Sustained levels of FGF2 maintain undifferentiated stem cell cultures with biweekly feeding.
An essential aspect of stem cell culture is the successful maintenance of the undifferentiated state. Many types of stem cells are FGF2 dependent, and pluripotent stem cells are maintained by replacing FGF2-containing media daily, while tissue-specific stem cells are typically fed every 3rd day. Frequent feeding, however, results in significant variation in growth factor levels due to FGF2 instability, which limits effective maintenance due to spontaneous differentiation. We report that stabilization of FGF2 levels using controlled release PLGA microspheres improves expression of stem cell markers, increases stem cell numbers and decreases spontaneous differentiation. The controlled release FGF2 additive reduces the frequency of media changes needed to maintain stem cell cultures, so that human embryonic stem cells and induced pluripotent stem cells can be maintained successfully with biweekly feedings.


X-linked dominant protoporphyria in a Chinese pedigree reveals a four-based deletion of ALAS2.
X-linked dominant protoporphyria (XLDPP) is a rare, hereditary disorder that leads to hepatobiliary and hematologic abnormalities including increased erythrocyte protoporphyrin, cutaneous photosensitivity, and decreased iron stores that is caused by a pathogenic mutation of ALAS2 gene. This study aimed to confirm the existence of XLDPP in a Chinese pedigree. We observed and described the dermatoscopic findings of this disorder under dermoscopy, and assessed photo damage in XLDPP patients using the Fotofinder system and very high frequency (VHF) skin ultrasonic system. We performed next generation sequencing and Sanger sequencing to detect and confirm genetic variants in DNA samples from the XLDPP family. Moreover, we monitored the hepatobiliary function as well as hematologic changes in related family members. As compared to unaffected control subjects, patients exhibited evidence of severe cutaneous photodamage, causing photoaging, an increase in the size of the gallbladder, increased levels of protoporphyrin in red blood cells, an increase in blood levels of uroporphyrin and hematoporphyrin, and iron deficiency. XLDPP was validated by the identification of a four-base-pair deletion (c.1706_1709delAGTG, p.E569fs) in ALAS2 (NM_000032.4) in the proband which segregated with the disease in an X-linked dominant pattern, with hemizygous males being more severely affected than heterozygous females. We also found a missense variant in GATA Binding Protein 1 (GATA1).


Reduction of the P5A-ATPase Spf1p phosphoenzyme by a Ca2+-dependent phosphatase.
P5 ATPases are eukaryotic pumps important for cellular metal ion, lipid and protein homeostasis; however, their transported substrate, if any, remains to be identified. Ca2+ was proposed to act as a ligand of P5 ATPases because it decreases the level of phosphoenzyme of the Spf1p P5A ATPase from Saccharomyces cerevisiae. Repeating previous purification protocols, we obtained a purified preparation of Spf1p that was close to homogeneity and exhibited ATP hydrolytic activity that was stimulated by the addition of CaCl2. Strikingly, a preparation of a catalytically dead mutant Spf1p (D487N) also exhibited Ca2+-dependent ATP hydrolytic activity. These results indicated that the Spf1p preparation contained a co-purifying protein capable of hydrolyzing ATP at a high rate. The activity was likely due to a phosphatase, since the protein i) was highly active when pNPP was used as substrate, ii) required Ca2+ or Zn2+ for activity, and iii) was strongly inhibited by molybdate, beryllium and other phosphatase substrates. Mass spectrometry identified the phosphatase Pho8p as a contaminant of the Spf1p preparation. Modification of the purification procedure led to a contaminant-free Spf1p preparation that was neither stimulated by Ca2+ nor inhibited by EGTA or molybdate. The phosphoenzyme levels of a contaminant-free Spf1p preparation were not affected by Ca2+. These results indicate that the reported effects of Ca2+ on Spf1p do not reflect the intrinsic properties of Spf1p but are mediated by the activity of the accompanying phosphatase.


[Assessment of the endangered status and conservation priorities for the rare and endangered plant species in Inner Mongolia, China].
With 127 rare and endangered plant species of Inner Mongolian in Catalogue of Rare and Endangered Plants in China, China Plant Red Book, National Key Protected Wild Plants List (List No. 1), China Species Red List, Red List of Biodiversity in China: Volume of Higher Plants, List of Rare and Endangered Plants in Inner Mongolia and Atlas of Rare and Endangered Plants in Inner Mongolia as objects, an evaluation system of the threatened grades and conservation priority of rare and endangered plants were built based on data collection and consultation with experts. We set the five criteria, including endangered coefficient, genetic coefficient, utilization coefficient, habitat coefficient, and reproduction coefficient, under which there were 17 subordinate indicators. The analytic hierarchy process was employed to determine the weight of indicators in the system and calculate the endangered grades and conservation priority grades for the rare and endangered species. According to the results of evaluations, two critically endangered (CR) species, 13 endangered (EN) species, 37 vulnerable (VU) species, 44 near threatened (NT) species, and 31 least concern (LC) species were identified, accounting for 1.6%, 10.2%, 29.1%, 34.7% and 24.4% of the total, respectively. Among those species, 52 species were threatened, namely CR, EN and VU species, accounting for 40.9% of the total. The evaluation results of conservation priority grades were: 35 species of Class 1 protected plants, 72 species of Class 2 protected plants, and 20 species of Class 3 protected plants, accounting for 27.6%, 56.7% and 15.8% of the total, respectively. According to the results of evaluation comparison between Red List of Biodiversity in China: Volume of Higher Plants and List of Rare and Endangered Plants in Inner Mongolia, endangered grades of 75 plant species and the protection classes of 62 plant species were calibrated. In this evaluation, the endangered grades of nine plant species and the protection classes of 32 plant species were newly added.


Lesions in four parts of the basal forebrain change basic behaviour in rats.
Subdivisions of the basal forebrain (BF) are postulated to be differently involved in the regulation of behavioural functions. We compared 16 parameters of rat open field behaviour before, 10 days and 60 days after lesions in four BF subdivisions, the anterior and the posterior nucleus basalis magnocellularis (NBMa, NBMp), the vertical and the horizontal diagonal band nucleus (VDB, HDB) with sham-operated control rats. Every lesion type caused a significantly different pattern of significant behavioural changes. VDB and NBMp lesions both produced different patterns of ambulatory and exploratory hyperactivity, whereas NBMa lesions caused hypoactivity. Changes after HDB lesions were relatively small. The rate of habituation also shared changes that differed according to the lesion site. These changes in basic unlearned behaviour are likely to interact with more complex functions such as cognitive processes, learning and memory.


Secretion of Alzheimer's disease Abeta amyloid peptide by activated human platelets.
Alzheimer's disease (AD) is characterized by the deposition of Abeta (betaA4) peptides of 39 to 43 amino acid residues, which are normal cellular metabolic products derived by proteolysis of the amyloid precursor protein (APP). The physiologic function of Abeta/APP in vivo is poorly understood. We analyzed human platelets for Abeta production by immunoprecipitation coupled to immunoblotting. A 4-kd Abeta fragment that comigrates with an Abeta40 synthetic peptide and reacts with several antibodies specific for the N- and C-termini of Abeta is detected. The majority of platelet Abeta appears to end at residue 40, as determined by immunoreactivity with an Abeta40-specific antibody. Furthermore, Abeta is secreted upon platelet stimulation with the physiologic agonists thrombin and collagen, together with secretion of soluble APP (sAPP). A comparison between serum and plasma shows a 1.6-fold increase in Abeta levels and a 2.4-fold increase in sAPP levels in serum. This is consistent with the view that platelets are the primary source of circulating Abeta and APP. The release of platelet Abeta by physiologic stimuli suggests that it may play a role in platelet aggregation and coagulation or in the repair mechanisms associated with injury.


PET scanning and measuring the impact of treatment.
Positron emission tomography (PET) scanning with F18-fluorodeoxyglucose or FDG is a becoming a standard method for tumor staging. The prediction and evaluation of therapy response are newer applications of FDG-PET. PET often offers an early readout of treatment efficacy and is an attractive alternative to conventional anatomic assessments of treatment response. This article reviews the methods available with PET to monitor therapy response. Disease specific applications of PET imaging are then reviewed. While FDG is the most commonly used radiotracer for PET, many other radioligands could be applied in the future.


Water-Solubilizing Hydrophobic ZnAgInSe/ZnS QDs with Tumor-Targeted cRGD-Sulfobetaine-PIMA-Histamine Ligands via a Self-Assembly Strategy for Bioimaging.
Exploring the organic-to-aqueous phase transfer of quantum dots (QDs) is significant for achieving their versatile applications in biomedical fields. In this thematic issue, surface modification, size control, and biocompatibility of QDs and QDs-based nanocomposites are core problems. Herein, the new highly fluorescent tumor-targeted QDs-clusters consisting of ZnAgInSe/ZnS (ZAISe/ZnS) QDs and sulfobetaine-PIMA-histamine (SPH) polymer with the ανβ3 integrin receptor cyclic RGD (c-RGD) were developed via ligand exchange and an accompanying self-assembly process. It was found that the structure of RGD-SPH QDs-clusters was propitious to reduce the capture of reticulo-endothelial system (RES) in virtue of external stealth ligands, and benefit to selectively accumulate at the tumor site after intravenous injection via active tumor targeting cooperated with the enhanced permeability and retention (EPR) effect. In the meantime, those clusters also recognized and enriched the cell surface when cocultured with the ανβ3 integrin receptor overexpressed malignant cells (U87MG tumor). On the basis of the results, fabricating mutil-functional nanocomposites integrated with the long-term circulation and dual-targeting effects should be an interesting strategy for imaging cancer in vitro and in vivo.


'Test-and-treat': the end of the HIV epidemic?
The aim is to review recent literature on 'test-and-treat', a prevention strategy that promotes high levels of HIV testing and initiating antiretroviral therapy upon diagnosis, regardless of CD4 cell count. Antiretroviral therapy (ART) has been shown to dramatically reduce viral load which is strongly associated with the risk of transmission, therefore there is the potential to reduce HIV transmissions with ART. Recent papers from observational studies on heterosexual sero-discordant couples found an overall rate of transmission of HIV-1 from ART-treated patients of 0.46 per 100 person-years, confirming the possibility of using ART as a prevention strategy. Several models have been used to predict the effect of this strategy and the potential risks of it. Randomized controlled trials are currently ongoing investigating the effect of ART on reducing infectiousness and the feasibility of this policy. More precise estimations of the transmission risk under virally suppressive ART (especially in MSM) and of change in sex risk behaviour at diagnosis and at start of ART are needed. Further, the benefit to individual health of very early ART initiation and the feasibility of this policy need to be evaluated. Achieving very high levels of testing should be a high priority due to the benefits of initiating ART in all those who are in need (CD4 cell count < 350 cells/μl) and potential benefits on incidence due to reductions in risk behaviour in those diagnosed. Use of ART immediately at diagnosis in those with high CD4 cell counts should await results from further studies.


