Cytolytic effector mechanisms and gene expression in autologous graft-versus-host disease: distinct roles of perforin and Fas ligand.
The administration of cyclosporin A (CsA) after autologous stem cell transplantation (SCT) paradoxically elicits a systemic autoimmune syndrome that resembles graft-versus-host disease (GVHD); this is termed autologous GVHD (autoGVHD). Although dominated by activated CD8+ cytotoxic T lymphocytes, the complex cellular reaction also includes CD4+ T cells and involves multiple effector mechanisms. To determine the temporal development and relative importance of these mechanisms in autoGVHD, perforin/granzyme, Fas ligand (FasL), interferon-gamma (IFN-gamma), tumor necrosis factor (TNF)-alpha, and interleukin-18 gene expression in peripheral blood mononuclear cells was examined in 36 patients treated with CsA after SCT. Quantitative real-time polymerase chain reaction analysis revealed that perforin/granzyme B, TNF-alpha, and interleukin-18 messenger RNA (mRNA) levels in peripheral blood mononuclear cells from patients in whom autoGVHD developed were markedly higher (and temporally associated with the onset of autoaggression) compared with the levels detected in healthy individuals and in control, non-CsA-treated SCT patients. It is interesting to note that patients in whom autoGVHD did not develop also demonstrated increased mRNA levels for these cytokines: however, expression was substantially lower compared with that in patients with autoGVHD. It is important to note that IFN-gamma mRNA levels were selectively increased in CD8+ cells only from patients in whom autoGVHD developed. The development of autocytolytic T cells in autoGVHD correlated with increased expression of perforin, IFN-gamma, and TNF-alpha mRNA. Furthermore, enhanced autoreactive T-cell activity and the induction of autoGVHD was also concordant with perforin and TNF-alpha mRNA upregulation in CD4+ cells. Surprisingly, FasL mRNA levels were significantly decreased, with a progressive loss of FasL mRNA expression as autocytolytic activity increased. These findings suggest that IFN-gamma/perforin-based CD8+ cytotoxic T lymphocytes seem to play a dominant role in autoGVHD and that TNF-alpha/perforin-based CD4+ cells may amplify this autoaggressive syndrome. The FasL pathway may play an important role in the regulation of this immune syndrome.


Hypotension induced by prostacyclin treatment during cardiopulmonary bypass does not increase the risk of cerebral complications.
High-dose prostacyclin treatment during cardiopulmonary bypass reduces platelet activation and possibly postoperative blood loss. A side-effect is arterial hypotension. We studied the incidence of cerebral complications in 79 patients requiring coronary bypass. Only patients without known cerebrovascular disease were studied. Thirty-nine patients received prostacyclin 50 ng/kg/min during cardiopulmonary bypass and 40 patients served as controls. Mean arterial blood pressure in the group given prostacyclin was below 30 mm Hg during the first 30 minutes of bypass but remained above 60 mm Hg in the control group. Postoperative neurological examination revealed transient cerebral dysfunction in six control patients and two prostacyclin-treated patients. Investigation of cerebrospinal fluid showed signs of blood-brain barrier damage in 12 control and seven prostacyclin-treated patients. Cytologic changes in cerebrospinal fluid consistent with brain tissue damage occurred in two control patients but in no patient given prostacyclin. Myelin basic protein and adenylate kinase in cerebrospinal fluid were assayed as being markers of brain damage. Myelin basic protein was within the normal range in all patients. Adenylate kinase was moderately increased (greater than 0.035 U/L) in five of 15 control patients and six of 13 prostacyclin-treated patients. We conclude that treatment with prostacyclin 50 ng/kg/min during cardiopulmonary bypass does not increase the risk of postoperative cerebral damage.


Moving Toward a Multimodal Analgesic Regimen for Acute Sickle Cell Pain with Non-Opioid Analgesic Adjuncts: A Narrative Review.
Sickle cell disease (SCD) is an inherited hemoglobinopathy with potential life-threatening complications that affect millions of people worldwide. Severe and disabling acute pain, referred to as a vaso-occlusive crisis (VOC), is a fundamental symptom of the disease and the primary driver for acute care visits and hospitalizations. Despite the publication of guidelines for VOC management over the past decade, management of VOCs remains unsatisfactory for patients and providers. Acute SCD pain includes pain secondary to VOCs and other forms of acute pain. Distinguishing VOC from non-VOC pain may be challenging for both patients and clinicians. Further, although opioids have been the gold-standard for VOC pain management for decades, the current highest standard of care for all acute pain is a multimodal approach that is less dependent on opioids, and, instead incorporates analgesics and adjuvants from different mechanistic pathways. In this narrative review, we focus on a multimodal pharmacologic approach for acute SCD pain management and explore the evidence for existing non-opioid pharmacological adjuncts. Moreover, we present an explanatory model of pain, which is not only novel in its application to SCD pain but also captures the multidimensional nature of the SCD pain experience and supports the need for such a multimodal approach. This model also highlights opportunities for new investigative and therapeutic targets - both pharmacological and non-pharmacological. Multimodal pain regimens that are less dependent on opioids are urgently needed to improve acute pain outcomes for individuals with SCD. The proposed explanatory model for SCD pain offers novel opportunities to improve acute pain management for SCD patients.


Airway luminal area and the resistive work of breathing during exercise in healthy young females and males.
We examined the relationship between the work of breathing (Wb) during exercise and in vivo measures of airway size in healthy females and males. We hypothesized that sex differences in airway luminal area would explain the larger resistive Wb during exercise in females. Healthy participants (n = 11 females and n = 11 males; 19-30 yr) completed a cycle exercise test to exhaustion where Wb was assessed using an esophageal balloon catheter. On a separate day, each participant underwent a bronchoscopy procedure for optical coherence tomography measures of seven airways. In vivo measures of luminal area were made for the fourth to eighth airway generations. A composite index of airway size was calculated as the sum of the luminal area for each generation, and the total area was calculated based on Weibel's model. We found that index of airway size (males: 37.4 ± 6.3 mm2 vs. females: 27.5 ± 7.4 mm2) and airway area calculated based on Weibel's model (males: 2,274 ± 557 mm2 vs. females: 1,594 ± 389 mm2) were significantly larger in males (both P = 0.003). When minute ventilation was greater than ∼60 L·min-1, the resistive Wb was higher in females. At the highest equivalent flow achieved by all subjects, resistance to inspired flow was larger in females and significantly associated with two measures of airway size in all subjects: index of airway size (r = 0.524, P = 0.012) and Weibel area (r = 0.525, P = 0.012). Our findings suggest that innate sex differences in luminal area result in a greater resistive Wb during exercise in females compared with males.NEW & NOTEWORTHY We hypothesized that the higher resistive work of breathing in females compared with males during high-intensity exercise is due to smaller airways. In vivo measures of the fourth to eighth airway generations made using optical coherence tomography show that females tend to have smaller airway luminal areas of the fourth to sixth airway generations. Sex differences in airway luminal area result in a greater resistive work of breathing during exercise in females compared with males.


Antisera to synthetic peptides of bovine rhodopsin: use as site-specific probes of disc membrane changes in retinal dystrophic dogs.
Based on the amino acid sequence of bovine rhodopsin, five peptides corresponding to the carboxyl terminus and one loop region have been synthesized. Rabbit antisera to these peptides recognize rhodopsin in whole bovine and dog retinas. Antisera were used to detect differences in specific regions of rhodopsin in dystrophic vs normal dog retinas. As detected on both "dot blots" and Western blots, rhodopsin from retinas of dystrophic dogs has a reduced reaction with antisera to peptides, Rhod-4 and Rhod-10 (# 341-348 and 232-239, respectively). Since these sites on rhodopsin are possible binding sites for transducin and rhodopsin kinase, an alteration in these regions would have profound effects in the dystrophic state.


Inhaled glutathione decreases PGE2 and increases lymphocytes in cystic fibrosis lungs.
Reduced glutathione (GSH), a major antioxidant and modulator of cell proliferation, is decreased in the bronchoalveolar lavage fluid (BALF) of cystic fibrosis (CF) patients. We previously have shown that GSH inhalation in CF patients significantly increased GSH levels in BALF and improved lung function (M. Griese et al., 2004, Am. J. Respir. Crit. Care Med.169, 822-828). GSH depletion in vitro enhances susceptibility to oxidative stress, increases inflammatory cytokine release, and impairs T cell responses. We therefore hypothesized that an increase in GSH in BALF reduces oxidative stress, decreases inflammation, and modulates T cell responses in lungs of CF patients. BALF from 17 CF patients (median FEV1 67% (43-105%) of predicted) was assessed before and after GSH inhalation for total protein, markers of oxidative stress (8-isoprostane, myeloperoxidase, and ascorbic and uric acid), pattern of protein oxidation, prostaglandin E2 (PGE2), and proinflammatory cytokines. BALF cells were differentiated using cytospin slides, and lymphocytes were further analyzed by flow cytometry. Inhalation of GSH decreased BALF levels of PGE2 and increased CD4+ and CD8+ lymphocytes in BALF significantly but had no effect on markers of oxidative stress. BALF lymphocytes correlated positively with lung function, whereas levels of PGE2 showed an inverse correlation. The patients with the greatest improvement in lung function after GSH treatment also had the largest decline in PGE2 levels. We conclude that GSH inhalation in CF patients increases lymphocytes and suppresses PGE2 in the bronchoalveolar space. Thus, GSH primarily affected the pulmonary immune response rather than the oxidative status in CF patients. The effect of GSH inhalation on PGE2 levels and lymphocytes in CF warrants further investigation.


Effect of surgical blood loss and volume replacement on antibiotic pharmacokinetics.
Antibiotic prophylaxis is used widely to prevent postoperative infection associated with cardiac operations. The influence of perioperative blood loss on antibiotic pharmacokinetics has not been well studied. Plasma cefazolin sodium samples from 8 patients undergoing a cardiac operation were collected postoperatively and analyzed. The results suggest that blood loss has a minimal effect on the elimination of cefazolin and that therapeutic concentrations are maintained without altering the dosage regimen.


Changes in healthcare utilisation after surgical treatment: mitigating risk through multidisciplinary and collaborative care.
A recent study by Fowler and colleagues identified increased utilisation of healthcare resources among patients, mostly from deprived social areas with chronic diseases, undergoing emergency and high-risk surgery. Reasons for these findings include the intrinsic risk of surgery, postoperative complications, and the need for chronically ill patients to have their usual treatment resumed after surgery. To improve the overall outcome of surgery in this category of patients, a number of elements in the process of care should be adjusted. This includes minimising the number of emergency procedures and enhancing collaboration between all healthcare professionals inside and outside hospitals.


