Urine derived cells are a potential source for urological tissue reconstruction.
Contemporary approaches to tissue engineering and cell therapy for urinary tract reconstruction require invasive tissue biopsies to obtain autologous cells. However, these procedures are associated with potential complications. We determined whether the cells present in urine have characteristics of normal bladder cells and investigated their potential uses for urological reconstructive procedures. A total of 55 urine samples were collected from 15 healthy individuals and 8 patients with vesicoureteral reflux. Urine derived cells were isolated, expanded and tested for progenitor and differentiated cell specific markers using flow cytometry, immunofluorescence and Western immunoblotting. The chromosomal stability of cultured urine derived cells was determined by karyotype analysis. Clones were successfully established from primary cultures of urine derived cells. Isolated cells showed 3 phenotypes, including fully differentiated, differentiating and progenitor-like cells. Some urine derived cells stained positive for the surface markers c-Kit, SSEA4, CD105, CD73, CD91, CD133 and CD44. Two to 7 cells per 100 ml urine were multipoint progenitors that could expand extensively in culture. Single progenitor cells had the ability to differentiate into the cell lineages expressing urothelial, smooth muscle, endothelial and interstitial cell markers. The expression of lineage markers was characterized by Western blot and immunofluorescence analysis. Urine derived cells also maintained a normal karyotype after serial culture. A subpopulation of cells isolated from urine had progenitor cell features and the potential to differentiate into several bladder cell lineages. Urine derived cells could serve as an alternative cell source for urinary tract tissue engineering and reconstruction.


Bilateral Pedicled Superficial Inferior Epigastric Artery Flap in the Treatment of Hidradenitis Suppurativa.
The authors present a new technique in surgically treating hidradenitis suppurativa (HS), a debilitating skin condition. In HS, surgical treatment is often the best option because of the high recurrence rates despite extensive medical treatment. A commonly successful surgical method is using skin flaps after excision of the affected area. A superficial inferior epigastric artery flap is demonstrated here as a new alternative approach to treating a case of extensive HS of the groin. By using the pedicled superficial inferior epigastric artery flap for groin reconstruction, inguinal HS can be widely excised and reconstructed with minimal donor-site morbidity and a good aesthetic outcome.


Modulation of Synovial Fluid-Derived Mesenchymal Stem Cells by Intra-Articular and Intraosseous Platelet Rich Plasma Administration.
The aim of this study was to evaluate the effect of intra-articular (IA) or a combination of intra-articular and intraosseous (IO) infiltration of Platelet Rich Plasma (PRP) on the cellular content of synovial fluid (SF) of osteoarthritic patients. Thirty-one patients received a single infiltration of PRP either in the IA space (n = 14) or in the IA space together with two IO infiltrations, one in the medial femoral condyle and one in the tibial plateau (n = 17). SF was collected before and after one week of the infiltration. The presence in the SF of mesenchymal stem cells (MSCs), monocytes, and lymphocytes was determined and quantified by flow cytometry. The number and identity of the MSCs were further confirmed by colony-forming and differentiation assays. PRP infiltration into the subchondral bone (SB) and the IA space induced a reduction in the population of MSCs in the SF. This reduction in MSCs was further confirmed by colony-forming (CFU-F) assay. On the contrary, IA infiltration alone did not cause variations in any of the cellular populations by flow cytometry or CFU-F assay. The SF of osteoarthritic patients contains a population of MSCs that can be modulated by PRP infiltration of the SB compartment.


Effects of cytokinin production under two SAG promoters on senescence and development of tomato plants.
Two promoters of senescence-associated ARABIDOPSIS genes, SAG12 and SAG13, were used in tomato plants to express IPT that catalyzes the rate-limiting step in cytokinin biosynthesis. Expression of these heterologous promoters in tomato plants was analyzed using the reporter gene beta-glucuronidase. Both promoters are expressed in tomato leaves in a manner similar to their expression in ARABIDOPSIS plants. The SAG12 promoter is very specific to senescing leaves, whereas the SAG13 promoter is expressed in mature leaves prior to the onset of visible senescence and its expression increases in senescing leaves. Expression of both promoters in tomato tissues other than leaves was very low . IPT expressed under the control of SAG12 and SAG13 promoters ( PSAG12::IPT and PSAG13::IPT, respectively) resulted in suppression of leaf senescence and advanced flowering, as well as in a slight increase in fruit weight and fruit total soluble solids (TSS). However, expression of PSAG13::IPT also led to stem thickening, short internodal distances and loss of apical dominance. In contrast to the autoregulation of PSAG12::IPT, PSAG13::IPT is expressed at higher levels in mature leaves. This difference is likely due to PSAG13::IPT exhibiting two phases of expression - a senescence-independent expression prior to the onset of senescence that is not subjected to autoregulation by cytokinin, and enhanced expression throughout senescence which is autoregualted by cytokinin. This moderate different autoregulated behavior of PSAG12::IPT and PSAG13::IPT markedly influenced plant development, emphasizing the biological effects of cytokinin in addition to senescence inhibition.


ABR in newborns: effects of electrode configuration, stimulus rate, and EEG rejection levels on test efficiency.
This study investigated the effect of electrode configuration, stimulus rate, and EEG rejection level on the efficiency of ABR testing in babies. ABR to click stimuli at 40 dB nHL were simultaneously recorded from two electrode configurations, ipsilateral mastoid to high forehead (Mi-Fh) and nape to high forehead (N-Fh), with two EEG rejection levels (± 5 μV and ± 10 μV). Stimulus rates were between 39.1 and 69.1 per second. Efficiency was measured by confidence in the ABR for a given test time. Thirty babies who had passed a targeted newborn hearing screen with ABR thresholds ≤ 40 dB nHL. The N-Fh configuration, as expected, gave on average a larger response amplitude compared to the Mi-Fh configuration but was only marginally significantly better in terms of test efficiency. There was no significant effect of stimulus rate on test efficiency between 39.1/s and 59.1/s. The lower ± 5 μV EEG rejection level was more test efficient. This study provides some evidence that, for ABR threshold testing in babies, alternatives of ipsilateral mastoid or nape electrode and a range of stimulus rates have little or no effect on test efficiency. The results support the use of low EEG rejection limits.


Histone Deacetylases Exert Class-Specific Roles in Conditioning the Brain and Heart Against Acute Ischemic Injury.
Ischemia-reperfusion (IR) injury comprises a significant portion of morbidity and mortality from heart and brain diseases worldwide. This enduring clinical problem has inspired myriad reports in the scientific literature of experimental interventions seeking to elucidate the pathology of IR injury. Elective cardiac surgery presents perhaps the most viable scenario for protecting the heart and brain from IR injury due to the opportunity to condition the organs prior to insult. The physiological parameters for the preconditioning of vital organs prior to insult through mechanical and pharmacological maneuvers have been heavily examined. These investigations have revealed new insights into how preconditioning alters cellular responses to IR injury. However, the promise of preconditioning remains unfulfilled at the clinical level, and research seeking to implicate cell signals essential to this protection continues. Recent discoveries in molecular biology have revealed that gene expression can be controlled through posttranslational modifications, without altering the chemical structure of the genetic code. In this scenario, gene expression is repressed by enzymes that cause chromatin compaction through catalytic removal of acetyl moieties from lysine residues on histones. These enzymes, called histone deacetylases (HDACs), can be inhibited pharmacologically, leading to the de-repression of protective genes. The discovery that HDACs can also alter the function of non-histone proteins through posttranslational deacetylation has expanded the potential impact of HDAC inhibitors for the treatment of human disease. HDAC inhibitors have been applied in a very small number of experimental models of IR. However, the scientific literature contains an increasing number of reports demonstrating that HDACs converge on preconditioning signals in the cell. This review will describe the influence of HDACs on major preconditioning signaling pathways in the heart and brain.


Genetic diversity of competence gene loci in clinical genotypes of Streptococcus mutans.
The frequencies of 21 competence genes were analyzed in 94 genotypes of Streptococcus mutans. These include those of a main regulatory system (comCDE), structural, and other regulatory orthologues identified in the genome of strain UA159. PCR and Southern blot analysis revealed that all genes are widespread within the species.


[A case of cerebral cryptococcosis, with special reference to computerized tomography findings].
A case of cryptococcosis simulating brain tumor was reviewed. A 66-year-old female was admitted to our hospital with chief complaint of vertigo, gait disturbance and dysarthria. These symptoms started about one year before admission and worsened. Vomiting and urinary incontinence appeared. Neurological examination revealed left cerebellar ataxia and dysarthria. In plain CT (computerized tomography) irregular ill-defined low density area was noted in the cerebellar vermis and bilateral cerebellar hemispheres. And slight ventricular dilatation was found. Irregular shape of ring-like enhancement corresponding to capsule and patchy or mottled enhancement inside the tumor were seen. Suboccipital craniectomy was performed and yellowish necrotic tumor with hard capsule was removed. Histological diagnosis was not neoplasm or tuberculoma. Postoperatively liver function progressively worsened. She died due to disseminated intravascular coagulation. Autopsy revealed typical liver cirrhosis without malignant change. 3.0 X 2.5 cm sized, slightly hard, yellowish lesion was found on upper part of cerebellar hemispheres. This had extremely necrotic tissue and a great number of cryptococcus neoformans were found. And other intracranial lesion was not confirmed. Finding of pulmonary cryptococcosis was not gained. Our case is very rare because of solitary cerebellar abscess and absence of meningitic episode or pulmonary cryptococcosis. There are three types of inflammation in cerebral cryptococcosis. The commonest manifestation is the meningitic type, the second mode is granulomatous lesion and the third and the least presentation is intracranial abscess formation. CT reveals various findings according to clinical stage. CT findings are those of meningitis, meningoencephalitis, granuloma and abscess. Cryptococcal granuloma or abscess often simulates brain abscess, glioma and metastatic brain tumor.(ABSTRACT TRUNCATED AT 250 WORDS)


Emergency department care and hospitalization as predictors of disability.
Duration of complete and partial disability for work, school, and home activities and activities of daily living during the first 18 months after injury were compared for 2,043 emergency department (ED) patients and 151 hospitalized patients from 22 northwestern Vermont communities who received their initial medical care for injury at the Medical Center Hospital of Vermont and were subsequently interviewed. Larger proportions of hospitalized patients than ED patients had any disability or prolonged disability. During almost all time frames, and even among patients who still had ongoing disability at 18 months, the majority of persons with disability had required ED treatment only. These data suggest that, based on disability, ED patients should not necessarily be considered to have merely minor injuries.