Tension and disordered eating behaviors in the daily lives of adolescents and young adults from the general population: Associations and moderating role of trait emotion regulation.
Experiences of tension and difficulties in emotion regulation have been linked to eating pathology in clinical samples and are targeted in respective treatment approaches. The present study aimed to investigate the impact of tension on engagement in disordered eating behaviors (DEBs) and potential moderating effects of trait emotion regulation in young people from the general population. A subsample of 971 adolescents and young adults from an epidemiological cohort study reported on levels of tension and four different DEBs (skipping eating, restrained eating, eating large amounts of food, loss-of-control-eating) via ecological momentary assessment (EMA), as well as on trait emotion regulation via questionnaire. In multilevel models, momentary tension did not predict levels of subsequent DEBs. However, higher average levels of tension across the EMA period predicted higher levels of all DEBs. No interactions with emotion regulation emerged. Individuals experiencing overall higher levels of tension appear to be more prone to engaging in DEBs.


[Fournier gangrene].
Since first described in 1883 by Fournier only 420 cases of Fournier's gangrene have been published worldwide during the last 100 years. The anatomic association between the fascies of penis, scrotum, perineum, groin and gluteal favors the fast spread of tissue necrosis. We report two cases in which the penis, scrotum and a great part of the trunk and extremities were affected. Extensive débridement, including removal of the scrotum, and antibiotic treatment permitted us to get rid of the symptoms. Reconstruction of the skin defects was done in a second step by applying mesh grafts. The histological findings correspond to those found in cases of necrotizing fasciitis. The basic difference between these two illnesses is their localization. While Fournier's gangrene, as a special form, is localized primarily in the anogenital area, fasciitis may arise in all locations. For successful treatment of Fournier's gangrene speedy radical débridement and local application of antibiotics to cover the entire area are required.


Investigation into the Photoluminescence Red Shift in Cesium Lead Bromide Nanocrystal Superlattices.
The formation of cesium lead bromide (CsPbBr3) nanocrystal superlattices (NC SLs) is accompanied by a red shift in the NC photoluminescence (PL). The values of the PL red shift reported in the literature range from none to ∼100 meV without unifying explanation of the differences. Using a combination of confocal PL microcopy and steady-state optical spectroscopies we found that an overall PL red shift of ∼96 meV measured from a macroscopic sample of CsPbBr3 NC SLs has several contributions: ∼ 10-15 meV from a red shift in isolated and clean SLs, ∼ 30 meV from SLs with impurities of bulklike CsPbBr3 crystals on their surface, and up to 50 meV or more of the red shift coming from a photon propagation effect, specifically self-absorption. In addition, a self-assembly technique for growing micron-sized NC SLs on the surface of perfluorodecalin, an inert perfluorinated liquid and an antisolvent for NCs, is described.


The magnetic penetration depth influenced by the proximity to the surface.
The effect of smooth inhomogeneities near a superconductor boundary on the magnetic penetration depth λ is studied with emphasis on the proximity-induced spatial dependence of the Cooper pair amplitude. The influence of surface pair breaking or pair formation on λ is described within the Ginzburg-Landau theory, with no model assumptions, for both strongly type-II and strongly type-I homogeneous superconductors. Generic values of λ, which can differ greatly from the London penetration depth, are identified and demonstrated to be induced by large-scale inhomogeneities, when superconductivity is strongly suppressed on the surface.


Mastoid and middle ear cavity obliteration for control of otitis media.
Total soft tissue obliteration of the mastoid-middle ear cavity and primary closure of the external auditory meatus were employed in six patients to control chronic suppurative otitis media. Previously, all these patients had unsuccessful surgery and all but one had profound loss of cochlear function. Thorough removal of cholesteatoma, granulation tissue, and ear canal skin is necessary before total obliteration can be performed safely. Wound healing was complete by one to two weeks postoperatively and all patients have been asymptomatic for two to seven years.


Evaluation of Overactive Bladder in Male Antidepressant Users: A Prospective Study.
In this study, we investigated overactive bladder (OAB) functions in male patients who used antidepressant drugs (ADs) that were previously examined in female patients, based on conflicting data in literature regarding the effects of AD on OAB and the differences between male and female urinary system physiologies (anatomical and hormonal). The study included 202 male patients (a control group of 90 healthy subjects, and an experimental group of 112 patients taking ADs for different disorders). All the patients completed the overactive bladder-validated 8 (OAB-V8) questionnaire, the International Consultation on Incontinence Questionnaire-Short Form (ICIQ-SF), and the Beck Depression Inventory (BDS). The OAB-V8, ICIQ-SF, and BDS scores for the antidepressant users were significantly higher than those of the control group. The highest prevalence of OAB symptoms was observed in patients taking venlafaxine (68.2%), and the lowest prevalence was in patients taking sertraline (28.0%). Moreover, the frequency of OAB between the antidepressant groups was statistically significant. The univariate logistic regression analyses showed a significant relationship between the presence of OAB, antidepressant usage, BDS score, and the age of a patient. In the multivariate logistic regression analyses, the association between the presence of OAB and antidepressant usage was statistically significant. The present study showed that the incidence of OAB and the severity of OAB symptoms increased in males using antidepressants for various disorders. This may have been due to unique pharmacological effects, on a molecular or individual level, of serotonin-norepinephrine reuptake inhibitors.


Effectiveness of telephone-based referral care management, a brief intervention to improve psychiatric treatment engagement.
This study examined the effectiveness of a telephone-based referral care management (TBR-CM) intervention for improving engagement in psychiatric treatment. From September 2005 to May 2006, 169 primary care patients at the Philadelphia Veterans Affairs Medical Center completed a psychiatric diagnostic interview and were identified as needing psychiatric care. From this total of eligible patients, 113 (67%) gave informed consent and were randomly assigned to receive either usual care or the intervention. Usual care consisted of participants' being schedule for a behavioral health care appointment, followed by a letter and reminder by telephone. The intervention group received the same, plus one or two brief motivational telephone sessions. Participant interviews and medical records provided study data. Research participants were primarily African American and 22-83 years old. In the sample, 40 patients (39%) had severe depression, 40 (39%) had substance use problems, and 33 (22%) had co-occurring severe depression and substance abuse. Overall, 40 participants (70%) in the intervention group compared with 18 (32%) in the usual care group engaged in at least one psychiatric treatment appointment (p<.001). Analyses also indicated that on average the intervention group attended more appointments (more than three) compared with the usual care group (less than two) (p=.008). The TBR-CM intervention program was effective at improving psychiatric treatment engagement. Future research is necessary to examine effectiveness of TBR-CM in more heterogeneous and larger samples and to evaluate economic benefits versus costs of intervention delivery.


Biomechanical Three-Dimensional Finite Element Analysis of Single Implant-Supported Prostheses in the Anterior Maxilla, with Different Surgical Techniques and Implant Types.
The aim of this study was to use three-dimensional finite element analysis to analyze the stress distribution transferred by single implant-supported prostheses placed in the anterior maxilla using different connections (external hexagon, internal hexagon, or Morse taper), inclinations of the load (0, 30, or 60 degrees), and surgical techniques for placement (monocortical/conventional, bicortical, or bicortical with nasal floor elevation). Nine models representing a bone block of this region were simulated by computer-aided design software (InVesalius, Rhinoceros, SolidWorks). Each model received one implant, which supported a cemented metalloceramic crown. Using FEMAP software, finite elements were discretized while simulating a 178-N load at 0, 30, and 60 degrees relative to the long axis of the implant. The problem was solved in NEi Nastran software, and postprocessing was performed in FEMAP. Von Mises stress and maximum principal stress maps were made. The von Mises stress analysis revealed that stress increased with increasing inclination of the load, from 0 to 30 to 60 degrees. Morse taper implants showed less stress concentration around the cervical and apical areas of the implant. The bicortical technique, associated or not with nasal floor elevation, contributed to decreasing the stress concentration in the apical area of the implant. Maximum principal stress analysis showed that the increase in inclination was proportional to the increase in stress on the bone tissue in the cervical area. Lower stress concentrations in the cortical bone were obtained with Morse taper implants and the bicortical technique compared with other connections and surgical techniques, respectively. Increasing the inclination of the applied force relative to the long axis of the implant tended to overload the peri-implant bone tissue and the internal structure of the implants. The Morse taper connection and bicortical techniques seemed to be more favorable than other connections or techniques, respectively, for restoring the anterior maxilla.


Physicochemical and sensory properties of milk supplemented with lactase microcapsules coated with enteric coating materials.
In this paper, we report the physicochemical and sensory properties of milk supplemented with a powder of microencapsulated lactase. The core material was lactase (β-galactosidase), the primary coating material was medium-chain triglyceride (MCT), and the secondary (enteric) coating material was either hydroxypropyl methylcellulose phthalate (HPMCP) or shellac, comparing both against market milk as a control. The physicochemical properties of both types of microcapsules were analyzed, including the particle size, zeta potential, and in vitro release behavior. To survey the stability of the microcapsules in milk during storage, we studied the residual lactose content and pH. Furthermore, to determine the properties of milk supplemented with the microcapsules, changes in color and sensory properties were evaluated during storage. The particle sizes (volume-weighted mean; D[4,3]) of the microcapsules coated with HPMCP or shellac were 2,836 and 7,834 nm, respectively, and the zeta potential of the capsules coated with shellac was higher than the zeta potential of those coated with HPMCP. The pH levels of milk supplemented with the lactase microcapsules were similar to those of the control (unsupplemented market milk); however, for milk supplemented with HPMCP-coated microcapsules, the pH was slightly lower. The core material, lactase, was released from the microcapsules during 12-d storage, and 18.82 and 35.09% of lactose was hydrolyzed in the samples for HPMCP- and shellac-coated microcapsules, respectively. The sensory characteristics of milk containing microcapsules coated with HPMCP did not show significant differences from the control, in terms of sweetness or off-taste, until 8 d of storage. However, shellac-coated microcapsules showed significant difference in sweetness and off-taste at d 8 and 6 of storage, respectively. The color of milk containing HPMCP-coated microcapsules did not show a significant difference during storage. However, that containing shellac-coated microcapsules was somewhat higher in color values than others. In particular, it showed significance from 0 to 4 d storage in L* and C* values. In conclusion, a powder of lactase microcapsules coated with HPMCP can be suitable as a supplement for milk.


