhibited the expression of glycogen synthase kinase-3β (GSK-3β) and TP53 through physically interacting with motifs in the promoters of the GSK-3β and TP53 genes. Additionally, blockage of the Wnt/β-catenin pathway resulted in significant inhibition of cell proliferation, and activation of the Wnt/β-catenin pathway resulted in significant enhancement of cell proliferation, as induced by EZH2. Taken together, our data demonstrate that EZH2 promotes cell proliferation and tumor formation in cervical cancer through activating the Wnt/β-catenin pathway by epigenetic silencing via GSK-3β and TP53.


Outside the box: will information technology be a viable intervention to improve the quality of cancer care?
The use of health information technology (IT) to resolve the crisis in communication inherent within the fragmented service environment of medical care in the United States is a strategic priority for the Department of Health and Human Services. Yet the deployment of health IT alone is not sufficient to improve quality in health service delivery; what is needed is a human factors approach designed to optimize the balance between health-care users, health-care providers, policies, procedures, and technologies. An evaluation of interface issues between primary and specialist care related to cancer reveals opportunities for human factors improvement along the cancer care continuum. Applications that emphasize cognitive support for prevention recommendations and that encourage patient engagement can help create a coordinated health-care environment conducive to cancer prevention and early detection. An emphasis on reliability, transparency, and accountability can help improve the coordination of activities among multiple service providers during diagnosis and treatment. A switch in emphasis from a transaction-based approach to one emphasizing long-term support for healing relationships should help improve patient outcomes during cancer survivorship and end-of-life care. Across the entire continuum of care, an emphasis on "meaningful use" of health IT-rather than on IT as an endpoint-should help put cancer on a path toward substantive continuous quality improvement. The accompanying research questions will focus on reducing the variance between the social and technical subsystems as IT is used to improve patient outcomes across the interfaces of care.


Transcriptomic responses to functional feeds in Atlantic salmon (Salmo salar).
Functional feeds are diets that have positive effects on both health and growth promoting performance of the animals ingesting them, by supplying additional compounds above and beyond the basic nutritional requirements for animal growth alone. The most common additives used in aquaculture diets are probiotics, prebiotics, immunostimulants, vitamins and nucleotides. Inclusion of these components to fish diets can increase feed conversion efficiency and growth, as well as having positive effects on the fish immune system. This review discusses the results from previous studies on fish nutrition and includes a novel genomic approach, using microarray analysis, to elucidate nutritional responses in Atlantic salmon (Salmo salar) fed a newly developed functional feed health premix diet. The transcriptome analysis demonstrated that compared to the standard diet feeding with the functional feed had significant effects on biological processes in the liver. This resulted in a reduction of the expression of genes related to protein turnover, reduced circulating plasma proteins and a down regulation of genes involved in the immune response. These results suggest that the functional feed may infer a decrease in whole body metabolic demands, suppressing both protein turnover and whole body oxygen demand, as well as down regulating several genes involved in the innate immune system. Together these changes appear to result in less energy wastage in fish and an enhanced growth and performance.


Warm heart surgery: experience with long cross-clamp times.
Although hypothermic cardioplegic arrest prolongs the period of ischemic arrest by reducing oxygen demands, it leaves the heart dependent solely on anaerobic metabolism for its energy demands and exposes it to the detrimental effects of hypothermia. Consequently, myocardial protection is compromised, and safe aortic occlusion time is limited to 120 minutes. As electromechanical arrest accounts for 90% of myocardial oxygen consumption, we hypothesized that an ideal state of the heart might be chemically arrested and perfused with warm blood, ie, aerobic arrest. We applied this approach to myocardial protection in 308 consecutive procedures. To assess the adequacy of this method, we reviewed the results in a group of 22 patients in whom the aortic cross-clamp time was, of necessity, greater than or equal to 3 hours (mean time, 204 minutes; range, 180 to 393 minutes). Nineteen of the patients represented a high operative risk with grade 3 or 4 left ventricular function and New York Heart Association class III or IV. All hearts resumed spontaneous normal sinus rhythm without defibrillation, and 21 patients were easily weaned from bypass within minutes of removal of the aortic cross-clamp without inotropic or intraaortic balloon pump support. Mortality was 4.5%, low-output syndrome occurred in 4.5%, and there were no perioperative myocardial infarctions. Our results suggest that warm aerobic arrest is safe and effective in prolonged high-risk procedures, virtually eliminating the period of ischemia, limiting the period and injury of reperfusion, and abolishing the detrimental effects of hypothermia.


