A neuropathologic feature of brain aging: multi-lumen vascular profiles.
Cerebrovascular pathologies other than frank infarctions are commonly seen in aged brains. Here, we focus on multi-lumen vascular profiles (MVPs), which are characterized by multiple vessel lumens enclosed in a single vascular channel. Little information exists on the prevalence, risk factors, and co-pathologies of MVPs. Therefore, we used samples and data from the University of Kentucky Alzheimer's Disease Research Center (n = 91), the University of Kentucky Pathology Department (n = 31), and the University of Pittsburgh Pathology Department (n = 4) to study MVPs. Age at death was correlated with MVP density in the frontal neocortex, Brodmann Area 9 (r = 0.51; p < 0.0001). Exploratory analyses were performed to evaluate the association between conventional vascular risk factors (e.g., hypertension, diabetes), cardiovascular diseases (e.g., heart attack, arrhythmia), and cerebrovascular disease (e.g., stroke); the only nominal association with MVP density was a self-reported history of brain trauma (Prevalence Ratio = 2.1; 95 CI 1.1-3.9, before correcting for multiple comparisons). No specific associations were detected between neuropathological (e.g., brain arteriolosclerosis) or genetic (e.g., APOE) variables and MVP density. Using a tissue clearing method called SeeDB, we provide 3-dimensional images of MVPs in brain tissue. We conclude that MVPs are an age-related brain pathology and more work is required to identify their clinical-pathological correlation and associated risk factors.


Consumption of vitamin and mineral supplements and its correlates among medical students in Eastern Province, Saudi Arabia.
Although vitamin and mineral supplements are popular in many countries, few studies have documented their use among college students. Moreover, there is not much national data on the use of supplements by Saudi medical students. The aim of this study was to determine the prevalence of the use of vitamin and mineral supplements by female medical students at Imam Abdulrahman Bin Faisal University (IAU) in Dammam, Saudi Arabia, and to identify characteristics associated with the use of supplements in this population. For this cross-sectional study, all female medical students at IAU in Dammam, Saudi Arabia, were approached during September to November 2016. Results were presented as frequency distribution. Chi-square test and logistic regression analysis were performed to determine factors associated with supplement use; P < 0.05 was considered statistically significant. Completed questionnaires were returned by 474 students (response rate of 77%). The prevalence of the use of vitamin and mineral supplements was 44.6%; Vitamin D was the most commonly used supplement. Association of supplement use with a higher family income and the habit of regular exercise was statistically significant. However, this use of supplements was not significantly associated with smoking or marital status. The use of supplements, particularly Vitamin D, by female medical students was 44.6%. The highest percentage of users belonged to families with higher incomes and had the habit of exercising regularly.


Cost effectiveness of interferon-γ release assay for TB screening of HIV positive pregnant women in low TB incidence countries.
To assess the cost effectiveness for tuberculosis (TB) screening of high-risk human immunodeficiency virus (HIV) positive pregnant women by using interferon-gamma release assays (IGRAs) compared to the tuberculin skin test (TST) in low TB incidence countries. We constructed Markov models using a public health payer perspective. The target population was a hypothetical cohort of 20 year-olds HIV positive pregnant women until age 50 years in three most common screening situations; close contacts, immigrants from high burden countries and occasional screenings. BCG vaccination status was considered. Five strategies; TST, QuantiFERON®-TB Gold In-Tube (QFT), T-SPOT®. TB (T-SPOT), TST followed by QFT and TST followed by T-SPOT were modeled. The main outcome measure of effectiveness was quality-adjusted life-years (QALYs) gained. The incremental cost effectiveness ratio (ICER) of each screening arm was applied and compared. In the base case analyses of close contacts, T-SPOT yielded the greatest benefits at the lowest cost. In the base case analyses of immigrants and occasional screenings, TST followed by QFT yielded the greatest benefits at the lowest cost. Using an IGRA for TB screening of high-risk HIV positive pregnant women is recommended on the basis of the cost effectiveness in low TB incidence countries.