A pilot study of anti-nephrin antibodies in podocytopaties among adults.
Minimal change disease (MCD) and focal segmental glomerulosclerosis (FSGS) are podocytopathies characterized by damage to the glomerular filtration barrier, leading to proteinuria and nephrotic syndrome. The production of anti-podocyte antibodies has been proposed as potential circulating factors contributing to the development of these conditions. The aim of the study is to evaluate the levels of anti-nephrin antibodies in patients with podocytopathies and healthy subjects. In this study, a total of 77 patients with active glomerulopathy and 11 healthy subjects were included. Forty one patients were diagnosed with FSGS, 11 with MCD, and 25 with MN. To measure the levels of anti-nephrin antibodies, enzyme-linked immunosorbent assay was used. The levels of antibodies to nephrin were significantly higher in patients with MCD 61.2 [28.9-66.3] ng/mL and FSGS 32.5 [17.2-58.4] ng/mL compared to MN 20.3 [14.4-38.4] and healthy individuals 15.3 [12-18.9] ng/mL, p < .05. In patients with primary FSGS, the levels of antibodies to nephrin were significantly higher 45.2 [20-64.3] ng/mL compared to patients with secondary FSGS 26.7 [11.2-44.1] ng/mL, p < .05. There were no significant differences in the remission rate between the anti-nephrin antibodies positive and negative groups (log-rank test: p = .158). The level of anti-nephrin antibodies was found to be significantly higher in patients with MCD and pFSGS compared to those with sFSGS, MN, and healthy subjects. Anti-nephrin antibodies in MCD and primary FSGS may be associated with the severity of podocytopathies, however they did not have an impact on the response to therapy.


Reduced Multidrug Susceptibility Profile Is a Common Feature of Opportunistic Fusarium Species: Fusarium Multi-Drug Resistant Pattern.
The resistance among various opportunistic Fusarium species to different antifungal agents has emerged as a cause of public health problems worldwide. Considering the significance of multi-drug resistant (MDR), this paper emphasizes the problems associated with MDR and the need to understand its clinical significance to combat microbial infections. The search platform PubMed/MEDLINE and a review of 32 cases revealed a common multidrug-resistant profile exists, and clinically relevant members of Fusarium are intrinsically resistant to most currently used antifungals. Dissemination occurs in patients with prolonged neutropenia, immune deficiency, and especially hematological malignancies. Amphotericin B displayed the lowest minimum inhibitory concentrarions (MICs) followed by voriconazole, and posaconazole. Itraconazole and fluconazole showed high MIC values, displaying in vitro resistance. Echinocandins showed the highest MIC values. Seven out of ten (70%) patients with neutropenia died, including those with fungemia that progressed to skin lesions. Clinical Fusarium isolates displayed a common MDR profile and high MIC values for the most available antifungal agents with species- and strain-specific differences in antifungal susceptibility. Species identification of Fusarium infections is important. While the use of natamycin resulted in a favorable outcome in keratitis, AmB and VRC are the most used agents for the treatment of fusariosis in clinical settings.


Impact of remuneration on guideline adherence: empirical evidence in general practice.
Changes in the Dutch GP remuneration system provided the opportunity to study the effects of changes in financial incentives on the quality of care. Separate remuneration systems for publicly insured patients (capitation) and privately insured patients (fee-for-service) were replaced by a combined system of capitation and fee-for-service for all in 2006. The effects of these changes on the quality of care in terms of guideline adherence were investigated. A longitudinal study from 2002 to 2009 using data from patient electronic medical records in general practice. A multilevel (patient and practice) approach was applied to study the effect of changes in the remuneration system on guideline adherence. 21 421 to 39 828 patients from 32 to 52 general practices (dynamic panel of GPs). Sixteen guideline adherence indicators on prescriptions and referrals for acute and chronic conditions. Guideline adherence increased between 2002 and 2008 by 7% for (formerly) publicly insured patients and 10% for (formerly) privately insured patients. In general, no significant differences in the trends for guideline adherence were found between privately and publicly insured patients, indicating the absence of an effect of the remuneration system on guideline adherence. Adherence to guidelines involving more time investment in terms of follow-up contacts was affected by changes in the remuneration system. For publicly insured patients, GPs showed a higher trend for guideline adherence for guidelines involving more time investment in terms of follow-up contacts compared with privately insured patients. The change in the remuneration system had a limited impact on guideline adherence.


An electrophoretic polymorphism in salivary amylases (Amy-1) of mastomys (Praomys coucha).
An electrophoretic polymorphism of salivary amylases (Amy-1) in mastomys (Praomys coucha) (MWC, MRJ and MCC strains) was detected. Amylase in MWC or MRJ saliva, which migrated fast toward the anode, was designated as AMY-1A, and that in MCC saliva migrating slowly as AMY-1B. Salivary amylases are controlled by a pair of codominant alleles at a single autosomal locus (Amy-1). No polymorphism was seen in pancreatic amylases (Amy-2). The frequencies of these phenotypes did not differ between the sexes. Some isoamylases were observed and these were different from those in mouse or rat.


Detection of Left Ventricular Regional Function in Asymptomatic Children with beta-Thalassemia Major by Longitudinal Strain and Strain Rate Imaging.
Cardiac failure due to iron overload remains the most common cause of death in patients with beta-thalassemia major. This study aimed to evaluate myocardial function in children with beta-thalassemia major using standard echocardiography technique and strain rate imaging. Conventional echocardiographic analysis, tissue velocity imaging, and strain/strain rate imaging of the left ventricle were evaluated in 48 children with beta-thalassemia major (19 girls, 29 boys; 8.39±4.05 years) and 22 healthy children (11 girls, 11 boys; 8±3.72 years). Conventional echocardiographic examinations revealed that beta-thalassemia patients had larger left ventricular end-systolic diameter, end-diastolic and end-systolic volume, left ventricular mass index, and mitral early/late diastolic flow velocity ratio (p<0.05). Strain and strain rate imaging study of the basal lateral wall of the left ventricle was higher in patients than in controls, at p=0.035 and p=0.008, respectively. We found that superior systolic strain and strain rate imaging of the left ventricle indicated the presence of regional systolic function in the left ventricular wall. We suggest that left ventricle volume and mass index parameters might be more sensitive than the other conventional and strain/strain rate imaging parameters during childhood. However, the adulthood strain and strain rate imaging values may be lower than controls, exceeding the critical level of iron overload. None declared.


Laparoscopic Versus Open Resection for Liver Cavernous Hemangioma: A Single Center of 2-Year Experience.
To evaluate the feasibility and safety of laparoscopic versus open resection for liver cavernous hemangioma (LCH). A total of 131 patients suffering from LCH operated in our department between January 2013 and December 2014 were divided into 2 groups: 31 for laparoscopic liver resection (LR) and 100 for open liver resection (OR). Age, sex, presence or absence of chronic liver disease, tumor size, tumor location, type of resection, estimated intraoperative blood loss, operative time, length of postoperative hospital stay, morbidity, and mortality were equivalent between the 2 groups. There were no significant differences in estimated intraoperative blood loss between the LR and OR groups. The operation time of the LR group was longer than the OR group and the hospitalization expenses less than the OR group. However, the time of postoperative hospital stay and time of oral intake were shorter in the LR group than the OR group. The tumor of the LR group was smaller than the OR group. In liver function, alanine aminotransferase after operation of the LR group was lower than the OR group, the same as aspartate transaminase after operation. But there were no significant differences in total bilirubin after operation. Laparoscopic resection for LCH is a safe and feasible procedure as OR.


Branchinotogluma bipapillata n. sp., a new branchiate scale worm (Annelida: Polynoidae) from two hydrothermal fields on the Southwest Indian Ridge.
A new species of Branchinotogluma, found at two hydrothermal vent fields on Southwest Indian Ocean Ridge, is described herein. It can be distinguished from its congeners by the small acicular lobe on the tentacular segment, the stout smooth notochaetae, 5 pairs of dorsal and ventral papillae surrounding the pharynx, 2 pairs of long ventral papillae and 4 pairs of ventral lamellae on males, and modified parapodia on posterior segments. Sexual dimorphism is also reported in the new species, as male and female individuals display differences in characters on the ventral papillae and posterior segments.


Serotonin and its place in the pathogenesis of depression.
The authors review the literature in an attempt to evaluate the relationship between serotonin and depression. Animal studies show that the administration of tryptophan (the precursor of serotonin) increased serotonin synthesis and influenced behavior. Low plasma tryptophan levels have been found in patients with endogeneous depression. Postmortem studies have shown an association between lowered hindbrain serotonin levels and suicide among depressed persons. The decreased serotonin levels in blood platelets during depression mirrored the neuronal changes. Tricyclic antidepressants inhibited platelet serotonin uptake and reduced imipramine binding sites on the platelets. A positive correlation between depression rating scores and platelet aggregatory response has been reported. Serotonin stimulated release of prolactin and growth hormone, although the prolactin response was less marked in depression. A marker for depressive illness is still sought, but it is likely to be related in some way to serotonin.


New Phenylthiazoles: Design, Synthesis, and Biological Evaluation as Antibacterial, Antifungal, and Anti-COVID-19 Candidates.
The combination of antibacterial and antiviral agents is becoming a very important aspect of dealing with resistant bacterial and viral infections. The N-phenylthiazole scaffold was found to possess significant anti-MRSA, antifungal, and anti-COVID-19 activities as previously published; hence, a slight refinement was proposed to attach various alkyne lipophilic tails to this promising scaffold, to investigate their effects on the antimicrobial activity of the newly synthesized compounds and to provide a valuable structure-activity relationship. Phenylthiazole 4 m exhibited the most potent anti-MRSA activity with 8 μg/mL MIC value. Compounds 4 k and 4 m demonstrated potent activity against Clostridium difficile with MIC values of 2 μg/mL and moderate activity against Candida albicans with MIC value of 4 μg/mL. When analyzed for their anti-COVID-19 inhibitory effect, compound 4 b emerged with IC50 =1269 nM and the highest selectivity of 138.86 and this was supported by its binding score of -5.21 kcal mol-1 when docked against SARS-CoV-2 M pro . Two H-bonds were formed, one with His164 and the other with Met49 stabilizing phenylthiazole derivative 4 b, inside the binding pocket. Additionally, it created two arene-H bonds with Asn142 and Glu166, through the phenylthiazole scaffold and one arene-H bond with Leu141 via the phenyl ring of the lipophilic tail.