Formal Synthesis of Indolizidine and Quinolizidine Alkaloids from Vinyl Cyclic Carbonates.
Cyclic carbonates have long been considered relatively inert molecules acting as protecting groups in complex multistep synthetic routes. This study shows that a concise, yet modular synthesis of indolizidine and quinolizidine alkaloids can be developed from vinyl-substituted cyclic carbonate (VCC) intermediates. Through a highly stereoselective palladium-catalyzed allylic alkylation reaction, these alkaloid motifs can be assembled in four synthetic and only two column purification steps. The combined results help to further advance functionalized cyclic carbonates as useful and reactive intermediates in natural product synthesis.


Effects of a Phytogenic Feed Additive Versus an Antibiotic Feed Additive on Oxidative Stress in Broiler Chicks and a Possible Mechanism Determined by Electron Spin Resonance.
Phytogenic feed additives are plant-derived products used in poultry feeding to improve overall performance of broilers. In this study, 588 one day-old Cobb 500 chicks were fed one of four diets and housed on either dirty or clean litter for 3wks. Treatments included: Group I: commercial diet with no additive and housed on clean litter; Group II: commercial diet with no additive and housed on dirty litter; Group III: commercial diet with a 0.05% inclusion of the anitobiotic, BMD (bacitracin methylene disalicylate); Group IV: commercial diet with a 0.05% inclusion of a phytogenic feed additive (PFA). The study was designed around a random block assignment of treatments allocated to groups of twenty-one birds per pen. Blood samples were obtained from chicks at 18 days of age for measurement of leukocyte oxidative activity by a bioluminescence technique. Results of the study showed that chicks in the treatment groups fed the PFA had significantly lower oxidative stress (p<0.02) when compared to the BMD treatment group. Once this was determined, electron spin resonance (ESR) spin trapping was used to detect and measure hydroxyl or superoxide radicals in. Fenton chemistry was utilized for production of hydroxyl radicals and a xanthine/xanthine oxidase reaction for the production of superoxide radicals in the diet and in RAW 264.7 mouse peritoneal monocytes exposed to the diet. Results from the reactions showed that the antibiotic scavenges hydroxyl and superoxide radicals more efficiently than the phytogenic. The results were comparable to those measured in the RAW 264.7 cells.


Clinical reasoning case studies as teaching tools.
Students are taught how to apply clinical reasoning methods through a variety of teaching methods, including the use of case studies. Various types of case studies have been described in the literature: paper cases, videotape cases, simulated client cases, and real client cases. This study examined the effectiveness of a new type of paper case study--the clinical reasoning case study--in teaching the clinical reasoning process to occupational therapy students. Four seniors in an undergraduate occupational therapy program completed intervention plans in response to both traditional medical model and clinical reasoning paper case studies. Qualitative methods were used to analyze intervention plans and videotaped discussion about this learning experience. Themes discovered in the data sources suggest that compared with traditional case studies, the clinical reasoning case studies increased the quality of participants' intervention plans, participants' confidence levels about their plans, and participants' understanding of the clinical reasoning process. Participants also reported preferring clinical reasoning case studies over traditional paper case studies. The clinical reasoning case studies are effective teaching tools because they provide students with a holistic picture of the client and his or her occupational therapy treatment. In addition, these case studies model the clinical reasoning process by organizing client information according to the types of clinical reasoning that would be used to gather that information. Occupational therapy educators may find this type of paper case study useful in introducing students to the intervention planning process.


