Evaluation of an in-house PCR for diagnosing scrub typhus along with a preliminary study of genotypic characterization of Orientia tsutshugamushi circulating in Chhattisgarh: A prospective clinico-microbiological study.
Scrub typhus, caused by Orientia tsutsugamushi (O. tsutsugamushi) present nonspecific clinical features during manifestation of acute undifferentiated febrile illness (AUFI) to render its early diagnosis difficult. Accordingly, this study was undertaken to assess an in-house groEL PCR versus IgM ELISA for the diagnosis of scrub typhus and to genotypically characterise the randomly selected scrub typhus positive cases. Blood samples, collected from two hundred twenty one (221) AUFI cases were subjected to groEL PCR and IgM ELISA for diagnosis of scrub typhus. Eleven randomly selected PCR positive cases were processed for DNA sequencing to determine the genetic diversity of O. tsutsugamushi in Chhattisgarh. Scrub typhus prevalence of 35.2% were detected among AUFI cases using both in-house groEL PCR and IgM ELISA. PCR alone showed sensitivity, specificity, positive and negative predictive values of 66.6% (CI: 55.08-76.94), 100% (CI: 90 to 100),100% (CI: 93.15 to 100) and 57.37% (CI: 44.05 to 69.96) while for IgM ELISA, these parameters were 62.8% (CI: 51.13-73.50), 100% (CI: 90 to 100), 100% (CI: 92.75 to 100) and 54.68% (CI: 41.75 to 67.18) respectively. PCR and ELISA could detect scrub typhus in 37.2% and 33.3% cases, when tested alone. groEL PCR detected the O. tsutsugamushi throughout the course of infection. Phylogenetic analysis depicted 5 of 11 positive cases belonged to Kuroki, Japan strain of O. tsutsugamushi, followed by Gilliam and Karp strain in 4 and 2 cases respectively. Scrub typhus should be considered in differential diagnosis of AUFI. groEL PCR may aid on to IgM ELISA test for optimum laboratory diagnosis of scrub typhus by its implementation especially in seronegative cases. Predominance of Kuroki-like strain followed by Gillian and Karp strains of O. tsutsugamushi in Chhattisgarh confirm variable geographical distribution of O. tsutsugamushi and provide the baseline epidemiological data which will eventually be used to help the researchers for developing better diagnostic tests and vaccine covering the predominant genotypes.


[Determination of copper, lead and zinc in fruit preserves].
Copper, lead and zinc content in canned fruit manufactured in Moldavia in 1979 and 1980 was determined by polarography and atomic-absorption spectrophotometry. The authors developed a method for determining the above trace elements in one sample using an atomic-absorption spectrophotometer. The errors of each method were calculated. It was ascertained that the data obtained by both the methods were identical. Analysis of the data indicates that the content of copper, lead and zinc is within the standards approved by the USSR Ministry of Health. The data obtained in the studies can be used in both toxicological work on setting standards of heavy metals content in canned food and in the tables of the chemical composition of canned food. The methods for copper, lead and zinc determination can be used as the basis for the development of standard procedures for heavy metals.


[Does the residual mediastinal mass have prognostic significance in children with Hodgkin's disease (HD)?].
Prognostic significance of residual mediastinal tumor mass in children treated for HD as well as the choice of the optimal management of these cases still remains unknown. In years 1994-2001 in 10 PPLLSG participating centers 480 children (age 2-19.7 years) were treated for HD (stages I-IV). In 338 cases initial mediastinal/lung hilus involvement was present. All patients with initial mediastinal/lung hilus involvement were treated with multidrug chemotherapy combined with involved field radiotherapy. In five cases remission was not achieved. Complete remission (CR) was achieved in 226 patients and uncertain complete remission (UCR) in 107 patients, in whom after completion of planned treatment residual changes in mediastinum/lung hilus were identified in radiological examinations. Twenty four children with persistent mediastinal tumor underwent thoracoscopy or thoracotomy. In only one case histopathological examination revealed the presence of neoplastic cells in mediastinal mass tissue, in 2 another cases cystic changes in mediastinum were present, in one case thymic tissue was identified, necrotic tissue was present in 20 cases. Among 107 children with residual mediastinal tumor and 226 patients with CR achieved, relapses occurred in 6 and 18 patients respectively. Over 5-year relapse-free survival was 92.4% and 91.3% respectively. Patients with the presence of mediastinal/lung hilus tumor after the completion of the treatment do not have an increased risk of relapse, but before the completion of therapy they require careful, clear-sighted and repeated examinations including computed tomography (CT), magnetic resonance imaging (MRI) and especially positron emission tomography (PET) to evaluate the nature of persistent lesions. Only in clinically and radiologically doubtful cases tumor biopsy with subsequent histopatological examination should be performed.


Deterioration of heart rate recovery index in patients with systemic lupus erythematosus.
Systemic lupus erythematosus (SLE) is an autoimmune disorder resulting in multisystemic inflammatory damage. It is reported that cardiovascular diseases (CVD) are responsible for 20%-30% of deaths in patients with SLE. Heart rate recovery after exercise is a function of vagal reactivation, and its impairment is an independent prognostic indicator for cardiovascular and all-cause mortality. The aim of our study was to evaluate the heart rate recovery index in patients with SLE. The study population included 48 patients with SLE (35 women, mean age 46.3 ± 12.8 yrs, mean disease duration 6.0 ± 2.3 yrs) and 44 healthy controls (30 women, mean age 45.7 ± 12.9 yrs). Basal electrocardiography, echocardiography, and treadmill exercise testing were performed on all patients and controls. The heart rate recovery index was defined as the reduction in the heart rate from the rate at peak exercise to the rate at the first minute (HRR(1)), second minute (HRR(2)), third minute (HRR(3)), and fifth minute (HRR(5)) after stopping exercise stress testing. There were significant differences in HRR(1) and HRR(2) indices between patients with SLE and the control group (24.1 ± 6.5 vs 33.3 ± 9.3; p < 0.001, and 44.6 ± 13.3 vs 53.7 ± 9.9; p < 0.001, respectively). Similarly, HRR(3) and HRR(5) indices of the recovery period were lower in patients with SLE, compared with indices in the control group (57.6 ± 13.0 vs 64.9 ± 11.7; p = 0.006, and 67.2 ± 12.3 vs 75.0 ± 15.4; p = 0.009, respectively). Effort capacity was markedly lower (9.0 ± 1.9 vs 11.1 ± 2.3 metabolic equivalents; p = 0.001, respectively) among the patients with SLE. The heart rate recovery index is deteriorated in patients with SLE. When the prognostic significance of the heart rate recovery index is considered, these results may contribute to explain the increased occurrence of cardiac death. It points to the importance of the heart rate recovery index in the identification of high-risk patients.


Technetium-99m-tetrofosmin as a new radiopharmaceutical for myocardial perfusion imaging.
A new cationic complex, [99mTc(tetrofosmin)2O2]+, where tetrofosmin is the ether functionalized diphosphine ligand 1,2-bis[bis(2-ethoxyethyl)phosphino]ethane, has been synthesized and evaluated for potential use in myocardial perfusion imaging. The structure of the complex has been determined by x-ray crystallography of the 99Tc analog. In comparison with previously reported 99mTc complexes of alkyl-phosphines, the tetrofosmin species shows substantially increased clearance from nontarget tissue, especially blood and liver. A freeze-dried kit formulation has been developed. The kit provides a product of high radiochemical purity up to 8 hr after reconstitution at room temperature.


Expression of a PKR dominant-negative mutant in myogenic cells interferes with the myogenic process.
In this study, we explored the possibility that PKR, a dsRNA-activated regulatory protein, is an essential component in the differentiation program of myogenic cells in vitro. For this purpose, we used a retroviral expression vector pMV7, harboring the PKR dominant-negative mutant PKRDelta6 (pMV7-p68Delta6). Murine C2C12 myogenic cells were transfected either with pMV7 or with pMV7-p68Delta6. Neomycin-resistant clones from both types were isolated and expanded and the results obtained with the representative clones C2-NEO (transfected with pMV7) and clone 17 and clone 22 (both transfected with pMV7-p68Delta6) are presented. In clone 17 and 22 cells, regardless of IFN treatment, a similar level of the transfected human p68 PKR mutant was detected. This protein was absent in C2-NEO cells. In parallel, in all types of cells, a low basal level of the endogenous murine p65 PKR protein was observed, which was further induced by IFN. However, PKR enzymatic activity was significantly induced by IFN only in C2-NEO cells, while it was hardly detected in both clones 17 and 22, even after IFN treatment. Furthermore, in contrast to C2-NEO cells, only a slight to moderate increase in enzymatic activity was observed in clone 17 and 22 differentiating cells. Next, cells were grown either in growth medium (GM) or differentiation medium (DM), and the progression of the myogenic program was studied. An inhibition in myotube formation in clone 17 versus C2-NEO cells cultivated in DM was clearly observed. Furthermore, while the growth rate and thymidine incorporation were reduced in C2-NEO cells grown in DM, both clone 17 and 22 cells were less affected under the same conditions. Similarly, a delay in the accumulation of the transcription factors MyoD and myogenin, as well as in creatine kinase activity and accumulation of troponin T, was detected in DM-cultivated clone 17 and clone 22 cells. Moreover, a delay in the induction of p21 (WAF1), in down-regulation of cyclin D1 and c-myc, and in the accumulation of the underphosphorylated form of pRb was also observed in clone 17 cells. We conclude that inhibition of endogenous PKR activity by a PKR dominant-negative mutant interferes with the myogenic program of murine C2C12 myogenic cells.


Effects of Ultrasound-Guided Stellate Ganglion Block on Postoperative Quality of Recovery in Patients Undergoing Breast Cancer Surgery: A Randomized Controlled Clinical Trial.
Surgery has been the primary treatment for breast cancer. However, instant postoperative complications, such as sleep disorder and pain, dramatically impair early postoperative quality of recovery, resulting in more extended hospital stays and higher costs. Recent clinical trials indicated that stellate ganglion block (SGB) could prolong sleep time and improve sleep quality in breast cancer survivors. Moreover, during the perioperative period, SGB enhanced the recovery of gastrointestinal functions in patients with laparoscopic colorectal cancer surgery and thoracolumbar spinal surgery. Furthermore, perioperative SGB decreased intraoperative requirements for anesthetics and analgesics in patients with complex regional pain syndrome. However, information is scarce regarding the effects of SGB on postoperative quality recovery in patients with breast cancer surgery. Therefore, we investigated the effects of SGB on the postoperative quality of recovery of patients undergoing breast cancer surgery. Sixty patients who underwent an elective unilateral modified radical mastectomy were randomized into two 30-patient groups that received either an ultrasound-guided right-sided SGB with 6 ml 0.25% ropivacaine (SGB group) or no block (control group). The primary outcome was the quality of postoperative recovery 24 hours after surgery, assessed with a Chinese version of the 40-item Quality of Recovery (QoR-40) questionnaire. Secondary outcomes were intraoperative requirements of propofol and opioids, rest pain at two, four, eight, and 24 hours after surgery, patient satisfaction score, and the incidence of postoperative abdominal distension. At 24 hours after surgery, global QoR-40 scores were higher in the SGB group than in the control group. Besides, in the SGB group, patients needed less propofol, had a lower incidence of postoperative abdominal bloating, and had higher satisfaction scores. Ultrasound-guided SGB could improve the quality of postoperative recovery in patients undergoing breast cancer surgery by less intraoperatively need for propofol and better postoperative recovery of sleep and gastrointestinal function.