Chiral separations using dipeptide polymerized surfactants: effect of amino acid order.
Chiral separations using various polymerized dipeptide surfactants in electrokinetic capillary chromatography (EKC) are investigated. The two main dipeptide surfactants used in this study were sodium N-undecylenyl-L-valine-L-leucine (L-SUVL), and sodium N-undecylenyl-L-leucine-L-valine (L-SULV). These studies were performed in order to determine if the order of amino acids in dipeptide surfactants is important in terms of chiral recognition and separations. Both the monomer and the polymer of these two surfactants were compared for the separation of two model atropisomers, (+/-)-1,1-bi-2-naphtol (BOH) and (+/-)-1,1'-bi-2-naphthyl-2,2'-diyl hydrogen phosphate (BNP). Some advantages and disadvantages of the polymer relative to the monomer are discussed. Four other surfactants, the polymers of sodium N-undecylenyl-L-leucine-L-leucine (L-SULL), sodium N-undecylenyl-L-valine-L-valine (L-SUVV), sodium N-undecylenyl-L-valine (L-SUV), and sodium N-undecylenyl-L-leucine (L-SUL), were also used in this study, and their performance was compared to that of poly(L-SULV). These data show conclusively that the order of amino acids in dipeptide surfactants has a dramatic effect on chiral recognition. Our investigations indicate that poly-(L-SULV) provides the best enantioselectivity among the four dipeptide and two single amino acid surfactants for the separation of BNP and BOH. The advantages of poly-(L-SULV) are demonstrated via the ultrafast separation of the enantiomers of BNP and BOH in less than 1 min.


Fibrinolytic and antibiotic treatment of prosthetic vascular graft infections in a novel rat model.
We developed a rat model of prosthetic vascular graft infection to assess, whether the fibrinolytic tissue plasminogen activator (tPA) could increase the efficacy of antibiotic therapy. Rats were implanted a polyethylene graft in the common carotid artery, pre-inoculated with approx. 6 log10 colony forming units (CFU) of methicillin resistant Staphylococcus aureus. Ten days after surgery, rats were randomized to either: 0.9% NaCl (n = 8), vancomycin (n = 8), vancomycin + tPA (n = 8), vancomycin + rifampicin (n = 18) or vancomycin + rifampicin + tPA (n = 18). Treatment duration was seven days. Approximately 36 hours after the end of treatment, the rats were euthanized, and grafts and organs were harvested for CFU enumeration. All animals in the control group had significantly higher CFU at the time of euthanization compared to bacterial load found on the grafts prior to inoculation (6.45 vs. 4.36 mean log10 CFU/mL, p = 0.0011), and both the procedure and infection were well tolerated. Vancomycin and rifampicin treatment were superior to monotherapy with vancomycin, as it lead to a marked decrease in median bacterial load on the grafts (3.50 vs. 6.56 log10 CFU/mL, p = 0.0016). The addition of tPA to vancomycin and rifampicin combination treatment did not show a further decrease in bacterial load (4.078 vs. 3.50 log10 CFU/mL, p = 0.26). The cure rate was 16% in the vancomycin + rifampicin group vs. 37.5% cure rate in the vancomycin + rifampicin + tPA group. Whilst interesting, this trend was not significant at our sample size (p = 0.24). We developed the first functional model of an arterial prosthetic vascular graft infection in rats. Antibiotic combination therapy with vancomycin and rifampicin was superior to vancomycin monotherapy, and the addition of tPA did not significantly reduce bacterial load, nor significantly increase cure rate.


Severe asthma guidelines in children and adolescents: A practical document for physicians.
Asthma is a common disease in childhood with a minority of affected children suffering from severe asthma. Patients with severe asthma require high dose inhaled glucocorticoids plus a second controller and/or systemic corticosteroids to be well-controlled or remain uncontrolled despite such treatment. Although only a small subset of children and adolescents falls in this category, the management of affected patients represents a major concern for pediatricians. Guidelines and recommendations have been designed to guide the management of this group of patients. Though the terms "recommendations" and "guidelines" are often used interchangeably, it should be noted that the first one should be used more narrowly to identify specific actions and the second one to broadly refer to the umbrella under which multiple recommendations for a specific condition are provided. Moreover, the availability of several and sometimes-conflicting documents on severe asthma management both in adult and pediatric age could generate confusion among health care professionals. The manuscript analyses seven papers addressing severe asthma, comparing any key aspects and differences. Finally, we tried to create a more practical document for physicians to simplify the interpretation of the several available documents on severe asthma management focusing the pediatric age.


Hippocampal damage and anterograde object-recognition in rats after long retention intervals.
Although several studies in rats have found that hippocampal damage has negligible effects on anterograde object-recognition memory, the findings are not entirely conclusive, because most studies have used retention intervals lasting only a few hours. We assessed the effects of neurotoxic hippocampal lesions on anterograde object recognition, using a novel-object preference test, with retention intervals that were considerably longer than in previous studies-24 h, 1 week, and 3 weeks. To promote object recognition after such long intervals, rats were familiarized with a sample object in an open field for 5 min/day for 5 consecutive days. Recognition was assessed by comparing the amount of time spent investigating the sample versus a novel object on a preference test at one of the postlearning intervals. The rats with hippocampal lesions displayed a normal novelty preference after a 3-week interval, and their performance across the three delay conditions was not significantly different from that of control rats. The findings indicate that extensive hippocampal damage spares anterograde object recognition in rats, even after retention intervals lasting days or weeks.


Cerebellopontine angle mass mimicking lingual nerve injury after dental implant placement: a case report.
This is a rare case report of a cerebellopontine angle (CPA) mass mimicking lingual nerve injury after a dental implant placement. Lingual nerve injury is a common complication following dental implant placement. CPA masses are likely to cause symptomatic trigeminal neuralgia, and thus can mimic and be easily confused with oral diseases. We experienced a case of CPA mass mimicking lingual nerve injury after dental implant placement. The patient was a 57-year-old Japanese female who complained of glossalgia. She underwent dental implant placement in the mandible before visiting our clinic. Panoramic x-ray radiography revealed no abnormalities; the salivary flow rate by gum test was 7.0 ml/10 min. She was diagnosed with lingual nerve injury and secondary burning mouth syndrome. Vitamin B12 and oral moisturizer did not provide relief; furthermore, numbness in the lower lip emerged. A Semmes Weinstein test demonstrated elevation of her sensitivity threshold. Finally, magnetic resonance imaging revealed a 20-mm diameter mass in the CPA. The patient is now being followed under conservative management. Our experience underscores the importance of including CPA mass in the differential diagnosis of dental diseases.


Production of glycosylated physiologically "normal" human alpha 1-antitrypsin by mouse fibroblasts modified by insertion of a human alpha 1-antitrypsin cDNA using a retroviral vector.
Alpha 1-Antitrypsin (alpha 1AT) deficiency is a hereditary disorder characterized by reduced serum levels of alpha 1AT, resulting in destruction of the lower respiratory tract by neutrophil elastase. As an approach to augment alpha 1AT levels in this disorder with physiologically normal human alpha 1AT, we have integrated a full-length normal human alpha 1AT cDNA into the genome of mouse fibroblasts. To accomplish this, the retroviral vector N2 was modified by inserting the simian virus 40 early promoter followed by the alpha 1AT cDNA. Southern analysis demonstrated that the intact cDNA was present in the genome of selected clones of the transfected murine fibroblasts psi 2 and infected NIH 3T3. The clones produced three mRNA transcripts (5.8, 4.8, and 2.4 kilobases) containing human alpha 1AT sequences, secreted an alpha 1AT molecule recognized by an anti-human alpha 1AT antibody, with the same molecular mass (52 kDa) as normal human alpha 1AT and that complexed with and inhibited human neutrophil elastase. The psi 2 produced alpha 1AT was glycosylated, and when infused intravenously into mice, it had a serum half-life similar to normal alpha 1AT purified from human plasma and markedly longer than that of nonglycosylated human alpha 1AT cDNA-directed yeast-produced alpha 1AT. These studies demonstrate the feasibility of using a retroviral vector to insert the normal human alpha 1AT cDNA into non-alpha 1AT-producing cells, resulting in the synthesis and secretion of physiologically "normal" human alpha 1AT.


Use of steroid and nonsteroidal anti-inflammatories in the treatment of rheumatoid arthritis: Systematic review protocol.
Rheumatoid arthritis affects 1% of the world's population and its current treatment options are costly. There are not enough studies that evaluated the efficacy and safety of anti-inflammatory drugs medications used to reduce rheumatoid arthritis's symptoms. This study will evaluate the effectiveness and the safety of steroid and nonsteroidal anti-inflammatory drugs for the treatment of patients with rheumatoid arthritis. Randomized clinical trials eligible for our systematic review will enroll adults with rheumatoid arthritis treated with anti-inflammatory drugs compared with a control group (placebo or active control) at any dose, duration, and route of administration and double blind studies. In order to include all forms of rheumatoid arthritis and anti-inflammatory drugs, we will search the following electronic databases: Cochrane Central Register of Controlled Trials, MEDLINE (via Ovid); ExcerptaMedica Database (via Ovid); Cumulative Index to Nursing and Allied Health Literature (via Ovid); Web of Science; ClinicalTrial.gov; and WHO International Clinical Trials Registry Platform. We will not impose any language restrictions or publication status. Outcomes of interest include are pain, physical function, swelling, stiffness, grip force, radiological image of the joint, quality of life, adverse events, discontinuation due to adverse events, satisfaction with the treatment, and rescue medication for pain. A team of reviewers will independently screen search results, extract data from eligible trials, and assess risk of bias. We will use the Grading of Recommendations Assessment, Development and Evaluation approach to rate overall certainty of the evidence by outcome. Dichotomous data will be summarized as risk ratios; continuous data will be given as standard average differences with 95% confidence intervals. The evidence derived by this study will increase awareness of the effectiveness and safety of steroid and nonsteroidal anti-inflammatory drugs for the treatment of rheumatoid arthritis. The results could guide patients and healthcare practitioners and help facilitate evidence-based shared care decision making.


[Effects of butylphthalide on extracellular 6-keto-PGF1 alpha, TXB2 and 6-keto-PGF1 alpha/TXB2 ratio in cultured rat cortical neurons].
The effects of 3-n-butylphthalide(NBP) on the levels of 6-keto-PGF1 alpha, TXB2 and 6-keto-PGF1 alpha/TXB2 ratio were studied with methods of RIA. d-NBP and l-NBP(0.1-100 mumol.L-1) concentration-dependently increased 6-keto-PGF1 alpha release, decreased TXB2 release from neuronal cells, and significantly enhanced extracellular 6-keto-PGF1 alpha/TXB2 ratio in primary cultured rat cortical neurons exposed to hypoxic-hypoglycemic media for 5 h or hypoxic-hypoglycemic media for 5 h following normal media for 3 h. Aspirin(0.1-100 mumol.L-1) was also shown to inhibit TXB2 release from cortical neurons in a dose-dependent manner. However aspirin only increased 6-keto-PGF1 alpha/TXB2 ratio at low dose because aspirin inhibited both 6-keto-PGF1 alpha and TXB2 release simultaneously at large dose(10-100 mumol.L-1). This suggests that the action of l-NBP, d-NBP and dl-NBP on the increase of 6-keto-PGF1 alpha/TXB2 ratio might be one of the mechanisms in which NBP enhanced focal cerebral blood flow and improved ischemic brain damage.