Macrocyclic G-quadruplex ligands.
G-quadruplex stabilizing compounds have recently received increased interest due to their potential application as anticancer therapeutics. A significant number of structurally diverse G-quadruplex ligands have been developed. Some of the most potent and selective ligands currently known are macrocyclic structures which have been modeled after the natural product telomestatin or from porphyrin-based ligands discovered in the late 1990s. These two structural classes of G-quadruplex ligands are reviewed here with special attention to selectivity and structure-activity relationships, and with focus on the recent developments.


Relationship between Self-Rated Health and Lifestyle and Food Habits in Japanese High School Students.
Self-rated health (SRH), a subjective assessment of health status, is extensively used in the field of public health. It is an important and valid measure that is strongly related to morbidity, mortality, longevity and health status. Adolescence is a crucial period for the formation of health status, because health-risk behaviours (e.g., skipping breakfast) are often established during this period. In this study, we investigated the relationship of SRH with lifestyle and eating habits in Japanese high school students. In this study, 1296 students aged 16-18 years from 11 high schools in Japan participated. A questionnaire was administered to these participants that included a question on SRH, five questions on demographic characteristics, six questions on lifestyle items (e.g., wake-up time), five questions on miscellaneous health issues (e.g., anorexia), and 25 questions on food habits and attitudes towards food. We examined the differences between self-rated healthy and unhealthy groups using logistic regression analysis adjusted for gender and age. A dichotomy regression analysis was performed using a stepwise elimination method. Of the 1296 respondents, 16.7% reported feeling unhealthy, 57.7% of whom were females. The self-rated healthy group had a higher frequency of eating breakfast (odds ratio (OR): 2.13; confidence interval (CI): 1.07-4.24) and liked home meals to a greater extent (OR: 3.12; CI: 1.27-7.65) than the self-rated unhealthy group. The two groups did not differ significantly in terms of other lifestyle factors or unidentified complaints. Our results suggest that liking home meals during adolescence may lead to the development of good eating habits, i.e., eating breakfast, and better SRH.


Imprinting along the Kcnq1 domain on mouse chromosome 7 involves repressive histone methylation and recruitment of Polycomb group complexes.
Imprinted genes are clustered in domains, and their allelic repression is mediated by imprinting control regions. These imprinting control regions are marked by DNA methylation, which is essential to maintain imprinting in the embryo. To explore how imprinting is regulated in placenta, we studied the Kcnq1 domain on mouse distal chromosome 7. This large domain is controlled by an intronic imprinting control region and comprises multiple genes that are imprinted in placenta, without the involvement of promoter DNA methylation. We found that the paternal repression along the domain involves acquisition of trimethylation at Lys27 and dimethylation at Lys9 of histone H3. Eed-Ezh2 Polycomb complexes are recruited to the paternal chromosome and potentially regulate its repressive histone methylation. Studies on embryonic stem cells and early embryos support our proposal that chromatin repression is established early in development and is maintained in the placenta. In the embryo, however, imprinting is stably maintained only at genes that have promoter DNA methylation. These data underscore the importance of histone methylation in placental imprinting and identify mechanistic similarities with X-chromosome inactivation in extraembryonic tissues, suggesting that the two epigenetic mechanisms are evolutionarily linked.


Rapid recovery of spontaneous baroreflex after sevoflurane anesthesia in ambulatory surgery.
Although sevoflurane seems to have fulfilled the criteria of ideal anesthetic agent for ambulatory surgery, its effects on intraoperative alteration and postoperative recovery of arterial baroreflex have not been well documented. This study assessed the time required for patients anesthetized with sevoflurane to regain their baseline baroreflex after ambulatory surgeries. Ten ASA class I female patients scheduled for minor gynecological operation (D&C) were enrolled in this study. Spontaneous baroreflex sensitivity (SBR) calculated by sequence analysis of beat-to-beat variations in systolic arterial pressure and R-R intervals was recorded before anesthesia (Baseline), during anesthesia (Anesthesia), five min after operation (Post-op 1) and ten min after operation (Post-op 2). Data obtained from Anesthesia, Post-op 1, and Post-op 2 was compared with the Baseline. SBR value determined in Baseline group (28.6 +/- 4.9 ms/mmHg) was significantly different from Anesthesia group (12.2 +/- 2.1 ms/mmHg) and Post-op 1 group (15.8 +/- 1.8 ms/mmHg). Patients regained their conscious baseline baroreflex sensitivity within ten min after sevoflurane was switched off. Sevoflurane anesthesia depresses SBR and provides a quick SBR recovery upon emergence.


Parents' and Early Childhood Educators' Perceptions on Movement and Learning Program Implementation.
There is currently limited evidence on parents' and early childhood educators' perspectives on implementing programs that combine cognitive and motor tasks in early childhood. An online survey was distributed across Australia through social network platforms and emails at preschool centres, asking 65 parents of preschool children and early childhood educators about their preferences on program delivery, duration, and mode. Responses from the survey were evaluated in order to develop and pilot a 4 week home-based (n = 5 parents) and a 6 week school-based program (n = 5 educators) including cognitively engaging physical activity, requesting parents' and educators' perspectives, respectively, about the program components. Results from the online survey showed a preference for programs with online (e.g., video-based) compared to traditional delivery (e.g., books), emphasising the potential benefits on children's physical activity levels, sleep, and cognitive function. However, after piloting the program, educators preferred to use the book version instead of the video. This program has the potential to become part of daily regular practice. Barriers reported include logistics issues (i.e., book size), connectivity issues with internet, and the need for varying activities.


Preventing driveline infection during left ventricular assist device support by the HeartMate 3: A survey-based study.
Driveline infection (DLI) is a significant source of morbidity and mortality during left ventricular assist device (LVAD) support yet limited studies are available to describe the center-level prevalence, preventive practices, and their potential effectiveness. We surveyed LVAD centers in the United States to determine program burden and preventive practices for DLI during HeartMate (HM) 3 support. An online, anonymous, question-based survey was sent to expert providers at implanting centers. Only a single respondent completed the survey for each center. As an exploratory analysis, we compared specific DLI preventive practices between centers with low (≤10%) and high (>10%) reported prevalence of DLI. Seventy-eight centers responded to the survey (response rate: 50%). Respondents were comprised of 37 (47%) heart failure cardiologists, 27 (35%) LVAD coordinators, and 14 (18%) cardiothoracic surgeons. The prevalence of DLI during HM3 was reported as ≤10% by 27 (35%), 11%-25% by 36 (46%), and >25% by 16 (19%) centers. Thirteen (17%) centers had a body mass index threshold for device placement, 29 (37%) utilized a counter incision, 66 (81%) placed an anchor stitch, and 69 (88%) used an external device to stabilize the DL. Proportionally, more centers with a low DLI prevalence used a wound vacuum 6 (22%) versus 3 (6%, p = 0.03) than those with high DLI. Variation exists in reported prevalence and practices of preventing and managing driveline infections across centers during HM3 support. Further studies are warranted to develop and assess the effectiveness of standardized preventive strategies.


Role of blocking ADAM10 hydrolysis site on N-cadherin by single-chain antibody in ventricular remodeling.
The present study aimed to investigate the roles of the hydrolytic process of N-cadherin by A disintegrin and metalloproteases 10 (ADAM10) in sustaining myocardial structure and integrity, and discuss the mechanisms of ventricular remodeling in dilated cardiomyopathy (DCM). Single chain variable fragment antibody (ScFv) with the ability to specifically block the ADAM10 hydrolysis site of N-cadherin was designed and constructed. Western blot analysis and flow cytometry were used to detect the expression of N-cadherin and its C-terminal fragment 1 (CTF1) on cardiomyocytes, and cells were also subjected to a cell adhesion assay. Furthermore, in a rat model of dilated cardiomyopathy (DCM), the effects of intracardiac injection of the recombinant adenovirus on cardiac structure and contractile function were observed by hematoxylin and eosin staining and color Doppler echocardiography. The recombinant ScFv-expressing adenoviral plasmid with the ability to block the ADAM10 hydrolysis site on N-cadherin was successfully constructed and efficiently transfected into H9C2 cells. After transfection, N-cadherin protein expression was significantly increased, CTF1 protein was significantly decreased and the adhesion capability of myocardial cells was significantly improved. In the in vivo experiment, N-cadherin expression was significantly increased in the treatment group compared with that in the model group, and the structure and function of the heart were significantly improved. In conclusion, blocking of the ADAM10 hydrolysis site on N-cadherin by ScFv increased N-cadherin expression and improved ventricular remodeling. The present study provided experimental evidence for a novel approach for the treatment and prevention of DCM.


Genistein, a natural phytoestrogen from soy, relieves neuropathic pain following chronic constriction sciatic nerve injury in mice: anti-inflammatory and antioxidant activity.
There is great interest in soy isoflavones as alternatives to endogenous estrogens not only in hormonal pathologies, but also in inflammatory, neurodegenerative diseases, and pain. We investigated the effect of the isoflavone genistein on neuropathic pain. Genistein binds estrogen receptors (ER) with higher affinity for the ERbeta particularly expressed in neuronal and immune cells. Neuropathy was induced in mice by means of chronic sciatic nerve constriction, and the subcutaneous administration of genistein from the third day after the lesion reversed pain hypersensitivity in a time- and dose-dependent manner. This effect may have been due to the activation of classical nuclear receptor and/or anti-oxidant, anti-inflammatory, and immunomodulating properties of genistein. The fact that a specific ERbeta antagonist prevented both its anti-allodynic and anti-hyperalgesic action, whereas a specific ERalpha antagonist was ineffective and a non-selective ER antagonist only reversed the anti-allodynic effect, suggests the involvement of ERbeta. Antioxidant effects are also involved as the anti-nociceptive dose reversed the increase in reactive oxygen species and malondialdehyde in injured paw tissues, and increased the activity of anti-oxidant enzymes. The phytoestrogen had immunomodulatory and anti-inflammatory activities as it reduced peripheral and central nuclear factor-kappaB, nitric oxide system and pro-inflammatory cytokine over-activation. Taken together, our results suggest that genistein could ameliorate painful neuropathy by multiple mechanisms.