Contribution to ligand binding by multiple carbohydrate-recognition domains in the macrophage mannose receptor.
The extracellular portion of the macrophage mannose receptor is composed of several cysteine-rich domains, including a fibronectin type II repeat and eight segments related in sequence to Ca(2+)-dependent carbohydrate-recognition domains (CRDs) of animal lectins. Expression of portions of the receptor in vitro, in fibroblasts and in bacteria, has been used to determine which of the extracellular domains are involved in binding and endocytosis of ligand. The NH2-terminal cysteine-rich domain and the fibronectin type II repeat are not necessary for endocytosis of mannose-terminated glycoproteins. CRDs 1-3 have at most very weak affinity for carbohydrate, so the carbohydrate binding activity of the receptor resides in CRDs 4-8. CRD 4 shows the highest affinity binding and has multispecificity for a variety of monosaccharides. However, CRD 4 alone cannot account for the binding of the receptor to glycoproteins. At least 3 CRDs (4, 5, and 7) are required for high affinity binding and endocytosis of multivalent glycoconjugates. In this respect, the mannose receptor is like other carbohydrate-binding proteins, in which several CRDs, each with weak affinity for single sugars, are clustered to achieve high affinity binding to oligosaccharides. In the mannose receptor, these multiple weak interactions are achieved through several active CRDs in a single polypeptide chain rather than by oligomerization of polypeptides each containing a single CRD.


Costimulation of the CD3 pathway by CD28 ligation in human intestinal lymphocytes.
A distinctive characteristic of human intestinal lymphocytes is their low responsiveness to stimuli of the CD3 pathway in vitro. This may be due to anergy resulting from CD3 stimulation without costimulation. Costimulatory molecules, such as HML-1, VLA-4, CD44, and CD28, may be lacking or unresponsive to ligation. Expression of these costimulatory markers was examined by immunofluorescent staining and flow cytometry. Their ability to costimulate intestinal lymphocytes was tested by measuring changes in proliferation, IL-2 production, and calcium ion mobilization. HML-1 was found on 87 +/- 8% of intraepithelial lymphocytes but only 52 +/- 10% of lamina propria lymphocytes; the density of expression was three times higher on the former. VLA-4 and CD44 were expressed on more than 80% of both lymphocyte types. CD28 was found on 31 +/- 26% of intraepithelial lymphocytes and 55 +/- 15% of lamina propria lymphocytes; virtually all CD4+ T cells and 15 to 26% of CD8+ T cells were CD28+. Of the four costimulatory molecules, ligation of only CD28 increased proliferation and IL-2 production; none affected calcium ion mobilization with stimulation through the CD3 pathway. The marker B7/BB1, a binding partner to CD28, was not present on lymphocytes or epithelial cells by flow cytometry. CD28 is the only one of these costimulatory molecules that enhanced CD3-induced functions of both intestinal lymphocyte types, but little ligand is available in vivo. Without coligation, CD3 activation may lead to anergy.


[Radiologic diagnosis of post-traumatic cerebrospinal rhinorrhea].
It is imperative to identify the cause of cerebrospinal fluid (CSF) leak due to the major risk of infectious complications. Mainly the consequence of a skull base or facial trauma, its most frequent symptom is rhinorrhea. It represents a real diagnostic, clinical and radiological challenge. Two extreme situations raise different problems: either CSF leak is active, with important facial damage involving frontal, ethmoidal or sphenoidal sinuses and petrous bone; in such cases, evaluation by CT is sufficient and easy. Or the leak has dried up, there is no evidence of bony fracture; it is imperative to localize the exact site of the bony or dural defect: CT in axial and coronal planes with thin slices, computerized tomographic cisternography with non ionic water soluble contrast media in order to exhibit the leak in the paranasal sinuses. In some cases however, pitfalls still occur in the determination of such meningeal fistulae.


Electrostatics, structure prediction, and the energy landscapes for protein folding and binding.
While being long in range and therefore weakly specific, electrostatic interactions are able to modulate the stability and folding landscapes of some proteins. The relevance of electrostatic forces for steering the docking of proteins to each other is widely acknowledged, however, the role of electrostatics in establishing specifically funneled landscapes and their relevance for protein structure prediction are still not clear. By introducing Debye-Hückel potentials that mimic long-range electrostatic forces into the Associative memory, Water mediated, Structure, and Energy Model (AWSEM), a transferable protein model capable of predicting tertiary structures, we assess the effects of electrostatics on the landscapes of thirteen monomeric proteins and four dimers. For the monomers, we find that adding electrostatic interactions does not improve structure prediction. Simulations of ribosomal protein S6 show, however, that folding stability depends monotonically on electrostatic strength. The trend in predicted melting temperatures of the S6 variants agrees with experimental observations. Electrostatic effects can play a range of roles in binding. The binding of the protein complex KIX-pKID is largely assisted by electrostatic interactions, which provide direct charge-charge stabilization of the native state and contribute to the funneling of the binding landscape. In contrast, for several other proteins, including the DNA-binding protein FIS, electrostatics causes frustration in the DNA-binding region, which favors its binding with DNA but not with its protein partner. This study highlights the importance of long-range electrostatics in functional responses to problems where proteins interact with their charged partners, such as DNA, RNA, as well as membranes.


The prevalence of atrial fibrillation in Greenland: a register-based cross-sectional study based on disease classifications and prescriptions of oral anticoagulants.
Previous studies of the prevalence of atrial fibrillation (AF) in Greenland are based on either single-point electrocardiograms (ECGs) or patients admitted with stroke. This study estimates the prevalence of AF based on disease classifications in the electronic medical record system (EMR) and prescriptions of oral anticoagulants (OACs). Patients given a diagnose classification code for AF or atrial flutter or prescribed the vitamin K antagonist Warfarin or the direct-acting oral anticoagulant Rivaroxaban were identified in the EMR. Descriptive data and selected laboratory values were extracted, and a minimum CHA2DS2-VASc score was calculated for the 790 patients identified in the EMR (66% men). A total prevalence of AF of 1.4% was found in the general population (1.8% among men and 1.0% among women), with a significantly lower prevalence among women younger than 70 years. There was a significant increase in AF-prevalence with advancing age (p<0.001) for both men and women. A minimum CHA2DS2-VASc was estimated and app. 10% of the patients may be undertreated with OACs. The prevalence of AF found in this study is higher than that found in previous studies in Greenland and comparable to the prevalence found in other Western countries, indicating that AF is common in Greenland.


Regulation of p38 MAP kinase by anastellin is independent of anastellin's effect on matrix fibronectin.
Anastellin is an angiogenesis inhibitor derived from the first type III repeat of fibronectin (FN). Anastellin binds to fibronectin and promotes the polymerization of soluble fibronectin into a highly polymerized form termed superfibronectin. In addition, anastellin also causes remodeling of pre-existing fibronectin matrix and modulates cell signaling pathways in both endothelial cells and fibroblasts. In the present study, we address the relationship of anastellin's effects on fibronectin matrix to its effects on p38 MAP kinase (MAPK) activation. Using a mutant form of anastellin which binds to fibronectin matrix, but does not stimulate formation of superfibronectin, we demonstrate that the activation of p38 MAPK by anastellin is not dependent on the formation of superfibronectin. The mutant form of anastellin does stimulate matrix remodeling, but experiments using FN(-/-) cells show that the effect of anastellin on p38-MAPK activation is completely independent of fibronectin. Anastellin was able to activate p38 MAPK on cells in suspension as well as on cells null for beta1 integrins, suggesting that anastellin activity did not require ligation of integrins. These data suggest that the activation of p38 MAPK by anastellin is independent of anastellin's effects on fibronectin matrix organization.


Toll-like receptor-4: renal cells and bone marrow cells signal for neutrophil recruitment during pyelonephritis.
The molecular mechanisms of pathogen recognition that initiate infective pyelonephritis are poorly understood. Toll-like receptor-4 (TLR4) mutant mice infected with uropathogenic Escherichia coli lack renal CXCL2 mRNA expression, subsequent neutrophil recruitment, and renal abscess formation. We used a bone marrow transplant approach in order to investigate the contribution of TLR4 in intrinsic renal cells or bone-marrow-derived immune cells to neutrophil recruitment during infective pyelonephritis. Both chimera either expressing mutant tlr4 in intrinsic renal cells and wild-type tlr4 in bone marrow-derived cells or vice versa showed an impaired response to uropathogenic E. coli infection in terms of leukocyturia and renal abscess formation when compared to tlr4 wild-type mice with congenic bone marrow transplants. These data suggest that TLR4 is required on both intrinsic renal cells (e.g., tubular epithelial cells) and bone marrow-derived immune cells for the control of ascending uropathogenic E. coli infection by initiating chemokine-driven renal neutrophil recruitment.


Impact of Gender and Relationship Status on Young People's Autonomy and Psychological Wellbeing.
This study uses scales of autonomy and psychological wellbeing to determine whether young people's gender and romantic relationship status give rise to differences in relation to a series of specific dimensions. To this end, we used Ryff's Model of Psychological Wellbeing, which comprises several dimensions: self-acceptance, positive relations with others, autonomy, environmental mastery, personal growth, and purpose in life; and our own Transition to Adulthood Autonomy Scale (EDATVA), whose dimensions are: self-organization, understanding context, critical thinking, and socio-political engagement. As a result, a quantitative study was performed with 1,148 young people aged 16-21 from Madrid, Spain and Bogotá, Colombia, of whom 60.2% were female and 39.8% were male. The findings show that in the gender variable there are differences between males and females in the dimensions of positive relations with others, personal growth (wellbeing questionnaire), and understanding context (autonomy questionnaire); the female sample obtained the highest scores. In the relationship variable, differences were found in environmental mastery and purpose in life; higher scores were obtained by young people in a romantic relationship. However, no differences were found in the different dimensions in the autonomy questionnaire between young people in a relationship and those not.