[Limited proteolysis of integrin alpha v beta3 from human placenta].
Purification of alpha v beta 3 integrin from human placenta with successive usage of two affinity sorbents--immobilized monoclonal antibodies to alpha v beta 3 integrin and immobilized RGD-containing decapeptide allowed to purify this integrin's partially degraded fraction, that was nevertheless able to interact with its ligand. During the incubation of partially degraded alpha v beta 3 integrin at 37 degrees C its further degradation went on. Addition of serine proteinase inhibitors: (phenylmethilsulfonyl fluoride, leupeptin and aprotinin) completely suppressed integrin further degradation of alpha v beta 3. In preparations of intact and partially degraded alpha v beta 3 integrin specific activity of two serine proteinases--urokinase and dipeptidilpeptidase IV--was discovered. alpha v beta 3 integrin, undergoing limited proteolysis, had lesser affinity towards RGD peptide, that intact integrin. The results show, that alpha v beta 3 integrin from human placenta co-purifies with serine proteinases. It is suggested that a definite part of functionally active alpha v beta 3 integrin, extracted from human placenta by triton X-100, forms a stable complex with serine proteinases.


The need for support services for family carers of people with motor neurone disease (MND): views of current and former family caregivers a qualitative study.
Family carers provide the majority of home-based care for people with motor neurone disease (MND). Carers' need for, and use of, support services are not fully understood; this study aimed to explore, from a qualitative perspective, the views of current and former family carers of people with MND. A qualitative study was undertaken in Northwest England, using narrative interviews with current (18) and former (10) carers of a family member with MND. An optional longitudinal element involving diary completion was offered to the current carers. Data were analyzed using a thematic framework approach. Carer's needs vary, but encompass the provision of information and training, availability of respite care, counselling, and access to trained paid-for carers. There is need for a range of support services to be made available from which carers can select those most appropriate for them. Some support services are not always available for carers of this client group. There is a need for carers to access greater manual handling and training for physical care. Without sufficient support, carer burden can be overwhelming which may impact on the place of care of the patient and ultimately has implications for health and social care services.


Activation of protein kinase C as a modulator of potentiated UK-14304-induced contractions in dog mesenteric artery and vein.
We assessed the role of protein kinase C (PKC) in the mechanism responsible for the potentiation of UK-14304-induced contractions produced when isolated dog mesenteric vascular rings were pretreated with threshold concentrations of 12-O-tetradecanoyl-phorbol-13-acetate (TPA), KCl, or endothelin-1 (ET-1). In dog mesenteric artery. UK-14304 produced a biphasic concentration-response curve in the presence of TPA, KCl, or ET-1, with the curve portion at lower concentrations being alpha 2-adrenoceptor dependent and the portion at higher concentrations being alpha 1-adrenoceptor dependent. Calphostin C (10(-6)M), a PKC inhibitor, abolished amplified UK-14304-induced contraction in the TPA-pretreated tissues. In the KCl- and ET-1-pretreated tissues. 10(-6)M calphostin C antagonized amplified UK-14304-induced contractions by approximately 20% in both parts of the concentration-response curve. In contrast, in dog mesenteric vein, amplified UK-14304-induced contractions by TPA, KCl, and ET-1 were entirely dependent on alpha 2-adrenoceptors. Calphostin C (10(-6)M), which in control experiments had no effect on KCl-induced contraction and antagonized responses to TPA by 60.1%, inhibited UK-14304-induced contraction by 18.3%. Amplified UK-14304-induced contraction was antagonized by 10(-6)M calphostin C by 21.8% in KCl-precontracted tissues, 58.1% in ET-1-precontracted tissues, and 66.3% in TPA-precontracted tissues. In the ET-1- and TPA-pretreated dog mesenteric veins, 10(-6)M calphostin C decreased maximal tensions of enhanced UK-14304-induced contractions to the same level as the UK-14304-induced maximal tension inhibited by 10(-6)M calphostin C in untreated dog mesenteric vein. Therefore, TPA can be a precontracting agent that amplifies UK-14304-induced contractions through PKC activation in both dog mesenteric artery and vein. PKC predominantly mediates the contraction amplification mechanisms after exposure to ET-1 in dog mesenteric vein and does not play a major role in the amplification of UK-14304-induced contraction by KCl in both dog mesenteric artery and vein. These data show that a common mechanism need not underlie amplification of adrenergic responses in mesenteric artery and vein.