Men's and women's reactions to sexually explicit films: a serendipitous finding.
We proposed that men and women would be "turned off" by watching men and women like themselves engaged in sexual activity and ""turned on" by watching someone of the opposite sex engaged in the same activity. We tested this hypothesis by showing college men and women eight films: films depicting male and female homosexuality, male and female masturbation, and males and females engaged in heterosexual intercourse. We assessed men and women's reactions via the Byrne-Sheffield (1965) Feeling Scale and Griffitt's (1975) Physiological Arousal Scale. We found support for our hypothesis. We also attempted to determine whether men and women differed in how easily they became aroused by sexually explicit films. We found that they did not.


Insulin requirement of human leukemic cell lines.
The growth characteristics of human leukemic cell lines in serum supplemented medium and in serum free medium with and without the addition of insulin were investigated. No relation was found between the insulin binding capacity of the cells and their hormone-dependence for growth.


Risk factors for lumbosacral radiculoplexus neuropathy.
Recently, our group found an association between diabetes mellitus (DM) and lumbosacral radiculoplexus neuropathy (LRPN) in Olmsted County, Minnesota; we found a higher risk (odds ratio [OR], 7.91) for developing LRPN in diabetic compared with nondiabetic patients. However, the influence of other comorbidities and anthropomorphic variables was not studied. Demographic and clinical data from 59 LRPN patients and 177 age/sex-matched controls were extracted using the Rochester LRPN epidemiological study. Differences between groups were compared by chi-square/Fisher exact test or Wilcoxon rank-sum test. Uni- and multivariate logistic regression analysis were performed. Factors predictive of LRPN on univariate analysis were DM (OR, 7.91; 95% confidence interval [CI], 4.11-15.21), dementia (OR, 6.36; 95% CI, 1.13-35.67), stroke (OR, 3.81; 95% CI, 1.32-11.01), dyslipidemia (OR, 2.844; 95% CI, 1.53-5.27), comorbid autoimmune disorders (OR, 2.72; 95% CI, 1.07-6.93), hypertension (OR, 2.25; 95% CI, 1.2-4.13), obesity (OR, 2.05; 95% CI, 1.11-3.8), body mass index (BMI) (OR, 1.1; 95% CI, 1.04-1.15), and weight (OR, 1.02; 95% CI, 1.009-1.037). On multivariate logistic regression analysis only DM (OR, 8.03; 95% CI, 3.86-16.7), comorbid autoimmune disorders (OR, 4.58; 95% CI, 1.45-14.7), stroke (OR, 4.13; 95% CI, 1.2-14.25), and BMI (OR, 1.07; 95% CI, 1.01-1.13) were risk factors for LRPN. DM is the strongest risk factor for the development of LRPN, followed by comorbid autoimmune disorders, stroke, and higher BMI. Altered metabolism and immune dysfunction seem to be the most influential factors in the development of LRPN.


[Anti-inflammatory drugs and community-acquired pneumonia].
Nonsteroidal anti-inflammatory drugs (NSAIDs) are commonly used in ambulatory medicine for their analgesic and antipyretic properties and are often used as self-medication. Their use in community-acquired pneumonia is associated with an increased risk of loco-regional complications, especially pleural empyema. Appropriate therapeutic care and hospital admissions are often delayed because of initial improvement of symptoms with NSAIDs. Despite worrying observational data, a causal link remains to be established. Currently, there is no recommendation cautioning against the use of NSAIDs in the management of community-acquired pneumonia.


Ion-induced fusion of phosphatidic acid vesicles and correlation between surface hydrophobicity and membrane fusion.
Divalent cation (Ca2+ and Mg2+) and hydrogen ion-induced fusions of small unilamellar phosphatidic acid vesicles were studied by the use of fluorescence fusion assay. These fusogenic ions also increased interfacial tension and reduced the surface dielectric constant of phosphatidic acid membranes as the ion concentration increased. A good correlation was found between the threshold of vesicle fusion and the degree of changes in such membrane properties induced by these fusogenic ions. At the fusion threshold point, the increase in interfacial tension and the decreased surface dielectric constant of the membrane were approximately 6 and 14 dyn/cm, respectively, regardless of Ca2+, Mg2+ or H+. Such changes correspond to the increases in surface hydrophobicity of the membrane. As these fusogenic ionic concentrations increased, the electrophoretic mobility of the phosphatidic acid vesicle decreased, because of the binding of ions to negatively charged sites of the membranes. However, the quantities of the reduced negative surface charges were not necessarily in good correlation with the threshold of vesicle fusion. It is suggested that the complexes of the phosphate group with these fusogenic ions are responsible for increases in the surface hydrophobicity of the membrane, which is an important factor for membrane fusion.


[Mechanism of inhibiting the proliferation of prostate cancer by finasteride: a study using cDNA microarray].
To screen the genes related to finasteride treatment that reduces the proliferation of prostate cancer cells. The prostate cancer cells of the line LNCaP sensitive to finasteride treatment were cultured. Finasteride was added into the culture fluid. A cDNA microarray consisting of 96 human genes was used to identify the genes showing differential expression. Finasteride significantly inhibited the proliferation of the prostate cancer cells of the line LNCaP. Twenty-nine genes out of the 96 genes showed differential expression under finasteride treatment 11 were upregulated and 18 were down-regulated, including those significant in cell metabolism, proliferation, apoptosis, and cell signal transduction, such as AKR1B1, PTEN, NKX3.1, PMEPA1, PSA, and XRCC2. Inhibiting the proliferation of prostate cancer cells by finasteride may involve the cooperative effect of multiple genes and pathways.


A case of metastatic treatment-emergent small cell/neuroendocrine prostate cancer with BRCA2 mutation diagnosed by liver biopsy.
Treatment-emergent small cell/neuroendocrine prostate cancer occurs predominantly in advanced or metastatic castration-resistant prostate cancer that arises when prostate adenocarcinoma is transformed after androgen deprivation therapy. The clinical course for the pathogenesis involved or associated genetic information have not been clearly elucidated. A Japanese male, 63-year-old, underwent a para-aortic lymph biopsy due to sudden severe bilateral leg edema, with a final diagnosis of stage IV prostate adenocarcinoma. He was initially responsive to upfront abiraterone with androgen deprivation therapy; however, relapse occurred in the liver and bone 10 months after initial treatment, with serum neuron-specific enolase elevation and without prostate-specific antigen elevation. Pathological findings of liver tumor revealed treatment-emergent small cell/neuroendocrine prostate cancer. FoundationOne® CDx was used for cancer-related gene profiling of liver tumor specimen; a BRCA2 mutation was identified. Early detection of this transformation and pathological diagnosis can improve patient survival when genetic mutations, including BRCA 1/2.


Neuropathic pain with features of complex regional syndrome in the upper extremity after herpes zoster.
We report a case of a 73-year-old female who developed unbearable neuropathic pain after a herpes zoster episode. The pain persisted and could not be controlled despite multimodal analgesia. In addition to postherpetic neuralgia, myelitis and complex regional pain syndrome were diagnosed during the evolution of neuropathic pain. This complex neuropathic pain was resolved after sympathetic ganglion block.


Lower extremity strength and hopping and jumping ground reaction forces in children with neurofibromatosis type 1.
The purpose of this study was to (1) extend the research findings of decreased muscular force production in grip strength to the lower extremity strength of children with NF1 and (2) to determine if there was a relationship between isometric strength and functional activities in children with NF1. Force production was assessed using a hand held dynamometer (HHD) and a functional task (hopping and jumping) on a force plate. Data from twenty-six children with NF1 were compared to data from 48 typically developing children of similar sex, weight and height. Children with NF1 demonstrated statistically significant lower force production with HHD (p<0.01) during hip extension, but similar force production for knee extension and ankle plantar flexion compared to the control group. A relationship existed between the ground reaction forces at take-off from both hopping and jumping and the force generated from knee extensor strength in the NF1 group. The addition of a functional task to hand held dynamometry is useful for determining a relationship between common clinical measures and functional activities.


Use of adjunct faculty members in classroom teaching in departments of pharmacy practice.
To determine trends among departments of pharmacy practice regarding use of adjunct faculty members for classroom-based teaching and to assess departmental support provided to these faculty members. Chairs of pharmacy practice departments in US colleges and school of pharmacy were contacted by e-mail and asked to complete an 11-item electronic survey instrument. Chair respondents reported an average of 5.7 adjunct faculty members hired to teach required courses and 1.8 adjunct faculty members hired to teach elective courses. Compensation averaged $108 per lecture hour and $1,257 per 1-credit-hour course. Twenty-five percent of the respondents expected to hire more adjunct faculty members to teach required courses in the upcoming year due to curricular changes, faculty hiring freezes, and the shortage of full-time faculty members. Only 7% of respondents reported that they provided a teaching mentor and 14% offered no support to their adjunct faculty members. Departments of pharmacy practice commonly use adjunct faculty members to teach required and elective courses. Given the pharmacy faculty shortage, this trend is expected to increase and may be an area for future faculty development.


Effects of acute and chronic administration of secretin and caerulein on rat duodenal and gastric growth.
We studied the effects of acute and chronic administration of secretin and caerulein, alone and in combination, on RNA and protein synthesis in the duodenum and oxyntic gland area as well as content of DNA, RNA and protein in rats. Secretin, 100 micrograms . kg-1, three times a day for 5 days, was associated with duodenal hypertrophy after the first 24 h of treatment and hyperplasia at the end of days 2 and 4; hypertrophy of the oxyntic gland area was observed only at 4 h after the first injection. Caerulein, 1 microgram . kg-1, also promoted duodenal hyperplasia after 2 and 4 days of treatment. The oxyntic gland area showed hypertrophy only at 4 h after the second injection of caerulein. These data indicate that both hormones can induce duodenal hyperplasia, probably by an amplification of the normal renewal cycle of the epithelial cells. They also demonstrated that growth of the oxyntic gland area is not promoted by these two peptides at the doses studied.


A history of asthma is associated with susceptibility to hidradenitis suppurativa: a population-based longitudinal study.
The association of hidradenitis suppurativa (HS) and asthma remains to be investigated. To assess the bidirectional association between HS and asthma. A population-based study was conducted to compare HS patients (n = 6779) with age-, sex-, and ethnicity-matched control subjects (n = 33,259) with regard to the incidence of new onset and the prevalence of preexisting asthma. Adjusted hazard ratios (HRs) and adjusted odds ratios (ORs) were calculated. The prevalence of preexisting asthma was higher in patients with HS relative to controls (9.6% vs. 6.9%, respectively; P < 0.001). The odds of HS were 1.4-fold greater in patients with a history of asthma (fully adjusted OR 1.41; 95% CI 1.27-1.55; P < 0.001). The incidence rate of new-onset asthma was estimated at 9.0 (95% CI 6.3-12.7) and 6.2 (95% CI 5.1-7.5) cases per 10,000 person-years among patients with HS and controls, respectively. The risk of asthma was not statistically different in patients with HS and controls (fully adjusted HR 1.53; 95% CI 0.98-2.38; P = 0.062). Relative to other patients with HS, those with HS and comorbid asthma were younger at the onset of HS (30.7 [14.7] vs. 33.3 [15.1], respectively; P < 0.001) and had a comparable risk of all-cause mortality (adjusted HR 0.86; 95% CI 0.44-1.68; P = 0.660). A history of asthma confers susceptibility to subsequent development of HS. This observation is of importance for clinicians managing both patients with HS and asthma. Further research is warranted to elucidate the pathomechanism underlying this finding.