Regional anesthesia for postoperative pain control in children: focus on continuous central and perineural infusions.
Regional anesthesia is widely employed for postoperative pain control in both adults and children. Central or perineural approaches can be performed as a bolus injection or as a continuous infusion of local anesthetics. However, bolus injections, even with the addition of adjuvants, are inadequate for prolonged surgery and long-term pain control. Continuous infusion remains the technique of choice when there is a prolonged operation or intense postoperative pain. This article reviews the safety and efficacy of central and perineural continuous infusions for postoperative pain control in children. The literature confirms the very low rate of complications and adverse effects of regional anesthesia in children. However, clinicians need to be aware of the key points for performing a block and placing a catheter in children: good knowledge of anatomic and physiologic differences between adults and children is necessary; the use of newer local anesthetics, such as ropivacaine and levobupivacaine, increases the therapeutic window; and that it is mandatory to work with dedicated pediatric equipment. Through the use of new techniques such as nerve mapping and/or ultrasound the success of blocks can be improved and the risks reduced.


Prolactin receptors in the ovary.
The binding of prolactin (PRL) to the plasma membranes of bovine and human ovaries was investigated using both homologous and heterologous 125I-prolactin. Saturation and Scatchard analysis demonstrated that human prolactin binds to human ovarian membranes with a Kd of 2 x 10(-10) M; to bovine ovarian membranes with a Kd of 1.9 x 10(-10) M; and to bovine corpora lutea membranes with a Kd of 1.9 x 10(-10) M. The concentrations of binding sites in bovine and human ovaries were 2.9 x 10(-15) moles/mg of protein and 2.0 x 10(-15) moles/mg of protein, respectively. The number of bindings sites in the bovine corpora lutea was 1.5 x 15(-15) moles/mg of protein. Specificity studies with bovine PRL, ovine PRL, human luteinizing hormone, human follicle-stimulating hormone, and bovine growth hormone showed this binding to be specific. Comparison of binding of PRL to membranes of other target and nontarget tissues suggests that the ovary is one of the primary target tissues. These data suggest that prolactin plays a role in the ovarian cycle.


Leukocyte response to somatostatin in normals and patients with bacterial leukocytosis.
In normals and patients with acute bakterial leukocytosis synthetic somatostatin decreases cell count of stab neutrophils, neutrophils and eosinophils. After termination of somatostatin infusion a rebound phenomenon occurred. In healthy normals lymphocyte count increased during somatostatin infusion. This effect outlasts the end of infusion for more than 24 h. No significant effect could be demonstrated in patients with a bacterial leukocytosis.


1,2-O-Iso-propyl-idene-β-d-lyxo-furan-ose.
In the title compound C8H14O5, the pento-furan-ose five-membered ring has a twisted conformation on two carbon atoms while the five-membered ring of the iso-propyl-idene group has an envelope conformation on an oxygen atom. Hy-droxy groups are involved an infinite network of O-H⋯O hydrogen bonds that leads to the formation of a layer parallel to the (001) plane. Only weak C-H⋯O contacts exist between neighboring layers.


Telephone consultations with otolaryngology - head and neck surgery reduced emergency visits and specialty consultations in northern Alberta.
RAAPID (Referral, Access, Advice, Placement, Information, and Destination) is a 24-h call center in Alberta, Canada, facilitating urgent telephone consultations between physicians and specialists. We evaluated the extent to which RAAPID calls to Otolaryngology-Head and Neck Surgery (OHNS) reduced visits to the emergency department and specialty clinics. This was a cross-sectional study evaluating all telephone consultations to OHNS from physicians in northern Alberta between 2013 and 2014 (T1) (where consultations by residents occurred) and 2015 to 2017 (T2) (where consultations were done by consultants during office hours and residents during after hours). Outcomes of the calls included medical advice, specialty clinic referrals, and emergency department (ED) referrals. Differences in the reduction of ED visits and costs, overall as well as in T1 and T2, were assessed using multivariate logistic regression. Overall, 62.3% (1064/1709) of telephone consultations reduced ED visits consisting of advice being provided (n = 884; 83.1%) and referral to specialty clinics (n = 180; 16.9%). The adjusted odds ratio of calls reducing emergency visits in T2 as compared to T1 was 2.47 (95% CI 1.99 to 3.08). The adjusted odds ratio of reducing ED visits during office hours compared to after-hours 2.54 (95% CI 1.77-3.64). The estimated direct costs avoided from ED visits in T1 and T2 were $42,224.22 and $114,393.86, respectively. RAAPID telephone consultations to OHNS were effective in reducing ED visits and healthcare costs. This model should be considered in other areas to improve efficiencies within the health system.