Studies on the chemical and intestinal metabolic profiles of Polygalae Radix by using UHPLC-IT-MSn and UHPLC-Q-TOF-MS method coupled with intestinal bacteria incubation model in vitro.
Polygala Radix (PR) is one of the most commonly used traditional Chinese medicines (TCM), but its effective materials are still not very clear. In order to solve the existing problems, the chemical and intestinal metabolic profiles of PR were analyzed via intestinal bacteria incubation model. An integrated analysis method combined ultra-high performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UHPLC-Q-TOF MS) method and ultra-high performance liquid chromatography with ion-trap multi-stage tandem mass spectrometry (UHPLC-IT-MSn) method was established. As a result, a total of 115 polygala compounds were identified from the PR extract. Moreover, 44 metabolites via intestinal microflora were characterized, of which three compounds were formed from xanthone C-glycosides, 25 compounds were formed from polygala saponins and 16 compounds were formed from oligosaccharide multi-esters. To our knowledge, the metabolites of the PR via intestinal microflora metabolism, especially polygala saponins, are the first reported in vitro. The obtained results would provide the methodological support for further studying the effective materials of the PR in vivo.


[The procedures and results of a quality control program in mammography carried out on a regional basis].
In 1990 the region of Tuscany instituted a quality control program for mammography, in order to check the working conditions of mammography centers in the region, to investigate the feasibility of applying some working standards in line with European recommendations, and to promote the training of personnel. The program involves the following: a test of newly acquired mammographic equipment to establish that it meets the required standards; an annual test of generators, X-ray beams, grid, AEC, screen-film system, film processing, dose and image quality; daily test (phantom radiography and film sensitometry) performed by the personnel of each mammographic Unit; and a sustained advisory service for the technical problems occurring between checks. Thirty-nine of 57 centers participated on a voluntary basis, and 135 checks were performed annually. The results of the program show a performance below the recommended standard in relation to AEC, film sensitometry and the adequacy of lights in dark rooms in 40%, 50% and 63%, respectively, of the centers. Entrance exposure was over 1500 mR, with a maximum value of 3450 mR, in 6 cases in the first round. 5 cases in second, and only 1 case in the third. In general, the results are slightly improving, but mammography optimization is still a long way off. It is necessary to carry out suitable programs for quality control in mammography on a regional basis and to establish precise guidelines for individual mammographic Units to follow in order to meet the European standards.


Questionnaire as diagnostic tool in chronic pelvic pain (CPP): a pilot study.
No standard screening instrument is available enabling physicians to assign the diagnosis chronic pelvic pain (CPP) to women with lower abdominal pain. Therefore, our aim was to evaluate an easy-to-use questionnaire, which can be applied as a validated primary screening test for diagnosing CPP. From the general female population, 577 women completed a questionnaire addressing chronic symptoms in the pelvic region. Included were (amongst others) questions on lower abdominal pain, low back pain, voiding symptoms, dyspareunia, pelveo-perineal dysesthetic feelings and evacuation problems. Serious chronic lower abdominal pain of unknown origin was considered as CPP. Three criteria were applied to validate the questionnaire: construct validity, comparison with results of a previous study and content validity. In addition, the internal consistency was checked to ascertain the reliability of the questionnaire. All items, with the exception of those concerning voiding symptoms and dyspareunia, withstood the validity tests applied, were interrelated and occurred significantly more often in women with CPP than those without. There were no significant differences in the frequency of the occurrence of low back pain, dyspareunia and evacuation problems between CPP women in the current community study and outpatients diagnosed with CPP in an earlier study performed at the University Hospital Rotterdam. Compared to our current study group, pelveo-perineal dysesthesia (PPD) and voiding symptoms were significantly more often reported by the CPP outpatients. The CPP questionnaire can be considered as a validated tool for primary screening of CPP.


Undifferentiated (embryonal) sarcoma of the liver. A tumor of uncertain histogenesis showing divergent differentiation.
Pathologic features of eight cases of undifferentiated (embryonal) sarcoma of the liver (USL) in childhood were studied. Light microscopic examination showed a diffuse growth of spindle cells with occasional polygonal cells and multinucleated giant cells and also revealed focal areas of storiform pattern in four tumors, cambium layer formation in one tumor, and alveolar arrangement in one tumor. Immunohistochemical study showed positive staining of proliferating cells for suggestive histiocytic markers (A1AT in 6/6, A1ACT in 5/6, lysozyme in 4/6, and KP1 in 4/6) and for muscle markers (desmin in 4/6 and HHF35 in 3/6). Ultrastructural examination demonstrated that the individual tumors were composed of a mixture of cells having fibroblastic, histiocytoid, fibrohistiocytoid, myofibroblastic, and undifferentiated (primitive mesenchymal) morphologies. Also identified were cells with definite myoblastic morphology in three tumors: leiomyoblastic in one and rhabdomyoblastic in two. In conclusion, the tumor cells in USL show phenotypical diversity comparable to those of malignant fibrous histiocytoma with or without additional rhabdomyosarcomatous or leiomyosarcomatous differentiation.


Epicardial off-pump pulmonary vein isolation and vagal denervation improve long-term outcome and quality of life in patients with atrial fibrillation.
The limited information available on thoracoscopic pulmonary vein isolation combined with ganglionated plexi ablation and the lack of studies regarding its effect on quality of life and physical capacity urged us to study its acute and long-term results in patients with atrial fibrillation. Forty-three patients (mean age 57.1 years) with symptomatic atrial fibrillation referred for thoracoscopic off-pump epicardial pulmonary vein isolation and ganglionated plexi ablation using radiofrequency energy were included. The physical capacity improved significantly at 6-month follow-up compared with baseline (mean +/- standard deviation, 165.2 +/- 65 Watt versus 155.9 +/- 57 Watt, P = .02). Quality of life (Short Form-36 health survey) significantly improved 12 months after surgery compared with baseline in all subscales except for bodily pain. The symptom severity questionnaire score decreased significantly from mean 15.2 +/- 4.0 points to 10.7 +/- 4.8 points (P = .02). Overall, 25 of 33 patients (76%) followed up for 12 months had no symptomatic atrial fibrillation recurrences or atrial fibrillation episodes on 24-hour Holter recordings. The corresponding figures were 79% (19/24) for patients with paroxysmal atrial fibrillation, 100% (2/2) for persistent atrial fibrillation, and 57% (4/7) for permanent atrial fibrillation. The most common complication was bleeding events (9%) during pulmonary vein dissection. Epicardial off-pump pulmonary vein isolation combined with ganglionated plexi ablation improved quality of life, symptoms, and exercise capacity and therefore may be considered for patients with atrial fibrillation who fail endocardial pulmonary vein ablation or as a first-line procedure if left atrial appendage exclusion is warranted.


Chronic malaria and hyper-reactive malarial splenomegaly: a retrospective study on the largest series observed in a non-endemic country.
Chronic malaria is usually defined as a long-term malarial infection in semi-immune subjects, usually without fever or other acute symptoms. The untreated infection may evolve to hyper-reactive malarial splenomegaly (HMS), a life-threatening complication. This paper describes the largest series of HMS ever observed outside endemic countries, and the clinical outcome after a single anti-malarial treatment. Contrarily to most authors, still reporting the traditional, long-term treatment, regardless possible further exposure, the patients in this series did not receive any further prophylaxis if they were not re-exposed to malaria infection. A retrospective, longitudinal study, describing all patients with HMS diagnosed at the Centre for Tropical Diseases of Negrar, Verona, took place over a 25-year period. HMS was defined by a longitudinal spleen diameter ≥16 cm, IgM ≥ 2.5 g/L, anti-malarial antibody titre ≥160, exclusion of other causes of splenomegaly. The short-term (≤6 months) clinical outcome after a single anti-malarial treatment was analysed and so was the long-term outcome of subjects re-exposed to malaria and submitted or not to anti-malarial prophylaxis or intermittent treatment. The association of the outcome with the main independent variables was first assessed with univariate analysis. Logistic regression was also performed. Forty-four subjects with HMS were retrieved. Of those with a short-term follow-up visit (<6 months, median 43 days) available before returning to endemic areas, 20/22 resulted improved/cured, two were unchanged. Of 22 expatriates seen at long-term follow-up after re-exposure, 18 were improved/cured, including eight out of nine who had followed an anti-malarial prophylaxis and 10/13 who had opted for the alternative of an intermittent treatment. HMS is the most severe form of chronic malaria. A single anti-malarial treatment is probably adequate to treat HMS in the absence of re-exposure, while an adequate prophylaxis is necessary for patients exposed again to malaria transmission. Intermittent treatment would probably be the only viable approach in endemic countries.


Effects of Thoratec pulsatile ventricular assist device timing on the abdominal aortic wave intensity pattern.
Arterial waves are seen as possible independent mediators of cardiovascular risks, and the wave intensity analysis (WIA) has therefore been proposed as a method for patient selection for ventricular assist device (VAD) implantation. Interpreting measured wave intensity (WI) is challenging, and complexity is increased by the implantation of a VAD. The waves generated by the VAD interact with the waves generated by the native heart, and this interaction varies with changing VAD settings. Eight sheep were implanted with a pulsatile VAD (PVAD) through ventriculoaortic cannulation. The start of PVAD ejection was synchronized to the native R wave and delayed between 0 and 90% of the cardiac cycle in 10% steps or phase shifts (PS). Pressure and velocity signals were registered, with the use of a combined Doppler and pressure wire positioned in the abdominal aorta, and used to calculate the WI. Depending on the PS, different wave interference phenomena occurred. Maximum unloading of the left ventricle (LV) coincided with constructive interference and maximum blood flow pulsatility, and maximum loading of the LV coincided with destructive interference and minimum blood flow pulsatility. We believe that noninvasive WIA could potentially be used clinically to assess the mechanical load of the LV and to monitor the peripheral hemodynamics such as blood flow pulsatility and risk of intestinal bleeding.


Tuning of Bistability, Thermal Stability of the Metastable States, and Application Prospects in the C3 -Symmetric Designs of Multiple Azo(hetero)arenes Systems.
Light-responsive molecular systems with multiple photoswitches in C3 -symmetric designs have enormous application potential. The design part of such molecular systems is critical due to its influence in several properties associated with the photoswitches. In order to tune, and in the evaluation of the design-property relationship, we synthesized 18 tripodal systems with variations in the core, linkers, connectivity, and azo(hetero)arene photoswitches. Through extensive spectroscopic and computational studies, we envisaged the factors controlling near-quantitative photoisomerization in both the directions (bistability) and the thermal stability of the metastable states. Furthermore, we also evaluated the impact of designs in obtaining reversible photo-responsive sol-gel phase transitions, solvatochromism, photo- and thermochromism.