[The health in Tunisia. Achievements and limitations].
Since the independence in 1956, the health sector has scored a great deal of progress. Such a development has been observed also in other developing nations. The country is now facing a crucial issue. On one hand the resources devoted to the health sector are ever increasing and are likely to reach the limits acceptable to the national economy. On the other hand the citizens are seeking for more and better care and are no longer satisfied with the care and health insurance provided to them. On this background, the inequalities of health resources between the regions and areas of the country add an other aggravating factor.


[Depolarization of primary afferents induced by evoked activation of intersegmental connections of the substantia gelatinosa of the spinal cord in the cat].
Primary afferent depolarization evoked by selective activation of intersegmental connections of the substantia gelatinosa was studied in nembutal-anesthetized cats. Intensive depolarization was observed in high-threshold muscle and cutaneous afferents, but it was very weak in group Ia muscle afferent fibres. It is supposed that intersegmental spreading of activity in substantia gelatinosa is transmitted via chains of successively excited neurons. Possible function of primary afferent depolarization is discussed as a factor stabilizing the coordinated activity of spinal neurons.


The evaluation of critically ill transplant patients with infectious diarrhea.
The review summarizes the epidemiology, risk factors, clinical presentation, diagnosis and management of the most important etiologic agents of infectious diarrhea in critically ill transplant recipients. Diarrhea, frequently caused by infectious pathogens, can cause significant morbidity and mortality in transplant recipients. Diarrhea can lead to severe dehydration, acute renal failure, medication toxicity, rejection, graft-versus-host disease and impairs patients' quality of life. Opportunistic infectious pathogens can pose significant diagnostic and therapeutic challenges in immunocompromised hosts. In transplant recipients, it is vital to differentiate infectious from noninfectious diarrhea to adequately manage their therapeutic approach. Supportive measures and reduction in immunosuppression are essential for the treatment management.


Erdheim-Chester disease: an uncommon cause of upper urinary tract obstruction.
Erdheim-Chester disease is a rare non-Langerhans form of systemic histiocytosis of unknown origin. We describe a 45-year-old man presenting with bilateral hydronephrosis suggestive of extrinsic urinary tract obstruction. Computed tomography revealed extensive hypodense soft tissue infiltration in the retroperitoneum surrounding the kidneys. Needle biopsy of the retroperitoneal soft tissue revealed aggregates of lipid-laden histiocytes expressing CD68 but negative for CD1a and S100 protein. The diagnosis of Erdheim-Chester disease was supported by typical radionuclide bone scinitigraphic findings. Treatment with prednisolone, sirolimus, and regular ureteric stent revision was initiated to achieve adequate urinary tract drainage. To our knowledge, this is the second patient with Erdheim-Chester disease reported in Hong Kong. A high index of suspicion is required to avoid delay in the diagnosis of this rare disease.


Role of Pial Microvasospasms and Leukocyte Plugging for Parenchymal Perfusion after Subarachnoid Hemorrhage Assessed by In Vivo Multi-Photon Microscopy.
Subarachnoid hemorrhage (SAH) is associated with acute and delayed cerebral ischemia. We suggested spasms of pial arterioles as a possible mechanism; however, it remained unclear whether and how pial microvasospasms (MVSs) induce cerebral ischemia. Therefore, we used in vivo deep tissue imaging by two-photon microscopy to investigate MVSs together with the intraparenchymal microcirculation in a clinically relevant murine SAH model. Male C57BL/6 mice received a cranial window. Cerebral vessels and leukocytes were labelled with fluorescent dyes and imaged by in vivo two-photon microscopy before and three hours after SAH induced by filament perforation. After SAH, a large clot formed around the perforation site at the skull base, and blood distributed along the perivascular space of the middle cerebral artery up to the cerebral cortex. Comparing the cerebral microvasculature before and after SAH, we identified three different patterns of constrictions: pearl string, global, and bottleneck. At the same time, the volume of perfused intraparenchymal vessels and blood flow velocity in individual arterioles were significantly reduced by more than 60%. Plugging of capillaries by leukocytes was observed but infrequent. The current study demonstrates that perivascular blood is associated with spasms of pial arterioles and that these spasms result in a significant reduction in cortical perfusion after SAH. Thus, the pial microvasospasm seems to be an important mechanism by which blood in the subarachnoid space triggers cerebral ischemia after SAH. Identifying the mechanisms of pial vasospasm may therefore result in novel therapeutic options for SAH patients.