[Surgical approaches of pheochromocytoma].
To evaluate the effects of surgery of Pheo. Pheochromocytoma was removed in 89 patients. In 39 patients, pheochromocytoma was removed by intercostal incision at the 11th rib and 15 at the 10th rib, 14 by transrectal incision, 9 by abdominothoracic incision, 5 by abdominal subcostal oblique incision, 3 by inferior abdominal incision, 3 by posterolateral incision. One patient received laparoscopic surgery. Pheochromocytoma definitely localized was removed by intercostal incision at the 10th or 11th rib. The suitable pathway for removal giant pheochromocytoma or pheochromocytoma related big vessels is transrectal incision or abdominothoracic incision. The incision of multiple pheochromocytoma or extra-adrenal pheochromocytoma may be in the middle of the abdomen.


Postexercise recovery of the spatial QRS/T angle as a predictor of sudden cardiac death.
Postexercise measurement of heart rate (HR) recovery and QT interval dynamics provides prognostic information in various patient populations. The purpose of this study was to assess whether the measurement of the spatial relationship between the depolarization and repolarization wavefronts (total cosine R-to-T [TCRT]) during the postexercise recovery phase would yield prognostic information. The population consisted of 1297 patients (56 ± 13 years; 67% men) who performed a clinically indicated bicycle stress test. The exercise-recovery hysteresis of TCRT was quantified from the 12-lead exercise electrocardiogram by measuring the TCRT/HR loop area bounded by the exercise and first 3-minute postexercise recovery curves. The HR-corrected TCRT/HR hysteresis was calculated by dividing the area with the HR decrement during the first 3 minutes of recovery. HR recovery was measured at 1 minute postexercise recovery. End points were cardiac death and sudden cardiac death. During an average follow-up of 45 ± 12 months, 74 patients died (5.7%); 35 (2.6%) were cardiac deaths and 24 (1.9%) were sudden cardiac deaths. Reduced TCRT/HR loop area and TCRT/HR hysteresis were associated with cardiac mortality (P <.001). After adjustments for clinical variables, including ejection fraction, TCRT/HR loop area remained an independent predictor of cardiac death (hazard ratio 5.6; 95% confidence interval 1.6-19.1; P = .007) and sudden cardiac death (10.7; 95% confidence interval 1.4-83.7; P = .024). HR recovery did not remain a significant predictor in the multivariate analysis. Attenuated hysteresis of the depolarization and repolarization wavefronts during postexercise recovery is associated with an increased risk of cardiac and sudden cardiac death. Analysis of repolarization dynamics from exercise electrocardiogram represents a promising tool for risk stratification.


Enzymatic hydrolysis of cellulosic materials using synthetic mixtures of purified cellulases bioengineered at N-glycosylation sites.
Mutant forms of recombinant endoglucanase II (EG II, N194A), cellobiohydrolase I (CBH I, N45A) and cellobiohydrolase II (CBH II, N219A) from Penicillium verruculosum with enhanced cellulase activities, achieved by engineering of enzyme N-glycosylation sites in our previous studies, were used as components of the binary and ternary mixtures of cellulases in hydrolysis of Avicel and milled aspen wood. Using the engineered forms of the enzymes at a dosage of 10 mg/g substrate resulted in significant boosting of the glucose release from cellulose in the presence of excess β-glucosidase relative to the performance of the corresponding wild-type mixtures at the same loading. The boosting effects reached 11-40% depending on the reaction time and substrate type. In hydrolysis of both cellulosic substrates by the binary mixtures of cellulases, all the enzyme pairs exhibited synergism. The magnitude of the synergistic effects (Ks) did not depend notably upon the induced mutations in the enzymes, and they were in the range of 1.3-1.8 for the combinations of EG II with CBH I (or CBH II), and 2.3-2.9 for the CBH I-CBH II pair. The results of this study should provide a basis for the development of a more effective fungal strain capable of producing cellulase cocktails with enhanced hydrolytic performance against lignocellulosic materials.