[Behavior associated with HIV and HCV infection in female prison inmates in São Paulo, Brazil].
An increase has been observed in AIDS and hepatitis C cases in women, including female prison inmates. This study focused on inmates' behavioral factors associated with risk of HIV and HCV transmission in a women's detention facility in São Paulo, Brazil. Behavioral questionnaires were applied and HIV and HCV serology were performed. The selected measure of association was odds ratio for both the bi and multivariate logistic regression analyses. 290 inmates participated in the study. HIV and HCV prevalence rates were 13.9% and 16.2%, respectively. Statistically significant associations were observed (p < 0.05) between HIV and the following variables: partner with AIDS OR = 6.9 (2.7-35.2); injection drug users (IDU) OR = 3.3 (1.6-14.7); regular partner OR = 3.7 (1.5-8.3), and between HCV and: IDU OR = 13.7 (4.4-42.7); IDU partner OR = 4.9 (1.9-12.2); previous arrest OR = 2.8 (1.2-6.5) adjusted for: partner with AIDS, IDU, IDU partner, drug user, and previous arrest. In conclusion, parenteral risk was associated with HIV and HCV infection and sexual risk with HIV. Appropriate and continuous preventive programs are recommended in the prison.


Parking: an effective strategy described.
A hospital car park needs to balance the demands of patients, visitors, and staff, without, for example, compromising emergency vehicles' access to the Accident & Emergency Department. However, with car ownership on the rise, the half a million car parking spaces across all Trust sites in England may not be enough to meet future demand. Philip Herring, managing director of VINCI Park UK, which designs, finances, builds, and operates, car parks for a variety of sectors, argues that having in place an effective strategy for future parking is vital to meet the growing needs of patients, visitors, and staff, as well as budgetary and environmental commitments.


Thrombotic thrombocytopenic purpura as an etiology of thrombocytopenia in systemic lupus erythematosus: case report.
Abstract Thrombotic thrombocytopenic purpura (TTP) is an unusual complication of systemic lupus erythematosus (SLE). Although the reported association between SLE and TTP is increasing, a few cases do improve without plasmatherapy. We report a case of TTP which was successfully treated without plasmatherapy, which might be underestimated as an etiology of thrombocytopenia in SLE. TTP should always be considered as a concomitant disease when Coombs' negative hemolytic anemia or thrombocytopenia is seen in SLE patients.


Reliability and validity of the short mast among psychiatric inpatients.
Examined the reliability and empirical validity of the Short Michigan Alcoholism Screening Test (SMAST) orally administered as an embedded scale to a sample of 120 male and female psychiatric inpatients, who reported a broad spectrum of current and lifetime drinking problems. Test results showed substantial reliability within and across administrations that spanned 1 to 3 days, but they were related only moderately to the psychiatric diagnosis.


Homing of neural stem cells from the venous compartment into a brain infarct does not involve conventional interactions with vascular endothelium.
Human neural stem cells (hNSCs) hold great potential for treatment of a wide variety of neurodegenerative and neurotraumatic conditions. Heretofore, administration has been through intracranial injection or implantation of cells. Because neural stem cells are capable of migrating to the injured brain from the intravascular space, it seemed feasible to administer them intravenously if their ability to circumvent the blood-brain barrier was enhanced. In the present studies, we found that interactions of hNSCs in vitro on the luminal surface of human umbilical vein endothelial cells was enhanced following enforced expression of cutaneous lymphocyte antigen on cell surface moieties by incubation of hNSCs with fucosyltransferase VI and GDP-fucose (fhNSCs). Interestingly, ex vivo fucosylation of hNSCs not only did not improve the cells homing into the brain injured by stroke following intravenous administration but also increased mortality of rats compared with the nonfucosylated hNSC group. Efforts to explain these unexpected findings using a three-dimensional flow chamber device revealed that transmigration of fhNSCs (under conditions of physiological shear stress) mediated by stromal cell-derived factor 1α was significantly decreased compared with controls. Further analysis revealed that hNSCs poorly withstand physiological shear stress, and their ability is further decreased following fucosylation. In addition, fhNSCs demonstrated a higher frequency of cellular aggregate formation as well as a tendency for removal of fucose from the cell surface. In summary, our findings suggest that the behavior of hNSCs in circulation is different from that observed with other cell types and that, at least for stroke, intravenous administration is a suboptimal route, even when the in vitro rolling ability of hNSCs is optimized by enforced fucosylation.


Mortality, patient-reported outcome measures, and the health economic burden of prosthetic joint infection.
Prosthetic joint infection (PJI) is one of the most devastating complications for a patient following arthroplasty. This scoping review aims to evaluate the burden of PJI on individual patients and the healthcare system regarding the mortality rate, patient-reported quality of life, and healthcare resource utilisation. Patients with PJI have up to a five-fold higher mortality rate than those who have undergone an uninfected primary arthroplasty. There is an increased use of ambulatory aids and reduced joint function scores in patients with PJI. Global quality of life is poorer, specifically measured by the EQ-5D. Direct hospitalisation costs are two- to five-fold higher, attributed to surgery and prostheses, antibiotics, and a prolonged inpatient stay. There is an immense clinical and health economic burden secondary to PJI worldwide. This is expected to rise exponentially due to the increasing number of primary procedures and an ageing population with comorbidities Improving preventative and treatment strategies is imperative for patients and the healthcare system.


Repeated Acute Exacerbations of Chronic Inflammatory Demyelinating Polyradiculoneuropathy Accompanied by Pain and Swelling in Distal Extremities.
An 81-year-old man experienced acute progression of weakness in the extremities accompanied by a fever, tenderness, and swelling in distal parts of the extremities. He had flaccid tetraparesis with fasciculations and general hyporeflexia. Nerve conduction studies indicated demyelinating sensorimotor neuropathy. A cerebrospinal fluid examination revealed elevated proteins without pleocytosis. Immunological treatments were effective, but his symptoms exhibited repeated relapse and remission phases. He was diagnosed with chronic inflammatory demyelinating polyradiculoneuropathy (CIDP) with an acute onset. The highlight of this case is pain with inflammatory reaction recognized as red flags of CIDP, with the clinical course and electrophysiological findings compatible with CIDP.


Heart Transplant Outcomes After Total Artificial Heart.
We sought to compare heart transplant (HTX) outcomes from patients with a total artificial heart (TAH), biventricular assist device (BiVAD), or left ventricular assist device (LVAD) as a bridge to transplant (BTT). Interagency Registry for Mechanically Assisted Circulatory Support (INTERMACS)-Scientific Registry of Transplant Recipients (SRTR) created a dataset with TAH or durable mechanical circulatory support (MCS) who reached HTX between 2006 and 2015. The retrospective analysis compared TAH outcomes with those with a BiVAD or LVAD before HTX. The primary outcome was posttransplant survival at 1, 36, and 60 months. Secondary outcomes included simultaneous heart-kidney transplants, donor characteristics, and mortality risk factors. INTERMACS-SRTR cohort had, at the time of HTX, 2762 patients with LVAD; 205 BiVAD (139 durable and 66 temporary RVAD); 176 TAH (6 prior HeartMate II). Sixty months after HTX, mortality rates were 16.5% in the total group: LVAD 15.2%, BiVAD 22.4%, and TAH 29%. Survival differed between the LVAD, the TAH, and BiVAD but not between the BiVAD and TAH groups. One-year survival and complication rates were similar across groups-there was no difference in survival by donor age in the overall cohort. There was a difference in TTD based on recipient age in the LVAD group but not in BiVAD or TAH groups. Occurrence of HTX-kidney and post-transplant dialysis were higher in the TAH versus LVAD and BiVAD groups. The TAH is an efficacious BTT. Refinements in technology and patient selection may improve outcomes.


Discontinuous atmospheric pressure interface for mass spectrometry using a solenoid pulse valve.
For the development of on-site mass spectrometry for security and safety, point-of-care analysis, etc., the gas volume introduced into the vacuum should be reduced to a minimum. To cope with this demand, a discontinuous atmospheric pressure interface using a solenoid pulse valve was developed. The sample gas was introduced discontinuously into the ionization cell with a volume of 0.17 cm(3) . The sampled gas in the cell was ionized by an alternating current (ac) corona discharge. The generated ions were sampled through a 0.25 mm i.d. and 12 mm long nickel capillary into the vacuum of a time-of-flight mass spectrometer. A gas flow rate of ~25 mL/min was achieved with the 1 Hz pulse valve operation and 20 ms valve opening time. Sub-ng limits of detection for less volatile compounds such as explosives and drugs were obtained. Due to its compact size and low gas load to the vacuum, this new interface may be useful for applications in miniaturized mass spectrometry. Copyright © 2016 John Wiley & Sons, Ltd.


Extracellular vesicles derived from Talaromyces marneffei contain immunogenic compounds and modulate THP-1 macrophage responses.
Pathogenic eukaryotes including fungi release extracellular vesicles (EVs) which are composed of a variety of bioactive components, including peptides, nucleic acids, polysaccharides, and membrane lipids. EVs contain virulence-associated molecules suggesting a crucial role of these structures in disease pathogenesis. EVs derived from the pathogenic yeast phase of Talaromyces (Penicillium) marneffei, a causative agent of systemic opportunistic mycoses "talaromycosis," were studied for their immunogenic components and immunomodulatory properties. Some important virulence factors in EVs including fungal melanin and yeast phase specific mannoprotein were determined by immunoblotting. Furthermore, fluorescence microscopy revealed that T. marneffei EVs were internalized by THP-1 human macrophages. Co-incubation of T. marneffei EVs with THP-1 human macrophages resulted in increased levels of supernatant interleukin (IL)-1β, IL-6 and IL-10. The expression of THP-1 macrophage surface CD86 was significantly increased after exposed to T. marneffei EVs. These findings support the hypothesis that fungal EVs play an important role in macrophage "classical" M1 polarization. T. marneffei EVs preparations also increased phagocytosis, suggesting that EV components stimulate THP-1 macrophages to produce effective antimicrobial compounds. In addition, T. marneffei EVs stimulated THP-1 macrophages were more effective at killing T. marneffei conidia. These results indicate that T. marneffei EVs can potently modulate macrophage functions, resulting in the activation of these innate immune cells to enhance their antimicrobial activity.