Modality of tumor endothelial VEGFR2 silencing-mediated improvement in intratumoral distribution of lipid nanoparticles.
The vascular endothelial growth factor (VEGF)-mediated enhancement in vascular permeability is considered to be a major factor in tumor-targeting delivery via the enhanced permeability and retention (EPR) effect. We previously reported that the silencing of the endothelial VEGF receptor (VEGFR2) by a liposomal siRNA system (RGD-MEND) resulted in an enhanced intratumoral distribution of polyethylene glycol (PEG)-modified liposomes (LPs) in a renal cell carcinoma, a type of hypervascularized cancer, although the inhibition of VEGF signaling would be expected to decrease the permeability of the tumor vasculature. We herein report that the enhancement in the intratumoral distribution of LPs by VEGFR2 inhibition was dependent on the vascular type of the tumor (stroma vessel type; SV and tumor vessel type; TV). In the case of TV-type tumors (renal cell carcinoma and hepatocellular carcinoma), inhibiting VEGFR2 improved intratumoral distribution, while no effect was found in the case of SV-type tumors (colorectal cancer). Moreover, through a comparison of the intratumoral distribution of LPs with a variety of physical properties (100nm vs 400nm, neutral vs negative vs positive), VEGFR2 inhibition was found to alter the tumor microenvironment, including heparan sulfate proteoglycans (HSPGs). In addition, the results regarding the effect of the size of nanoparticles indicated that VEGFR2 inhibition improved the penetration of nanoparticles through the vessel wall, but not via permeability, suggesting the involvement of an unknown mechanism. Our findings suggest that a combination of anti-angiogenic therapy and delivery via the EPR effect would be useful in certain cases, and that altering the tumor microenvironment by VEGFR2 blockade has a drastic effect on the intratumoral distribution of nanoparticles.


Effects of Creating Awareness Through Photographs and Posters on Skin Self-Examination in Nursing Students.
Nurses can have an influence on primary and secondary prevention of skin cancer. Therefore, it is important to help them acquire knowledge and skills necessary to modify behavior in relation to skin cancers. The aim of this study was to examine effects of creating awareness through photographs and posters on knowledge and skills of skin self-examination in nursing students. The study had a quasi-experimental design with a pretest and a posttest in a single group. The study population included 249 last-year nursing students in Turkey. Non-probability sampling was used to reach the population. The study sample was composed of 201 students volunteering to participate in the study, and the response rate was 81 %. Of the students, 85.1 % (n = 171) were female and the mean age of the students was 22.18 ± 0.78 years. Of the students, 94.5 % did not know the asymmetry, border, color, diameter, evolution (ABCDE) criteria on the pretest, but this rate decreased to 20.9 % on the posttest. The mean score for knowledge of symptoms of skin cancer was 10.95 ± 1.37 on the pretest and 11.48 ± 0.90 on the posttest. There was a significant linear increase in the scores for knowledge of the symptoms (F = 7.874, p < 0.001) after the intervention with photographs and posters. The students were observed to learn the ABCDE criteria and had increased knowledge of skin cancer symptoms after the intervention using photographs and posters. Photographs and posters are effective tools which can be used to increase awareness of skin self-examination.


Female sterilization using tubal coagulation.
3 different techniques have been described for laparoscopic sterilization using using tubal coagulation. Our experience using all 3 different techniques with a 3-year follow-up is evaluated. Unipolar electrocoagulation (UPE) was used in only 17 cases and subsequently replaced. 1 ectopic pregnancy was encountered (5.88%). There was no pregnancy when bipolar electrocautery (BPE) was applied to 94 women. Endothermic coagulation (EC) was used in 166 patients. 9 pregnancies (5.42%) were noted. An analysis of the possible causes of such high pregnancy rate did not answer our questions. 200 patients were followed by hysterosalpingography (HSG) after tubal coagulation. 4 if them (2%) showed radiologic signs of tubal patency. 1/2 of the clinical pregnancies had a negative HSG performed. BPE is the method of choice for tubal coagulation. EC has a high failure rate. UPE should be abandoned. HSG is not a reliable method for control after sterilization.


The carboxyl modifier 1-ethyl-3-[3-(dimethylamino)propyl]carbodiimide (EDC) inhibits half of the high-affinity Mn-binding site in photosystem II membrane fragments.
The diphenylcarbazide(DPC)/Mn2+ assay [Hsu, B.-D., Lee, J.-Y., & Pan, R.-L. (1987) Biochim. Biophys. Acta 890, 89-96] was used to assess the amount of the high-affinity Mn-binding site in manganese-depleted photosystem II (PS II) membrane fragments from spinach and Scenedesmus obliquus. The assay mechanism at high DPC concentration was shown to involve noncompetitive inhibition of only half of the control level of DPC donation to PS II by micromolar concentrations of Mn at pH 6.5 (i.e., one of two DPC donation sites is inhibited). At low DPC concentration both DPC and Mn2+ donate to PS II additively. Treatment with the carboxyl amino acid modifier 1-ethyl-3-[3-(dimethylamino)propyl]carbodiimide (EDC) inhibited half of the high-affinity Mn-binding site in spinach and Scenedesmus WT PS II membranes and all of the available site in Scenedesmus LF-1 mutant PS II membranes. A similar EDC concentration dependence was observed in all cases. Addition of 2 mM MnCl2 to the 10 mM EDC modification buffer provided complete protection for the Mn-binding site from modification. This protection was specific for Mn2+; six other divalent cations were ineffective. We conclude that EDC modifies that half of the high-affinity Mn-binding site that is insensitive to the histidine modifier diethyl pyrocarbonate (DEPC) [Seibert, M., Tamura, N., & Inoue, Y. (1989) Biochim. Biophys. Acta 974, 185-191] and directly affects ligands that bind Mn. The effects of EDC and DEPC that influence the high-affinity site are mutually exclusive and are specific to the lumenal side of the PS II membrane. Removal of the two more loosely bound of the four functional Mn from PS II membranes uncovers that part of the high-affinity site associated with carboxyl but not histidyl residues. We suggest that carboxyl residues on reaction center proteins are associated with half of the high-affinity Mn-binding site in PS II and are involved along with histidine residues in binding Mn functional in the O2-evolving process.


Femoral nailing to treat fractures after lengthening for congenital femoral deficiency in young children.
Congenital femoral deficiency in children can be treated with femoral lengthening. A common complication is fracture soon after removal of the external fixator, often despite prophylactic hip spica cast application. These fractures present special challenges because the patients have tight soft tissues and sclerotic intramedullary canals. We treated nine such fractures in eight children (average age, 5.4 years). Most were 'spontaneous' events resulting in transverse fracture through regenerate bone or pin sites. All were stabilized with intramedullary Rush pins using special insertion techniques. Union was achieved (average, 6 weeks); no significant complications occurred. We describe details of the surgical technique.


P2Y2 Receptor and EGFR Cooperate to Promote Prostate Cancer Cell Invasion via ERK1/2 Pathway.
As one member of G protein-coupled P2Y receptors, P2Y2 receptor can be equally activated by extracellular ATP and UTP. Our previous studies have proved that activation of P2Y2 receptor by extracellular ATP could promote prostate cancer cell invasion and metastasis in vitro and in vivo via regulating the expressions of some epithelial-mesenchymal transition/invasion-related genes (including IL-8, E-cadherin, Snail and Claudin-1), and the most significant change in expression of IL-8 was observed after P2Y2 receptor activation. However, the signaling pathway downstream of P2Y2 receptor and the role of IL-8 in P2Y2-mediated prostate cancer cell invasion remain unclear. Here, we found that extracellular ATP/UTP induced activation of EGFR and ERK1/2. After knockdown of P2Y2 receptor, the ATP -stimulated phosphorylation of EGFR and ERK1/2 was significantly suppressed. Further experiments showed that inactivation of EGFR and ERK1/2 attenuated ATP-induced invasion and migration, and suppressed ATP-mediated IL-8 production. In addition, knockdown of IL-8 inhibited ATP-mediated invasion and migration of prostate cancer cells. These findings suggest that P2Y2 receptor and EGFR cooperate to upregulate IL-8 production via ERK1/2 pathway, thereby promoting prostate cancer cell invasion and migration. Thus blocking of the P2Y2-EGFR-ERK1/2 pathway may provide effective therapeutic interventions for prostate cancer.


Towards an Ecosystem Approach to Cheese Microbiology.
Cheese is an ideal environment to serve as a model for the behavior of microbes in complex communities and at the same time allow detailed genetic analysis. Linking organisms, and their genes, to their role in the environment becomes possible in the case of cheese since cheese microbial communities have been "in culture" for thousands of years, with the knowledge of how to grow these organisms passed down by generations of cheesemakers. Recent reviews have described several emerging approaches to link molecular systems biology to ecosystem-scale processes, known as ecosystems biology. These approaches integrate massive datasets now available through high-throughput sequencing technologies with measurements of ecosystem properties. High-throughput datasets uncover the "parts list" (e.g., the species and all the genes within each species) of an ecosystem as well as the molecular basis of interactions within this parts list. Novel computational frameworks make it possible to link species and their interactions to ecosystem properties. Applying these approaches across multiple temporal and spatial scales makes it possible to understand how changes in the parts lists over space and time lead to changes in ecosystems processes. By manipulating the species present within model systems, we can test hypotheses related to the role of microbes in ecosystem function. Due to the tractability of cheese microbial communities, we have the opportunity to use an ecosystems biology approach from the scale of individual microbial cells within a cheese to replicated cheese microbial communities across continents. Using cheese as a model microbial ecosystem can provide a way to answer important questions concerning the form, function, and evolution of microbial communities.


Rare causes of anemia in liver diseases.
Anemia is a common finding among patients with liver diseases. Patients who suffer from anemia are at a higher risk of liver function decompensation and hospitalization. It affects significantly their quality of life and contributes to mortality. Anemia is present in 70% of patients with liver cirrhosis and with varying incidence accompanies other liver disorders. As the etiology of anemia in liver diseases is multifactorial, various cases represent different clinical entities. Anemia accompanying hepatic disorders can be broadly divided into several types, such as anemia associated with blood loss, as well as aplastic, hemolytic and micronutrient deficiency anemia. However, it is sometimes difficult to delineate between those types in the clinical practice, as several pathophysiological causes can be present in one patient. It is reported that the most common cause of anemia in liver disease is blood loss and iron deficiency. Still, the incidence of unclear cases reaching over 50% suggests that other types of anemia can be underdiagnosed. This review comprehensively describes less frequent types of anemia associated with liver disease, namely hemolytic and aplastic anemia (AA). Hemolytic anemia can complicate autoimmune liver diseases or be a manifestation of membranopathy of red blood cells, dependent on severe hepatic function impairment or alcoholic liver disease. Aplastic anemia is best known as a sequela of viral hepatitis, but some degree of bone marrow inhibition can complicate virtually all advanced liver diseases.