Multivariate hierarchical frameworks for modeling delayed reporting in count data.
In many fields and applications, count data can be subject to delayed reporting. This is where the total count, such as the number of disease cases contracted in a given week, may not be immediately available, instead arriving in parts over time. For short-term decision making, the statistical challenge lies in predicting the total count based on any observed partial counts, along with a robust quantification of uncertainty. We discuss previous approaches to modeling delayed reporting and present a multivariate hierarchical framework where the count generating process and delay mechanism are modeled simultaneously in a flexible way. This framework can also be easily adapted to allow for the presence of underreporting in the final observed count. To illustrate our approach and to compare it with existing frameworks, we present a case study of reported dengue fever cases in Rio de Janeiro. Based on both within-sample and out-of-sample posterior predictive model checking and arguments of interpretability, adaptability, and computational efficiency, we discuss the relative merits of different approaches.


[Classification of cell-based medicinal products and legal implications: An overview and an update].
In general, cell-based medicinal products do not represent a uniform class of medicinal products, but instead comprise medicinal products with diverse regulatory classification as advanced-therapy medicinal products (ATMP), medicinal products (MP), tissue preparations, or blood products. Due to the legal and scientific consequences of the development and approval of MPs, classification should be clarified as early as possible. This paper describes the legal situation in Germany and highlights specific criteria and concepts for classification, with a focus on, but not limited to, ATMPs and non-ATMPs. Depending on the stage of product development and the specific application submitted to a competent authority, legally binding classification is done by the German Länder Authorities, Paul-Ehrlich-Institut, or European Medicines Agency. On request by the applicants, the Committee for Advanced Therapies may issue scientific recommendations for classification.


Dacomitinib overcomes afatinib-refractory carcinomatous meningitis in a lung cancer patient harbouring EGFR Ex.19 deletion and G724S mutation; a case report.
It has been reported that the efficacy of EGFR-TKI is predicted, not by which exon of the EGFR gene is mutated, but by the structural change in the EGFR protein due to the mutation. Here, we present an EGFR-mutated lung cancer patient with a 13-year history of anticancer treatment, in which EGFR ex.19 deletion (E746_S752 > V) and G724S mutations were detected by liquid biopsy during 12th line afatinib treatment, and switching to dacomitinib showed improvement of cancerous meningitis. We choose dacomitinib as 14th line chemotherapy based on ex.19 deletion and G724S mutant EGFR structure and its penetration rate to cerebral fluid, which successfully prolonged her life by 6 months. The optimal EGFR-TKI may be selected by understanding the EGFR compound mutation profile by next generation sequencing and predicting the effect based on the structure. Dacomitinib may be effective choice in afatinib-refractory carcinomatous meningitis harboring G724S mutation. This is the first case report showing that a change to dacomitinib responded to afatinib refractory cancerous meningitis.


Immuno-modulation effect of sulphated polysaccharide (porphyran) from Porphyra vietnamensis.
Our investigation explores the immuno-efficiency of sulphated polysaccharides enriched Porphyra vietnamenis. Isolated polysaccharide fraction (17.1-25.8%) was characterized by FTIR and NMR spectroscopy which showed the presence of typical linear backbone structure called as porphyran. Oral administration of porphyran (200-500 mg/kg) evoked a significant (P ≤ 0.05) increase in weight of the thymus, spleen and lymphoid organ cellularity. The total leucocyte and lymphocyte count was increased significantly (P<0.005). The increase in the percent neutrophil adhesion to nylon fibres as well as a dose-dependent increase in antibody titre values was observed. A decreased response to DTH reaction induced by SRBC was recorded. A potential phagocytic response was seen and significant changes were observed in the formation of formazone crystals. It also prevented myelosuppression in cyclophosphamide drug treated rats. The results indicated that P. vietnamenis possesses potential immunomodulatory activity and has therapeutic potential for the prevention of autoimmune diseases.