Increased expression of HIF-1A and its implication in the hypoxia pathway in primary advanced uterine cervical carcinoma.
The development of cervical cancer exhibits some unique differences compared to other solid tumors. Normal cervical stratified epithelia have characteristics of hypoxic tissue. Lack of oxygen (hypoxia) induces the HIF-1 (hypoxia-inducible factor-1) transcription factor, which is a heterodimer composed of a constitutively expressed β subunit and a hypoxia-inducible α-subunit. HIF-1A targets the transcription of over 70 genes involved in many aspects of cancer biology. In well-oxygenated environments, the HIF-1A subunit is hydroxylated, recognized and marked for proteosomal destruction by an E3 ubiquitin ligase, the von Hippel-Lindau protein (pVHL) complex. Under hypoxic stress, proline hydroxylase (PHD) activity is diminished, and stabilized HIF-1A is involved in the activation of the tissue response to hypoxia. Here, we examined the HIF-1A and VHL transcript levels and HIF-1A protein levels in cancerous tissue (n=30) and non-cancerous, normal uterine cervical tissue (n=30). We also compared the methylation status of HIF-1A and of the VHL promoter regions in cancerous and normal tissue samples. Significantly higher levels of HIF-1A and VHL transcripts (p<0.0001 and p=0.0042, respectively) and of HIF-1A protein (p=0.0037) were detected in cancerous tissue compared to normal samples. We did not observe DNA methylation in the HIF-1A and VHL promoter region in either control or cancerous tissue samples. VHL has a functional hypoxia response element (HRE) in the promoter region, and the induction of this HRE acts within a negative feedback loop to limit the hypoxic HIF-1A response. Our findings may suggest that HIF-1A could promote its own degradation by the induction of VHL gene expression (Spearman correlation coefficient, 0.515; p=0.003). Our study shows for the first time that this increase in VHL expression could be HIF-1A-dependent and serves within a negative feedback pathway during hypoxia to regulate the cell-specific oxygen threshold for HIF-1A activation.


Anti-HLA antibodies after cryopreserved allograft valve implantation does not predict valve dysfunction at three-year follow up.
As the cause of allograft heart valve degeneration is poorly understood, the study aim was to investigate the host antibody response to allograft valve implantation. Sera were obtained from 92 recipients of allograft heart valves (61 pulmonary, 31 aortic). Sera were tested for anti-HLA class I antibodies by ELISA and complement-dependent cytotoxicity (CDC) methods, and anti-HLA class II antibodies by ELISA. Specificities of recipient anti-HLA class I antibodies were defined by standard CDC testing against a panel of T lymphocytes from 80 blood donors. Donor valve HLA typing was performed on stored donor DNA samples using molecular methods. The presence of donor-specific anti-HLA class I antibodies was hence defined in recipient sera. The presence of anti-HLA antibodies and donor-specific anti-HLA class I antibodies were correlated with function of allograft valves at the most recent echocardiographic follow up. At a mean of 3.0 years (range: 0.3-5.4 years) after allograft implantation, 96% (87/92) and 82% (75/92) of patients were positive for anti-HLA class I and II antibodies, respectively, by ELISA testing. Some 68% (61/90) of patients were positive for anti-HLA class I antibody (PRA > 5%) by CDC testing. PRA levels decreased with greater postoperative interval (r = -0.31, p = 0.003). In 68 recipients where donor HLA type was defined, 54% (37/68) of patients had antibodies specific to at least one donor HLA class I antigen. In 87 patients with a recent echocardiographic examination available for analysis (at a mean of 3.5 +/- 1.6 years postoperatively), there was no association between valve dysfunction and antibody status. Anti-HLA class I and II antibodies were detected by ELISA methods in most patients after allograft implantation extending to 5.4 years. The clinical significance of these findings is unclear, as no correlation was found between the prevalence of anti-HLA antibody and echocardiographic parameters of valve dysfunction at a mean of 3.5 years follow up.


Spermatogenesis associated 4 promotes Sertoli cell proliferation modulated negatively by regulatory factor X1.
Spermatogenesis associated 4 (Spata4), a testis-specific and CpG island associated gene, is involved in regulating cell proliferation, differentiation and apoptosis. To obtain insight into the role of Spata4 in cell cycling control, we characterized the promoter region of Spata4 and investigated its transcriptional regulation mechanism. The Spata4 promoter is unidirectional transcribed and possesses multiple transcription start sites. Moreover, we present evidence that regulatory factor X1 (RFX1) could bind the typical 14-bp cis-elements of Spata4 promoter, modulate transcriptional activity and endogenous expression of Spata4, and further regulate the proliferation of Sertoli cells. Overexpression of RFX1 was shown to down-regulate both the promoter activity and mRNA expression of Spata4, whereas knockdown of RFX1 demonstrated the opposite effects. Our studies provide insight into Spata4 gene regulation and imply the potential role of RFX1 in growth of Sertoli cells. RFX1 may have negative effect on cell proliferation of Sertoli cells via modulating Spata4 expression levels by binding the conserved 14-bp cis-elements of Spata4 promoter.


Anti-inflammatory drug delivery from hyaluronic acid hydrogels.
Two different types of hyaluronic acid (HA) hydrogels were synthesized by crosslinking HA with divinyl sulfone (DVS) and poly(ethylene glycol)-divinyl sulfone (VS-PEG-VS). Vitamin E succinate (VES), an anti-inflammatory drug, and bovine serum albumin (BSA), a model of anti-inflammatory protein drugs, were loaded into the gels and their release kinetics were measured in vitro. VES and BSA released with a burst from both HA hydrogels during the first few hours, and release continued gradually for several days. The rate of release from HA-VS-PEG-VS-HA hydrogels was faster than that from HA-DVS-HA hydrogels, presumably due to the lower crosslink density in the former. The anti-inflammatory action of released VES was tested by incubating peripheral blood mononuclear cells (PBMC) on HA hydrogels with and without VES in the gel. The number of cells adhering on HA hydrogels was very low compared to that on tissue culture polystyrene (TCPS), which might be one of the important advantages of using HA hydrogels for implant coatings or tissue engineering applications. ELISA test results showed that the tumor necrosis factor-alpha (TNF-alpha) concentration was very low in the supernatant of the wells containing the HA hydrogel with VES in contact with the activated macrophages compared to that without VES. This is probably the effect of the released VES reducing the production of anti-inflammatory cytokine, TNF-alpha. HA hydrogels containing anti-inflammatory drugs may have potential for use in tissue engineering and also as biocompatible coatings of implants.


Maximum a posteriori local histogram estimation for image registration.
Image similarity measures for registration can be considered within the general context of joint intensity histograms, which consist of bin count parameters estimated from image intensity samples. Many approaches to estimation are ML (maximum likelihood), which tends to be unstable in the presence sparse data, resulting in registration that is driven by spurious noisy matches instead of valid intensity relationships. We propose instead a method of MAP (maximum a posteriori) estimation, which is well-defined for sparse data, or even in the absence of data. This estimator can incorporate a variety of prior assumptions, such as global histogram characteristics, or use a maximum entropy prior when no such assumptions exist. We apply our estimation method to deformable registration of MR (magnetic resonance) and US (ultrasound) images for an IGNS (image-guided guided neurosurgery) application, where our MAP estimation method results in more stable and accurate registration than a traditional ML approach.


Comparative effectiveness of topical calcineurin inhibitors in adult patients with atopic dermatitis.
Atopic dermatitis (AD) is a chronic inflammatory skin disease characterized by extreme pruritis and lichenified papules and plaques that may begin in or persist into adulthood. Topical corticosteroids are first-line prescription therapy for AD; they are efficacious and have a well established safety profile. The topical calcineurin inhibitors tacrolimus and pimecrolimus were approved by the US FDA in 2000 and 2001, respectively, as second-line topical therapy for AD. This review evaluates the available studies on the comparative effectiveness, safety, cost, and impact on quality of life of topical corticosteroids and topical calcineurin inhibitors for the treatment of adult AD. Tacrolimus was found to be as effective as class III-V topical corticosteroids for AD of the trunk and extremities, and more effective than low-potency class VI or VII corticosteroids for AD of the face or neck. Pimecrolimus was less effective than both tacrolimus and low-potency topical corticosteroids for moderate to severe AD. The short-term safety studies found that, compared with topical corticosteroid-treated adults, patients treated with topical calcineurin inhibitors had an increased frequency of application-site reactions, an equivalent infection risk, and a decreased risk of skin atrophy. The long-term safety of topical calcineurin inhibitors remains under investigation. Currently published studies that evaluated the comparative cost and quality-of-life effects compared tacrolimus with less potent topical corticosteroids despite the availability of equivalent potency corticosteroids. Further cost and quality-of-life studies are needed that compare topical calcineurin inhibitors with stronger classes of topical corticosteroids over longer time periods. The available clinical trials data do not suggest an efficacy advantage for topical calcineurin inhibitors over topical corticosteroids in adults with AD of the trunk and extremities, and there is not yet adequate evidence to support topical calcineurin inhibitors as first-line therapy for adult AD.


Current and Investigational Medication Delivery Systems for Treating Attention-Deficit/Hyperactivity Disorder.
To review currently available formulations and emphasize unmet needs in the pharmacologic management of attention-deficit/hyperactivity disorder (ADHD). Publications and clinical trials were identified through PubMed and ClinicalTrials.gov, respectively. A Web-based search identified prescribing information for approved agents for treating ADHD, along with relevant guidelines and diagnostic criteria. The following search terms were used: (1) ADHD or attention-deficit/hyperactivity disorder or ADD or attention deficit hyperactivity disorder and/or (2) amphetamine or methylphenidate or atomoxetine or guanfacine or clonidine. Additional searches were performed using product brand names, and clinical trial was applied as a filter. Relevant studies were only included if published in English-language peer-reviewed journals and if involved agents were currently available (or pursuing approval) in the United States. Reviews of literature prior to 2005 and from 2005 to 2008 have been published previously; therefore, the present search focused on studies published from January 2009 through May 2016. In addition, reference lists of review articles and relevant studies were also examined to help identify additional studies. A total of 578 publications were identified from the PubMed search, from which 426 publications were initially excluded based on review of the title and abstract. Reasons for exclusion included a focus on comorbid disorders, specific subpopulations, endpoints unrelated to improving ADHD symptomatology (eg, executive function, cognition, substance use), or quality of life measures. A more thorough assessment of the remaining citations, including publications and prescribing information, produced the final 219. Pharmacotherapy with stimulant and nonstimulant options is the most common approach for treating ADHD in adults and children. Stimulants are mostly formulated as either tablets or capsules; however, the newer generation includes transdermal patches, oral suspensions (liquids), chewable tablets, and orally disintegrating tablets. Nonstimulants are available in oral capsule (atomoxetine) and tablet (guanfacine and clonidine) formulations. Despite the broad range of treatment options currently available, nonadherence remains a significant problem in ADHD. While evidence is currently lacking, the availability of new formulations may improve adherence.