Interleukin 6 stimulates the production of immunoreactive endothelin 1 in human breast cancer cells.
To investigate the potential regulation of endothelin 1 production in human breast cancer cells, we measured the release of immunoreactive endothelin 1 (ir-ET-1) from the MCF-7 and ZR-75-1 breast cancer cell lines in response to various agents including estrogen and tamoxifen as well as several cytokines. ir-ET-1 was detected in conditioned medium of MCF-7 cells and ZR-75-1 cells by specific radioimmunoassay. Among the agents tested, estrogen, tamoxifen, tumor necrosis factor, gamma-interferon, interleukin (IL) 1, and transforming growth factor beta had no effect on ir-ET-1 secretion by these breast cancer cells. However, IL-6 (20 ng/ml) treatment of MCF-7 cells and ZR-75-1 cells caused maximal increases in the amount of ir-ET-1 secreted into the culture medium to 206 and 314% of basal values after 6 h, respectively. This effect of IL-6 on ir-ET-1 secretion was inhibited by actinomycin D and cycloheximide, indicating that IL-6 stimulates de novo synthesis of ir-ET-1 at a transcriptional level. Reverse-phase high performance liquid chromatography coupled with radioimmunoassay in the conditioned medium from IL-6-treated cells revealed one major ir-ET-1 component corresponding to human standard ET-1. The present study demonstrates the potential for IL-6 to stimulate ir-ET-1 production in human breast cancer cells, which may participate in the process of acute phase reactant-like expression of this peptide and/or in the process of IL-6 enhanced breast cancer cell motility, the latter being recently clarified.


Rodent population cycle as a determinant of gastrointestinal nematode abundance in a low-arctic population of the red fox.
We analyzed an 11-year time series (2005-2015) of parasite abundance for three intestinal nematode species in the red fox (Vulpes vulpes) as a function of the multi-annual rodent population cycle in low-arctic Norway, while correcting for other potential covariates that could influence prevalence and abundance. Rodents are paratenic and facultative intermediate hosts for the two Ascarididae species Toxascaris leonina and Toxocara canis, respectively and key prey for the red fox. Still the relative importance of indirect transmission through rodents and direct transmission through free-living stages is unclear. Abundance of these Ascarididae species in individual red foxes (N = 612) exhibited strongly cyclic dynamics that closely mirrored the 4-year rodent cycle. Negative binomial models provided evidence for a direct proportional increase in Ascarididae abundance with rodent density suggesting that predator functional response to rodent prey is the key transmission mechanism. In contrast, no cycles and constantly very low abundance were apparent for Uncinaria stenocephala - a third nematode species recorded without paratenic or intermediate stages.


Exercise stress echocardiography for detection of pulmonary arterial hypertension in a patient with systemic sclerosis.
The authors report the case of a 57-year-old patient, diagnosed with systemic sclerosis 16 years ago. She had been followed in the rheumatology outpatient clinic since 2003 and complained of marked fatigue and dyspnea on exertion. In addition to full medical evaluation, she also underwent standard Doppler echocardiography at rest, which was unremarkable, and exercise stress echocardiography. In the latter examination, she was diagnosed with pulmonary arterial hypertension, later confirmed during right heart catheterization. The authors discuss the potential value of this methodology for the study of patients with systemic sclerosis.


Protein interaction-based genome-wide analysis of incident coronary heart disease.
Network-based approaches may leverage genome-wide association (GWA) analysis by testing for the aggregate association across several pathway members. We aimed to examine if networks of genes that represent experimentally determined protein-protein interactions (PPIs) are enriched in genes associated with risk of coronary heart disease (CHD). Genome-wide association analyses of approximately ≈700,000 single-nucleotide polymorphisms in 899 incident CHD cases and 1823 age- and sex-matched controls within the Nurses' Health and the Health Professionals Follow-up Studies were used to assign genewise P values. A large database of PPIs was used to assemble 8351 unbiased protein complexes and corresponding gene sets. Superimposed genewise P values were used to rank gene sets based on their enrichment in genes associated with CHD. After correcting for the number of complexes tested, 1 gene set was overrepresented in CHD-associated genes (P=0.002). Centered on the β1-adrenergic receptor gene (ADRB1), this complex included 18 protein interaction partners that have not been identified as candidate loci for CHD. Of the 19 genes in the top complex, 5 are involved in abnormal cardiovascular system physiological features based on knockout mice (4-fold enrichment; Fisher exact test, P=0.006). Ingenuity pathway analysis revealed that canonical pathways, especially related to blood pressure regulation, were significantly enriched in the genes from the top complex. The integration of a GWA study with PPI data successfully identifies a set of candidate susceptibility genes for incident CHD that would have been missed in single-marker GWA analysis.


Safety, tolerability and pharmacokinetics of the N-methyl-D-aspartate antagonist Ro-01-6794/706 in patients with acute ischemic stroke. The Dextrorphan Study Group and Hoffmann-La Roche.
Dextrorphan HCl (Ro 01-6794/706) is an NMDA receptor antagonist with clinical potential for administration in an elderly population of acute ischemic stroke patients. In vivo experience with such patients demonstrated a consistent pharmacologic effect/adverse experience profile that is typical of an NMDA receptor antagonist (e.g., nystagmus, nausea, vomiting, agitation, somnolence, hallucinations and hypertension). For the most part, these pharmacologic effects were mild to moderate in severity; short-lived; reversible; not life-threatening and subjectively tolerated. The most serious pharmacologic effect produced by dextrorphan administration was hypotension, which occurred within a well-defined window of 90 minutes from the start of the loading dose infusion in patients who received 200 mg/hr or greater loading dose infusions. In all cases it was reversible without neurologic sequelae. Careful review of demographic and pharmacokinetic parameters did not demonstrate any overriding factor(s) to the production of hypotension other than the rate of the loading dose infusion. Severe hypotension, severe decreased levels of consciousness and respiratory depression should not be generally expected at loading doses less than 200 mg/hr. In summary, dextrorphan can be safely given to an elderly population of ischemic stroke patients as a loading dose rate below 200 mg/hr and as a maintenance dose rate between 50-90 mg/hr for 24 hours when patients are monitored carefully for pharmacologic effects.


Reduction of Marek's Disease Virus Infection by Toll-Like Receptor Ligands in Chicken Embryo Fibroblast Cells.
Evolutionarily conserved pattern recognition receptors, including Toll-like receptors (TLRs) recognize pathogen-associated molecular patterns (PAMPs) that are present in microbes. PAMPs induce several pathways downstream of TLRs that lead to induction of antiviral responses. The objective of this study was to investigate the stimulatory effect of various PAMPs (in the form of TLR ligands) in reducing Marek's disease virus (MDV) infection in chicken embryo fibroblast cells (CEFs). To this end, CEFs were pretreated with Pam3CSK4, Poly(IC), lipopolysaccharide (LPS), and CpG ODN as TLR2, TLR3, TLR4, and TLR21 ligands, respectively for 24 h followed by infection with MDV. The results indicated that pretreatment with Poly(IC) resulted in a robust reduction (by about 81%) of MDV infection in CEFs at 96 h postinfection while a moderate reduction was observed with treatment of Pam3CSK4 (35%), LPS (26%), and CpG ODN (23%) PAMPs. Transcriptional analysis of gene expression in CEFs demonstrated that all TLR ligand treatments and MDV infection significantly increased the expression of type I interferons, interleukin (IL)-1β, interferon regulatory factor 7 (IRF7), interferon induced protein with tetratricopeptide repeats 5 (IFIT5), and myxoma-resistance protein (Mx). Further studies are needed to explore the mechanism by which PAMPs, particularly the TLR3 ligands could reduce MDV infection in CEFs, which may play an important role in controlling the replication of MDV in chicken.


An audit of fresh frozen plasma usage in a tertiary referral centre in a developing country.
This paper evaluates the practice of fresh frozen plasma (FFP) transfusion at the University Hospital, Kuala Lumpur, and analyses its usage by the various clinical departments. The aim of this study is to identify where it is inappropriately used and the clinical indications in which such misuse is common. A retrospective analysis of the blood bank request forms and work sheets during a 6-month period between January 1998 and June 1998 formed the basis of this study. Overall, 40% of 2665 units transfused were considered appropriate. However, out of the 931 episodes of FFP transfusions only 31% were for appropriate indications. The average FFP requirement when used for appropriate indication was about 4 units per episode, whereas for inappropriate indication it was 2.5 units per episode. Inappropriate use in terms of the number of units was highest by the surgical services (68%) and Orthopaedics (64%), while the Department of Paediatrics had the lowest incidence of inappropriate use (40%). When Paediatrics was used as the benchmark, the incidence of inappropriate use by other departments was significantly higher (p < 0.01). As for FFP usage in common clinical indications, there was a high incidence of inappropriate use in burns (82%), perioperative period (73%), cardiac surgery (68%), massive bleeding (62%) and trauma (60%). The findings in this study, specifically the use of FFP for volume support in trauma, massive bleeding and burns, routine requests without identified indication in cardiac bypass surgery, and prophylactic use in the perioperative period can be the basis for recommendations to minimize the inappropriate use of FFP in the future.


Subcutaneous tumoral seeding from a glioblastoma following stereotactic biopsy: case report and review of the literature.
Extracranial metastases from glioblastoma are uncommon, likely because short patient survival time prevent them to occur. Most of the few previously reported cases occurred after invasive surgical procedures. We describe a case of glioblastoma with concomitant seeding along the stereotactic biopsy trajectory and subcutaneous metastasis. A 60-year-old woman presented with severe headache. Neuroradiological work-up (including cranial computed tomographic scan and magnetic resonance imaging) showed a heterogeneous hyperdensity, suggestive of malignant glioma, in the left parietal region. A computed tomographic-guided stereotactic biopsy was performed and microscopic examination attested a diagnosis of glioblastoma. Radiotherapy and chemotherapy were administered. Eight months later, the patient presented with a subcutaneous tumor in the left occipital region. A cranial computed tomographic scan revealed a large enhancement of the initial tumor, intracranial tumor seeding along the stereotactic biopsy trajectory, and a subcutaneous tumor. Partial resection of the subcutaneous lesion was performed, and histological examination identified an extracranial metastasis from the glioblastoma. Although uncommon, this observation points to the risk of tumor seeding following stereotactic biopsy, and to the close connection between this intracranial seeding and subcutaneous metastasis.


Morphological pattern elicited by agents affecting spermatogenesis by stimulation.
A variety of agents (clomiphene citrate, cyproterone acetate, estradiol 17-beta, medrogestone, medroxyprogesterone, and methoxychlor) thought to disrupt the normal pathway by which hormones stimulate the testis were tested in rats to determine the early morphologic changes in the testis. After initial trial experiments at which dose and sacrifice intervals were determined, the earliest morphological abnormalities were assessed quantitatively and related to the particular spermatogenic stage in which these defects were initiated. All agents tested produced an identical morphological pattern of response displaying a significant increase in Stage VII degenerating cells (pachytene spermatocytes, step 7 and step 19 spermatids) as compared with vehicle-administered rats. The number of degenerating cells in other stages did not significantly change. The rise in degenerating cells in Stage VII, as seen in this study, was similar to that reported in Russell and Clermont (1972) utilizing hypophysectomized rats, and that reported by other investigators who have used a variety of agents to disrupt the hormonal stimulation of the seminiferous tubules. The data indicate that no matter how, or at what level the hormonal stimulation of the testis is interrupted, the morphological pattern of response is the same, and that such a pattern is useful in predicting the mechanism of action of agents suspected of interfering with hormone action.