OL-Protocadherin is essential for growth of striatal axons and thalamocortical projections.
The ventral telencephalon in the embryonic brain is thought to provide guidance cues for navigation of thalamocortical axons, but the mechanisms involved remain largely elusive. OL-protocadherin (OL-pc), a member of the cadherin superfamily, is highly expressed by striatal neurons in the developing ventral telencephalon. Here we show that OL-pc-deficient (Pcdh10(-/-)) mice have defects in axon pathways through the ventral telencephalon; for example, thalamocortical and corticothalamic projections cannot cross the ventral telencephalon. In the ventral telencephalon, striatal axons fail to grow out, and, concomitantly, the caudal portion of the globus pallidus and the associated 'corridor' thought to be important for thalamocortical fiber navigation do not form. The inability of the striatum to extend axons is also observed in vitro. These results show that OL-pc is essential for both elongation of striatal axons and patterning of the putative guidance cues for thalamocortical projections.


Occurrence of ²¹⁰Po and biological effects of low-level exposure: the need for research.
Polonium-210 (²¹⁰Po) concentrations that exceed 1 Bq/L in drinking-water supplies have been reported from four widely separated U.S. states where exposure to it went unnoticed for decades. The radionuclide grandparents of ²¹⁰Po are common in sediments, and segments of the public may be chronically exposed to low levels of ²¹⁰Po in drinking water or in food products from animals raised in contaminated areas. We summarized information on the environmental behavior, biokinetics, and toxicology of ²¹⁰Po and identified the need for future research. Potential linkages between environmental exposure to ²¹⁰Po and human health effects were identified in a literature review. ²¹⁰Po accumulates in the ovaries where it kills primary oocytes at low doses. Because of its radiosensitivity and tendency to concentrate ²¹⁰Po, the ovary may be the critical organ in determining the lowest injurious dose for ²¹⁰Po. ²¹⁰Po also accumulates in the yolk sac of the embryo and in the fetal and placental tissues. Low-level exposure to ²¹⁰Po may have subtle, long-term biological effects because of its tropism towards reproductive and embryonic and fetal tissues where exposure to a single alpha particle may kill or damage critical cells. ²¹⁰Po is present in cigarettes and maternal smoking has several effects that appear consistent with the toxicology of ²¹⁰Po. Much of the important biological and toxicological research on ²¹⁰Po is more than four decades old. New research is needed to evaluate environmental exposure to ²¹⁰Po and the biological effects of low-dose exposure to it so that public health officials can develop appropriate mitigation measures where necessary.


Urinary beta-glucuronidase in screening and followup of primary urinary tract malignancy.
The usefulness of urinary beta-glucuronidase in screening and followup of urinary tract malignancy was tested. The activity and concentration of urinary beta-glucuronidase were determined in 80 healthy subjects, 113 urology patients with no cancer, and 60 with transitional cell and 23 with renal cell carcinoma. Patients with cancer were followed for 3 to 33 weeks. The enzyme activity and concentration were significantly greater in the cancer group, including residual tumor or recurrence after therapy, than in the controls with excellent discrimination. Urinary beta-glucuronidase was indeed useful in screening and followup of urinary tract malignancy.