The impact of butylphthalide on the hypothalamus-pituitary-adrenal axis of patients suffering from cerebral infarction in the basal ganglia.
Butylphthalide sodium chloride injection for patients with acute cerebral infarction has a certain effect. Although there are several proposed mechanisms of drug action, no related research on improving the inflammatory cytokines that regulate the body's immune system through the hypothalamus-pituitary-adrenal axis has been published. To determine the impact of butylphthalide and sodium chloride injection on the hypothalamus-pituitary-adrenal (HPA) axis after acute cerebral infarction in the basal ganglia. Patients were randomly divided into treatment and control groups; the treatment group received intravenous drips of butylphthalide, while the control group did not. The levels of adrenocorticotropic hormone (ACTH) and cortisol (COR), along with the National Institutes of Health Stroke Scale (NIHSS) scores of both groups were detected using the radioimmunoassay method. This was done at regular intervals after cerebral infarction in the basal ganglia was detected. Fourteen days after treatment, the levels of serum ACTH and COR in both groups were higher than normal. The NIHSS score and levels of ACTH and COR of the treatment group were significantly lower than those of the control group (p<0.05). The data was computed and analyzed using SPSS17.0 software. Butylphthalide treatment for patients suffering from acute basal ganglia infarction can reduce the adverse effects on the HPA axis, thus improving patient prognosis.


Rapid biodegradation of organophosphorus pesticides by Stenotrophomonas sp. G1.
Organophosphorus insecticides have been widely used, which are highly poisonous and cause serious concerns over food safety and environmental pollution. A bacterial strain being capable of degrading O,O-dialkyl phosphorothioate and O,O-dialkyl phosphate insecticides, designated as G1, was isolated from sludge collected at the drain outlet of a chlorpyrifos manufacture plant. Physiological and biochemical characteristics and 16S rDNA gene sequence analysis suggested that strain G1 belongs to the genus Stenotrophomonas. At an initial concentration of 50 mg/L, strain G1 degraded 100% of methyl parathion, methyl paraoxon, diazinon, and phoxim, 95% of parathion, 63% of chlorpyrifos, 38% of profenofos, and 34% of triazophos in 24 h. Orthogonal experiments showed that the optimum conditions were an inoculum volume of 20% (v/v), a substrate concentration of 50 mg/L, and an incubation temperature in 40 °C. p-Nitrophenol was detected as the metabolite of methyl parathion, for which intracellular methyl parathion hydrolase was responsible. Strain G1 can efficiently degrade eight organophosphorus pesticides (OPs) and is a very excellent candidate for applications in OP pollution remediation.


Utility of cytokeratin subsets for distinguishing poorly differentiated synovial sarcoma from peripheral primitive neuroectodermal tumour.
Poorly differentiated synovial sarcoma (PDSS) is a round cell sarcoma that may be difficult to distinguish from other round cell sarcomas, such as peripheral primitive neuroectodermal tumour (pPNET), on histological examination alone. Furthermore, these two tumours may show overlapping immunophenotypes, as some cases of PDSS express CD99, and, on the other hand, pPNET may express epithelial markers. The goal of this study was to determine the utility of cytokeratin (CK) subsets in distinguishing between these two lesions. We evaluated 13 cases of pPNET with RT-PCR detected EWS/FLI-1 fusion transcript and 21 cases of PDSS for the immunohistochemical expression of CK7 and CK19. All cases of PDSS had areas of recognizable monophasic or biphasic synovial sarcoma that expressed at least one epithelial marker. Thirteen of 21 (62%) PDSS stained with AE1/AE3. CK7 and 19 were expressed in 11 (52%) and nine (43%) cases, respectively. Although six of 19 (32%) PDSS demonstrated cytoplasmic staining for CD99, none showed a membranous pattern of immunoreactivity. In contrast, 12 of 13 (92%) pPNET showed strong membranous immunoreactivity for CD99. Four tumours (31%) showed focal staining with AE1/AE3, two of which (15%) stained for CK19. CK7 was not detected in any of the pPNETs. Although AE1/AE3 may be found in up to 31% of cases of pPNET, the expression of CK7 makes this diagnosis less likely.


Fathers and sons: their relationship and personality characteristics associated with the son's smoking behavior.
The purpose of the present study was to examine the interrelationship of the domains of father personality attributes, father socialization techniques, and adolescent personality attributes, and the son's use of tobacco. A sample of 246 male adolescents and their fathers from intact homes was administered questionnaires. Two main findings emerged. First, each of the three domains was significantly associated with the son's tobacco use despite control on the remaining domains. Second, the effectiveness of the father may interact synergistically with or be mitigated by the son's personality attributes in its association with the son's tobacco use. In concert, the findings support the importance of examining the father-son relationship for a greater understanding of the son's tobacco use.


Efficacy evaluation of a protocol for safe hip surgery (total hip arthroplasty).
To propose a multidisciplinary protocol to standardize the care of patients undergoing total hip arthroplasty (THA) and evaluate it effectiveness after implementation. Retrospective evaluation of 95 consecutive patients undergoing THA divided into two groups, one group of 47 patients operated before the protocol implementation and 48 after. Assessing the re-admission rate, among 47 patients evaluated prior to implementation of the protocol, seven (14.9%) were re-admitted, and when observing the 48 patients evaluated after implementation, one (2.1%) was re-admitted, showing statistical significance (p < 0.05). The chance of re-admission before the protocol was eight times the chance of hospitalization after implementation (95% CI: 1.01 to 377.7). By comparing the clinical complications among the groups, it was observed that there was a lower rate of complications following implementation of the protocol (p = 0.006). The introduction of a multidisciplinary protocol to standardize the management of patients undergoing THA decreased the rates of rehospitalization and clinical complications after the procedure.


Nanoparticles that target the mitochondria of tumor cells to restore oxygen supply for photodynamic therapy: Design and preclinical validation against breast cancer.
Photodynamic therapy, in which photosensitizers locally generate cytotoxic reactive oxygen species, can treat tumor tissue with minimal effects on surrounding normal tissue, but it can be ineffective because of the anoxic tumor microenvironment. Here we developed a strategy to inactivate the mitochondria of tumor cells in order to ensure adequate local oxygen concentrations for photodynamic therapy. We conjugated the photosensitizer 5-aminolevulinic acid to the lipophilic cation triphenylphosphine, which targets mitochondria. Then we packaged the conjugate into nanoparticles that were based on biocompatible bovine serum albumin and coated with folic acid in order to target the abundant folate receptors on the tumor surface. In studies in cell culture and BALB/c mice bearing MCF-7 xenografts, we found that the nanoparticles helped solubilize the cation-photosensitizer conjugate, prolong its circulation, and enhance its photodynamic antitumor effects. We confirmed the ability of the nanoparticles to target tumor cells and their mitochondria using confocal laser microscopy and in vivo assays of pharmacokinetics, pharmacodynamics, and tissue distribution. Our results not only identify a novel nanoparticle system for treating cancer, but they demonstrate the feasibility of enhancing photodynamic therapy by reducing oxygen consumption within tumors.


South Carolina Partners for Preterm Birth Prevention: a regional perinatal initiative for the reduction of premature birth in a Medicaid population.
The objective of the study was to improve the distribution of preterm deliveries in a Medicaid population through a regional perinatal risk assessment and case management initiative. An innovative public/private partnership was initiated in the 8 county Lowcountry (LC) perinatal region to reduce preterm birth (PTB) among Medicaid recipient women. Eligible women were identified and underwent telephonic risk assessment, education, and access to a 24 hours, 7 days per week perinatal hotline. Women with predetermined risk factors for PTB were offered patient-centered case management. Medicaid claims and birth certificate data were used to compare obstetric outcomes for 2006 (intervention) and 2004 (control) in both the Lowcountry (LC; program) and Midlands (ML; nonprogram) perinatal regions. There were 6356 Medicaid deliveries in the LC in 2006. Of these, 2111 were referred for telephonic risk assessment; 317 had identifiable PTB risk factors and consented to case management. Compared with 2004, there was a significant improvement in the distribution of preterm birth (P = .05) in the LC region, primarily confined to deliveries less than 28 weeks (1.6% vs 1.1%; P = .029, relative risk [RR] 0.75, 95% confidence interval [CI], 0.51-0.96). There were also reductions in the frequency (6.7% vs 5.8%; RR 0.86, 95% CI, 0.75-0.98; P = .04) and mean duration (25.0 vs 20.6 days; 95% CI, 1.03-7.77; P = .01) of neonatal intensive care unit (NICU) admissions. No changes were identified in the ML region. A regional initiative of telephonic risk assessment and case management of Medicaid recipient women significantly reduced deliveries less than 28 weeks and NICU care.


Using a geographic information system to enhance patient access to point-of-care diagnostics in a limited-resource setting.
Rapid and accurate diagnosis drives evidence-based care in health. Point-of-care testing (POCT) aids diagnosis by bringing advanced technologies closer to patients. Health small-world networks are constrained by natural connectivity in the interactions between geography of resources and social forces. Using a geographic information system (GIS) we can understand how populations utilize their health networks, visualize their inefficiencies, and compare alternatives. This project focuses on cardiac care resource in rural Isaan, Thailand. A health care access analysis was created using ArcGIS Network Analyst 10.1 from data representing aggregated population, roads, health resource facilities, and diagnostic technologies. The analysis quantified cardiac health care access and identified ways to improve it using both widespread and resource-limited strategies. Results indicated that having diagnostic technologies closer to populations streamlines critical care paths. GIS allowed us to compare the effectiveness of the implementation strategies and put into perspective the benefits of adopting rapid POCT within health networks. Geospatial analyses derive high impact by improving alternative diagnostic placement strategies in limited-resource settings and by revealing deficiencies in health care access pathways. Additionally, the GIS provides a platform for comparing relative costs, assessing benefits, and improving outcomes. This approach can be implemented effectively by health ministries seeking to enhance cardiac care despite limited resources.