Invasive Mechanisms of One of the World's Worst Alien Plant Species Mimosa pigra and Its Management.
Mimosa pigra is native to Tropical America, and it has naturalized in many other countries especially in Australia, Eastern and Southern Africa and South Asia. The species is listed in the top 100 of the world's worst invasive alien species and is listed as Least Concern in the IUCN Red List of Threatened Species. M. pigra forms very large monospecific stands in a wet-dry tropical climate with conditions such as floodplains, riverbanks, grasslands, forests and agricultural fields. The stands expand quickly and threaten the native flora and fauna in the invasive ranges. Possible mechanisms of the invasion of the species have been investigated and accumulated in the literature. The characteristics of the life history such as the high reproduction and high growth rate, vigorous mutualism with rhizobia and arbuscular mycorrhizal fungi, very few natural enemies, and allelopathy, and certain secondary metabolites may contribute to the invasiveness and naturalization of M. pigra. Herbicide application, such as aerial spraying, foliar, cut-stump and soil treatments, is the primary control methods of M. pigra. The investigation of the natural enemies of M. pigra has been conducted in its native ranges since 1979, and biological control agents have been selected based on host specificity, rearing and availability. Mechanical control practices, such as hand weeding, bulldozing, chaining and fire, were also effective. However, the species often regrow from the remaining plant parts. Integration of multiple weed control practices may be more effective than any single practice. This is the first review article focusing on the invasive mechanism of M. pigra.


Association between altered lipid profile, body mass index, low plasma adiponectin and varied blood pressure in Trinidadian type 2 diabetic and non-diabetic subjects.
The obesity and hypertension have become the causes for the development type 2 diabetes. There is a limited study done on the contribution of body mass index (BMI) to blood pressure (BP) in the Caribbean population. Aim of our study was to determine the associations between lipid profile, BMI, adiponectin, and BP in Trinidadian type 2 diabetic patients with regards to age and ethnicity. This was a cohort study comprised of 266 subjects (85 males and 181 females) attending primary and tertiary healthcare settings in central Trinidad. Of which, 126 diabetic subjects were matched with 140 non-diabetic subjects. Along with clinical history and anthropometry, adiponectin and lipid profile were measured in fasting blood samples. The diabetic group had higher triglycerides, very low density lipoprotein (VLDL), and BP values which were statistically significant (P < 0.05) when compared to non-diabetic subjects. The high-density lipoprotein cholesterol (HDL-c) and adiponectin were lower in diabetic subjects. HDL-c showed significant changes for ethnicity (P = 0.013) and gender (P = 0.043). The mean adiponectin concentrations were found to be significantly different among the ethnic groups (P = 0.001). Systolic pressure varied significantly with age (P = 0.018). As age increased, BP also increased. Ethnic groups had a significant difference in diastolic pressure (P = 0.027). East Indians had the highest mean diastolic pressure (80.74 ± 10.29) when compared to all other ethnic groups. HDL-cholesterol, low levels of adiponectin, and varied BP are associated in Trinidadian type 2 diabetic subjects with regards to age, gender, and ethnicity.


Glyceraldehyde 3-Phosphate:NADP Reductase of Spinach Leaves : Steady State Kinetics and Effect of Inhibitors.
The steady state kinetics of glyceraldehyde 3-phosphate:NADP(+) oxidoreductase (GNR) (EC 18.104.22.168) have been investigated. The enzyme exhibits hyperbolic behavior over a wide range of substrate concentrations. Double-reciprocal plots are nearly parallel or distantly convergent with limiting K(m) values of 2 to 5 micromolar for NADP(+) and 20 to 40 micromolar for D-glyceraldehyde 3-phosphate (G3P). The velocity response to NADP(+) as the varied substrate is however sigmoidal if G3P concentration exceeds 10 micromolar, whereas the response to G3P may show inhibition above this concentration. This ;G3P-inhibited state' is alleviated by saturating amounts of NADP(+) or NADPH. Product inhibition patterns indicate NADPH as a potent competitive inhibitor to NADP(+) (K(i) 30 micromolar) and mixed inhibitor towards G3P, and 3-phosphoglycerate (3PGA) as mixed inhibitor to both NADP(+) and G3P (K(i) 10 millimolar). The data, and those obtained with dead-end inhibitors, are consistent with a nonrapid equilibrium random mechanism with two alternative kinetic pathways. Of these, a rapid kinetic sequence (probably ordered with NADP(+) binding first and G3P binding as second substrate) is dominant in the range of hyperbolic responses. A reverse reaction with 3PGA and NADPH as substrates is unlikely, and was not detected. Of a number of compounds tested, erythrose 4-phosphate (K(i) 7 micromolar) and Pi (K(i) 2.4 millimolar) act as competitive inhibitors to G3P (uncompetitive towards NADP(+)) and are likely to affect the in vivo activity. Ribose 5-phosphate, phosphoenolpyruvate, ATP, and ADP are also somewhat inhibitory. Full GNR activity in the leaf seems to be allowed only under high photosynthesis conditions, when levels of several inhibitors are low and substrate is high. We suggest that a main function of leaf GNR is to supply NADPH required for photorespiration, the reaction product 3PGA being cycled back to chloroplasts.


Diels-Alder reactions of graphene: computational predictions of products and sites of reaction.
The cycloaddition reactions and noncovalent π interactions of 2,3-dimethoxybutadiene (DMBD), 9-methylanthracene (MeA), tetracyanoethylene (TCNE), and maleic anhydride (MA) with graphene models have been investigated using density functional theory (DFT) calculations. Reaction enthalpies have been obtained to assess the reactivity and selectivity of covalent and noncovalent functionalization. Results indicate that graphene edges may be functionalized by the four reagents through cycloaddition reactions, while the interior regions cannot react. Noncovalent complexation is much more favorable than cycloaddition reactions on interior bonds of graphene. The relative reactivities of different sites in graphene are related to loss of aromaticity and can be predicted using Hückel molecular orbital (HMO) localization energy calculations.


Omic analyses unravels global molecular changes in the brain and liver of a rat model for chronic Sake (Japanese alcoholic beverage) intake.
The effects of chronic administration of Sake (Japanese alcoholic beverage, Nihonshu) on brain and liver of female F334 (Fisher) rats were surveyed via global omic analyses using DNA microarray, 2-DE, and proton nuclear magnetic resonance. Rats weaned at 4 wk of age were given free access to Sake (15% alcohol), instead of water. At 13 months of age, and 24 h after withdrawal of Sake supply, rats were sacrificed, and the whole brain and liver tissues dissected for analyses. In general, molecular changes in brain were found to be less than those in liver. Transcriptomics data revealed 36 and 9, and 80 and 62 up- and down-regulated genes, in the brain and liver, respectively, with binding and catalytic activity gene categories the most prominently changed. Results suggested Sake-induced fragility of brain and liver toxicity/damage, though no significant abnormalities in growth were seen. At protein level, a striking decrease was found in the expression of NADH dehydrogenase (ubiquinone) Fe-S protein 1 in brain, suggesting attenuation of mitochondrial metabolism. In liver, results again suggested an attenuation of mitochondrial function and, in addition, glycoproteins with unknown function were induced at protein and gene levels, suggesting possible changes in glycoprotein binding in that organ. Metabolomic analysis of brain revealed significant increases in valine, arginine/ornithine, alanine, glutamine, and choline with decreases in isoleucine, N-acetyl aspartate, taurine, glutamate, and gamma aminobutyric acid. Our results provide a detailed inventory of molecular components of both brain and liver after Sake intake, and may help to better understand effects of chronic Sake drinking.


Knee cartilage: efficient and reproducible segmentation on high-spatial-resolution MR images with the semiautomated graph-cut algorithm method.
This HIPAA-compliant study was exempt from institutional review board approval because the 10 image data sets were deidentified in the Osteoarthritis Initiative database, and they were processed and analyzed without any clinical information being accessed. The purpose of this study was to prospectively evaluate the efficiency and reproducibility of the semiautomated graph-cut method (SA method) in the segmentation of knee cartilage and to compare its performance with that of the conventional manual delineation segmentation method (M method). Two radiologists independently performed segmentation with each method in two separate sessions: They performed the M method (M1 and M2 for the first and second sessions, respectively) for every third section and the SA method (SA1 and SA2 for the first and second sessions, respectively) for every section. The SA method was significantly more efficient (mean processing time, 53 minutes vs 156 minutes for SA1 vs M1 and 53 minutes vs 118 minutes for SA2 vs M2; P < .001) and reproducible (mean volume overlap, 94.3% vs 87.8% for the SA method vs the M method; P < .001) than the M method.


Comparison of agar dilution, microdilution, and disk elution methods for measuring the synergy of cefotaxime and its metabolite against anaerobes.
The activities of cefotaxime (CTX) and desacetyl cefotaxime (des-CTX) were tested both singly and in combination against 173 anaerobic clinical isolates. The MIC of CTX for 50% of 60 Bacteroides fragilis isolates was 22.4 micrograms/ml in broth, compared with 47.4 micrograms/ml in agar. This reduced efficacy in agar was seen with all species tested and is in apparent conflict with reported clinical efficacy of the drug. Synergy between CTX and des-CTX was observed with 70 to 100% of the isolates, including 60% of all Bacteroides spp. tested. The susceptibility results in a synergy system correlated well with those noted in a broth-disk elution method incorporating 32 micrograms of CTX and 8 micrograms of des-CTX per ml. The correlation was poorer when the broth-disk method contained 16 micrograms of CTX and 8 micrograms of des-CTX per ml.


[Early and prolonged peritoneal lavage with laparoscopy in severe acute pancreatitis].
The evaluation of precocious and prolonged lavage and drainage by laparoscopic approach in the treatment of severe acute pancreatitis. This therapeutic prospective study was conducted during 01.01.2006-31.03.2008 on a sample consisting of 35 subjects with the severe acute pancreatitis has been divided into two lots. A lot is formed by 16 patients whom were applied the method mentioned ahead and in the B lot--19 patients, treated by conventional and known methods of treatment. The method proposed, completed about laparoscopic approach in a third day of admission, consists, after setting lesional balance, in lavage of peritoneal space and mounting two tubes drainage, one subhepatic space and the other in Douglas space. Post operator for 7 days performed peritoneal lavage discontinuous with physiological serum. For an accurate assessment may were determined serum and peritoneal concentrations of interleukin 6. The duration of organic disfunction was 8 days for A lot subjects and 17,7 days for B lot subjects. The complications occurred and the needs of surgical treatment was net disadvantage at the patients treated conventionally. The mortality at A lot was 12.5% (n=2) and at lot B 36.8% (n=7). The tardive mortality by sepsis was null at subjects with lavage and was 15,8% (n=3) at subjects without lavage. The precocious mortality was 12,5% (n=2) in the A lot and 21% (n=4) in the lot B. The average hospitalization was 25,8 days in subjects with peritoneal lavage and 35,8 days in those treated conservatively, the difference is statistically significant. The method has proved not generating of morbidity and supplementary mortality. Study of the variation of serum and peritoneal concentrations of interleukin 6 shows the modulator effect of the peritoneal lavage early and prolonged of the inflammatory response and offers an argument of the inefficiency of short peritoneal lavage. The data obtained indicate early and extended peritoneal lavage by laparoscopic method as a useful therapy in the management of severe acute pancreatitis.