Purple acid phosphatases in coffee-genome-wide identification and the transcriptional profile of selected members in roots upon Pi starvation.
Phosphorus (P) availability is determinant for crop productivity and, despite the sufficient amount of this nutrient in arable land, most of it remains insoluble, leading to the need of high fertilizer input. To cope with P scarcity forecasts and also for cropping costs alleviation, genotypes better adapted to promote P solubilization and absorption are required, especially for perennial crops. Coffee is one of these important perennial crops cultivated in soils with low P availability, and thus we aimed to study adaptive strategies to coffee genotypes in acquire phosphorus. In this study, we focused on rhizosphere phosphatase activity, a major characteristic related to P solubilization from organic pools. To explore the genetic basis of this characteristic, we firstly identified 29 Purple Acid Phosphatases (PAP) genes in C. canephora genome and selected five candidates with higher potential to encode secreted PAPs. We found that C. arabica genotypes have diverse profiles of rhizosphere phosphatase activity, as well as microbial biomass carbon, which we measured to explore the impact of the plant on microbiota and also to discriminate the phosphatase activity origin (plant or microorganism-derived). We selected two C. arabica cultivars with contrasting phosphatase activity and found that one PAP gene has a correlated gene expression profile with this characteristic. This work explores coffee adaptative responses to P starvation conditions, and the information provided can further contribute to breeding programs aiming better adapted genotypes for sustainable agriculture in face of current challenges. The online version contains supplementary material available at 10.1007/s13205-022-03399-6.


[Spontaneous recovery in perforated gastroduodenal ulcer. The indications for conservative treatment].
Two observations of patients, having perforated duodenal ulcer, not operated, where spontaneous recovery was achieved, have been reported. The said opportunity has been interpreted in connection with the accepted method for conservative treatment of gastroduodenal ulcer. The indications and the contraindications for the conservative treatment have been stated.


Long-term biocompatibility evaluation of 0.5 % zinc containing hydroxyapatite in rabbits.
This study investigates the long-term biocompatibility of 0.5 % zinc-containing hydroxyapatite compared with hydroxyapatite. Spheres (425 < ∅ < 550) of both materials were produced by extrusion of ceramic slurry in calcium chloride and characterized by FTIR, XRD, XRF and SEM. Fifteen White New Zealand rabbits were submitted to general anesthesia, and an perforation (2 mm), was made in each tibia, one for zinc-containing hydroxyapatite sphere implantation and one for hydroxyapatite sphere implantation. After 26, 52 and 78 weeks, the animals were euthanized, and the fragment containing the biomaterial was harvested. A 30-50 μm section was obtained for histological analysis in bright field and polarized light. SEM images revealed similar morphologies between the tested biomaterials. Histological analysis showed that there was no difference between the test groups. The morphometric analysis, however, indicates that there was a greater absorption. The materials are biocompatible, promote osteogenesis and that the zinc-containing hydroxyapatite microspheres were absorbed more quickly.


Employing DNA binding dye to improve detection of Enterocytozoon hepatopenaei in real-time LAMP.
Enterocytozoon hepatopenaei (EHP) is a pathogen in the pancreatic tissue of prawn, as reported in recent years. Enterosporidiosis caused by EHP in Penaeus genus is spreading widely, which seriously threatens the sustainable development of shrimp aquaculture in the world. Empolying the DNA binding dye of SYTO-16, a real-time quantitative loop-mediated isothermal amplification (LAMP) method has been established herein to detect EHP. The primer sequences used in the LAMP reaction were according to the SSU rRNA gene. The LAMP assay has reached a sensitivity of 101 copies/µL and no cross-reaction with other aquatic pathogens. Compared with normal PCR, the efficiency of the LAMP technique is more sensitive, which has a shorter detection time. The use of fluorescent nucleic acid dye (SYTO-16) reveals a more satisfactory performance relative to calcein. The quantitative LAMP assay can be considered as a valid tool for rapid detection of microsporidian in prawns. Our study provides a scientific basis to follow the effect of the pathogen infection on growth of cultured penaeid shrimp.