An improved primer set and amplification protocol with increased specificity and sensitivity targeting the Symbiodinium ITS2 region.
The Internal Transcribed Spacer 2 (ITS2) rRNA gene is a commonly targeted genetic marker to assess diversity of Symbiodinium, a dinoflagellate genus of algal endosymbionts that is pervasively associated with marine invertebrates, and notably reef-building corals. Here we tested three commonly used ITS2 primer pairs (SYM_VAR_5.8S2/SYM_VAR_REV, ITSintfor2/ITSReverse, and ITS-DINO/ITS2Rev2) with regard to amplification specificity and sensitivity towards Symbiodinium, as well as sub-genera taxonomic bias. We tested these primers over a range of sample types including three coral species, coral surrounding water, reef surface water, and open ocean water to assess their suitability for use in large-scale next generation sequencing projects and to develop a standardised PCR protocol. We found the SYM_VAR_5.8S2/SYM_VAR_REV primers to perform superior to the other tested ITS2 primers. We therefore used this primer pair to develop a standardised PCR protocol. To do this, we tested the effect of PCR-to-PCR variation, annealing temperature, cycle number, and different polymerase systems on the PCR efficacy. The Symbiodinium ITS2 PCR protocol developed here delivers improved specificity and sensitivity towards Symbiodinium with apparent minimal sub-genera taxonomic bias across all sample types. In particular, the protocol's ability to amplify Symbiodinium from a range of environmental sources will facilitate the study of Symbiodinium populations across biomes.


Vital pulp therapy with new materials for primary teeth: new directions and treatment perspectives.
Vital pulp therapy aims to treat reversible pulpal injury and includes 2 therapeutic approaches: (1) indirect pulp treatment for deep dentinal cavities and (2) direct pulp capping or pulpotomy in cases of pulp exposure. Indirect pulp treatment is recommended as the most appropriate procedure for treating primary teeth with deep caries and reversible pulp inflammation, provided that this diagnosis is based on a good history, a proper clinical and radiographic examination, and that the tooth has been sealed with a leakage-free restoration. Formocresol has been a popular pulpotomy medicament in the primary dentition and is still the most universally taught pulp treatment for primary teeth. Concerns have been raised over the use of formocresol in humans, and several alternatives have been proposed. Controlled clinical studies have been critically reviewed, and mineral trioxide aggregate and ferric sulfate have been considered appropriate alternatives to formocresol for pulpotomies in primary teeth with exposed pulps. In most of the studies reviewed, the caries removal method has not been described. The use of a high-speed handpiece or laser might result in an exposure of a "normal" pulp that would otherwise not be exposed.


Plasma anti-Müllerian hormone in adult dairy cows and associations with fertility.
The objectives of this study were to identify factors associated with concentrations of anti-Müllerian hormone (AMH) in plasma of dairy cows and to investigate the relationships between plasma AMH and fertility responses during a 100-d breeding season. Lactating cows, 1,237 in 2 seasonally calving herds, had estrous cycles presynchronized and were enrolled in a timed artificial insemination (AI) protocol. All cows were inseminated on the first day of breeding season, considered study d 0. Blood was sampled on d -8 and analyzed for concentrations of AMH and progesterone. From d 19 to 35, detection of estrus was performed daily and cows detected in estrus were reinseminated. On d 36, bulls were placed with cows for 65 d of natural service breeding. Factors identified to be associated with concentrations of AMH in plasma were breed of the cow and lactation number. Concentrations of AMH were greater for Jerseys, followed by crossbreds, and then Holsteins. Cows on lactations 2 and 3 had greater concentrations of AMH than those on lactations 1 and ≥ 4. Although pregnancy per AI at the timed AI was not associated with concentrations of AMH, cows with low AMH had greater detection of estrus at timed AI, and the latter benefited pregnancy per AI, particularly in cows that had low progesterone at the beginning of the synchronization protocol. Pregnancy loss between gestation d 30 and 65 was greater in cows with low AMH compared with those with intermediate or high AMH. Return to estrus in cows that failed to become pregnant from the timed AI was not associated with AMH, but pregnancy rate in cows bred on estrus (reinsemination + natural service) was associated positively with AMH. In conclusion, breed and lactation number were identified to be associated with concentrations of AMH in plasma. Concentration of AMH was associated positively with maintenance of pregnancy at the first postpartum AI and with pregnancy rate in cows inseminated after detection of spontaneous estrus. Synchronization of ovulation might override positive associations between AMH and fertility.