Experiences With Involuntary Nursing Home Relocation: The Staff Perspective.
Although involuntary nursing home closures for operational and care-related reasons occur infrequently, few studies have examined the centrality of the actions of staff to facilitate residents' relocation adjustment. We interviewed 18 administrators and 43 staff working at 27 facilities about the relocation process for residents from two facilities that lost their Medicare and State Medicaid certification due to ongoing care deficiencies. Thematic analysis revealed three major themes throughout each aspect of the relocation process: (a) staff expectations were not met, (b) barriers/challenges were persistent, and (c) resident/family involvement was minimal. We offer proactive participant-derived recommendations for policy and practice to strengthen involuntary relocation processes to ensure a cooperative work environment and residents' well-being, regardless of the cause for involuntary closure.


A data-driven approach to build a predictive model of cancer patients' disease outcome by utilizing co-expression networks.
Next Generation Sequencing (NGS) technologies have revolutionized genomics data research over the last decades by facilitating high-throughput sequencing of genetic material such as RNA Sequencing (RNAseq). A significant challenge is to explore innovative methods for further exploitation of these large-scale datasets. The approach described in this paper utilizes the results of RNAseq analysis to identify biomarkers related to the disease and deploy a disease outcome predictive model. Chronic Lymphocytic Leukemia (CLL) was used as an example in the implementation of this approach. The approach proposed follows this methodology: (1) Analysis of RNAseq raw data, (2) Construction of a gene correlation network, (3) Identification of modules and hub genes in this network, which constitute the features for the classification algorithm, (4) Deployment of an efficient predictive model, with the use of state-of-the-art machine learning techniques and the association of the indicators with the clinical information. The features/hub genes finally selected were 25 in total and were used as the input to the classifiers. The models, then, were validated leading to very satisfactory results, with the best performing of them achieving 95% cross-validation and 93,75% external validation accuracy. Concluding, this exploratory data-driven approach attempts to make use of big genomic data by summarizing them in a way that is more understandable and facilitates their use by other techniques, such as Machine Learning. This method manages to extract a gene set that can predict the disease progression. The validation results of the proposed data-driven predictive models are very promising and constitute a significant contribution to medical research and personalized medicine.


High-resolution melting PCR analysis for genotyping the gene polymorphism of TNF-α, TGF-β1, IL-10, and IFN-γ in lung transplant recipients.
High-resolution melting (HRM) analysis is a genotyping method which has the advantages of simple, rapid, low-cost and closed-tube operation. This study evaluated HRM analysis as an option for detecting the single nucleotide polymorphism (SNP) of cytokine, and profiled the distribution of cytokine gene polymorphism in the lung transplant recipients (LTRs). High-resolution melting-polymerase chain reaction (HRM-PCR) assays for genotyping tumor necrosis factor alpha (TNF-α) (-308 A/G), tumor growth factor beta 1 (TGF-β1) (+869 T/C), interleukin 10 (IL-10) (-592 C/A, -819 T/C, -1082 G/A), and interferon gamma (IFN-γ) (+874 T/A) SNPs were developed on the LightCycler® 480. The SNPs of the aforementioned cytokine genes in 322 LTRs and 266 normal controls were detected using HRM-PCR approach. To confirm the accuracy of the HRM-PCR assay, we randomly selected 100 samples from the LTRs and detected the aforementioned SNPs with sequence-specific primer-polymerase chain reaction (SSP-PCR) method, using a commercial kit. The data show that the HRM-PCR assay can distinguish all the cytokine SNPs, and the results of HRM-PCR analysis are in complete concordance to the genotyping results obtained using a commercial kit (κ = 1.0). Our data also show that the allele and genotype frequencies of the abovementioned cytokine are not significantly different between the LTRs and the control groups (p > 0.05). In addition, we found the genotypes of TGF-β1 +869 associated with high expression phenotype were prevalent in the LTRs. On the contrary, for TNF-α -308, IL-10 and IFN-γ, the genotypes associated with low expression phenotype were most common in the LTRs. In this study, we described a rapid, low-cost and high-throughput HRM-PCR technology for genotyping cytokine SNPs. Our data may be utilized for future studies examining the associations of cytokine gene polymorphisms with the prognosis of the LTRs.


Proteinase inhibitors: a promising drug class for treating leishmaniasis.
This review presents and discusses the current status and perspectives of leishmaniasis treatment, with a special focus on the use of proteinase inhibitors. The history of treatment development, the first- and second-choice modern drugs and the advantages and disadvantages of using proteinases inhibitors as leishmanicidal treatments are presented and discussed. The reports gathered herein confirm the potential usefulness of proteinases inhibitors as an alternative or complement to the current leishmaniasis treatments. They also support the hypothesis that a combined treatment with multiple proteinase inhibitors may be efficient against Leishmania infections in vertebrate hosts.


Overlapping and unique contributions of two conserved polysaccharide loci in governing distinct survival phenotypes in Vibrio vulnificus.
As an aetiological agent of bacterial sepsis and wound infections, Vibrio vulnificus is unique among the Vibrionacea. Its continued environmental persistence and transmission are bolstered by its ability to colonize shellfish and form biofilms on various marine biotic surfaces. We previously identified a polysaccharide locus, brp, which contributes to the survival phenotypes of biofilm formation, rugose colony formation and stress resistance. Here, we describe a second polysaccharide locus, rbd (regulation of biofilm development), which also enhanced biofilm formation when expressed. Despite this functional overlap, the development of stress resistance and rugosity could be uniquely attributed to brp expression, whereas rbd expression augmented aggregate formation. Simultaneous expression of both loci led to the formation of a dramatic pellicle and maximum biofilm formation. Unlike the brp locus, transcription of the rbd locus was regulated not by c-di-GMP, but by a response regulator (RbdG) that was encoded within the locus. We propose that the ability to regulate the expression of polysaccharides with overlapping and unique characteristics in response to different environmental cues enables V. vulnificus to 'fine tune' its biofilm lifestyle to the prevailing environmental conditions and maximally benefit from the characteristics associated with each polysaccharide.


Studies on a circulating anticoagulant inhibiting factor XI in a patient with congenital deficiency and carcinoma of the prostate.
An inhibitor of plasma thromboplastin antecedent (PTA, factor XI), measured in coagulant and radioimmunoassays, was detected in a 60-year-old man with carcinoma of the prostate who had no evidence of a bleeding tendency. Family studies indicated that the patient was either a homozygote or a heterozygote for hereditary factor XI deficiency. In contrast to earlier described patients with factor XI deficiency in whom inhibitors were detected, the patient was unaware of having been transfused with blood or blood products at any time before the discovery of the inhibitor. The inhibitor of factor XI in the patient's plasma appeared to be predominantly in the IgG4 fraction and to be directed at a locus on the factor XI molecule other than the active site; it did not block the amidolytic properties of activated factor XI (XIa). Rather, it appeared to block adsorption of factor XI to negatively charged surfaces. The inhibitor interfered with measurement of other components of the intrinsic pathway of thrombin formation, perhaps explaining the low titres of other coagulation factors of the intrinsic system reported in patients with strong inhibitors directed against factor XI.


Pandemic peril.
As avian flu has slipped from the headlines, funding for preparedness efforts has lagged. But public health experts say the threat remains, and the needed actions are expensive. Last year, $398 million was disbursed to 62 states and territories to address all hazards, and that's "not a lot of money," says Gregg Pane, left, director of national healthcare preparedness programs. And that leaves little for flu programs.


The use of an intermediate resin in the acid-etch procedure: retentive strength, microleakage, and failure mode analysis.
Three composite resin systems, one conventional (Concise) and two microfilled (IsoPast and Silar), used with and without an intermediate resin (Concise Enamel Bond), were evaluated with and without thermal stress for in vitro retentive strength and sealing ability to acid-etched human enamel. All three materials yielded significantly different shear strength values. The use of an intermediate resin significantly improved both the shear strength and resistance to microleakage of Silar, while no differences were observed for Concise and IsoPast. The thermocycling procedure did not generally or consistently affect the results. The failure mode of the three acid-etched composite resins was studied by evaluation of the failure surface and by describing the percentage of enamel, composite, and interface which were present. The use of an intermediate resin did not significantly influence the failure mode, and the correlation between the failure mode and retentive strength, although significant was low. The study confirmed the importance of adequate wetting action of a resin on the etched enamel to ensure proper bonding.


A case of external iliac arteriovenous fistula and high-output cardiac failure after endovenous laser treatment of great saphenous vein.
Valvular incompetence in the great saphenous vein (GSV) is the most common cause of superficial venous insufficiency and symptomatic varicose vein development. Recently, less invasive modalities such as foam sclerotherapy, radiofrequency ablation (RFA), and endovenous laser treatment (EVLT) have gained popularity in the treatment of saphenofemoral junction and saphenous truncal incompetence over the traditional approach of surgical ligation and stripping. Here, we present the case of a 32-year-old woman who underwent EVLT and was diagnosed subsequently with ipsilateral external iliac arteriovenous (AV) fistula and high-output cardiac failure. She was stabilized medically and treated surgically with a covered stent placed in the external iliac artery with complete resolution of the fistula and cardiac failure. We reviewed the literature and discuss the complications of AV fistulae after EVLT.