Registry-Based Mortality Analysis Reveals a High Proportion of Patient Decrees and Presumed Limitation of Therapy in Severe Geriatric Trauma.
The treatment of severely injured patients, especially in older age, is complex, and based on strict guidelines. We conducted a retrospective study by analyzing our internal registry for mortality risk factors in deceased trauma patients. All patients that were admitted to the trauma bay of our level-1-trauma center from 2014 to 2018, and that died during the in-hospital treatment, were included. The aim of this study was to carry out a quality assurance concerning the initial care of severely injured patients. In the 5-year period, 135 trauma patients died. The median (IQR) age was 69 (38-83) years, 71% were male, and the median (IQR) Injury Severity Score (ISS) was 25 (17-34) points. Overall, 41% of the patients suffered from severe traumatic brain injuries (TBI) (AIShead ≥ 4 points). For 12.7%, therapy was finally limited owing to an existing patient's decree; in 64.9% with an uncertain prognosis, a 'therapia minima' was established in consensus with the relatives. Although the mortality rate was primarily related to the severity of the injury, a significant number of deaths were not exclusively due to medical reasons, but also to a self-determined limitation of therapy for severely injured geriatric patients. The conscientious documentation concerning the will of the patient is increasingly important in supporting medical decisions.


Comparison of causal forest and regression-based approaches to evaluate treatment effect heterogeneity: an application for type 2 diabetes precision medicine.
Precision medicine requires reliable identification of variation in patient-level outcomes with different available treatments, often termed treatment effect heterogeneity. We aimed to evaluate the comparative utility of individualized treatment selection strategies based on predicted individual-level treatment effects from a causal forest machine learning algorithm and a penalized regression model. Cohort study characterizing individual-level glucose-lowering response (6 month reduction in HbA1c) in people with type 2 diabetes initiating SGLT2-inhibitor or DPP4-inhibitor therapy. Model development set comprised 1,428 participants in the CANTATA-D and CANTATA-D2 randomised clinical trials of SGLT2-inhibitors versus DPP4-inhibitors. For external validation, calibration of observed versus predicted differences in HbA1c in patient strata defined by size of predicted HbA1c benefit was evaluated in 18,741 patients in UK primary care (Clinical Practice Research Datalink). Heterogeneity in treatment effects was detected in clinical trial participants with both approaches (proportion predicted to have a benefit on SGLT2-inhibitor therapy over DPP4-inhibitor therapy: causal forest: 98.6%; penalized regression: 81.7%). In validation, calibration was good with penalized regression but sub-optimal with causal forest. A strata with an HbA1c benefit > 10 mmol/mol with SGLT2-inhibitors (3.7% of patients, observed benefit 11.0 mmol/mol [95%CI 8.0-14.0]) was identified using penalized regression but not causal forest, and a much larger strata with an HbA1c benefit 5-10 mmol with SGLT2-inhibitors was identified with penalized regression (regression: 20.9% of patients, observed benefit 7.8 mmol/mol (95%CI 6.7-8.9); causal forest 11.6%, observed benefit 8.7 mmol/mol (95%CI 7.4-10.1). Consistent with recent results for outcome prediction with clinical data, when evaluating treatment effect heterogeneity researchers should not rely on causal forest or other similar machine learning algorithms alone, and must compare outputs with standard regression, which in this evaluation was superior.


Ultrasonographic Imaging Protocol and Sonoanatomy of the Lumbar Spine in Healthy Dogs.
Ultrasound is an imaging technique commonly used in veterinary medicine. Ultrasound devices are widely available, their means of examination are relatively short and cheap, and they do not generate ionizing radiation. In addition, ultrasound generally does not need to be performed under general anesthesia. This study was performed on 23 canine cadavers with full clinical histories and with no confirmed pathological changes in the spine region. The imaging modalities were established in dogs in lateral recumbency, with the selected side being the uppermost angle, in a neutral position. All dogs were examined in the transverse and longitudinal planes. Sacral crest, intertransverse ligament, vertebral canal floor, vertebral body, and intervertebral discs were only visible in the longitudinal plane. Vertebral arch, supraspinal ligament, dorsal wall of the vertebral canal and muscles were visualized only in the transverse plane. This article provides a brief and relatively easy-to-perform protocol for ultrasound imaging of the lumbar spine of dogs. In addition, it presents a detailed description of the sonoanatomy of the area under investigation.


KSHV strategies for host dsDNA sensing machinery.
The innate immune system utilizes pattern recognition receptors cyclic GMP-AMP synthase (cGAS) to sense cytosolic double-stranded (ds) DNA and initiate type 1 interferon signaling and autophagy pathway, which collaborate to limit pathogen infections as well as alarm the adaptive immune response. The genomes of herpesviruses are large dsDNA, which represent a major class of pathogen signatures recognized by cellular DNA sensor cGAS. However, to successfully establish the persistent infection, herpesviruses have evolved their viral genes to modulate different aspects of host immune signaling. This review summarizes the evasion strategies of host cGAS DNA sensing pathway by Kaposi's Sarcoma-associated Herpesvirus (KSHV) and their contributions to KSHV life cycles.


Omeprazole: a study of its inhibition of gastric pH and oral pharmacokinetics after morning or evening dosage.
Pharmacodynamic and pharmacokinetic studies of omeprazole, a new gastric antisecretory agent, were undertaken in 8 healthy subjects. The drug was administered orally as an encapsulated enteric-coated granulate (40 mg daily at 9 am or 9 pm for 5 days), and its effect on the integrated 24-h gastric pH was determined, together with its apparent bioavailability. The pretreatment 24-h median pH was 1.9 (interquartile range 1.4-2.9). After 5 days of treatment, the median pH had risen to 5.0 (3.7-6.0) (p less than 0.01) with morning dosage and 4.5 (3.0-5.6) (p less than 0.01) with evening dosage. This corresponded to a greater than 99% reduction in 24-h median hydrogen ion activity, with morning dosage having a greater effect (from 9 am to 8 pm) (p less than 0.01) than evening dosage. The relative bioavailability of omeprazole increased twofold from day 1 to day 5 of treatment with morning dosage (p less than 0.02) and threefold with evening dosage (p less than 0.02), suggesting that increased absorption of this acid-labile drug occurs with increasing inhibition of acid secretion. We conclude that this formulation of omeprazole presently being used in clinical trials is a highly potent antisecretory agent in humans, although its optimal effect may not be observed for several days.


[Value of left atrial dilation in the diagnosis of silent myocardial ischemia in diabetes mellitus patients].
Accelerated atherothrombosis is a common feature in diabetes mellitus patients (DM), which can be related to abnormalities in vascular cell apoptosis and activation leading to the release of procoagulant microparticles (MPs). In DM patients, we hypothesized that circulating levels of biomarkers involved in atherothrombosis processes as well as cardiac and carotid echocardiography variables could be useful in the detection of silent myocardial diagnosed by myocardial perfusion imaging. We investigated, in 55 patients with diabetes (mean age 62+/-10 years) and 15 nondiabetics (46+/-14 years) patients the prevalence of silent myocardial ischemia (SMI) detected by a treadmill exercise or dipyridamole (99m)Tc-sestamibi stress test. Echocardiographic and -carotid variables were obtained using standardized methods. Biomarkers assessing endothelial apoptosis or activation (CD31+-MPs, CD62+-MPs, VCAM-1), inflammatory status (CD11a +/- MPs, MCP-1, CRP), platelet activation (GPIb+/-MPs, CD40-L, P-selectin, GPV) ventricular stretch (BNP) were measured in the plasma. SMI was diagnosed in 23/55 (42%) diabetics patients and in 3/15 (20%) nondiabetics patients. Enhanced inflammatory status and leukocyte damage (CD11a+-MPs) were evidenced in diabetic patients. Within the diabetic population, biomarkers levels of atherothrombosis were not significantly associated to the detection of SMI. In multivariable analyses adjusted for LV hypertophy, left atrial surface (LA) remained independent predictor of silent myocardial ischemia (OR 4.14; IC [1.7-16.13]; P=0.039). In diabetes mellitus patients, LA surface independently predicted silent myocardial ischemia after adjustment for established echocardiographic, and inflammatory risk factors. This simple measure of LA dilation could be helpful in the identification of diabetes mellitus patients at heightened cardiovascular risk.


Transfection of neuroprogenitor cells with iron nanoparticles for magnetic resonance imaging tracking: cell viability, differentiation, and intracellular localization.
Magnetic resonance imaging (MRI) can track labeled cells in the brain. The use of hemagglutinating virus of Japan envelopes (HVJ-Es) to effectively introduce the contrast agent to neural progenitor cells (NPCs) is limited to date despite their high NPC affinity. HVJ-Es and Lipofectamine 2000 were compared as transfection vehicles of superparamagnetic iron oxide (SPIO). Labeled NPCs were examined for iron content, MRI signal change, and fundamental cell characteristics. Prussian Blue staining was used after differentiation to determine SPIO localization. HVJ-Es transfected up to 12.5 +/- 8.8 times more SPIO into NPCs. HVJ-Es do not affect cell viability or differentiation capability. Superparamagnetic iron oxide was disseminated in both the soma and neurites. These findings indicate that HVJ-Es are an effective vehicle for SPIO transfection of NPCs. The intracellular localization after differentiation raises the question as to the capability of MRI to distinguish cell migration from axonal or dendritic growth in vivo.


Synchronous early primary adenocarcinoma of both rectum and gallbladder. Report of a case.
Synchronous early primary cancers are rare and in addition synchronous adenocarcinoma of both rectum and gallbladder is extremely rare. We report an unusual case of synchronous early primary adenocarcinoma of rectum and gallbladder. The patient was a 72-year-old woman with complaints of bloody stools and constipation. An endoscopy revealed adenocarcinoma of the lower rectum. A through preoperative investigation showed also cholelithiasis. The patient underwent abdominoperineal resection and cholecystectomy. The histopathological diagnosis was well to middle differentiate adenocarcinoma of the gallbladder (T2, N0, M0; stage II) and middle differentiate adenocarcinoma of the rectum (T2, N0, M0; stage II). For the cases of extracolonic primary cancer associated with colorectal primary carcinoma, Warren and Gates' diagnostic criteria are used. All patients with colorectal carcinoma, should undergo a throughout preoperative examination to exclude the possibility of synchronous early primary cancers.


Culturing and robotic harvesting of bone marrow, lymph nodes, peripheral blood, fibroblasts, and solid tumors with in situ techniques.
We have modified our in situ method for culturing amniocytes and have successfully used this new procedure to culture cells from bone marrow, lymph nodes, peripheral blood, fibroblasts, and solid tumors. We have been so satisfied with this method that we have used it in our routine practice for the past 2.5 years. A convenient feature of the in situ method is that the cultures can be processed with a commercially available robotic harvesting system (modified Tecan Robotic Sample Processor Model 505). This system also works well for in situ cultures of amniotic fluids. Robotic harvesting of in situ cultures provides a consistent and accurate procedure that saves about 10-15 minutes of technologist time per case.