Continuing education for pathology laboratory technologists: a needs analysis in a Singapore teaching hospital.
Continuing education is important to laboratory technologists. It helps them keep pace with the advances in medicine and pathology and thus to provide quality service. A survey was conducted to assess the attitudes of pathology laboratory technologists towards different educational activities. Relevant continuing education activities include lectures, case discussions, journal clubs, short courses, scientific conferences and higher educational courses. Technologists were asked for their views on these various activities, scoring each item 1-5 out of 5. The response rate was 66% (19/29); respondents' work experience ranged from <3 years to >20 years. Short courses was rated the most highly; 89% of respondents considered them good/excellent. General medical knowledge and the methods and rationale of key histopathology procedures were the preferred lecture topics. Most respondents (58%) were interested (4-5/5) to have their own journal club. 84% of respondents were fairly keen/keen (4-5/5) to participate in conferences. 74% responded that they were fairly keen/keen to consider pursuing higher education. 84% were fairly keen/keen to upgrade themselves and to assume higher responsibilities. The results indicate a positive outlook of technologists towards continuing education. The planning of future lecture topics has taken into account their preferences, and talks on the pursuit of higher education have been organised. It is hoped that these measures and others, will help boost the usefulness of continuing education and enhance the work environment.


Pregnancy risk among black, white, and Hispanic teen girls in New York City public schools.
Disparities in teen pregnancy rates are explained by different rates of sexual activity and contraceptive use. Identifying other components of risk such as race/ethnicity and neighborhood can inform strategies for teen pregnancy prevention. Data from the 2005 and 2007 New York City Youth Risk Behavior Surveys were used to model demographic differences in odds of recent sexual activity and birth control use among black, white, and Hispanic public high school girls. Overall pregnancy risk was calculated using pregnancy risk index (PRI) methodology, which estimates probability of pregnancy based on current sexual activity and birth control method at last intercourse. Factors of race/ethnicity, grade level, age, borough, and school neighborhood were assessed. Whites reported lower rates of current sexual activity (23.4%) than blacks (35.4%) or Hispanics (32.7%), and had lower predicted pregnancy risk (PRI = 5.4% vs. 9.0% and 10.5%, respectively). Among sexually active females, hormonal contraception use rates were low in all groups (11.6% among whites, 7.8% among blacks, and 7.5% among Hispanics). Compared to white teens, much of the difference in PRI was attributable to poorer contraceptive use (19% among blacks and 50% among Hispanics). Significant differences in contraceptive use were also observed by school neighborhood after adjusting for age group and race/ethnicity. Interventions to reduce teen pregnancy among diverse populations should include messages promoting delayed sexual activity, condom use and use of highly effective birth control methods. Access to long-acting contraceptive methods must be expanded for all sexually active high school students.


Effect of vitamin B-6 (pyridoxine) deficiency on lung elastin cross-linking in perinatal and weanling rat pups.
Weanling and perinatal rats were rendered vitamin B-6 (pyridoxine)-deficient. The rat pups were nursed from vitamin B-6-deficient or -sufficient dams and were killed at day 15 after parturition. The weanling rats were fed vitamin B-6-deficient or -sufficient diets and were killed after 5 weeks of treatment. Lung elastin from the groups of rats was then studied with respect to its content of lysine-derived cross-linking amino acids. Lung lysyl oxidase activity was also measured. B-6 deficiency decreased the number of lysine residues in elastin that were converted into the cross-linking amino acid precursor allysine. However, a more significant defect in cross-link formation was an apparent block in the condensation steps leading to the formation of desmosine. Desmosine was decreased, with an increase in the amounts of aldol condensation products (aldol CP) in elastin. It is proposed that the elevation in aldol CP results from the formation of thiazines, which are produced from the reaction between aldehyde and homocysteine. The concentration of homocysteine is significantly elevated in vitamin B-6-deficient rats.