Toward a Biology-Driven Treatment Strategy for Peripheral T-cell Lymphoma.
T-cell and natural killer-cell lymphomas are a relatively rare and heterogeneous group of diseases that are difficult to treat and usually have poor outcomes. To date, therapeutic interventions are of limited efficacy and there is a pressing need to find better treatments. In recent years, advances in molecular biology have helped to elucidate the underlying genetic complexity of this group of diseases and to identify mutations and signaling pathways involved in lymphomagenesis. In this review, we highlight the unique biological characteristics of some of the different subtypes and discuss how these may be targeted to provide more individualized and effective treatment approaches.


Regular blood donation improves endothelial function in adult males.
Endothelial dysfunction, secondary to systemic inflammation and oxidative stress, is known to play a major role in the development and progression of atherosclerosis. It is hypothesized that the lower incidence of coronary artery disease in the premenopausal period in females when compared with males is associated with regular menstrual blood loss. We investigated whether regular blood donation (BD) is associated with improved endothelial function in healthy adult males. Fifty young healthy male volunteers volunteers with a mean age of 30 ± 6 years without overt cardiovascular disease were enrolled to participate in serial consecutive BDs. Serum iron levels as oxidative stress parameters, flow-mediated dilatation (FMD) for endothelial function, 24-h mean diastolic blood pressure for peripheral vascular resistance identification, and high-sensitivity C-reactive protein (hs-CRP) levels as systemic inflammatory markers were evaluated before and after BD. This study used a prospective observational cohort design. Patients with cardiovascular and inflammatory diseases were excluded. BD was found to improve FMD steadily and significantly when compared with the baseline (mean ± SD: 9.9% ± 3.8%, 10.44% ± 3.9%, 10.65% ± 3.9%, and 10.75 ± 3.9%, respectively, p=0.15, p=0.02, p=0.006 as compared with the baseline). A steady decrease was identified in hs-CRP levels after serial BDs, although this decrease was not statistically significant in the all phases (2.96 ± 3.3 mg/L, 2.26 ± 1.5 mg/L, and 2.12 ± 1.5 mg/L, respectively, p=0.829, p=0.558). The 24-h mean diastolic blood pressures were significantly lower in the chronic phase (77 ± 9 mm Hg, 75 ± 7 mm Hg, and 72 ± 8 mm Hg, respectively, p=0.50, p=0.003), whereas there was no significant change in iron levels in the acute and chronic phases (66 ± 32 mg/dL, 72 ± 43 mg/dL, and 68 ± 33 mg/dL, respectively, p=1.000, p=1.000). The results of the study indicate that regular BD improves endothelial function.


Hysterectomy with and without oophorectomy and all-cause and cause-specific mortality.
Hysterectomy is one of the most commonly performed gynecologic surgeries, with an estimated 30% of women in Australia undergoing the procedure by age of 70 years. In the United States, about 45% of women undergo hysterectomy in their lifetime. Some studies have suggested that this procedure increases the risk of premature mortality. With many women making the decision to undergo hysterectomy for a benign indication each year, additional research is needed to clarify whether there are long-term health consequences of hysterectomy. This study aimed to examine the association between hysterectomy for benign indications, with or without removal of the ovaries, and cause-specific and all-cause mortality. Our cohort of 666,588 women comprised the female population of Western Australia with linked hospital and health records from 1970 to 2015. Cox regression models were used to assess the association between hysterectomy and all-cause, cardiovascular disease, cancer, and other mortality by oophorectomy type (categorized as none, unilateral, and bilateral), with no hysterectomy or oophorectomy as the reference group. We repeated these analyses using hysterectomy without oophorectomy as the reference group. We also investigated whether associations varied by age at the time of surgery, although small sample size precluded this analysis in women who underwent hysterectomy with unilateral salpingo-oophorectomy. In our main analysis, women who underwent hysterectomy or oophorectomy as part of cancer treatment were retained in the analysis and considered unexposed to that surgery. For a sensitivity analysis, we censored procedures performed for cancer. Compared with no surgery, hysterectomy without oophorectomy before 35 years was associated with an increase in all-cause mortality (hazard ratio, 1.29; 95% confidence interval, 1.19-1.40); for surgery after 35 years of age, there was an inverse association (35-44 years: hazard ratio, 0.93; 95% confidence interval, 0.89-0.97). Similarly, hysterectomy with bilateral salpingo-oophorectomy before 45 years of age was associated with increased all-cause mortality (35-44 years: hazard ratio, 1.15; 95% confidence interval, 1.04-1.27), but decreased mortality rates after 45 years of age. In our sensitivity analysis, censoring gynecologic surgeries for cancer resulted in many cancer-related deaths being excluded for women who did not have surgery for benign indications and thus increased the hazard ratios for the associations between both hysterectomy without oophorectomy and hysterectomy with bilateral salpingo-oophorectomy and risk of all-cause and cancer-specific mortality. The sensitivity analysis therefore potentially biased the results in favor of no surgery. Among women having surgery for benign indications, hysterectomy without oophorectomy performed before 35 years of age and hysterectomy with bilateral salpingo-oophorectomy performed before 45 years of age were associated with an increase in all-cause mortality. These procedures are not associated with poorer long-term survival when performed at older ages.


Modified apple polysaccharide prevents colitis through modulating IL-22 and IL-22BP expression.
Chronic intestinal inflammation enhances cell proliferation, angiogenesis, and migration, then promotes the development of colorectal cancer (CRC). Many ingredients of apples have been proven to have anti-inflammatory properties, and show benefits for colitis treatment. In our previous studies, we found modified apple polysaccharide (MAP) could prevent colitis associated colorectal carcinogenesis effectively. Herein, we further our study to observe the effect of MAP on dextran sodium sulfate (DSS)-induced colitis and to investigate the possible mechanisms. IL-22 has both pathogenic and protective effects during intestinal tissue damage. It could be neutralized by the soluble IL-22 receptor, known as the IL-22 binding protein (IL-22BP). A DSS-induced colitis mouse model, a mouse CRC cell line MCA-38 and a mouse dendritic cell line DC2.4 were treated with MAP. Western blot, ELISA, BrdU staining and a co-culture system were used to detect the expression of IL-22 and IL-22BP. MAP significantly protected ICR mice against DSS-induced colitis, and inhibited the growth of MCA-38 cells. The mechanisms may be that MAP down-regulated IL-22 level and up-regulated expression of IL-22BP. These data may provide another molecular basis for understanding how apples act to prevent colitis and suggest that MAP has a potential to treat colitis and prevent CRC.


Exploring the human lacrimal gland using organoids and single-cell sequencing.
The lacrimal gland is essential for lubrication and protection of the eye. Disruption of lacrimal fluid production, composition, or release results in dry eye, causing discomfort and damage to the ocular surface. Here, we describe the establishment of long-term 3D organoid culture conditions for mouse and human lacrimal gland. Organoids can be expanded over multiple months and recapitulate morphological and transcriptional features of lacrimal ducts. CRISPR-Cas9-mediated genome editing reveals the master regulator for eye development Pax6 to be required for differentiation of adult lacrimal gland cells. We address cellular heterogeneity of the lacrimal gland by providing a single-cell atlas of human lacrimal gland tissue and organoids. Finally, human lacrimal gland organoids phenocopy the process of tear secretion in response to neurotransmitters and can engraft and produce mature tear products upon orthotopic transplantation in mouse. Together, this study provides an experimental platform to study the (patho-)physiology of the lacrimal gland.


Does the type of prosthesis influence the incidence of permanent pacemaker implantation following isolated aortic valve replacement.
The incidence of conduction disorders requiring permanent pacing (PPM) in patients operated on for aortic valve replacement (AVR) has been reported to be 5.7%. However, perioperative risk predictors for PPM following AVR are not well characterized and debate exists regarding selection of the prosthesis-type most likely to minimize this incidence. The aim of the study was to assess whether the type of the prosthesis used influences the prevalence of PPM following aortic valve replacement. A total of 782 consecutive patients with predominant aortic stenosis accepted for isolated non-emergent AVR were studied over a 3 year period; of which 305 patients (Group A) received mechanical prostheses, 335 received stented tissue prostheses (Group B), and the remaining 142 received stentless tissue valves (Group C). A stepwise logistic regression analysis was used to identify the independent predictors for PPM and statistical significance was accepted at a level of P < .05. Univariate and multivariate analyses showed a significant relationship between the preoperative factors (poor ejection fraction < 35%; P < .001), left atrial enlargement (LAE; P < .001) and left bundle branch block (LBBB; P < .001), the perioperative variables (bypass time > 100 minutes with x-clamp time > 70 minutes; P < .001) and the incidence of PPM. The proposed predictive model correlated highly with actual pacemaker use, suggesting that the requirement for PPM results from either operative trauma or increased ischemic burden and the incidence of PPM is independent of prosthesis-type implanted.


Deoxyuridine triphosphate nucleotidohydrolase as a potential antiparasitic drug target.
This paper describes a structure-activity study to identify novel, small-molecule inhibitors of the enzyme deoxyuridine 5'-triphosphate nucleotidohydrolase (dUTPase) from parasitic protozoa. The successful synthesis of a variety of analogues of dUMP is described in which the substituents are introduced at the 3'- and 5'-positions, together with variation in the heteroatom at the 5'-position. The compounds were assayed against recombinant Plasmodium falciparum and Leishmania major enzymes and the human enzyme to give a measure of selectivity. The compounds were also tested in vitro against the intact parasites P. falciparum and L. donovani. A number of potent and selective inhibitors of the P. falciparum dUTPase that show drug-like properties and represent good leads for future development were identified. The best inhibitors included the compounds 5'-tritylamino-2',5'-dideoxyuridine (2j) (Ki = 0.2 microM) and 5'-O-triphenylsilyl-2',3'-didehydro-2',3'-dideoxyuridine (5h) (Ki = 1.3 microM), with selectivity greater than 200-fold compared to the human enzyme. Structural features important for antiplasmodial activity were determined. The correlation observed between the inhibition of the enzyme and the inhibition of the parasite growth in vitro demonstrates that the P. falciparum dUTPase constitutes a valid and attractive novel target for the development of much-needed new antimalarial drugs.