[A comparison of the management of severe high blood pressure between 1999 and 2003 in France].
To measure the change in blood-pressure control in two populations of patients with severe high blood pressure between 1999 and 2003. The French National Healthcare Fund performed two observational, cross-sectional surveys on the medical management of high blood pressure, the first in 1999 and the second in 2003. Each survey enrolled patients aged between 20 and 80 years old who filed a first-time request for exemption from co-payments for this long-term disorder. Study data was collected by the health fund's salaried physician advisors who directly examined the patients and from information gathered from each patient's attending physician. Adequate blood pressure control was defined according to the guidelines set out by the ANAES in 1997 and determined by calculating the average of the last three blood pressure figures recorded by the patient's attending physician (systolic [SBP] and diastolic [DBP] arterial blood pressures). 10,665 patients were enrolled in the 1999 survey and 2,584 were enrolled in the 2003 survey. The average age was 63.1 +/- 0.2 years in 1999 and 64.4 +/- 0.4 years in 2003. The proportion of diabetic patients in the two surveys remained stable: 27.5% +/- 1.0 in 1999 and 28.4% +/- 1.7 in 2003. Between 1999 and 2003, the percentage of patient who were treated for hypercholesterolemia increased from 44.0% +/- 1.1 to 54.3% +/- 1.9. Between 1999 and 2003, the proportion of patients who were considered well-controlled (SBP and DBP < 140/90 mmHg) or, if older than 60 years with isolated systolic high blood pressure (DBP < 90 mmHg and SBP < or = 160 mmHg) increased almost 5% points, going from 40.8% +/- 1.1 in 1999 to 45.5% +/- 1.9 in 2003. In diabetic patients, in whom the cut-off threshold is 130/85 mmHg, 6.7% +/- 1.1 were considered well-controlled in 1999 while 5.4% +/- 1.7 were deemed well-controlled in 2003. The proportion of well-controlled patients in the sub-group of hypertensive patients with renal failure (cut-off thresholds: 125/75 mmHg) remained relatively stable between 1999 and 2003: 5.2% [2.5; 7.9] versus 2.8% [0.5; 8.6]. There was a significant increase in the proportion of well-controlled hypertensive patients between 1999 and 2003. This increase occurred at the same time as a number of initiatives (scientific societies, federal government as well as the National Health Fund) intended to sensitize physicians to the need to obtain adequate blood pressure control.


Low overhead slipless carrier phase estimation scheme.
Two slipless schemes are compared with application to single carrier 30 Gbaud quadrature phase shift keying (QPSK) system. An equivalent linewidth model considering the phase noise induced by both the laser linewidth and fiber nonlinearity is applied in the performance analysis. The simulation results show that it is possible to mitigate cycle slip (CS) using only 0.39% pilot overhead for the proposed blind carrier phase recovery (CPR) + pilot-symbols-aided phase unwrapping (PAPU) scheme within 1 dB signal-to-noise ratio (SNR) penalty limit at the bit error ratio (BER) of 10(-3) with 4 MHz equivalent linewidth.


Octave-spanning supercontinuum generation from off-axis Raman oscillation in a monolithic KTP crystal.
We report generation of a visible and near-infrared supercontinuum from a high-gain, ultra-broadband, and mirrorless Raman oscillator in a monolithic KTP crystal. The plane transverse to the pump axis resonates and traps off-axis Stokes waves and their frequency-upconverted components bouncing between two crystal surfaces via total internal reflection. The Raman gain is maximized with the Stokes polarization perpendicular to the plane of reflections. When pumped by a Q-switched Nd:YAG laser, the monolithic oscillator generates quasi-mode-locked Stokes pulses with octave-spanning spectral groups across the visible and near-infrared spectra between 540 and 1800 nm.


Isolated dissection of the superior mesenteric artery: conservative approach in a selected patient.
The authors present the case report of effective conservative treatment in a patient with spontaneous, self-limiting, non-atherosclerotic dissection of the superior mesenteric artery (SMA) without fixed obstruction of the vessel lumen and signs of intestinal ischemia. Treatment with both anti-coagulant and anti-hypertensive agents succeeded in limiting the progression of intimal dissection and in preventing the potential dramatic sequelae of this rare clinical condition. Conservative treatment of spontaneous SMA dissection may be an alternative to surgery, if residual blood flow is maintained.


Test-Retest Reliability of the Genetics and Genomics in Nursing Practice Survey Instrument.
Assessment of nursing genomic competency is critical given increasing genomic applications to health care. The study aims were to determine the test-retest reliability of the Genetics and Genomics in Nursing Practice Survey (GGNPS), which measures this competency, and to revise the survey accordingly. Registered nurses (n = 232) working at 2 Magnet-designated hospitals participating in a multiinstitutional genomic competency study completed the GGNPS. Cohen's kappa and weighted kappa were used to measure the agreement of item responses between Time 1 and Time 2. Survey items were revised based on the results. Mean agreement for the instrument was 0.407 (range = 0.150-1.000). Moderate agreement or higher was achieved in 39% of the items. GGNPS test-retest reliability was not optimal, and the instrument was refined based on the study findings. Further testing of the revised instrument is planned to assess the instrument performance.


Different circulating lymphocyte profiles in patients with different subtypes of juvenile idiopathic arthritis.
To determine the immunophenotypic profiles of circulating lymphocytes in patients with different disease types of Juvenile Idiopathic Arthritis (JIA). Peripheral blood lymphocyte subsets from 19 patients with oligoarticular JIA (o-JIA), 10 patients with polyarticular JIA (p-JIA), 12 patients with systemic JIA (s-JlA) andfrom 41 age-matched healthy controls were characterized by two color immunofluorescence flow cytometry analysis. Patients with o-JIA and p-JIA had increased numbers of HLA-DR+ Tcells and Tcells co-expressing CD57 and CD16/56, indicating T cell activation and terminal differentiation of CD8+ T cells respectively. By contrast, in patients with s-JIA there was no increase in the activation or differentiation markers on T cells, but a profound decrease in circulating NK cells. All patients had hypergammaglobulinemia consistent with B cell hyperactivity, but increased numbers of CD5+ B cells were found only in o-JIA and p-JIA. Distinct immunophenotypic lymphocyte profiles in patients with o-JIA and p-JIA compared to patients with s-JIA as demonstrated in this study, are consistent with afundamental heterogeneity of the disease.


Antibodies to collagen IV in the serum of workers exposed to hydrocarbons and volatile organic chemicals.
Significant numbers of workers (14%) chronically exposed to volatile organic chemicals commonly found in spray paints had elevated levels of uncharacterized antibodies to collagen IV, a basement membrane protein. No increased frequency of subjects with positive results for anti-glomerular basement membrane antibodies (anti-GBM) was found in this group. These anti-GBM antibodies are directed against a specific epitope on the non-collagenous domain (NC1) of the α3 chain of collagen IV. Anti-GBM antibodies are diagnostic for Goodpasture's syndrome that has been reported to occur following acute inhalation of volatile substances. In another group of workers, exposed both dermally and by inhalation to petroleum-based oil mists, 5% had positive results for antiGBM antibodies. We conclude that the measurement of general, uncharacterized antibodies to collagen IV may be a useful indicator of basement membrane damage in workers occupationally exposed to volatile organic chemicals.


Dexamethasone recruitment of self-renewing osteoprogenitor cells in chick bone marrow stromal cell cultures.
Bone marrow stromal cells are a mixed population that contribute to the formation of the hematopoietic microenvironment. The osteogenic lineage includes populations of cells that, in culture, form discrete nodules of mineralized tissue when grown in the presence of ascorbic acid and sodium beta-glycerophosphate. We have used nodule formation to assay for the self-renewal capacity of osteoprogenitor cells in chick bone marrow cultures. To examine the regulatory influence of dexamethasone (Dx), first subcultures were grown continuously or split 1:1 at repeated subculture. Cells in continuous culture exhibited less than two population doublings, while cellular proliferation and alkaline phosphatase area were inhibited by 10(-8) mol/L Dx. Cells in split (redistributed) cultures exhibited up to 14 population doublings and cellular proliferation was also inhibited by Dx. In contrast with continuous cultures, redistributed cultures treated with Dx had increased alkaline phosphatase area and 15-fold larger amounts of mineralized tissue formation than controls. Osteogenesis was sustained for up to four subcultures and the ratio of mineralized tissue area to alkaline phosphotase positive cell area was at most 0.55. These data indicate that the osteogenic lineage of bone marrow stromal cells contains self-renewing progenitors that are recruited by Dx in culture and that at a maximum, only 55% of the alkaline phosphatase-positive cell population contributes to osteogenesis.


Identification of four novel human genes amplified and overexpressed in breast carcinoma and localized to the q11-q21.3 region of chromosome 17.
We have performed differential screening of a human metastatic lymph lymph node cDNA library to identify genes possibly involved during breast cancer progression. We have identified four novel genes overexpressed in malignant tiddues. They were all located on the long arm of chromosome 17, in loci located between q11 and q21.3, a region known to contain the c-erbB-2 oncogene and the BRCA1 breast carcinomas, and overexpression of three of them was dependent on gene amplification in breast cancer cell lines. These findings further support the concept that human chromosome 17 specifically carries genes possibly involved in breast cancer progression.


Anti-CCR4 treatment depletes regulatory T cells and leads to clinical activity in a canine model of advanced prostate cancer.
Targeting regulatory T cell (Treg) infiltration is an emerging strategy for cancer immunotherapy. However, its efficacy in advanced prostate cancer remains unclear. Here, we showed the therapeutic efficacy of anti-Treg treatment in a canine model of advanced prostate cancer. We used dogs with naturally occurring prostate cancer to study the molecular mechanism underlying Treg infiltration and the effect of anti-Treg treatment. Tumor-infiltrating Tregs was evaluated by immunohistochemistry, and the association with prognosis was examined in dogs with spontaneous prostate cancer. The molecular mechanism of Treg infiltration was explored by RNA sequencing and protein analyses. A non-randomized canine clinical trial was conducted to define the therapeutic potential of anti-Treg treatment for advanced prostate cancer. Human prostate cancer datasets were analyzed to compare gene expression in dogs and humans. Tumor-infiltrating Tregs were associated with poor prognosis in dogs bearing spontaneous prostate cancer. RNA sequencing and protein analyses showed a possible link between the CCL17-CCR4 pathway and the increase of tumor-infiltrating Tregs. Dogs with advanced prostate cancer responded to mogamulizumab, a monoclonal antibody targeting CCR4, with decreased circulating Tregs, improved survival, and low incidence of clinically relevant adverse events. Urinary CCL17 concentration and BRAFV595E mutation were independently predictive of the response to mogamulizumab. Analysis of a transcriptomic dataset of human prostate cancer showed that the CCL17-CCR4 axis correlated with Foxp3. In silico survival analyses revealed that high expression of CCL17 was associated with poor prognosis. Immunohistochemistry confirmed that tumor-infiltrating Tregs expressed CCR4 in human patients with prostate cancer. Anti-Treg treatment, through CCR4 blockade, may be a promising therapeutic approach for advanced prostate cancer in dogs and some population of human patients.


Prevention of cisplatin nephrotoxicity.
Cisplatin has a well-established role in the treatment of broad spectrum of malignancies; however its use is limited because of cisplatin-induced nephrotoxicity (CIN) which can be progressive in more than 50% of cases. The most important risk factors for CIN include higher doses of cisplatin, previous cisplatin chemotherapy, underlying kidney damage and concurrent treatment with other potential nephrotoxin agents, such as aminoglycosides, nonsteroidal anti-inflammatory agents, or iodinated contrast media. Different strategies have been offered to diminish or prevent nephrotoxicity of cisplatin. The standard approach for prevention of CIN is the administration of lower doses of cisplatin in combination with full intravenous hydration prior and after cisplatin administration. Cisplatin-induced oxidative stress in the kidney may be prevented by natural antioxidant compounds. The results of this review show that many strategies for prevention of CIN exist, however, attention to the administration of these agent for CIN is necessary.