Characterization and identification of antimicrobial peptides with different functional activities.
In recent years, antimicrobial peptides (AMPs) have become an emerging area of focus when developing therapeutics hot spot residues of proteins are dominant against infections. Importantly, AMPs are produced by virtually all known living organisms and are able to target a wide range of pathogenic microorganisms, including viruses, parasites, bacteria and fungi. Although several studies have proposed different machine learning methods to predict peptides as being AMPs, most do not consider the diversity of AMP activities. On this basis, we specifically investigated the sequence features of AMPs with a range of functional activities, including anti-parasitic, anti-viral, anti-cancer and anti-fungal activities and those that target mammals, Gram-positive and Gram-negative bacteria. A new scheme is proposed to systematically characterize and identify AMPs and their functional activities. The 1st stage of the proposed approach is to identify the AMPs, while the 2nd involves further characterization of their functional activities. Sequential forward selection was employed to extract potentially informative features that are possibly associated with the functional activities of the AMPs. These features include hydrophobicity, the normalized van der Waals volume, polarity, charge and solvent accessibility-all of which are essential attributes in classifying between AMPs and non-AMPs. The results revealed the 1st stage AMP classifier was able to achieve an area under the receiver operating characteristic curve (AUC) value of 0.9894. During the 2nd stage, we found pseudo amino acid composition to be an informative attribute when differentiating between AMPs in terms of their functional activities. The independent testing results demonstrated that the AUCs of the multi-class models were 0.7773, 0.9404, 0.8231, 0.8578, 0.8648, 0.8745 and 0.8672 for anti-parasitic, anti-viral, anti-cancer, anti-fungal AMPs and those that target mammals, Gram-positive and Gram-negative bacteria, respectively. The proposed scheme helps facilitate biological experiments related to the functional analysis of AMPs. Additionally, it was implemented as a user-friendly web server (AMPfun, http://fdblab.csie.ncu.edu.tw/AMPfun/index.html) that allows individuals to explore the antimicrobial functions of peptides of interest.


Structure and biochemical characterization of l-2-hydroxyglutarate dehydrogenase and its role in the pathogenesis of l-2-hydroxyglutaric aciduria.
l-2-hydroxyglutarate dehydrogenase (L2HGDH) is a mitochondrial membrane-associated metabolic enzyme, which catalyzes the oxidation of l-2-hydroxyglutarate (l-2-HG) to 2-oxoglutarate (2-OG). Mutations in human L2HGDH lead to abnormal accumulation of l-2-HG, which causes a neurometabolic disorder named l-2-hydroxyglutaric aciduria (l-2-HGA). Here, we report the crystal structures of Drosophila melanogaster L2HGDH (dmL2HGDH) in FAD-bound form and in complex with FAD and 2-OG and show that dmL2HGDH exhibits high activity and substrate specificity for l-2-HG. dmL2HGDH consists of an FAD-binding domain and a substrate-binding domain, and the active site is located at the interface of the two domains with 2-OG binding to the re-face of the isoalloxazine moiety of FAD. Mutagenesis and activity assay confirmed the functional roles of key residues involved in the substrate binding and catalytic reaction and showed that most of the mutations of dmL2HGDH equivalent to l-2-HGA-associated mutations of human L2HGDH led to complete loss of the activity. The structural and biochemical data together reveal the molecular basis for the substrate specificity and catalytic mechanism of L2HGDH and provide insights into the functional roles of human L2HGDH mutations in the pathogeneses of l-2-HGA.