[Bioadhesives: rationale, state of the art and therapeutic potential].
Pharmaceutical research is going on the way to formulate drugs in dosage forms and delivery systems able to improve their biopharmaceutical properties. For some routes of administration, among them the transmucosal ones, such improvements may be reached by increasing the time and the nature of the contact between mucosal tissues and drug dosage forms via chemical or physical bioadhesive links. Biopharmaceutical AFI Study Group efforts were oriented in analyzing the actual state of the art in the field of bioadhesive drug delivery. Rationals of use of drug dosage forms with bioadhesive properties, analytical method of control of bioadhesion (both in vitro and in vivo) and therapeutical potentialities are the matter of this review.


Extraordinary increase of lifetime of localized cold clouds by the viscous effect in thermally unstable two-phase interstellar media.
We numerically examine the influence of the viscosity on the relaxation process of localized clouds in thermally unstable two-phase media, which are locally heated by cosmic ray and cooled by radiation. Pulselike stationary solutions of the media are numerically obtained by a shooting method. In one-dimensional direct numerical simulations, localized clouds are formed during the two-phase separation and sustained extraordinarily. Such long-lived clouds have been recently observed in interstellar media. We demonstrate that the balance of the viscosity with a pressure gradient remarkably suppresses the evaporation of the clouds and controls the relaxation process. This balance fixes the peak pressure of localized structures and then the structure is attracted and trapped to one of the pulselike stationary solutions. While the viscosity has been neglected in most of previous studies, our study suggests that the precise treatment of the viscosity is necessary to discuss the evaporation of the clouds.


Transplantation of bioencapsulated bone marrow stem cells improves hepatic regeneration and survival of 90% hepatectomized rats: a preliminary report.
We transplanted bioencapsulated bone marrow stem cells intraperitoneally into 90% hepatectomized rats and found that this increases both the rates of hepatic regeneration and survival of the animals. Bone marrow cells isolated from Wistar rats were bioencapsulated using alginate-polylysine-alginate method. These bioencapsulated bone marrow cells were transplanted intraperitoneally into 90% hepatectomized syngeneic wistar rats. Control groups included 90% hepatectomized group receiving intraperitoneal injection of either empty microcapsules or free bone marrow cells. Unlike the control groups, transplantation of bioencapsulated bone marrow cells improved the survival of 90% hepatectomized rats, with an efficacy similar to that of bioencapsulated hepatocytes or free hepatocytes. These results suggest that syngeneic bioencapsulated bone marrow stem cells can increase the survival rates of 90% hepatectomized rats. We also discuss the potential for a new alternative to hepatocyte transplantation for cellular therapy of acute liver failure. In particular, bone marrow stem cells can be obtained from the same patient with no immunorejection, whereas in hepatocyte transplant, immunosuppressant will be needed to prevent immunorejection of the donor hepatocytes.


Simple non-invasive fibrosis scoring systems can reliably exclude advanced fibrosis in patients with non-alcoholic fatty liver disease.
Accurate evaluation of liver fibrosis in patients with non-alcoholic fatty liver disease (NAFLD) is important to identify patients who may develop complications. The aim of this study was to compare the diagnostic performance of simple non-invasive tests in identifying advanced fibrosis among patients with biopsy-proven NAFLD. Consecutive patients with biopsy proven NAFLD were recruited from the Newcastle Hospitals Fatty Liver Clinic from 2003 to 2009. The AST/ALT ratio, AST to platelet ratio index, BARD (weighted sum of BMI>28=1 point, AST/ALT ratio>0.8=2 points, diabetes=1 point), FIB-4 (agexAST (IU/l)/platelet count (x10(9)/litre)x radicalALT (IU/l)) and NAFLD fibrosis scores were calculated from blood tests taken at time of biopsy. 145 patients (82 male (61%), mean age 51+ or -12 years) were included. The mean body mass index was 35+ or -5 kg/m(2). 73 subjects (50%) had diabetes. 93 patients (64%) had non-alcoholic steatohepatitis. 27 (19%) had advanced fibrosis (Kleiner stage 3-4). The FIB-4 score had the best diagnostic accuracy for advanced fibrosis (area under receiver operator characteristic curve (AUROC) 0.86), followed by AST/ALT ratio (AUROC 0.83), NAFLD fibrosis score (AUROC 0.81), BARD (AUROC 0.77) and AST to platelet ratio index (AUROC 0.67). The AST/ALT ratio, BARD score, FIB-4 and NAFLD fibrosis scores had negative predictive values greater than 90% (93%, 95%, 95% and 92% respectively). Positive predictive values were modest. In order to exclude advanced fibrosis liver biopsy could potentially be avoided in 69% with AST/ALT ratio, 62% with FIB-4, 52% with NAFLD fibrosis score and 38% with BARD. The ALT/AST ratio, FIB-4 and NAFLD fibrosis scores can reliably exclude advanced fibrosis in a high proportion of patients with NAFLD, allowing liver biopsy to be used in a more directed manner.


Are Heptazine-Based Organic Light-Emitting Diode Chromophores Thermally Activated Delayed Fluorescence or Inverted Singlet-Triplet Systems?
Two chromophores derived from heptazine, HAP-3MF and HAP-3TPA, were synthesized and tested as emitters in light-emitting diodes (OLEDs) by Adachi and co-workers. Both emitters were shown to exhibit quantum efficiencies which exceed the theoretical maximum of conventional fluorescent OLEDs. The enhanced emission efficiency was explained by the mechanism of thermally activated delayed fluorescence (TADF). In the present work, the electronic excitation energies and essential features of the topography of the excited-state potential-energy surfaces of HAP-3MF and HAP-3TPA have been investigated with a wave function-based ab initio method (ADC(2)). It is found that HAP-3MF is an inverted singlet-triplet (IST) system; that is, the energies of the S1 and T1 states are robustly inverted in violation of Hund's multiplicity rule. Notably, HAP-3MF presumably is the first IST emitter which was implemented in an OLED device. In HAP-3TPA, on the other hand, the vertical excitation energies of the S1 and T1 states are essentially degenerate. The excited states exhibit vibrational stabilization energies of similar magnitude along different relaxation coordinates, resulting in adiabatic excitation energies which also are nearly degenerate. HAP-3TPA is found to be a chromophore at the borderline of TADF and IST systems. The spectroscopic data reported by Adachi and co-workers for HAP-3MF and HAP-3TPA are analyzed in light of these computational results.


Isoflavonoid Formation as an Indicator of UV Stress in Bean (Phaseolus vulgaris L.) Leaves : The Significance of Photorepair in Assessing Potential Damage by Increased Solar UV-B Radiation.
Induction of the isoflavonoid pigment, coumestrol (3,9-dihydroxy-6H-benzofuro-[3,2-c][1] benzopyran-6-one), in primary leaves of beans (Phaseolus vulgaris L. var Saxa) by ultraviolet (UV) radiation was used as a quantifiable marker for UV damage to a plant system. Coumestrol was induced only by wavelengths below 300 nanometers and its formation could be reversed by treatment with white, but not red light after the UV irradiation period. Formation of coumestrol by UV could also be prevented over a period of 14 hours by simultaneous irradiation with blue light provided that the blue fluence rate was high enough. The results suggest that coumestrol formation is mediated via UV-induced pyrimidine dimer formation in the plant DNA and the photorepair properties of blue light are discussed with respect to possible increases in solar UV due to stratospheric ozone depletion.


Pharmacological classification of centrally acting drugs using EEG in freely moving rats: an old tool to identify new atypical dopamine uptake inhibitors.
Atypical dopamine uptake inhibitors (DUIs) bind to the dopamine transporter and inhibit the reuptake of dopamine but have lower abuse potential than psychostimulants. Several atypical DUIs can block abuse-related effects of cocaine and methamphetamine, thus making them potential medication candidates for psychostimulant use disorders. The aim of the current study is to establish an in-vivo assay using EEG for the rapid identification of atypical DUIs with potential for medication development. The typical DUIs cocaine and methylphenidate dose-dependently decreased the power of the alpha, beta, and gamma bands. The atypical DUI modafinil and its F-analog, JBG1-049, decreased the power of beta, but in contrast to cocaine, none of the other frequency bands, while JHW007 did not significantly alter the EEG spectrum. The mu-opioid receptor agonists heroin and morphine dose-dependently decreased the power of gamma and increased power of the other bands. The effect of morphine on EEG power bands was antagonized by naltrexone. The NMDA receptor antagonist ketamine increased the power of all frequency bands. Therefore, typical and atypical DUIs and drugs of other classes differentially affected EEG spectra, showing distinctive features in the magnitude and direction of their effects on EEG. Comparative analysis of the effects of test drugs on EEG indicates a potential atypical profile of JBG1-049 with similar potency and effectiveness to its parent compound modafinil. These data suggest that EEG can be used to rapidly screen compounds for potential activity at specific pharmacological targets and provide valuable information for guiding the early stages of drug development. This article is part of the issue entitled 'Special Issue on Neurotransmitter Transporters'.


Elastic coupling of silica gel dynamics in a liquid-crystal-aerosil dispersion.
The dynamics of a thixotropic silica aerosil gel dispersed in an octylcyanobiphenyl liquid crystal were directly probed by x-ray intensity fluctuation spectroscopy. For all samples, the time-autocorrelation function of the gel was well described by a modified-exponential function over the q range studied. Compared to a pure gel sample, a dilute (0.06 g cm(-3)) gel embedded within the liquid crystal displayed more complex and temperature dependent dynamics. Near the second-order smectic-A-to-nematic phase transition of the liquid crystal the gel relaxation became significantly more complex and slower (tau approximately 2150 s) compared to relaxations observed well within either phase. This clearly demonstrates coupling between the dynamics of the gel and the host liquid crystal, consistent with critical slowing down of smectic and director fluctuations. A random dampening field, elastically coupled to the liquid crystal, would explain the earlier observed crossover of this transition towards 3d-XY behavior.