Inhibition of complement-mediated killing of Brucella abortus by fluid-phase immunoglobulins.
Bovine immunoglobulin preparations from normal serum and from sera containing antibodies against Brucella abortus interfered with the brucellacidal action of bovine serum, whereas unfractionated normal serum and antisera were not inhibitory. The inhibitory property of immunoglobulin appeared to be attributable to some anticomplementary property because it also interfered with serum-mediated hemolysis of antibody-coated erythrocytes. The supernatant phase obtained after ultracentrifugation of bovine anti-B abortus immunoglobulin did not inhibit brucellacidal activity of normal bovine serum. Results of this study indicate that bovine anti-B abortus immunoglobulin preparations contain microaggregates of protein that can inhibit the ability of bovine serum to kill B abortus. The most likely mechanism is nonspecific activation of complement by microaggregated immunoglobulin, which consumes complement and makes it unavailable for bactericidal activity.


Optimizing 1-μs-Resolution Single-Molecule Force Spectroscopy on a Commercial Atomic Force Microscope.
Atomic force microscopy (AFM)-based single-molecule force spectroscopy (SMFS) is widely used to mechanically measure the folding and unfolding of proteins. However, the temporal resolution of a standard commercial cantilever is 50-1000 μs, masking rapid transitions and short-lived intermediates. Recently, SMFS with 0.7-μs temporal resolution was achieved using an ultrashort (L = 9 μm) cantilever on a custom-built, high-speed AFM. By micromachining such cantilevers with a focused ion beam, we optimized them for SMFS rather than tapping-mode imaging. To enhance usability and throughput, we detected the modified cantilevers on a commercial AFM retrofitted with a detection laser system featuring a 3-μm circular spot size. Moreover, individual cantilevers were reused over multiple days. The improved capabilities of the modified cantilevers for SMFS were showcased by unfolding a polyprotein, a popular biophysical assay. Specifically, these cantilevers maintained a 1-μs response time while eliminating cantilever ringing (Q ≅ 0.5). We therefore expect such cantilevers, along with the instrumentational improvements to detect them on a commercial AFM, to accelerate high-precision AFM-based SMFS studies.


[Benign pre-sacral teratoma and vestigial retrorectal cysts in the adult].
Benign mature presacral teratomas or dermoid cysts occur rarely in the adult; the incidence is estimated at less than 1/40,000. They are considered to be retrorectal vestigial growths from embryonal rests; they are usually asymptomatic and discovered by chance. However, since they have a 20-30% risk of secondary infection and a 1-12.5% risk of malignant degeneration, they should undergo complete surgical resection. We present a case of presacral dermoid cyst in a young male and summarize the diagnostic and therapeutic management.


HPLC analysis of serotonin, tryptamine, tyramine, and the hydroxycinnamic acid amides of serotonin and tyramine in food vegetables.
Biogenic monoamines such as serotonin, tryptamine, and tyramine function as neurotransmitters and mitogenic factors in animals and are involved in flowering, morphogenesis, and protection from and adaptation to environmental changes in plants. In plants, serotonin and tyramine are conjugated to form phenolic compounds via thioester linkages during the synthesis of hydroxycinnamic acid amides, including p-coumaroylserotonin (CS), feruloylserotonin (FS), p-coumaroyltyramine (CT), and feruloyltyramine (FT). In this study, we determined the amounts of the biogenic monoamines CS, FS, CT, and FT in commonly consumed vegetables using high-performance liquid chromatography. Serotonin, tryptamine, and tyramine were detected in all vegetables tested. The serotonin levels ranged from 1.8 to 294 microg/g of dry weight, the tryptamine levels ranged from 0.8 to 372 microg/g of dry weight, and the tyramine levels ranged from 1.4 to 286 microg/g of dry weight. The highest serotonin and tryptamine contents were found in tomato and cherry tomato (140.3-222 microg/g of dry weight), while paprika and green pepper had higher tyramine contents than the other vegetables (286 and 141.5 microg/g of dry weight, respectively). Overall, the levels of CS, FS, CT, and FT ranged from 0.03 to 13.8 microg/g of dry weight, with green onion possessing the highest levels of CS (0.69 microg/g of dry weight), FT (1.99 microg/g of dry weight), and CT (13.85 microg/g of dry weight).


Detection of a bromoperoxidase in Streptomyces phaeochromogenes.
A bromoperoxidase could be detected after fractionation in the chloramphenicol producing actinomycete, Streptomyces phaeochromogenes. This enzyme is capable of catalyzing the bromination of the antifungal antibiotic pyrrolnitrin [3-chloro-4-(2-nitro-3-chlorophenyl)pyrrole] in the 2-position of the pyrrole ring. The enzyme had a pH optimum of 5.0. This procaryotic bromoperoxidase requires the presence of H2O2 and can also brominate monochlorodimedone, but cannot catalyze chlorination. This enzyme is the first haloperoxidase described from procaryotic sources.


Lectin pathway proteins of the complement system in normotensive pregnancy and pre-eclampsia.
The lectin pathway of the complement system may be involved in the pathogenesis of pre-eclampsia. We aimed to investigate changes in serum concentrations of a broad range of lectin pathway proteins during normal pregnancy and their association with pre-eclampsia, placental infarctions and intrauterine growth restriction (IUGR). We included 51 women with normotensive pregnancies and 54 women with pregnancies complicated by pre-eclampsia. Blood samples were obtained at gestational weeks 16, 33, 37, and after delivery for the normotensive pregnant women and before and after delivery for women with pre-eclampsia. Mannose-binding lectin (MBL), H- and M-ficolin, collectin liver-1 (CL-L1), MBL-associated serine protease (MASP)-1, MASP-2 and MASP-3 and MBL-associated proteins of 19 (MAp19) and 44 (MAp44) kDa were analysed. Clinical information was obtained from medical records. The placentae were examined by two experienced perinatal pathologists. Lectin pathway protein concentrations generally increased during normal pregnancy and decreased after delivery in both normotensive pregnant women and women with pre-eclampsia. Exceptions were MASP-3 which increased after delivery in both groups (P < 0.0001) and H-ficolin which increased after delivery in pre-eclampsia (P < 0.0001). H-ficolin (P < 0.0001), M-ficolin (P = 0.005) and MASP-3 (P = 0.03) concentrations were lower in women with pre-eclampsia than in normotensive pregnant women. Low MASP-3 concentrations were associated with placental infarction (P = 0.03) and IUGR (P = 0.04). Low H-ficolin concentrations were associated with IUGR (P < 0.01). In general, lectin pathway protein serum concentrations increased during normal pregnancy. H-ficolin and MASP-3 may be involved in the pathophysiology of pre-eclampsia and IUGR and could be potential future pre-eclampsia biomarkers.


Dynamic quantum crystallography: lattice-dynamical models refined against diffraction data. II. Applications to L-alanine, naphthalene and xylitol.
In the first paper of this series [Hoser & Madsen (2016). Acta Cryst. A72, 206-214], a new approach was introduced which enables the refinement of frequencies of normal modes obtained from ab initio periodic computations against single-crystal diffraction data. In this contribution, the performance of this approach is tested by refinement against data in the temperature range from 23 to 205 K on the molecular crystals of L-alanine, naphthalene and xylitol. The models, which are lattice-dynamical models derived at the Γ point of the Brillouin zone, are able to describe the atomic vibrations of L-alanine and naphthalene to a level where the residual densities are similar to those obtained from the independent atom model. For the more flexible molecule xylitol, larger deviations are found. Hydrogen ADPs (anisotropic displacement parameters) derived from the models are in similar or better agreement with neutron diffraction results than ADPs obtained by other procedures. The heat capacity calculated after normal mode refinement for naphthalene is in reasonable agreement with the heat capacity obtained from calorimetric measurements (to less than 1 cal mol-1 K-1 below 300 K), with deviations at higher temperatures indicating anharmonicity. Standard uncertainties and correlation of the refined parameters have been derived based on a Monte Carlo procedure. The uncertainties are quite small and probably underestimated.


Late presentation of hepatic pseudoaneurysm after liver trauma: a case of misdiagnosed cholecystitis and successful endovascular embolization.
Hepatic artery pseudoaneurysms are a significant health concern, often incidentally discovered during computed tomography (CT) scans, as their exact incidence rate remains unknown. The most common symptoms of a pseudoaneurysm are hematemesis, abdominal pain, anemia and jaundice. A triad of right upper quadrant pain, jaundice and overt upper gastrointestinal bleeding is a classic presentation of the condition, which occurs in only a third of patients with hemobilia. Patients have a high risk of rupture, and an endovascular approach with coil embolization is recommended. A case of a 28-year-old woman with a post-traumatic hepatic pseudoaneurysm is presented in which she was initially misdiagnosed and treated for cholecystitis. After the development of jaundice, hematemesis and melena, a CT angiography confirmed diagnosis. Endovascular embolization was performed successfully, and the patient recovered without complications. This case highlights the importance of prompt diagnosis of hepatic pseudoaneurysms, as a delayed diagnosis can result in significant morbidity and mortality.


Stream ecosystem functioning under reduced flow conditions.
Assessments of flow reduction in streams often focus on changes to biological communities and in-stream physical characteristics, with little consideration for changes in ecosystem functioning. It is unclear whether functional indicators of ecosystem condition may be useful for assessing the impacts of reduced discharge on small streams. Using weirs and diversions to reduce stream discharge during summer baseflow conditions, we tested the response of leaf breakdown, coarse particulate organic matter (CPOM) retention, and primary production to one month of water abstraction in before-after, control-impact (BACI) designed experiments. Discharge at impact (downstream) reaches decreased by over 85% in each of three small New Zealand streams compared to controls (upstream). There also were decreases in velocity, depth, and wetted width. Sediment cover increased at impact reaches, but there were only small changes to conductivity, pH, and surface water temperature. We installed mesh bags filled with willow leaves in-stream for one month to measure leaf breakdown. Reduced discharge had little influence on leaf breakdown rate in these streams. Travel distances and retention structures for CPOM were evaluated using releases of paper strips and wooden dowelling over a range of discharges. The distance traveled by released CPOM increased with increasing discharge, and the importance of riffles as retention structures increased at lower discharges. We measured the accumulation of chlorophyll a after one month on artificial substrates as an estimate of the relative primary production of control and impact reaches. The differences in chlorophyll a concentrations between control and impact reaches were inconsistent among streams. These ecosystem functions have responded inconsistently to water removal in these streams. However, the strong response of CPOM retention to reduced discharge could complement measures of biological community structure when the influence of reduced discharge is assessed. We recommend further investigation in a wide range of streams to assess the utility of these processes as functional indicators of reduced discharge.