Epidemiology of lameness in dairy cattle: the influence of cubicles and indoor and outdoor walking surfaces.
A survey of cubicles and indoor and outdoor walking surfaces on 37 farms served by four veterinary practices in Somerset, Cheshire, Wirral and west. Wales was carried out in 1989 to 1991. A study of the space requirements of Friesian/Holstein cows at pasture showed that they required approximately 240 cm x 120 cm lying space and a further 60 cm lunging space for rising. By these standards, 87 per cent of the cubicles were too short and 50 per cent were too wide or too narrow. Over 1500 observations on cows lying down, rising and standing indicated that only 12 per cent of the cubicles permitted real freedom of movement; 91 per cent of top partition rails were judged to be too low and 70 per cent of bottom rails too low or too high. In addition, the kerb was very high in 76 per cent of the cubicles. As a result, 10 per cent of cows appeared moderately or severely restricted when lying down, 33 per cent when rising and 55 per cent when standing. Over 2000 cubicle beds were also studied; 75 per cent had a concrete base and of those, 63 per cent were judged to have too little bedding and 11 per cent next to none. Higher incidences and prevalences of lameness were associated with limited borrowing space (P < 0.01) low bottom rails (P < 0.05), high kerbs (P < 0.05) and inadequate bedding (P < 0.01). Of 3190 walking surfaces, only 25 per cent were classified as satisfactory in the first winter and 34 per cent in the second. In general, surfaces in silage bays were too rough and those in other sites were too smooth. The farms with the smoothest indoor walking surfaces had a significantly higher incidence of lameness (P < 0.01). Of 3335 outdoor walking surfaces only 25 per cent were classified as satisfactory, and 70 per cent were too rough. The incidence of lameness was not significantly related to these findings.


Erosive loss of tooth substance is dependent on enamel surface structure and presence of pellicle - An in vitro study.
Aim of this in vitro study was to investigate erosive tooth loss in dependence of the enamel surface structure and presence of an acquired pellicle. Enamel specimens from 19 bovine incisors (4 specimens/incisor) were allocated to four experimental groups (n = 19). The surfaces of half of the specimens were polished (two groups), while the other half was left native (two groups). Specimens of one polished and one native group were placed in pooled human saliva (30 min) for the formation of an acquired pellicle. Thereafter, all specimens were demineralized by superfusion with hydrochloric acid (17 min, pH 2.3) with collection of the superfluent. Erosive substance loss was determined by measuring the dissolved calcium content using a colorimetric assay with Arsenazo III reagent. Differences in erosive substance loss were statistically analyzed with respect to enamel surface and pellicle. A linear mixed effects model was fitted to the data and pairwise differences between groups were evaluated (significance level α= 0.05). Enamel surface structure (p < 0.001) and presence of pellicle (p = 0.01) had a significant effect on erosive substance loss. Polished surfaces with pellicle showed the lowest cumulative calcium release [nmol Ca/mm2] (means ± standard deviation: 48+/-5), followed by polished specimens without (51+/-9) and native specimens with pellicle (54+/-10). No significant differences were found between these groups. Highest cumulative calcium release was found for native specimens without pellicle (61+/-9; p < 0.05). Both enamel surface structure and the acquired pellicle are important determinants of the susceptibility to erosive tooth loss.


Delimiting species using DNA and morphological variation and discordant species limits in spiny lizards (Sceloporus).
Haplotype phylogenies based on DNA sequence data are increasingly being used to test traditional species-level taxonomies based on morphology. However, few studies have critically compared species limits based on morphological and DNA data, and the methods used to delimit species using either type of data are only rarely explained. In this paper, we review three approaches for species delimitation (tree-based with DNA data and tree-based and character-based with morphological data) and propose explicit protocols for each. We then compare species limits inferred from these approaches, using morphological and mtDNA data for the Yarrow's spiny lizard (Sceloporus jarrovii), a traditionally polytypic species from the southwestern United States and Mexico. All three approaches support division of S. jarrovii into five species, but only two species are the same among the three approaches. We find the greatest support for the five species that are delimited based on mtDNA data, and we argue that mtDNA data may have important (and previously unappreciated) advantages for species delimitation. Because different data and approaches can disagree so extensively, our results demonstrate that the methodology of species delimitation is a critical issue in systematics.