Physiologically based toxicokinetic models and in silico predicted partition coefficients to estimate tetrachlorodibenzo-p-dioxin transfer from feed into growing pigs.
Tetrachlorodibenzo-p-dioxin (TCDD) is a ubiquitous, toxic, persistent and bioaccumulative organic pollutant. TCDD can potentially enter the food chain through contaminated food of animal origin as a consequence of feed contamination. Prediction of the TCDD transfer from feed into animal products is thus important for human health risk assessment. Here, we develop several physiologically based toxicokinetic (PBTK) models of TCDD transfer from contaminated feed into growing pigs (Sus scrofa) exposed to doses ranging from 24.52 to 3269.25 ng of TCDD. We test the consequences of explicit dose-dependent absorption (DDA) versus the indirect effects of a self-induced liver metabolism (SIM). The DDA and SIM models showed similar fit to experimental data, although currently it is not possible to unequivocally make statement on a mechanistic preference. The performance of both toxicokinetic models was successfully evaluated using the 1999 Belgian case of contaminated fats for feeding. In combination with toxicokinetic models of other dioxin congeners, they can be used to formulate maximum allowance levels of dioxins in feedstuffs for pigs. Additionally, the implementation of in silico-predicted partition coefficients was explored as a useful alternative to predict TCDD tissue distribution in low-dose scenarios without recurring to animal experiments.


Separation and colorimetric detection of Escherichia coli by phage tail fiber protein combined with nano-magnetic beads.
A colorimetric detection method for Escherichia coli (E. coli) in water was established based on a T7 phage tail fiber protein-magnetic separation. Firstly, the tail fiber protein (TFP) was expressed and purified to specifically recognize E. coli, which was verified by using fusion protein GFP-tagged TFP (GFP-TFP) and fluorescence microscopy. Then TFP conjugated with magnetic beads were applied to capture and separate E. coli. The TFP was covalently immobilized on the surface of magnetic beads and captured E. coli as verified by scanning electron microscopy (SEM). Finally, polymyxin B was used to lyse E. coli in solution and the released intracellular β-galactosidase (β-gal) could hydrolyze the colorimetric substrate chlorophenol red-β-D-galactopyranoside (CPRG), causing color change from yellow to purple. The high capture efficiencies of E. coli ranged from 88.70% to 95.65% and E. coli could be detected at a concentration of 102 CFU/mL by naked eyes. The specificity of the chromogenic substrate was evaluated using five different pathogen strains as competitors and tests with four kinds of real water samples showed recoveries of 86.00% to 92.25%. The colorimetric changes determined by visual inspection can be developed as an efficient platform for point-of-care detection of E. coli in resource-limited regions.


[Robot-assisted head and neck surgery].
Robot-assisted surgery (RAS) has already been approved for several clinical applications in head and neck surgery. In some Anglo-American regions, RAS is currently the common standard for treatment of oropharyngeal diseases. Systematic randomized studies comparing established surgical procedures with RAS in a large number of patients are unavailable so far. Experimental publications rather describe how to reach poorly accessible anatomical regions using RAS, or represent feasibility studies on the use of transoral robotic surgery (TORS) in established surgical operations. With general application of RAS in clinical practice, the question of financial reimbursement arises. Furthermore, the technical applications currently on the market still require some specific improvements for routine use in head and neck surgery.


Effects of Dietary Supplementation with Mulberry Leaf Powder on the Growth Performance, Lipid Metabolism Parameters, Immunity Indicators, and Gut Microbiota of Dogs.
Overfeeding and a lack of exercise are increasingly causing obesity in dogs, which has become a big problem threatening the health of dogs. Therefore, it is necessary to investigate how dietary regulations can help to improve dogs' body conditions and minimize obesity. This study was carried out to investigate the effects of dietary mulberry leaf powder (MLP) supplementation on the growth performance, lipid metabolism parameters, and gut microbiota of Chinese indigenous dogs. Fifteen Chinese indigenous dogs (6.34 ± 0.56 kg) were randomly assigned to three treatment groups and received either the control diet (CON), high-fat diet (HF), or high-fat diet containing 6% Mulberry leaf powder (MLP) for four weeks. The CON group received a basal diet, the HF group received a basal diet supplemented with 10% lard, and the MLP group received a basal diet supplemented with 10% lard and 6% MLP. The trial lasted for four weeks. The growth performance, lipid metabolism parameters, immune globulins, cytokines, and fecal microbiota were measured. Results showed that there was no significant difference in growth performance. The MLP group appeared to have decreased (p < 0.05) the serum level of low-density lipoprotein cholesterol (LDL-C) and apoliprotein-A1(APO-A1) in serum. The MLP group appeared to have higher (p < 0.05) serum immune globulin A (IgA) levels. UPGMA results showed that the MLP group was closer to the CON group than to the HF group. LEfSe analysis showed that dietary supplementation with MLP contributed to an alteration in the genus Alloprevotella, Sarcina, and species belonging to the Bacteroides and Lactobacillus genus. Overall, the dietary supplementation of 6% MLP can improve lipid metabolism conditions and immunity in high-fat-diet-fed dogs, and can alter the gut microbial composition of dogs.


Challenges and opportunities: what we are learning from the clinical natalizumab experience.
The approval of natalizumab for relapsing forms of multiple sclerosis, and the subsequent voluntary suspension of its use due to an unexpected viral infection, is a cautionary tale of how much we have to learn about how to prioritize and perform the necessary research and development of novel therapeutics for human diseases, the ethics of placebo-controlled trials and the relationships between researchers, regulatory authorities and the pharmaceutical industry.


The Use of Aspirin in Contemporary Primary Prevention of Atherosclerotic Cardiovascular Diseases Revisited: The Increasing Need and Call for a Personalized Therapeutic Approach.
The use of aspirin has been widely accepted for the secondary prevention of atherosclerotic cardiovascular disease (ASCVD) in all patient populations, as the benefits linked to the reduction of clinical events outweigh the risk of major bleeding. However, despite the undisputable, though modest, potential of aspirin to reduce atherothrombotic events, its overall efficacy and safety in primary ASCVD prevention remains debatable, despite being used for this purpose for decades. The net clinical benefit of aspirin was brought into question by three recent large contemporary randomized controlled trials evaluating its role in various primary prevention populations (individuals with diabetes [ASCEND], an elderly population [ASPREE], and middle-aged adults at high estimated cardiovascular risk [ARRIVE]) and numerous large meta-analyses published during the past year. As a result, the usual generalized recommendations for the use of aspirin in patients with estimated intermediate to high ASCVD risk but without overt ASCVD have already been removed from most international guidelines. Since the primary prevention framework encompasses heterogenous groups of subjects with variable absolute ASCVD risk, a more individualized approach based on the best possible estimated ratio between the potential health benefits from fewer atherothrombotic events and harms because of potential increases in major bleeding is warranted in clinical practice. With this compromise, clinicians can better decide on the personalized use of aspirin in patients at high risk of major adverse cardiovascular events.


Glutamatergic transmission in the rat brain and gravitational stress.
The effects of hypergravity stress on L-[14C]-glutamate release from synaptosomes obtained from the rat brain and on the kinetic parameters of high-affinity glutamate transport activity were investigated. We found that hypergravity stress affected only the Ca(2+)-dependent component of L-[14C]-glutamate release. It did not modify the transporter affinity, but the maximum rate of uptake dropped from 12.5 +/- 3.2 to 5.6 +/- 0.9 nmol/min/mg of protein (in control rats and in animals subjected to hypergravity, respectively).


Cap snatching of yeast L-A double-stranded RNA virus can operate in trans and requires viral polymerase actively engaging in transcription.
Eukaryotic mRNA bears a cap structure (m(7)GpppX-) at the 5' terminus crucial for efficient translation and stability. The yeast L-A double-stranded RNA virus furnishes its mRNA with this structure by a novel cap-snatching mechanism in which the virus transfers an m(7)Gp moiety from host mRNA to the diphosphorylated 5' terminus of the viral transcript, thus forming on it an authentic cap structure (referred to as cap0) in the budding yeast. This capping reaction is essential for efficient viral expression. His-154 of the capsid protein Gag is involved in the cap transfer. Here we show that the virus can utilize an externally added viral transcript as acceptor in the capping reaction. The acceptor needs to be 5' diphosphorylated, consistent with the fact that the viral transcript bears diphosphate at the 5' terminus. A 5' triphosphorylated or monophosphorylated transcript does not function as acceptor. N7 methylation at the 5' cap guanine of mRNA is essential for cap donor activity. We also demonstrate that the capping reaction requires the viral polymerase actively engaging in transcription. Because the cap-snatching site of Gag is located at the cytoplasmic surface of the virion, whereas Pol is confined inside the virion, the result indicates coordination between the cap-snatching and polymerization sites. This will allow L-A virus to efficiently produce capsid proteins to form new virions when Pol is actively engaging in transcription. The coordination may also minimize the risk of accidental capping of nonviral RNA when Pol is dormant.


Vanadate-based transition-state analog inhibitors of Cre-LoxP recombination.
Cre recombinase exchanges DNA strands at the LoxP recognition site via transphosphorylation reactions that involve pentacoordinate transition states. We demonstrate that meta-vanadate ion (VO(3)(-)) and appropriate DNA substrates assemble a transition-state analog-like complex in the Cre active site. Meta-vanadate inhibits recombination of LoxP-derived oligonucleotide substrates that contain a gap at either or both scissile phosphates, but does not inhibit reactions with intact LoxP. The 3(')-hydroxyl group of the gapped substrate is required for inhibition, suggesting that vanadate is ligated by three oxo ligands. Assembly of the inhibited complex is slow (t(1/2)=19min at 4mM NaVO(3)) and requires Cre, substrates, and meta-vanadate. Holliday junction intermediates accumulated at lower meta-vanadate concentrations, suggesting that the second strand exchange is inhibited more readily than the first. The apparent K(D) for meta-vanadate is 1.5-2mM and binding shows positive cooperativity. This methodology may have general application for mechanistic studies of recombinase/topoisomerase-mediated strand exchange reactions.


Generation and characterization of induced pluripotent stem (iPS) cell line (SDUCTi001-A) using fibroblasts derived from male-healthy donor.
Induced pluripotent stem (iPS) cell lines have wide valuable applications in experimental research, including developmental, pathological, and drug screening studies. Using integration-free episomal plasmids, we have generated a new iPS cell line from a 26-year-old healthy male donor. Characterization of the established cell line confirmed the expression of pluripotency markers, differentiation into the three germ layers, and absence of chromosomal abnormalities.


Thyroid hormones increase Na+-Pi co-transport activity in intestinal brush border membrane: role of membrane lipid composition and fluidity.
In the present study, we documented the promising role of thyroid hormones status in animals in modulation of Na+-Pi transport activity in intestinal brush border membrane vesicles (BBMV) which was accompanied with alterations in BBM lipid composition and fluidity. Augmentation of net Pi balance in hyperthyroid (Hyper-T) rats was fraternized with accretion of Pi transport across BBMV isolated from intestine of Hyper-T rats as compared to hypothyroid (Hypo-T) and euthyroid (Eu-T) rats while Na+-Pi transport across BBMV was decreased in Hypo-T rats relative to Eu-T rats. Increment in Na+-Pi transport in intestinal BBMV isolated from Hyper-T rats was manifested as an increase in the maximal velocity (Vmax) of Na+-Pi transport system. Furthermore, BBMV lipid composition profile in intestinal BBM from Hyper-T was altered to that of Hypo-T rats and Eu-T rats. The molar ratio of cholesterol/phospholipids was higher in intestinal BBM from Hypo-T rats. Fluorescence anistropy of diphenyl hexatriene (rDPH) and microviscosity were significantly decreased in the intestinal BBM of Hyper-T rats and decreased in Hypo-T rats as compared to Eu-T rats which corroborated with the alteration in membrane fluidity in response to thyroid hormone status of animals. Therefore, thyroid hormone mediated change in membrane fluidity might play an important role in modulating Na+-Pi transport activity of intestinal BBM.