Expansion of cooperatively growing populations: Optimal migration rates and habitat network structures.
Range expansion of species is driven by the interactions among individual- and population-level processes and the spatial pattern of habitats. In this work we study how cooperatively growing populations spread on networks representing the skeleton of complex landscapes. By separating the slow and fast variables of the expansion process, we are able to give analytical predictions for the critical conditions that divide the dynamic behaviors into different phases (extinction, localized survival, and global expansion). We observe a resonance phenomenon in how the critical condition depends on the expansion rate, indicating the existence of an optimal strategy for global expansion. We derive the conditions for such optimal migration in locally treelike graphs and numerically study other structured networks. Our results highlight the importance of both the underlying interaction pattern and migration rate of the expanding populations for range expansion. We also discuss potential applications of the results to biological control and conservation.


Current surgical management of ulcerative colitis.
Inflammatory bowel disease is divided into 2 major disease entities: Crohn's disease and ulcerative colitis. Ulcerative colitis is characterized by contiguous inflammation of the colorectal mucosa, always beginning in and involving the rectum and progressing for variable distances proximally within the colon. In ulcerative colitis, medical therapy, which is not curative, is directed at controlling symptoms and reducing the underlying inflammatory process. However, emergent or elective removal of the colon and rectum does cure the disease and also eliminates the possibility of developing a malignancy. Here we present the current surgical treatment of ulcerative colitis and issues in the management of ulcerative colitis. We discuss indications for surgical treatment, elective and emergent operative management, early and late complications of surgery, and functional results.


Changes in phospholipid metabolism during B lymphocyte activation.
We have examined phospholipid metabolism in murine B lymphocytes stimulated with anti-Ig bound to Sepharose. T cell-depleted splenic B lymphocytes cultured with Sepharose-coupled, affinity-purified goat anti-mouse Ig (GAMIg) increased the incorporation of 32PO4 into phosphatidic acid and phosphatidylinositol within 3 hr and increased [3H]-thymidine uptake at 48 hr. No increase in labeling was observed in phosphatidylethanolamine, phosphatidylcholine, or phosphatidylserine. Based on both negative and positive selection procedures, it was demonstrated that these responses occurred in B lymphocytes. In contrast to the thymidine uptake response of the GAMIg-stimulated B lymphocytes, the phospholipid response did not require the presence of accessory cells or exogenous cytokines. The same selective changes in phospholipid metabolism were observed in neoplastic B lymphocytes (BCL1) after treatment with Sepharose anti-mu, but not with Sepharose anti-Ia or Sepharose normal Ig. The dose-response relationships of 32PO4 incorporation into phosphatidic acid and phosphatidylinositol and [3H] thymidine uptake were nearly identical in BCL1 cells. The results of these experiments indicate that interaction of B lymphocytes with insolubilized anti-Ig results in prompt and selective changes in phospholipid metabolism that appear to be correlated with B lymphocyte proliferation.


Dual Targeting of Cell Growth and Phagocytosis by Erianin for Human Colorectal Cancer.
To investigate the effect of erianin on tumor growth and immune response in human colorectal cancer cells (CRC). The effect of erianin on tumor growth was determined by CCK8 and colony formation assay. Western blotting was used to evaluate the expression levels of relevant proteins and qRT-PCR was used to evaluate the mRNA level of the relevant gene. The transcriptional activity of β-catenin was determined by dual-luciferase reporter assay. Cellular thermal shift assay was used to quantify drug-target interactions. The cell surface CD47 was assessed by flow cytometry. The enrichment of H3K27 acetyl marks on CD47 promoter was evaluated by chromatin immunoprecipitation assay. Phagocytosis assay was used to determine the phagocytic activity of macrophage. In vivo role of erianin was studied on xenograft models. We found that erianin significantly decreased cell survival, colony formation, induced cell cycle arrest, and led to cell apoptosis in SW480 and HCT116 cells. Mechanism analysis demonstrated that erianin inhibited the nuclear translocation and transcriptional activity of β-catenin, which might result from erianin-β-catenin interaction. In addition, the downstream gene expressions, such as c-Myc and cyclin D1, was decreased. More interestingly, erianin decreased the expression of CD47 by regulating H3K27 acetyl marks enrichment on CD47 promoter. Consequently, macrophage-mediated phagocytosis was increased. Our in vivo experiments further confirmed the inhibitory effect of erianin on tumor growth. In summary, erianin could inhibit CRC cells growth and promoted phagocytosis, which suggested erianin as a potential therapeutic strategy for CRC patients.


Protein-polymer conjugation via ligand affinity and photoactivation of glutathione S-transferase.
A photoactivated, site-selective conjugation of poly(ethylene glycol) (PEG) to the glutathione (GSH) binding pocket of glutathione S-transferase (GST) is described. To achieve this, a GSH analogue (GSH-BP) was designed and chemically synthesized with three functionalities: (1) the binding affinity of GSH to GST, (2) a free thiol for polymer functionalization, and (3) a photoreactive benzophenone (BP) component. Different molecular weights (2 kDa, 5 kDa, and 20 kDa) of GSH-BP modified PEGs (GSBP-PEGs) were synthesized and showed conjugation efficiencies between 52% and 76% to GST. Diazirine (DA) PEG were also prepared but gave conjugation yields lower than for GSBP-PEGs. PEGs with different end-groups were also synthesized to validate the importance of each component in the end-group design. End-groups included glutathione (GS-PEG) and benzophenone (BP-PEG). Results showed that both GSH and BP were crucial for successful conjugation to GST. In addition, conjugations of 5 kDa GSBP-PEG to different proteins were investigated, including bovine serum albumin (BSA), lysozyme (Lyz), ubiquitin (Ubq), and GST-fused ubiquitin (GST-Ubq) to ensure specific binding to GST. By combining noncovalent and covalent interactions, we have developed a new phototriggered protein-polymer conjugation method that is generally applicable to GST-fusion proteins.


Total lung capacity in asbestosis: a comparison of radiographic and body plethysmographic methods.
The effect of asbestosis on lung volumes was studied in 46 workers by measuring total lung capacity (TLC), forced vital capacity, vital capacity, and residual volume using two standard methods: body plethysmography and radiographic lung area. Nine men had neither irregular opacities of any International Labour Office profusion category nor pleural abnormalities; 9 had pleural abnormalities only, 19 had irregular opacities only; and 9 had pulmonary asbestosis with pleural plagues. Of the 28 with irregular opacities, 18 had moderate or advanced asbestosis (category 2/1 or greater). The radiographic method gave a mean TLC of 8.11 L and the body plethysmographic mean was 8.09 L. Thus, the radiographic and plethysmographic methods produced virtually identical mean values for TLC and for residual volume 3.84 L and 3.8 L and residual volume/TLC, 47.3 and 47.3, respectively. Lung volumes measured by body plethysmography and by x-ray area were the same in each of the four radiographic categories of asbestosis. The air trapping and normal or slightly elevated TLC, which characterize asbestosis in men who have smoked cigarettes, are revealed by both methods. The radiographic method, when adequate measures are taken to obtain full inspirations, effectively duplicates the body plethysmographic method but is quicker and simpler, particularly for field studies.


[Differences of age and gender in orthostatic hypertension-a single-center study].
To explore the differences of Chinese people's age and gender in orthostatic hypertension (OHT) by a single-center experience study. A total of 2 994 patients with unexplained syncope and dizziness, who were outpatients or had been hospitalized in Second Xiangya Hospital, Central South University from January 2000 to August 2012, were chosen and subjected to the head-up tilt table test (HUTT). Their ages ranged from 2.00 to 78.00 years old, with an average at 19.07±14.78 years old. There were 1 406 and 1 588 cases for male and female, respectively. A total of 745 patients were OHT, who were divided into a adult group (≥18 years old, 247/904 cases) and a children group (<18 years old, 498/2 090 cases). The difference of the occurrence rate, classification, and 3 min quantity of blood pressure were analyzed. 1) The incidence of OHT in HUTT was 24.88% (745/2 994 cases). There was no statistical difference between the males and the females(25.75% vs 24.12%, χ2=1.058, P>0.05), but it was higher in the adult group compared with that in the children group (27.05% vs 23.83%, χ2=4.125, P=0.042). There were 52 cases (6.98%) of high systolic and high diastolic blood pressure (sOHT merge dOHT), 16 cases (2.15%) of simple high systolic blood pressure OHT (sOHT), and 677 cases (90.87%) of simple high diastolic blood pressure OHT (dOHT) in the 745 patients with OHT. The incidence of dOHT and sOHT merge sOHT in the adults group were significantly higher than those in the children group (1.11% vs 0.29%, χ2=7.965, P<0.01; 2.88% vs 1.24%, χ2=9.849, P<0.01, respectively); there was no statistical difference in simple dOHT between males and females (23.56% vs 22.20%, χ2=0.668, P>0.05); there was also no statistical difference in sOHT plus dOHT and dOHT between males and females (sOHT merge dOHT: 1.71% vs 1.76%, χ2=0.014, P>0.05; dOHT: 23.68% vs 21.66%, χ2=1.742, P>0.05; 1.71% vs 1.76%, χ2=0.014, P>0.05, respectively); but the dOHT was significantly higher in the males than that in the females in the children group (24.53% vs 19.74%, χ2=6.933, P>0.05). 2) There was no difference in the increment of systolic blood pressure in sOHT plus dOHT [(25.62±4.96) mmHg vs (23.54±5.83) mmHg, t=1.385, P>0.05] and the increment of diastolic blood pressure in dOHT [(13.46±3.49) mmHg vs (13.23±3.22) mmHg, t=0.840, P>0.05] between the adults group and the children group. There was no difference in the increment of systolic blood pressure in sOHT [(25.44±4.96) mmHg vs (23.68±5.35) mmHg, t=1.411, P>0.05] and the increment of diastolic blood pressure in dOHT [(14.09±4.28) mmHg vs (13.05±3.82) mmHg, t=1.887, P>0.05] between the adults group and the children group. The incidence of OHT is higher in the adults group than that in the children group. There is no difference in systolic and diastolic blood pressure increment within 3 minutes during HUTT between the adults and children.