Piezo1 plays a role in erythrocyte volume homeostasis.
Mechanosensitivity is an inherent property of virtually all cell types, allowing them to sense and respond to physical environmental stimuli. Stretch-activated ion channels represent a class of mechanosensitive proteins which allow cells to respond rapidly to changes in membrane tension; however their identity has remained elusive. The piezo genes have recently been identified as a family of stretch-activated mechanosensitive ion channels. We set out to determine the role of piezo1 during zebrafish development. Here we report that morpholino-mediated knockdown of piezo1 impairs erythrocyte survival without affecting hematopoiesis or differentiation. Our results demonstrate that piezo1 is involved in erythrocyte volume homeostasis, disruption of which results in swelling/lysis of red blood cells and consequent anemia.


Role of the major antigen of Mycobacterium tuberculosis in cell wall biogenesis.
The dominant exported proteins and protective antigens of Mycobacterium tuberculosis are a triad of related gene products called the antigen 85 (Ag85) complex. Each has also been implicated in disease pathogenesis through its fibronectin-binding capacities. A carboxylesterase domain was found within the amino acid sequences of Ag85A, B, and C, and each protein acted as a mycolyltransferase involved in the final stages of mycobacterial cell wall assembly, as shown by direct enzyme assay and site-directed mutagenesis. Furthermore, the use of an antagonist (6-azido-6-deoxy-alpha, alpha'-trehalose) of this activity demonstrates that these proteins are essential and potential targets for new antimycobacterial drugs.


ASSOCIATIONS BETWEEN ALLOZYME FREQUENCIES AND SOIL CHARACTERISTICS IN GAILLARDIA PULCHELLA (COMPOSITAE).
Selection favoring different alleles in different environments frequently has been suggested as an explanation for allozyme variation within and among populations. This hypothesis predicts that allozyme frequencies will be correlated with environmental variables. Previous studies on allozyme frequency-environment covariation in plants often have relied on qualitative assessments of the environment and have emphasized highly autogamous species. We have examined allozyme frequency-soil associations in Gaillardia pulchella, an obligately outcrossed annual plant, by regressing the frequencies of 15 common allozymes representing six polymorphic enzyme loci on principal components from a set of 20 quantitative soil variables. Fifty-one populations, representing four taxonomic varieties, were included in the analysis. Among the 26 populations representing the var. pulchella, allozymes Adh-2f and Pgm-1c were significantly associated with a block of highly inter-correlated soil characteristics which serve to discriminate between soils derived from calcareous vs. non-calcareous rock types. This geographically complex pattern of allozyme frequency-soil covariation is not likely to be spurious and, thus, indicates the presence of adaptively differentiated soil races, or ecotypes. However, these results are not sufficient to conclude that the allozyme frequency divergence between ecotypes was mediated by selection, either directly or through genetic hitchhiking. The pattern of allozyme frequency-soil covariation within var. pulchella was not found among the other taxonomic varieties. Patterns of genotype-environment covariation often may be recognizable only within geographically or environmentally restricted groups of populations because of the confounding influences of other environmental variables.


Apoptosis and expression of Bcl-2, Bcl-XL, and Bax in renal cell carcinomas.
There are at present disparate published results with regard to the relevance of the Bcl-2 gene family, levels of apoptosis, and cell proliferation in the development and progression of renal cell carcinoma (RCC). The present study analyses the inter-relationship between the expression of representatives of the anti-apoptotic (Bcl-2, Bcl-XL) or pro-apoptotic (Bax) Bcl-2 proteins, incidence of apoptosis, and mitosis in a selected small group of 22 graded RCCs that had paired normal renal tissue, or non-neoplastic tissue in the renal biopsy specimen. The cases were chosen to determine the feasibility of measuring these parameters as potential surrogate markers of progression or treatment failure of the cancers. The results showed that in approximately 50% of the RCCs, where Bcl-2 and/or Bcl-XL expression was high, apoptosis was not detected, and when expression of these proteins was low or not found, increased levels of apoptosis were seen. In most of the remaining 50% of samples, high levels of Bcl-XL but not Bcl-2 were negatively correlated with low levels of apoptosis (Bcl-XL: r = -0.437, P = 0.07 and Bcl-2: r = +0.560, P = 0.02). For the same group of samples, high Bax expression was found in association with apoptosis (r = +0.578, P = 0.02). A novel finding was an association between low expression of Bcl-2 and/or Bcl-XL in normal tissue and the level of expression of these proteins in the RCCs, an intrinsic variation that may be an individual patient factor. The results indicate that, in RCCs with increased expression of Bcl-2 and/or Bcl-XL, levels of apoptosis are minimal and these combined factors may assist in progression of the cancers and resistance to treatments.


Anomalous origin of the circumflex coronary artery in a patient with atrial and ventricular septal defects.
We report a patient in whom the circumflex coronary artery arose anomalously from the right aortic sinus. The patient also had atrial and ventricular septal defects in the setting of Eisenmenger physiology. We recommended transplantation of the heart and lungs.


Reactive oxygen and nitrogen species: friends or foes?
Chemical and physiological functions of molecular oxygen and reactive oxygen species (ROS) and existing equilibrium between pools of pro-oxidants and anti-oxidants providing steady state ROS level vital for normal mitochondrial and cell functioning are reviewed. The presence of intracellular oxygen and ROS sensors is postulated and few candidates for this role are suggested. Possible involvement of ROS in the process of fragmentation of mitochondrial reticulum made of long mitochondrial filaments serving in the cell as "electric cables", as well as the role of ROS in apoptosis and programmed mitochondrial destruction (mitoptosis) are reviewed. The critical role of ROS in destructive processes under ischemia/reoxygenation and ischemic preconditioning is discussed. Mitochondrial permeability transition gets special consideration as a possible component of the apoptotic cascade, resulting in excessive "ROS-induced ROS release".


4,300 steps per day prior to surgery are associated with improved outcomes after pancreatectomy.
Decreased preoperative physical fitness and low physical activity have been associated with preoperative functional reserve and surgical complications. We sought to evaluate daily step count as a measure of physical activity and its relationship with post-pancreatectomy outcomes. Patients undergoing pancreatectomy were given a remote telemonitoring device to measure their preoperative levels of physical activity. Patient activity, demographics, and perioperative outcomes were collected and compared in univariate and multivariate logistic regression analysis. 73 patients were included. 45 (61.6%) patients developed complications, with 17 (23.3%) of those patients developing severe complications. These patients walked 3437.8 (SD 1976.7) average daily steps, compared to 5918.8 (SD 2851.1) in patients without severe complications (p < 0.001). In logistic regression analysis, patients who walked less than 4274.5 steps had significantly higher odds of severe complications (OR = 7.5 (CI 2.1, 26.8), p = 0.002). Average daily steps below 4274.5 before surgery are associated with severe complications after pancreatectomy. Preoperative physical activity levels may represent a modifiable target for prehabilitation protocols.


Trial-and-error ethics: experimenting with non-heartbeating cadaver organ donation.
It seems commendable that Michael A. DeVita and James V. Snyder, and the physicians, nurses, and Ethics Committee of the University of Pittsburgh Medical Center (UPMC) on whose behalf they write, continually and publicly reflect on the policy they have enacted to obtain organs from non-heartbeating cadaver donors, and that they are willing to change their procedures in the light of their experiences with their protocol for this type of transplant, and the criticisms of it they have invited. It is also disquieting because, in effect, the Medical Center is engaged in trial-and-error experimentation with some features of the protocol that, from the outset, should not have been a part of it, and with certain revisions of the protocol that are as questionable as what has been modified. Is it really as virtuous as it appears, then, for UPMC health professionals and Ethics Committee members to constantly reevaluate and alter their policy for non-heartbeating organ donation, and for their spokespersons to issue one report after another containing their latest thoughts and actions in this regard?


Impact of Initial CSF Findings on Outcome Among Patients With National Cancer Institute Standard- and High-Risk B-Cell Acute Lymphoblastic Leukemia: A Report From the Children's Oncology Group.
Purpose To determine the prognostic significance of blasts, and of white and red blood cells, in CSF samples at diagnosis of acute lymphoblastic leukemia (ALL), a uniform CSF and risk group classification schema was incorporated into Children's Oncology Group B-cell ALL (B-ALL) clinical trials. Methods CSF status was designated as follows: CNS1, no blasts; CNS2a to 2c, < 5 WBCs/μL and blasts with/without ≥ 10 RBCs/μL or ≥ 5 WBCs/μL plus blasts, with WBCs ≥ 5 times the number of RBCs; CNS3a to 3c, ≥ 5 WBCs/μL plus blasts with/without ≥ 10 RBCs/μL or clinical signs of CNS disease. CNS2 status did not affect therapy; patients with CNS3 status received two extra intrathecal treatments during induction and augmented postinduction therapy with 18 Gy of cranial radiation. Results Among 8,379 evaluable patients enrolled from 2004 to 2010, 7,395 (88.3%) had CNS1 status; 857 (10.2%), CNS2; and 127 (1.5%), CNS3. The 5-year event-free and overall survival rates were, respectively, 85% and 92.7% for CNS1, 76% and 86.8% for CNS2, and 76% and 82.1% for CNS3 ( P < .001). In multivariable analysis that included age, race/ethnicity, initial WBC, and day-29 minimal residual disease < 0.1%, CSF blast, regardless of cell count, was an independent adverse predictor of outcome for patients with standard- or high-risk disease according to National Cancer Institute criteria. The EFS difference reflected a significant difference in the incidence of CNS, not marrow, relapse in patients with CNS1 versus CNS2 and/or CNS3 status. Conclusion Low levels of CNS leukemia, regardless of RBCs, predict inferior outcome and higher rates of CNS relapse. These data suggest that additional augmentation of CNS-directed therapy is warranted for CNS2 disease.


Co-precipitation of Ni, Cr, Mn, Pb and Zn in industrial wastewater and sediment samples with copper(II) cyclo-hexylmethyldithiocarbamate for their flame atomic absorption spectrometric determination.
A co-precipitation technique for nickel(II), chromium(II), manganese(II), lead(II) and zinc(II) with the aid of copper(II) cyclo-hexylmethyldithiocarbamate was established. The influences of some analytical parameters such as pH, sample volume, amounts of cyclo-hexylmethyldithiocarbamate and copper(II) on the recovery of metal ions were investigated. The heavy metals in the precipitate were determined by flame atomic absorption spectrophotometry. The range of detection limits for the heavy metals was 0.003-0.005 mg/L. The atomic spectrometric technique with co-precipitation procedure was successfully applied for the determination of Ni, Cr, Mn, Pb and Zn in industrial wastewater and sediment samples from Ladipo stream in Lagos, Nigeria. The mean concentrations for these metals using co-precipitation procedure were not significantly different from corresponding concentrations obtained using spectrometric techniques without co-precipitation procedure.