Modern management of the breech delivery.
Retrospective review of 457 breech deliveries failed to reveal a significant improvement in neonatal mortality and morbidity rates for those delivered by cesarean section vs. vaginal delivery. A slight but insignificant improvement in salvage was found for the 1,000 to 1,500 gram weight category when delivery was by cesarean section. Most of the deaths in the less than 1,500 gram breech presentation were associated with hyaline membrane disease or congenital defects incompatible with life and sepsis rather than directly related to the manner of breech delivery.


CD25 expression on donor CD4+ or CD8+ T cells is associated with an increased risk for graft-versus-host disease after HLA-identical stem cell transplantation in humans.
Graft-versus-host disease (GVHD) occurs in an unpredictable fashion after 30% to 50% of matched-related transplantations. The presence of increased frequencies of CD4(+)CD25(+) regulatory T cells in donor grafts has been shown to ameliorate GVHD after allogeneic transplantation in murine models. To determine whether a similar relationship exists in humans, we quantitated the coexpression of CD25 on CD4(+) and CD8(+) T cells within 60 donor grafts infused into matched siblings and examined GVHD incidence in the respective recipients. Recipients in whom GVHD developed received donor grafts containing significantly higher frequencies of CD4(+) T cells coexpressing CD25 than those who did not (median, 9.26% vs 2.22%; P =.004). Frequencies of donor graft CD8(+) T cells coexpressing CD25 were also higher (0.65% vs 0.14%; P =.002). Furthermore, transplant recipients who received grafts containing fewer CD4(+)CD25(+) and CD8(+)CD25(+) T cells were less likely to acquire acute GVHD, even though these donor-recipient pairs were similar to others with respect to relevant clinical variables. These data suggest that the coexpression of CD4 and CD25 may be insufficient to identify regulatory T cells in humans and that increased frequencies and numbers of CD25(+) T cells in donor grafts is associated with GVHD in transplant recipients.


Third-degree heart block developing in an HLA-B27-positive individual with a family history of ankylosing spondylitis.
A case of complete heart block is presented in a patient whose brothers were known to have ankylosing spondylitis. Uveitis and sacro-iliitis are well recognised among HLA-B27-positive siblings of ankylosing spondylitis. However, heart block is a rare occurrence as a sole presentation in siblings of patients with ankylosing spondylitis.


Antimicrobial susceptibilities of uropathogen Escherichia coli in renal transplant recipients: dramatic increase in ciprofloxacin resistance.
The urinary tract is the most common site of bacterial infections in renal transplant recipients. The management of urinary tract infections (UTI) in renal transplant recipients is becoming more difficult because of drug-resistant bacteria. The antimicrobial susceptibilities of uropathogen bacteria isolated from 398 patients who underwent renal transplantation between 2007 and 2011 were obtained from medical records. At least 1 UTI episode was diagnosed in 172 (43.2%) patients. Among the 703 bacteria isolated from these patients, Exherichia coli the most common pathogen, was isolated from 407/703 episodes (57.8%). Ciprofloxacin, co-trimoxazole, ceftriaxone, and gentamicin resistance rates were 59.4%, 85.7%, 40.7%, and 36.6%, respectively. Ninty six of 407 E. coli isolates (23.5%) were ESBL positive. Analysis of resistance rates in our center demonstrated ciprofloxacin resistance rate in uropathogenic E. coli to have increased gradually from 30.4% in 2003, 41.3% in 2007, and 59.4% in 2012. Instutional data regarding the etiologic agents and antimicrobial susceptibility results are important for proper management of patients with UTI.


Overlapping, complementary and site-specific expression pattern of genes of the EMILIN/Multimerin family.
The EDEN gene superfamily comprises genes that contain the EMI domain, a structural motif recently identified in proteins of the extracellular matrix. We report here the detailed expression pattern of genes of the EMILIN/Multimerin family, the most numerous group of EDEN superfamily, during mouse development. In situ hybridization has revealed that the EMILIN/Multimerin genes are particularly expressed in the cardio-vascular system and in mesenchymal cells. In general, the territories of expression of each gene are partially overlapping or complementary with that of other members of the family and, usually, more than one gene of the family is active in different tissues, consistent with the possibility of functional compensation. The analysis is particularly relevant in the interpretation of gene targeting experiments.


Cardiovascular tissue regeneration system based on multiscale scaffolds comprising double-layered hydrogels and fibers.
We report a technique to reconstruct cardiovascular tissue using multiscale scaffolds incorporating polycaprolactone fibers with double-layered hydrogels comprising fibrin hydrogel surrounded by secondary alginate hydrogel. The scaffolds compartmentalized cells into the core region of cardiac tissue and the peripheral region of blood vessels to construct cardiovascular tissue, which was accomplished by a triple culture system of adipose-derived mesenchymal stem cells (ADSCs) with C2C12 myoblasts on polycaprolactone (PCL) fibers along with human umbilical vein endothelial cells (HUVECs) in fibrin hydrogel. The secondary alginate hydrogel prevented encapsulated cells from migrating outside scaffold and maintained the scaffold structure without distortion after subcutaneous implantation. According to in vitro studies, resultant scaffolds promoted new blood vessel formation as well as cardiomyogenic phenotype expression of ADSCs. Cardiac muscle-specific genes were expressed from stem cells and peripheral blood vessels from HUVECs were also successfully developed in subcutaneously implanted cell-laden multiscale scaffolds. Furthermore, the encapsulated stem cells modulated the immune response of scaffolds by secreting anti-inflammatory cytokines for successful tissue construction. Our study reveals that multiscale scaffolds can be promising for the remodeling and transplantation of cardiovascular tissue.


Enantiomeric separation of norgestrel by reversed phase high-performance liquid chromatography using eluents containing hydroxypropyl-beta-cyclodextrin in stereoselective skin permeation study.
The chiral separation of norgestrel enantiomers using reversed-phase high-performance liquid chromatography (RP-HPLC) was studied with hydroxypropyl-beta-cyclodextrin (HP-beta-CD) as chiral mobile phase additive. The effect of mobile phase composition, concentration of HP-beta-CD and column temperature on enantioselective separation were investigated. The quantification properties of the developed RP-HPLC method were examined. A baseline separation of norgestrel enantiomers was achieved on a Agilent ZORBAX Eclipse XDB-C8 column (150 mm x 4.6 mm i.d., 5 microm). The mobile phase was a mixture of acetonitrile and phosphate buffer (pH 5.0, 20 mM) containing 25 mM HP-beta-CD (30:70, v/v) with a flow rate of 1.0 ml/min. The UV detector was set at 240 nm. Calibration curves were linear (n=8) in the range of 0.2-25 microg/ml, the limit of detection and quantitation were 0.10 and 0.20 microg/ml, respectively, for racemic norgestrel. The values of RSD of repeatability and intermediate precision for spiked sample were less than 4.8%. The method was successfully applied to the enantioselective determination of this drug in stereoselective skin permeation study.


A longitudinal study of maternal endothelial function, inflammatory response and uterine artery blood flow during the second half of pregnancy.
The objective of this study was to investigate serial changes in maternal endothelial function, inflammatory response and uterine artery blood flow in normal pregnancy, and to explore their inter-relation. In this prospective longitudinal observational study, 53 women with uncomplicated pregnancies were examined at 4-weekly intervals (248 observations) during 22-40 weeks of gestation. Uterine artery blood flow was measured using Doppler ultrasonography. Maternal endothelial function was assessed by flow-mediated vasodilatation (FMD) of the brachial artery. Circulating endothelial progenitor cells (EPC), defined as CD34(+) CD133(+) VEGFR2(+) cells, were quantified by flow cytometry. Biomarkers of inflammation, such as leptin and high sensitivity C-reactive protein (hsCRP), were measured in plasma samples. Multilevel modeling was used to investigate gestational-age-associated serial changes. The EPC increased from 6.5 to 12.3 per million mononuclear cells (p < 0.01) and FMD decreased from 16.3 to 13.4% (p = 0.20). Leptin increased from 18 to 22 ng/mL (p < 0.01), and hsCRP did not change significantly (p = 0.61). There was no significant association between FMD and EPC (p = 0.66). FMD was significantly associated with hsCRP (p = 0.002) and leptin (p = 0.003), but the EPC were not. Neither FMD nor EPC were significantly associated with uterine artery blood flow. Changes in FMD were significantly associated with inflammatory biomarkers, suggesting that the reduced nitric oxide-dependent vasodilatation in late gestation is related to maternal inflammatory response. As EPC and FMD did not correlate, mechanisms other than mobilization of EPC to repair endothelial damage must be responsible for the gestational-age-associated increase in EPC.


[Spectrophotometric examinations of the chlorhexidine content in the disinfected synthetic material of prosthesis].
The investigation showed that each acrylic plate adsorbs on its surface 41.62-46.78 micrograms of chlorhexidine.


Increase of glomerular filtration rate and renal plasma flow by insulin-like growth factor-I during euglycaemic clamping in anaesthetized rats.
Renal function was studied in anaesthetized rats receiving i.v. infusions of recombinant human insulin-like growth factor-I (IGF-I) under euglycaemic clamp conditions. IGF-I increased glomerular filtration rate up to 35% and renal plasma flow up to 100%, this increase being dose dependent with half-maximal stimulation at serum IGF-I concentrations of about 24 pmol ml-1. Renal vascular resistance was reduced up to 50%, filtration fraction decreased up to 30% and urine flow increased up to three fold while arterial blood pressure was unchanged. Renal haemodynamics were affected at serum IGF-I concentrations that did not stimulate total body glucose disposal during euglycaemic clamping. IGF-I seemed to regulate renal function through IGF-I receptors apparently independent of acute changes of glucose metabolism.


Alkyl-linked diglycerides inhibit protein kinase C activation by diacylglycerols.
Alkylacylglycerols are synthesized when choline-phospholipids are degraded by a phospholipase C. This class of compounds has been shown to have biological activities; however, the mechanism of action is unknown. A series of alkyl-linked diglycerides were synthesized and tested for activity in an in vitro assay for protein kinase C. When protein kinase C activity was stimulated with the synthetic diacylglyceride analog 1-oleoyl-2-acetyl-sn-glycerol, the addition of alkyl glycerides caused a concentration-dependent inhibition of protein kinase C activity. Comparison of the protein kinase C inhibition by this series of 1-O-alkyl-2-acyl analogs revealed that both saturated and unsaturated long-chain groups in position 1 were effective and that dietherglycerols with short-chain moieties in position 2 were also effective. It is concluded from these studies that the biological activity of alkyl-linked glycerides may be expressed through protein kinase C inhibition.