Identification of malnutrition-related diabetes mellitus among hospitalized patients in Taiwan.
A retrospective review of 1,089 diabetes mellitus admitted to the metabolic ward of Chang Gung Memorial Hospital was performed to screen for malnutrition-related diabetes mellitus (MRDM). None of the patients fulfilled the MRDM criteria, although 20 patients with a body mass index < or = 15 kg/m2 were further analyzed. These patients presented with poor glycemic control and most had associated chronic diseases. C-peptide levels were < 0.8 ng/mL in 17 patients and undetectable in six patients. Two patients experienced diabetic ketoacidosis (DKA). Insulin treatment was required by 18 patients with a mean (+/- SD) dosage of 30 +/- 9 U/day. During follow-up of 14 of the patients for 9.8 +/- 4.3 months, HbA1c decreased significantly. Their mean body weight and body mass index increased significantly from 36.6 +/- 4.6 kg to 47.2 +/- 6.2 kg and 13.9 +/- 0.9 to 18.0 +/- 1.9 kg/m2, respectively. Careful attention should be paid to diabetic control and associated chronic illness in patients with severe undernutrition.


VATER/VACTERL Association: Evidence for the Role of Genetic Factors.
The VATER/VACTERL association is typically defined by the presence of at least 3 of the following congenital malformations: Vertebral anomalies, Anal atresia, Cardiac malformations, Tracheo-Esophageal fistula, Renal anomalies, and Limb abnormalities. The involvement of genetic factors in the development of this rare association is suggested by reports of familial occurrence, the increased prevalence of component features among first-degree relatives of affected individuals, high concordance rates among monozygotic twins, chromosomal (micro-)aberrations or single gene mutations in individuals with the VATER/VACTERL phenotype, as well as murine knock-out models. Despite substantial efforts over the past decade, the genetic etiology of the VATER/VACTERL association in most instances remains elusive. The application of new genomic technologies such as high-resolution copy number variation studies or next-generation exome sequencing might lead to the identification of some of these causes.


Dopamine receptor subtypes in the human coronary vessels of healthy subjects.
Dopamine D(1)-D(5) receptors subtypes were studied in human coronary vessels of healthy subjects to assess their localization and their expression. Samples of intraparenchymal and extraparenchymal branches of human coronary arteries and veins were harvested from four normal native hearts explanted from four young brain dead heart donors in case of orthoptic transplant, not carried out for technical reasons. In all the samples morphological, biochemical, immunochemical, and morphometrical studies were performed including quantitative analysis of images and evaluation of data. Microanatomical section showed healthy coronary vessels, which expressed all dopamine receptors (from D(1) to D(5)) with a different pattern of distribution between the different layers, in the intra and in the extraparenchymal branches.D(1) and D(5) (with a prevalence D(1) over D(5)) were distributed in the adventitia and to a lesser extent in the outer media but they were absent in arterioles, capillaries and venules. Endothelial and the middle layer showed D(2), D(3) and D(4) receptors, with a greater expression of D(2). Immunoblot analysis of dopamine monoclonal antibodies and dopamine receptors showed a different migration band for each receptor: D(1) (45 KDa); D(2) (43 KDa); D(3) (42 kDa); D(4) (40-42 KDa); D(5) (38-40 KDa) These findings demonstrate the presence of all dopamine receptor subtypes in the wall of human coronary vessels of healthy subjects. Dopamine D(1) and D(2) receptor subtypes are the most expressed, suggesting their prominent role in the coronary vasoactivity.


Comparative genotoxicity of silver nanoparticles in human liver HepG2 and lung epithelial A549 cells.
With the rapid expanding of human exposure to silver nanoparticles (AgNPs), genotoxicity screening of nanosilver is necessary to ensure consumer safety. Here, we assessed one key DNA damage responsive pathway activated by GADD45a gene after 24 h of AgNPs exposure in stable luciferase reporter cell systems based on two widely used in vitro cell models, human liver HepG2 and lung epithelial A549 cells. The comet assay and micronucleus test were also conducted to confirm the genetic damage induced by AgNPs. Our results showed that AgNPs produced a strong dose-dependent increase in transcriptional activation of GADD45a promoter indicated by luciferase activity accompanying by the significant decreases in cell viability. Surprisingly, in HepG2-luciferase cells, the relative luciferase activity was greater than 4.5× the control level after being treated with 200 μg ml-1 AgNPs. These results were generally in line with the positive and dose-dependent responses in cytotoxicity, DNA strand breaks indicated by Olive tail moment, tail DNA (%) and tail length, and chromosome damage indicated by induction of micronuclei, nucleoplasmic bridges, and nuclear buds. Additionally, compared with the A549-luciferase cells, the HepG2-luciferase cells seemed to be more susceptible to AgNPs as higher levels of genotoxicity were induced. We concluded that our GADD45a promoter-driven luciferase reporter gene cell system, together with the comet assay and micronucleus test, can be used as valuable tools for rapid screening of genotoxic potential of nanosilver. Copyright © 2016 John Wiley & Sons, Ltd.


Manage Your Life Online (MYLO): a pilot trial of a conversational computer-based intervention for problem solving in a student sample.
Computerized self-help that has an interactive, conversational format holds several advantages, such as flexibility across presenting problems and ease of use. We designed a new program called MYLO that utilizes the principles of METHOD of Levels (MOL) therapy--based upon Perceptual Control Theory (PCT). We tested the efficacy of MYLO, tested whether the psychological change mechanisms described by PCT mediated its efficacy, and evaluated effects of client expectancy. Forty-eight student participants were randomly assigned to MYLO or a comparison program ELIZA. Participants discussed a problem they were currently experiencing with their assigned program and completed measures of distress, resolution and expectancy preintervention, postintervention and at 2-week follow-up. MYLO and ELIZA were associated with reductions in distress, depression, anxiety and stress. MYLO was considered more helpful and led to greater problem resolution. The psychological change processes predicted higher ratings of MYLO's helpfulness and reductions in distress. Positive expectancies towards computer-based problem solving correlated with MYLO's perceived helpfulness and greater problem resolution, and this was partly mediated by the psychological change processes identified. The findings provide provisional support for the acceptability of the MYLO program in a non-clinical sample although its efficacy as an innovative computer-based aid to problem solving remains unclear. Nevertheless, the findings provide tentative early support for the mechanisms of psychological change identified within PCT and highlight the importance of client expectations on predicting engagement in computer-based self-help.


Patient-derived C-terminal mutation of FANCI causes protein mislocalization and reveals putative EDGE motif function in DNA repair.
Fanconi anemia (FA) is a rare familial genome instability syndrome caused by mutations in FA genes that results in defective DNA crosslink repair. Activation of the FA pathway requires the FA core ubiquitin ligase complex-dependent monoubiquitination of 2 interacting FA proteins, FANCI and FANCD2. Although loss of either FANCI or FANCD2 is known to prevent monoubiquitination of its respective partner, it is unclear whether FANCI has any additional domains that may be important in promoting DNA repair, independent of its monoubiquitination. Here, we focus on an FA-I patient-derived FANCI mutant protein, R1299X (deletion of 30 residues from its C-terminus), to characterize important structural region(s) in FANCI that is required to activate the FA pathway. We show that, within this short 30 amino acid stretch contains 2 separable functional signatures, a nuclear localization signal and a putative EDGE motif, that is critical for the ability of FANCI to properly monoubiquitinate FANCD2 and promote DNA crosslink resistance. Our study enable us to conclude that, although proper nuclear localization of FANCI is crucial for robust FANCD2 monoubiquitination, the putative FANCI EDGE motif is important for DNA crosslink repair.


[Three-dimensional CT study of the carpus under pronation-supination constraints].
By studying 3D imaging of the wrist under pronation-supination strain, we found that the simple comparison of a series of two corresponding cuts may provide a great deal of useful information on how the carpus transmits the longitudinal torque from the forearm to the hand. A special wooden trestle was made to fix the subject in the CT scanner in a permanent effort of pronation or supination. In the first group of scans, this effort was said to be "free" because the hand was simply maintained in a fixed window without any muscular contraction, except pronation or supination muscles. In the second group of scans, this effort was said to be "constrained" because the hand gripped a fixed bar with contraction of the flexor muscles. The thickness of the cuts was 1.2 millimeters and they were separated by 1.5 millimeters. Four levels were specially studied: the lower radio-ulnar joint (LRUJ), the proximal row of the carpus, the distal one and the metacarpal bases. Many elementary movements occur in the carpus in constrained supination: the triquetrum "supinates" (7 degrees), the scaphoid flattens and "pronates" (2 degrees) around the capitum the ridges of the carpal anterior concavity approximate (3 mm). In constrained pronation, the anterior concavity of the carpus flattens emphasizing the role of the anterior retinaculum. The LRUJ is very unstable: in free pronation, the ulnar head moves dorsally, firmly pressing the posterior part of the sigmoid notch, responsible for fracture of a postero-medial fragment in Colles fracture. The quadratus pronatus is a very important muscle to coapt this joint. We propose the "screwing (or unscrewing) test" in the diagnosis of arthrosis or instability of the LRUJ. We define the notion of "rotational shift" to appreciate the quality of the pronation/supination torque transmission. In constrained pronation/supination, this rotational shift is 5 degrees in the radio-carpal joint. This is very important to appreciate the quality of the wrist prosthesis. In free pronation/supination, the rotational shift is 45 degrees between radius and metacarpal bases. In constrained pronation/supination, it becomes 10 degrees. The wrist ligaments are unable to resist the wrist rotational shift and favor the torque transmission. The tendinous caging of the wrist is the main factor for maintaining rigidity of the carpus and transmitting the torque as muscles are contracted. The wrist can be compared with a fluid drive clutch, whose pedal is muscular contraction.


A sodium calcium arsenate, NaCa(AsO(4)).
The title compound, NaCa(AsO(4)), was synthesized using a hydro-thermal method at 633-643 K. It has a dense structure composed of alternating layers of distorted [CaO(6)] octa-hedra and layers of [AsO(4)] tetra-hedra and distorted [NaO(6)] octa-hedra, stacked along the a axis. The As, Ca and two O atoms lie on the mirror plane at y = 1/4 (i.e. 4c), while the Na atom lies on an inversion centre (1/2, 1/2, 0) (i.e. 4b). Each distorted [CaO(6)] octa-hedron shares four equatorial common O vertices with four neighboring octa-hedra, forming a layer parallel to (100), whereas each distorted [NaO(6)] octa-hedron shares two opposite edges with two neighboring ones, forming a chain running along [010]. Each isolated [AsO(4)] tetra-hedron shares two edges with two different [NaO(6)] octa-hedra in one [NaO(6)] chain and a vertex with another chain. Simultaneously the above [AsO(4)] tetra-hedron located in a four-membered [CaO(6)] ring shares one edge of its base facet with one [CaO(6)] octa-hedron and three corners with three other [CaO(6)] octa-hedra of one [CaO(6)] layer, and the remaining apex is shared with another [CaO(6)] layer. [NaO(6)] octa-hedra and [CaO(6)] octa-hedra are linked to each other by sharing edges and vertices.