CD134 plus CD137 dual costimulation induces Eomesodermin in CD4 T cells to program cytotoxic Th1 differentiation.
Cytotoxic CD4 Th1 cells are emerging as a therapeutically useful T cell lineage that can effectively target tumors, but until now the pathways that govern their differentiation have been poorly understood. We demonstrate that CD134 (OX40) costimulation programs naive self- and virus-reactive CD4 T cells to undergo in vivo differentiation into cytotoxic Th1 effectors. CD137 (4-1BB) costimulation maximized clonal expansion, and IL-2 was necessary for cytotoxic Th1 differentiation. Importantly, the T-box transcription factor Eomesodermin was critical for inducing the cytotoxic marker granzyme B. CD134 plus CD137 dual costimulation also imprinted a cytotoxic phenotype on bystanding CD4 T cells. Thus, to our knowledge, the current study identifies for the first time a specific costimulatory pathway and an intracellular mechanism relying on Eomesodermin that induces both Ag-specific and bystander cytotoxic CD4 Th1 cells. This mechanism might be therapeutically useful because CD134 plus CD137 dual costimulation induced CD4 T cell-dependent tumoricidal function in a mouse melanoma model.


Double dissociation between hippocampal and prefrontal lesions on an operant delayed matching task and a water maze reference memory task.
The hippocampus and prefrontal cortex have both been implicated in various aspects of the acquisition, retention and performance of delayed matching to position (DMTP) tasks in the rat, although their precise respective contributions remain unclear. In the present study, rats were trained preoperatively on DMTP before receiving excitotoxic bilateral lesions of either the entire hippocampus or the medial prefrontal cortex. Rats with lesions of the prefrontal cortex exhibited a significant delay-dependent impairment on retention of the DMTP task, whereas hippocampal lesions were without effect. Rats were also exposed to a switch in the contingencies to a 'non-matching' rule, as an analogue of switching between decision rules in the human Wisconsin Card Sorting Test, in which human patients with prefrontal damage are impaired. Both lesion groups acquired the new contingency at control levels, providing no evidence towards a role for either of these areas in this type of rule-switching. The same rats were also assessed in a spatial reference memory task in the water maze, which revealed an impairment in escape latencies and path length that was specific to the hippocampal lesions. The results corroborate previous evidence that the hippocampus is not necessary for at least some aspects of working memory performance in the DMTP task, whereas the delay-dependent deficit in the prefrontal lesion group support this task as a potentially powerful tool for assessing the cognitive changes associated with frontal damage and repair.


Plasmodium interspersed repeats: the major multigene superfamily of malaria parasites.
Functionally related homologues of known genes can be difficult to identify in divergent species. In this paper, we show how multi-character analysis can be used to elucidate the relationships among divergent members of gene superfamilies. We used probabilistic modelling in conjunction with protein structural predictions and gene-structure analyses on a whole-genome scale to find gene homologies that are missed by conventional similarity-search strategies and identified a variant gene superfamily in six species of malaria (Plasmodium interspersed repeats, pir). The superfamily includes rif in P.falciparum, vir in P.vivax, a novel family kir in P.knowlesi and the cir/bir/yir family in three rodent malarias. Our data indicate that this is the major multi-gene family in malaria parasites. Protein localization of products from pir members to the infected erythrocyte membrane in the rodent malaria parasite P.chabaudi, demonstrates phenotypic similarity to the products of pir in other malaria species. The results give critical insight into the evolutionary adaptation of malaria parasites to their host and provide important data for comparative immunology between malaria parasites obtained from laboratory models and their human counterparts.


Computational and experimental study of the structure, binding preferences, and spectroscopy of nickel(II) and vanadyl porphyrins in petroleum.
We present a computational exploration of five- and six-coordinate Ni(II) and vanadyl porphyrins, including prediction of UV-vis spectroscopic behavior and metalloporphyrin structure as well as determination of a binding energy threshold between strongly bound complexes that have been isolated as single crystals and weakly bound ones that we detect by visible absorption spectroscopy. The excited states are calculated using the tandem of the time-dependent density functional theory (TD-DFT) and the conductor-like polarizable continuum model (CPCM). The excited-state energies in chloroform solvent obtained by using two density functionals are found to correlate linearly with the experimental Soret and alpha-band energies for a known series of five-coordinate vanadyl porphyrins. The established linear correction allows simulation of the excited states for labile octahedral vanadyl porphyrins that have not been isolated and yields Soret and alpha-band bathochromic shifts that are in agreement with our UV-vis spectroscopic results. The PBE0 and PW91 functionals in combination with DNP basis set perform best for both structure and binding energy prediction. The reactivity preferences of Ni(II) and vanadyl porphyrins toward aromatic fragments of large petroleum molecules are explored by using the density functional theory (DFT). Analysis of electrostatic potentials and Fukui functions matching shows that axial coordination and hydrogen bonding are the preferred aggregation modes between vanadyl porphyrins and nitrogen-containing heterocycle fragments. This investigation improves our understanding on the cause for broadening of the Ni and V porphyrin Soret band in heavy oils. Our findings can be useful for the development of metals removal methods for heavy oil upgrading.


Iatrogenic spinal cord ischemia: A patient level meta-analysis of 74 case reports and series.
We seek to characterize the features of iatrogenic spinal ischemia, determine which spinal levels are affected, and evaluate the efficacy of management strategies. We performed a meta-analysis of case reports and series of spinal ischemia in the past 10 years. 343 full-length case reports and case series were screened against predefined inclusion/exclusion criteria. 89 patients were included for our final meta-analysis using PRISMA guidelines. Mean age of patients was 59.62 years (range: 9 months-88 years). 66% of all cases were male. Endovascular surgery (32.6%) and aortic surgery (36.0%) were most common causes of iatrogenic injury, followed by non-aortic surgery (32.6%), and non-surgical procedures (22.47%). A- and B-level ASIA Impairment was found in 66% of all patients. Rehabilitation was the most common management (49.44% of cases), followed by blood pressure management (40.45%). Non-aortic surgeries had the poorest overall outcomes (OR = 0.28, p = 0.016), whereas aortic and endovascular surgeries saw significant improvement in outcomes measured at discharge (OR = 2.6, OR = 2.3, respectively, p < 0.05). Therapeutic surgical infarctions were found to be associated with improved outcomes (OR = 5.33, p = 0.032). Ischemic injury to T4-T7, and T10 were associated with significantly poorer outcomes. Autonomic impairment was associated with a likelihood of infarction at T10 (OR = 4.54, p = 0.0183). In this paper, we compare outcomes following iatrogenic spinal ischemia. We demonstrate the need for more comprehensive randomized controlled trials to test effective treatment strategies.


Potential placenta previa: definition, frequency, and significance.
Sixty-three (5%) of 1239 women studied by sonography during the second trimester of their pregnancies had a diagnosis of placenta previa. Follow-up was available for 51 of the 63 patients; in three of these, the original diagnosis was complete placenta previa, and in the other 48, the first diagnosis was partial or marginal placenta previa. At term, placenta previa was seen in only four patients for an overall frequency of 0.3% (4/1227). In all three of the patients with complete placenta previa, the condition persisted from the second trimester to term; previa persisted in only one of 48 patients with marginal/partial previa. Because of the infrequent persistence of marginal or partial placenta previa to term, we recommend using the term "potential placenta previa" in the second trimester, with follow-up sonography indicated only for vaginal bleeding.


Predicting Radioresistant Biomarkers in Nasopharyngeal Carcinoma Patients Via Protein-Protein Interaction Network Analysis.
Introduction: Radiotherapy as the first-line nasopharyngeal carcinoma (NPC) treatment provides different responses including radioresistant and radiosensitive states. In order to investigate the molecular basis of radioresistancy, protein-protein interaction network analysis of proteome data prior to therapy was performed. Methods: 20 dysregulated proteins of the patients who were radioresistant were extracted from the literature. Cytoscape and its plug-ins were used for the resistant network construction and its centrality analysis. Furthermore, ClueGO+ CluePedia application determined the most statistically significant biological processes (BP) related to the hubs. Results: Fourteen hubs were concluded and no differentially expressed protein (DEP) was among these agents. Among the hubs, albumin (ALB) and fibronectin (FN1) were the hub-bottlenecks, and the Serpin family was present. What is more, SERPIND1 was the highest degree-valued DEP in the network. Conclusion: It can be concluded that the central elements of the NPC network could be noteworthy for improving the radiotherapy outcome and overcoming its limitations. However, complementary studies are required for a better understanding of their major role.


[Discussions of the Ministry of Health, Labour and Welfare's Study Group on Training and Quality Improvement of Pharmacists].
The Ministry of Health, Labour and Welfare's "Study Group on the Training and Qualification Improvement of Pharmacists" met 10 times from July 2000 to June 2021, and its "Summary" was published on June 30, 2021. The purpose of this study group was to examine the future of pharmacists in Japan, based on the results of a survey on the supply and demand of pharmacists, in the face of the declining birth rate, aging population, and regional differences in population. The study group also assessed changes in the roles and duties of pharmacists, such as the promotion of family pharmacists and pharmacies, the promotion of team medicine in medical institutions, and the response of pharmacists to the regional comprehensive care system. The purpose of this symposium was to discuss how to train and improve the quality of pharmacists in the future, while taking into account the results of the survey on the estimated supply and demand of pharmacists. In this symposium, we would like to introduce what was actively discussed in the study group and what was compiled based on the discussion.


Perfluorododecanoic Acid Blocks Rat Leydig Cell Development during Prepuberty.
Perfluorododecanoic acid (PFDoA) has been used as a surfactant and may have reproductive toxicity. However, whether PFDoA influences Leydig cell development during prepuberty remains unknown. In the present study, 21-day-old male Sprague-Dawley rats were gavaged 0, 5, or 10 mg/kg PFDoA from postnatal day 21 to 35. PFDoA decreased the serum concentrations of testosterone, luteinizing hormone, and follicle-stimulating hormone at doses of 5 and 10 mg/kg without influencing Leydig cell number and proliferation. However, PFDoA down-regulated the expression of Leydig cell genes ( Lhcgr, Scarb1, Star, Cyp11a1, Cyp17a1, and Hsd11b1) or their proteins. PFDoA dose-dependently reduced SIRT1 and PGC-1α levels. PFDoA did not affect AMPK and AKT2 levels but decreased their phosphorylation. We also treated primary progenitor Leydig cells purified from prepubertal rat testes with PFDoA for 24 h. It in vitro lowered viability and decreased mitochondrial membrane potential of progenitor Leydig cells, but it stimulated the generation of the intracellular reactive oxygen species and induced Leydig cell apoptosis at 10 μM. In conclusion, PFDoA blocks rat Leydig cell development during the prepubertal period possibly via targeting AMPK/SIRT1/PGC-1α and AKT2 signaling pathways.