Enhanced orbital magnetic moments in magnetic heterostructures with interface perpendicular magnetic anisotropy.
We have studied the magnetic layer thickness dependence of the orbital magnetic moment in magnetic heterostructures to identify contributions from interfaces. Three different heterostructures, Ta/CoFeB/MgO, Pt/Co/AlOx and Pt/Co/Pt, which possess significant interface contribution to the perpendicular magnetic anisotropy, are studied as model systems. X-ray magnetic circular dichroism spectroscopy is used to evaluate the relative orbital moment, i.e. the ratio of the orbital to spin moments, of the magnetic elements constituting the heterostructures. We find that the relative orbital moment of Co in Pt/Co/Pt remains constant against its thickness whereas the moment increases with decreasing Co layer thickness for Pt/Co/AlOx, suggesting that a non-zero interface orbital moment exists for the latter system. For Ta/CoFeB/MgO, a non-zero interface orbital moment is found only for Fe. X-ray absorption spectra shows that a particular oxidized Co state in Pt/Co/AlOx, absent in other heterosturctures, may give rise to the interface orbital moment in this system. These results show element specific contributions to the interface orbital magnetic moments in ultrathin magnetic heterostructures.


Biochemical characterization of chitinase 2 expressed during the autolytic phase of the inky cap, Coprinellus congregatus.
Fungal cell walls consist of various glucans and chitin. The inky cap, Coprinellus congregatus, produces mushrooms at 25°C in a regime of 15 h light/9 h dark, and then the mushroom is autolyzed rapidly to generate black liquid droplets in which no cell walls are detected by microscopy. Chitinase cDNA from the mature mushroom tissues of C. congregatus, which consisted of 1,622 nucleotides (chi2), was successfully cloned using the rapid amplification of cDNA ends polymerase chain reaction technique. The deduced 498 amino acid sequence of Chi2 had a conserved catalytic domain as in other fungal chitinase family 18 enzymes. The Chi2 enzyme was purified from the Pichia pastoris expression system, and its estimated molecular weight was 68 kDa. The optimum pH and temperature of Chi2 was pH 4.0 and 35°C, respectively when 4-nitrophenyl N,N'-diacetyl-β-D-chitobioside was used as the substrate. The K m value and V max for the substrate A, 4-nitrophenyl N,N'-diacetyl-β-D-chitobioside, was 0.175 mM and 0.16 OD min(-1)unit(-1), respectively.


The Kager's fat pad radiological anatomy revised.
The aim of the study was to map connections within the Kager's fat pad between the structures which limit it. A retrospective re-review of 200 ankle magnetic resonance imaging (MRI) examination was conducted. Connections within the Kager's fat pad between the superior peroneal retinaculum, the fibulotalocalcaneal ligament, the posterior talocalcaneal ligament, the flexor hallucis longus, the paratenon of the Achilles tendon, the flexor retinaculum and bones were studied and a model of the connections was constructed. The superior peroneal retinaculum was directly connected with the fibulotalocalcaneal ligament in 85.5% of cases, the lateral part of the paratenon in 82.5%, the processus posterior tali in 78.5%, the posterior talofibular ligament in 32%, the flexor retinaculum in 29.5% and the anterior talofibular ligament in 9%. The fibulotalocalcaneal ligament was connected with the paratenon (on the medial side 88.5%, on the lateral side 68.5%), the flexor retinaculum in 70%, the posterior process of the talus in 79%, the osteofibrosus tunnel for the flexor hallucis longus in 53%, the posterior talofibular ligament in 43.5% and the calcaneofibular ligament in 10.5%. The posterior talocalcaneal ligament was connected with the fibulotalocalcaneal ligament in 71%, with the osteofibrosus tunnel for the flexor hallucis longus in 76.5%, with the flexor retinaculum in 70%. The plantaris tendon showed projection to the crural fascia in 34 of % cases. In the Kager's fat pad there are present more connections than previously reported. All the connections unit at the level of the posterior process of the talus.


Biocompatible photolithographic process for the patterning of biomolecules.
A new approach for the patterning of biomolecule layers is introduced based on the design of a new photoresist material with biocompatible lithographic processing requirements. The photoresist is based on poly(t-butyl acrylate), which allows positive imaging with very dilute basic solutions, tolerable by selected biomolecules used in immunoanalysis. Sensitivity at lambda>300 nm is obtained using a suitable sulfonium salt photoacid generator. Thermal steps also take place under conditions tolerable by biomolecules. Lithographic results on Si wafer substrates show resolution capabilities for equal lines/spaces, down to the range of 5-10 microm under biocompatible conditions. The process is also used on substrates of different geometries, including inner capillary surfaces. The patterning of the inner surface of a polystyrene capillary with mouse IgG is reported to demonstrate the principles of the above approach.


On the taxonomy of Afrotropical Coleophoridae (II) (Lepidoptera, Coleophoridae).
Eight new species of the genus Coleophora Hübner, 1822 are described: Coleophora kenyaensis sp. nov., C. vansoni sp. nov., C. elegans sp. nov., C. namaqua sp. nov., C. kamiesella sp. nov., C. kunenensis sp. nov., C. creola sp. nov., and C. hirsutella sp. nov. The genitalia of three previously described species are described and illustrated for the first time: Coleophora niphocrossa Meyrick, 1920, C. enchitis Meyrick, 1920 and C. textoria Meyrick, 1921. The larval case of C. enchitis is illustrated and data about its biology are given.


High versus low training frequency in cardiac rehabilitation using a systems model of training.
No study has reported the long term effects of cardiac rehabilitation, concerning the duration of beneficial effects of training program. The present study analyzed the influence of training session frequency on long-term beneficial effects in patients with coronary artery disease (CAD) undergoing phase 2 cardiac rehabilitation. Four patients with CAD completed 20 training sessions. Two patients were assigned to low training frequency (LTF) and two to high training frequency (HTF): three and five sessions per week(-1), respectively. The method was based on the systems model of training and required training quantification and the assessment of real exercise tolerance. Convolution of training quantity with real exercise tolerance provided the model exercise tolerance for every patient. The model parameters, the magnitude factor (k), and the time constant of decay (tau), were fitted from real and model exercise tolerances by the least squares method. LTF and HTF resulted in similar increases in exercise tolerance (12-14%). A model with one-component (fitness) allowed fitting exercise tolerance in all patients with r (2) = 0.77, 0.79, 0.84, and 0.91, respectively (p < 0.05). The addition of a second component did not improve the fit in any patient (p > 0.05). The k value was about twice as high with LTF (0.13 and 0.16 AU) than with HTF (0.05 AU for the two patients), whereas the tau value was about twice as low with LTF (37 and 41 days) than with HTF (72 and 89 days). The long term beneficial effects estimated by 4tau, were twice as long with HTF (288 and 356 days) than with LTF (148 and 164 days). We concluded that exercise tolerance was similarly increased with HTF and LTF but HTF training induced beneficial effects which were sustained twice as long.


Aquaporin gating.
An acceleration in the rate at which new aquaporin structures are determined means that structural models are now available for mammalian AQP0, AQP1, AQP2 and AQP4, bacterial GlpF, AqpM and AQPZ, and the plant SoPIP2;1. With an apparent consensus emerging concerning the mechanism of selective water transport and proton extrusion, emphasis has shifted towards the issues of substrate selectivity and the mechanisms of aquaporin regulation. In particular, recently determined structures of plant SoPIP2;1, sheep and bovine AQP0, and Escherichia coli AQPZ provide new insights into the underlying structural mechanisms by which water transport rates are regulated in diverse organisms. From these results, two distinct pictures of 'capping' and 'pinching' have emerged to describe aquaporin gating.


Age-associated changes in the effects of amphetamine on the blood-brain barrier of rats.
The effects of varying doses of amphetamine on the permeability of the blood-brain barrier (BBB) to albumin were studied in young and aged Fischer 344 rats. There was no change in the permeability of the BBB associated with aging as revealed by saline-treated control experiments. The changes in the permeability of the BBB did not correlate with the changes in blood pressure produced by corresponding doses of amphetamine in the young or aged animals. In the dose range reported the young and aged rats showed similar pressor responses to amphetamine. The aged rats showed a greater increase in the permeability of the BBB than the young, and this effect was seen with a lower dose of amphetamine in the aged rats. At the highest doses tested the permeability was restored to control levels in both the young and the aged rats. The enhanced vulnerability of the BBB of the aged rats may be related to a deterioration of the noradrenergic control of cerebrovascular permeability.


Does the 'high risk' patient with asymptomatic carotid stenosis really exist?
Recent evidence indicates that the risk of stroke symptoms in non-operated medically managed patients with asymptomatic severe carotid stenosis has fallen significantly over the last 25 years. This suggests concurrent improvements in vascular disease medical intervention efficacy. If the latest estimates of average annual stroke rate for non-operated patients are reflective of contemporary medical intervention and surgical stroke/death rates match those of the randomised trials, the current implication is that carotid surgery will not offer a stroke prevention advantage over medical intervention alone. Furthermore, it is still not possible to identify patients with asymptomatic severe carotid stenosis with a higher than average ipsilateral stroke risk despite current medical intervention. Even if such patients were one day reliably identified, they could also be at higher risk of stroke/death from instrumental intervention (surgery, angioplasty or stenting) and randomised trials will be required before being justification in routine clinical practice.


Sexual conflict and evolution in Trinidadian guppies.
Recent investigations have highlighted the importance of sexual conflict in the evolution of reproductive isolation. Examination of the Trinidadian guppy (Poecilia reticulata) shows how geographic variation in sexual conflict can mediate the emergence of isolating mechanisms. On the basis of pre-mating behaviour guppy females appear to be winning the battle of the sexes in low risk localities whereas males are apparently ahead in high-predation environments. However, the conclusion that sexual selection (through female choice) is replaced by sexual coercion of females (as a consequence of sneaky mating) in predator rich (and productive) assemblages takes no account of post-copulatory mechanisms. Recent work on sperm competition in guppies suggests coevolution between males and females may also occur in the post-mating, pre-zygotic arena. The potential for the evolution of reproductive isolation at each stage of the mating sequence is assessed.


Rapid diagnosis of lymphocytic meningitis by frequency-pulsed electron capture gas-liquid chromatography: differentiation of tuberculous, cryptococcal, and viral meningitis.
Cerebrospinal fluid specimens from patients with tuberculous (17 cases), cryptococcal (15 cases), and viral (14 cases) meningitis were analyzed by frequency-pulsed electron capture gas-liquid chromatography and mass spectrometry. Compounds that disappeared after therapy were found to be present in each of these specimens and were not detected in controls. They occurred in repetitive patterns such that these three types of meningitis could be rapidly distinguished. The compound associated with tuberculous meningitis has been tentatively identified. These finding have implications for rapid diagnosis, pathophysiological studies, and possible new therapeutic approaches.