Role of o-acetylserine in hydrogen sulfide emission from pumpkin leaves in response to sulfate.
In the presence of excess sulfate, cysteine synthesis in pumpkin (Cucurbita pepo) leaves is not limited by sulfate reduction, but by the availability of O-acetylserine. Feeding of O-acetylserine or its metabolic precursors S-acetyl-coenzyme-A and coenzyme A to leaf discs enhanced the incorportion of [(35)S]sulfate into reduced sulfur compounds, mainly into cysteine, at the cost of lowered H(2)S emission; the uptake and reduction of sulfate is not affected by these treatments. beta-Fluoropyruvate, an inhibitor of the generation of S-acetyl-coenzyme A via pyruvate dehydrogenase, stimulated H(2)S emission in response to sulfate. This stimulation is overcompensated by addition of O-acetylserine, S-acetyl-coenzyme A, or coenzyme A. These results indicate that, in the presence of high amounts of sulfate, excess sulfur is reduced and emitted as H(2)S into the atmosphere. The H(2)S emitted seems to be produced by liberation from a precursor of cysteine rather than by cysteine desulfhydration.


9-O-acetylated sialoglycoproteins are important immunomodulators in Indian visceral leishmaniasis.
Overexpression of disease-associated 9-O-acetylated sialoglycoproteins (9-O-AcSGPs) on peripheral blood mononuclear cells (PBMC) of visceral leishmaniasis (VL) patients (PBMC(VL)) compared to their levels of expression in healthy individuals has been demonstrated using a lectin, achatinin-H, with specificity toward 9-O-acetylated sialic acid derivatives alpha2-6 linkage with subterminal N-acetylgalactosamine (9-O-AcSAalpha2-6GalNAc). The decreased presence of disease-associated 9-O-AcSGPs on different immune cells of parasitologically cured individuals after successful treatment relative to the levels in patients with active VL prior to treatment was demonstrated. However, their contributory role as immunomodulatory determinants on PBMC(VL) remained unexplored. Accordingly, 9-O-AcSGPs on PBMC(VL) were sensitized with achatinin-H, leading to their enhanced proliferation compared to that observed with different known mitogens or parasite antigen. This lymphoproliferative response was characterized by evaluation of the TH1/TH2 response by intracellular staining and enzyme-linked immunosorbent assay for secreted cytokines, and the results were corroborated by their genetic expression. Sensitized PBMC(VL) evidenced a mixed TH1/TH2 cellular response with a predominance of the TH1 response, indicating the ability of 9-O-AcSGPs to modulate the host cell toward a favorable response. Interestingly, the humoral and cellular responses showed a good correlation. Further, high levels of anti-9-O-AcSGP antibodies with an order of distribution of immunoglobulin M (IgM) > IgG1 = IgG3 > IgG4 > IgG2 > IgE could be explained by a mixed TH1/TH2 response. A good correlation of enhanced 9-O-AcSGPs with both the cell-mediated (r = 0.98) and humoral (r = 0.99) response was observed. In summary, it may be concluded that sensitization of 9-O-AcSGPs on PBMC(VL) may provide a basis for the modulation of the host's immune response by their controlled expression, leading to a beneficial immune response and influencing the disease pathology.


Psychological aspects of staleness and dependence on exercise.
This article discusses the psychological symptoms of overtraining and the relationship between staleness and exercise dependence. Staleness may be prevented by monitoring the mood state of the individual so that, if necessary, appropriate action may be taken to taper the volume of training and maximize performance. The criterion for exercise dependence is difficult to define and some suggestions are made on clarifying the diagnosis and future research in this area.


Effect of Lipid Composition on the Membrane Orientation of the G Protein-Coupled Receptor Kinase 2-Gβ1γ2 Complex.
Interactions between proteins and cell membranes are critical for biological processes such as transmembrane signaling, and specific components of the membrane may play roles in helping to organize or mandate particular conformations of both integral and peripheral membrane proteins. One example of a signaling enzyme whose function is dependent on membrane binding and whose activity is affected by specific lipid components is G protein-coupled receptor (GPCR) kinase 2 (GRK2). Efficient GRK2-mediated phosphorylation of activated GPCRs is dependent not only on its recruitment to the membrane by heterotrimeric Gβγ subunits but also on the presence of highly negatively charged lipids, in particular phosphatidylinositol 4',5'-bisphosphate (PIP2). We hypothesized that PIP2 may favor a distinct orientation of the GRK2-Gβγ complex on the membrane that is more optimal for function. In this study, we compared the possible orientations of the GRK2-Gβγ complex and Gβγ alone on model cell membranes prepared with various anionic phospholipids as deduced from sum frequency generation vibrational and attenuated total reflectance Fourier transform infrared spectroscopic methods. Our results indicate that PIP2 affects the membrane orientation of the GRK2-Gβ1γ2 complex but not that of complexes formed with anionic phospholipid binding deficient mutations in the GRK2 pleckstrin homology (PH) domain. Gβ1γ2 exhibits a similar orientation on the lipid bilayer regardless of its lipid composition. The PIP2-induced orientation of the GRK2-Gβ1γ2 complex is therefore most likely caused by specific interactions between PIP2 and the GRK2 PH domain. Thus, PIP2 not only helps recruit GRK2 to the membrane but also "fine tunes" the orientation of the GRK2-Gβγ complex so that it is better positioned to phosphorylate activated GPCRs.


Cutaneous infiltrates and peripheral blood immune responses in dogs with immunomodulatory-responsive lymphocytic-plasmacytic pododermatitis.
This study characterizes T- and B-lymphocyte responses in the peripheral blood and lesional skin of dogs with immunomodulatory-responsive lymphocytic-plasmacytic pododermatitis (ImR-LPP), a term previously proposed to denote a subpopulation of dogs with idiopathic pododermatitis. T-cell (CD3+, CD4+ and CD8+ ) and B-cell (CD21+) counts were significantly increased in both the epidermis and dermis of lesional ImR-LPP skin compared with that in pedal skin from healthy controls. CD3+ , CD4+, CD8+ and CD21+ cells were commonly observed in perivascular sites in the superficial dermis, periadnexally, beneath the dermal-epidermal (DE) junction and in the epidermis of lesional ImR-LPP skin. The CD8+ /CD3+ T-cell ratio in peripheral blood was significantly increased in the ImR-LPP group (0.42 versus 0.35 in controls). Serum IgA, IgG and IgM concentrations were all significantly elevated in affected dogs. Lymphocyte stimulation indices in ImR-LPP dogs were comparable with control levels except for a lower response to ionomycin (6.0 versus 11.1). Dogs with ImR-LPP had a higher incidence and mean (semi-quantitative) score for IgA, IgG and IgM deposits in the epidermis, and a significantly increased incidence of dermal IgA+, IgG+ and IgM+ mononuclear inflammatory cells. The results indicate that upregulated T- and B-lymphocyte responses may contribute to the pathogenesis of the skin lesions observed in dogs with ImR-LPP.


Towards Replacing Titanium with Copper in the Bipolar Plates for Proton Exchange Membrane Water Electrolysis.
For proton exchange membrane water electrolysis (PEMWE) to become competitive, the cost of stack components, such as bipolar plates (BPP), needs to be reduced. This can be achieved by using coated low-cost materials, such as copper as alternative to titanium. Herein we report on highly corrosion-resistant copper BPP coated with niobium. All investigated samples showed excellent corrosion resistance properties, with corrosion currents lower than 0.1 µA cm-2 in a simulated PEM electrolyzer environment at two different pH values. The physico-chemical properties of the Nb coatings are thoroughly characterized by scanning electron microscopy (SEM), electrochemical impedance spectroscopy (EIS), X-ray photoelectron spectroscopy (XPS), and atomic force microscopy (AFM). A 30 µm thick Nb coating fully protects the Cu against corrosion due to the formation of a passive oxide layer on its surface, predominantly composed of Nb2O5. The thickness of the passive oxide layer determined by both EIS and XPS is in the range of 10 nm. The results reported here demonstrate the effectiveness of Nb for protecting Cu against corrosion, opening the possibility to use it for the manufacturing of BPP for PEMWE. The latter was confirmed by its successful implementation in a single cell PEMWE based on hydraulic compression technology.


Bandwidth-tunable narrowband rectangular optical filter based on stimulated Brillouin scattering in optical fiber.
We propose a rectangular optical filter based on stimulated Brillouin scattering (SBS) in optical fiber with bandwidth tuning from 50 MHz to 4 GHz at less than 15-MHz resolution. The rectangular shape of the filter is precisely achieved utilizing digital feedback control of the comb-like pump spectral lines. The passband ripple is suppressed to ~1 dB by mitigating the nonlinearity influences of the comb-like pump lines generated in electrical and optical components and fibers. Moreover a fiber with a single Brillouin peak is employed to further reduce the in-band ripple and the out-of-band SBS gain at the same time. Finally, we analyze the noise performance of the filter at different bandwidth cases and demonstrate the system performance of the proposed filter with 2.1-GHz bandwidth and 19-dB gain by amplifying a 2-GHz orthogonal frequency-division-multiplexing (OFDM) signal with quadrature-phase-shift-keying (QPSK) and 16-quadrature-amplitude-modulation (16-QAM) on each subscriber.


Expression of denitrification genes in response to a waterlogging event in a Fluvisol and its relationship with large nitrous oxide pulses.
The contributions of large N2 O pulses following waterlogging to the annual cumulative N2 O productions were significant in a Fluvisol. To uncover the mechanisms underlying these large N2 O pulses, a Fluvisol sampled from an agricultural field in Japan was subjected to waterlogging during incubation. Larger N2 O emissions were observed in intact soil cores when compared to emissions from sieved soils, indicating the importance of soil properties. The most important factor controlling the magnitude of the N2 O pulses after waterlogging was the soil moisture prior to waterlogging. The major pathway for N2 O production was denitrification. Quantitative PCR and quantitative RT-PCR analyses showed that the denitrification genes (nirS, nirK, and nosZ) correlated with N2 O emissions at the mRNA level but not at the DNA level. The change in denitrification gene mRNA levels was more prominent in the 0- to 1-cm soil compared with the 1- to 3-cm soil. Water-soluble and hot-water-soluble carbon contents also showed the highest amount in the 0- to 1-cm soil. These indicate that there was a strong variation in soil microbial properties over very small changes in soil depth, and this variation is important in determining the magnitude of N2 O emissions.