The effect of sub-Tenon's anaesthesia on intraocular pressure.
To assess the effect of sub-Tenon's anaesthesia on intraocular pressure (IOP) prior to cataract surgery. Fifty consecutive patients undergoing phacoemulsification of cataract were recruited. Routine sub-Tenon's anaesthesia was administered with 5 ml unpreserved 2% lignocaine. IOPs were measured immediately prior to and at 1, 3, 5 and 10 min after injection. Efficacy was assessed subjectively by the operating surgeon. No ocular compression was used. Pre- and post-injection IOPs were compared using the Wilcoxon signed rank test, whereas all other results were compared with baseline using Student's two-tailed paired t-tests. All patients achieved good analgesia and akinesis. There was no significant difference between the IOP prior to and 1 min after injection. At all time intervals after 3 min there was a significant reduction in IOP compared with the pre-injection measurement. At 5 min, the mean IOP reduction was 2.72 mmHg and at 10 min IOP was lowered by 2.92 mmHg. Both reductions were statistically significant compared with baseline. Sub-Tenon's anaesthesia does not cause any significant rise in IOP, thereby possibly making it the anaesthetic technique of choice when an increase in IOP is undesirable. There is no indication for the use of an ocular pressure-reducing device when sub-Tenon's anaesthesia is employed.


Sleep disturbances in a case of brain-stem lesions; pharmacological study.
A pharmacological study was carried out of a case of severe insomnia following brain-stem lesions; several polygraphic controls were used. Initially total duration of sleep was brief (less than 4 h) with a high REM/NREM ratio and a short paradoxical sleep (PS) latency. In addition, periodic breathing and tremor were observed. Slow injection of delta-sleep-inducing peptide (DSIP) improved sleep both quantitatively and qualitatively, although PS latency remained short. These effects were reversible. The effects of 5-HTP + benzerazide, of L-DOPA + benzerazide (Modopar) and of clonazepam (Rivotril) were compared.


Factors influencing lion movements and habitat use in the western Serengeti ecosystem, Tanzania.
Protected areas that restrict human activities can enhance wildlife habitat quality. Efficacy of protected areas can be improved with increased protection from illegal activities and presence of buffer protected areas that surround a core protected area. Habitat value of protected areas also can be affected by seasonal variation in anthropogenic pressures. We examined seasonal space use by African lions (Panthera leo) within a core protected area, Serengeti National Park, Tanzania, and surrounding buffer protected areas with varying protection strengths. We used lion locations in logistic regression models during wet and dry seasons to estimate probability of use in relation to protection strength, distance to protected area edge, human and livestock density, distance to roads and rivers, and land cover. Lions used strongly protected buffer areas over the core protected area and unprotected areas, and moved away from protected area boundaries toward the core protected area when buffer protected areas had less protection. Lions avoided high livestock density in the wet season and high human density in the dry season. Increased strength of protection can decrease edge effects on buffer areas and help maintain habitat quality of core protected areas for lions and other wildlife species.


Long-term follow-up of ulcerative colitis in Taiwan.
The incidence of ulcerative colitis (UC) has been increasing in Asia recently, but little long-term follow-up data is available. We aimed to understand the clinical characteristics of UC patients in the National Taiwan University Hospital (NTUH), a tertiary referral center in Taiwan. A retrospective study was conducted to review data from January 1, 1988 through December 31, 2008 compiled at NTUH. Patients' clinical information, demographic data, endoscopic pictures, treatment regimens, pathologic, and outcome details were reviewed, recorded, and analyzed. A total of 406 patients were included (233 males and 173 females; median age at diagnosis was 36 years). The follow-up period ranged from 0.25 to 40.8 (mean, 7.3) years. The prevalence of UC in Taiwan was at least 7.4/100,000 in 2008. Bloody stool was the most common presentation (77.3%). Total colon was the most common (41.0%) disease involvement and proctitis the least common (21.1%). Six patients (1.5%) died during the follow up. Most of the UC patients (72.4%) could be controlled with 5-aminosalicylic acid alone, but about one third (30.9%) were admitted for treating the UC or UC-related complications. Twenty-three patients (5.5%) were treated surgically. Extra-gastrointestinal tract manifestations were noted in 4.5% of the UC patients, with primary sclerosing cholangitis (6 in 406, 1.5%) the most common. Colon cancer/severe dysplasia occurred in six (1.5%) of the patients. The incidence of UC has increased in Taiwan. Interestingly, CRC/dysplasia and PSC occur more frequently here than in other Asian nations.


Jejunoileal atresia: a 27-year experience.
In this study, the authors review cases of jejunoileal atresia (JIA) to evaluate their surgical treatment strategy. Eighty-eight neonates who underwent surgical repair for JIA were divided into four groups for the type of lesion: group 1, membranous (n = 23), group II, interrupted (n = 49), group III, multiple (n = 9), and group IV, apple-peel (n = 7). Group I patients were treated with membranectomy or bowel resection and anastomosis, group II with resection of the dilated bowel and one anastomosis, group III with two to six multiple anastomoses to preserve bowel length, and group IV with minimal bowel resection and bowel anastomosis. During surgery a uniform protocol was used to minimize bowel resection and to perform an end-to-end single layer anastomosis using either Halsted horizontal mattress or conventional interrupted sutures. Mortality, morbidity, days for functional recovery, and central venous nutrition (CVN) were included in the review. Of 88 patients, three died of causes unrelated to operation for JIA. Nine patients underwent an additional laparotomy for leakage (n = 4) and obstruction (n = 5). Oral feeding was allowed on day 5.4+/-4.3 and full caloric intake via the enteric route on day 12.5+/-10.0. Twenty-one patients required CVN for 32.4+/-19.1 days. None required a long-term treatment for the short bowel syndrome. This study concludes that efforts to preserve bowel length are laudable to avoid the short bowel syndrome and that an end-to-end single layer anastomosis contributes to early recovery of bowel function.


Innovative methods of biliary tract diagnosis: intraductal ultrasound and tissue acquisition.
Tissue can be acquired from the bile and pancreatic duct with either brush cytology or biopsy forceps, often without the need for sphincterotomy. Although the diagnosis of malignancy with these sampling techniques is often specific, the sensitivity is limited, and a wide range of diagnostic accuracies has been reported. A combination of biopsy and brush cytology along with some newer techniques in the development stage may increase the sensitivity. Intraductal ultrasound is a newer technique offers the endoscopist the ability to image the bile and pancreatic duct wall, adjacent organs, and vascular structures with a high degree of detail. This visualization can be accomplished during the course of an ERCP or percutaneously under fluoroscopic guidance. Indications for the procedure include detection of choledocholithiasis, differentiation of benign and malignant ductal strictures, pancreaticobiliary tumor staging, and detection of various pancreatic tumors. Continued research into the design of the probes, which could improve durability and extend the depth of penetration, may promote more widespread use of this novel technology.


Myoglobinuria in two patients with Duchenne muscular dystrophy after treatment with zoledronate: a case-report and call for caution.
Rhabdomyolysis with myoglobinuria is a recognized complication of dystrophinopathies. It can be triggered by infections, exercise or volatile anesthetics. To our knowledge, it has never been reported in boys with Duchenne muscular dystrophy (DMD) after the administration of bisphosphonates. We report two patients with DMD who presented an apparent transient rhabdomyolysis with myoglobinuria after zoledronate administration. Possible mechanisms could involve hypophosphatemia, a known dose-dependent side effect of bisphosphonates, and/or direct myotoxicity of biphosphonates. Physicians and families should be aware of rhabdomyolysis with myoglobinuria as a potential uncommon side effect of bisphosphonates in DMD, in particular of zoledronate.


Acute effect of various doses of isosorbide-5-mononitrate on hemodynamic and exercise performance in coronary artery disease.
The optimal dose of nitrates is still controversial, as chronic usage of too high a dose may result in a decrease in vascular response. The dose should be determined to reduce preload acutely, and not to decrease the activity during chronic therapy. To determine this dose, 50 patients with angiographically confirmed coronary artery disease were studied. The effects on heart rate and mean systemic and pulmonary arterial pressures at rest and during exercise, and work capacity and duration were measured. The patients were classified into 5 groups, receiving placebo or isosorbide-5-mononitrate, 5, 10, 20 and 50 mg, respectively, as a single oral dose. Placebo had no effect on the measured parameters. All doses of isosorbide-5-mononitrate reduced mean pulmonary arterial pressure: 10 mg--by 16% at rest and 24% during exercise; 20 mg--24% and 34%, respectively (a near maximal effect); and 50 mg--27% and 38%. Similar results were found also for work capacity: 10 mg increased work capacity by 33%; 20 mg--79%; and 50 mg--56%. Thus, the therapeutically optimal single dose is about 20 mg. Higher doses produce no additional benefit and increase the risk for tolerance development.


Characterization of lymphocyte subpopulations in the blood of the cynomolgus monkey using flow cytometry.
This study was performed to assess the usefulness of flow cytometry in comparison with acid alpha-naphthyl acetate esterase (ANAE) staining for the enumeration of lymphocyte subpopulations in cynomolgus monkeys. For flow cytometry, use was made of mouse antihuman leukocyte monoclonal antibody (T11), recognized to undergo cross-reaction with monkey T lymphocytes or antimonkey IgG serum labelled with fluoroscein isothiocyanate. The percentages of lymphocyte subpopulations in mononuclear cells of thirty healthy female cynomolgus monkeys were 72.1 +/- 2.4% for T cells and 24.6 +/- 2.4% for B cells as assayed by flow cytometry evaluation, and 63.8 +/- 2.5% for T cells and 15.8 +/- 2.2% for B cells as determined by ANAE staining. Although the percentage of T cells shown by ANAE staining was significantly lower than that seen with flow cytometry, the coefficient of correlation indicated a close correlation in both T and B cell subsets between these two methods. In monkeys receiving 20 mg kg-1 cyclophosphamide daily for 14 days, counts of all leukocytes and T and B cells were decreased, whereas animals treated with 1.0 mg kg-1 muroctasin daily for 14 days showed higher monocyte and neutrophil counts without changes in T or B cells. These results suggest that flow cytometry evaluation has several advantages over ANAE staining with respect to rapidity and precision in toxicological studies using large numbers of monkeys.


Deracemization in a Complex Quaternary System with a Second-Order Asymmetric Transformation by Using Phase Diagram Studies.
Invited for the cover of this issue is the group of Gérard Coquerel at Université de Rouen Normandie. The image depicts a pyramid-like tetrahedron of the quaternary phase diagram showing where symmetry breaking can take place. Read the full text of the article at 10.1002/chem.201903338.