Neuropsychological and positron emission tomography correlates in idiopathic environmental intolerances.
It has been hypothesized that people with subjective hypersensitivity to chemicals may indeed suffer from neuronal damage due to widely distributed environmental toxins and that such deficits of diagnostic importance can be demonstrated with the help of functional neuroimaging even in single cases. In this study, a small group of well-characterized patients with idiopathic environmental intolerance were examined in order to identify such changes. Twelve patients with idiopathic environmental intolerance were investigated neuropsychologically and underwent cerebral F-18 fluorodeoxyglucose (F-18 FDG) positron emission tomography (PET). The imaging results were compared with findings from 17 healthy controls. Six patients showed deficits in verbal learning and memory, three of them also had a reduced information processing speed. In the individual analyses, 11 patients showed normal cerebral glucose metabolism. In the group analysis of the patients, no areas with significantly reduced glucose metabolism could be found. No consistent pathological cognitive performance and functional imaging pattern was found. It appears premature to claim specific neuropsychological or neuroimaging findings characteristic of idiopathic environmental intolerance. Therefore cerebral F-18 FDG PET should not be used to corroborate or rule out suspected idiopathic environmental intolerance, a syndrome whose potential biological underpinnings still need to be clarified.


Pre-B lymphocytes with intracytoplasmic mu chains in the peripheral blood of rheumatoid arthritis patients.
A dysregulation of B-cell differentiation and activation has long been evidenced in rheumatoid arthritis (RA). Such analyses have, however, usually focused on the latest stages of B-cell development. Using a classical technique of immunofluorescence labeling on cytospins, we investigated the presence of peripheral pre-B lymphocytes in 92 RA patients and 23 controls. Cells with intracytoplasmic mu chains were evidenced in 58.7% of the RA patients studied, ranging between 0 and 30%, while small numbers of c-mu cells, never exceeding 6% of peripheral blood lymphocytes, were observed in 9 controls. Relationships between this feature and clinical or laboratory data were investigated, showing a negative correlation between the percentage of c-mu + lymphocytes and Ritchie's index (P = 0.05), the number of tender or swollen joints (P = 0.05), erythrocyte sedimentation rate (P = 0.005), seropositivity (P = 0.05), and disease duration (P = 0.01).


Immediate postoperative non-invasive positive pressure ventilation following midface microvascular free flap reconstruction.
There is a rare need for postoperative non-invasive positive pressure ventilation (NIPPV) following microvascular reconstruction of the head and neck. In midface reconstruction, the free flap vascular pedicle is especially vulnerable to the compressive forces of positive pressure delivery. A 60 year old female with Amyotrophic Lateral Sclerosis (ALS) presented with squamous cell carcinoma of the anterior maxilla, for which she underwent infrastructure maxillectomy and fibula free flap reconstruction. To avoid tracheotomy, the patient was extubated postoperatively and transitioned to NIPPV immediately utilizing a full-face positive pressure mask with a soft and flexible sealing layer. The patient was successfully transitioned to NIPPV immediately after extubation. The free flap exhibited no signs of vascular compromise postoperatively, and healed very well. Postoperative non-invasive positive pressure ventilation can be successfully applied following complex microvascular midface reconstruction to avoid tracheotomy in select patients without vascular compromise of the free flap.


Implementation of Instrument Based on Eight Health Related Quality of Life Domains for Measuring of Willingness to Pay for Psoriasis Treatment.
Psoriasis vulgaris (PsV) is a chronic skin condition that has a major impact on health-related quality of life (HRQOL). To determine the individual burden of PsV on HRQOL using willingness to pay (WTP) instrument. Fifty-one consecutive PsV patients were asked to evaluate their overall health and psoriasis affected health by visual analogue scale (VAS), and interviewed on 8 domains (physical, emotional, sleep, work, social, self-care, intimacy, and concentration) of HRQOL and WTP for a hypothetical cure in each domain. Two additional questions proposing 6 alternatives for therapy were also asked. The analysis is performed with descriptive and frequency statistics, Mann-Whitney and Kruskal-Wallis tests. The domains ranked highly were: physical comfort (90%), social comfort (77%), emotional health (75%) and work (53%). The following tendencies concerning WTP for top four impacted domains were found: the median WTP were the highest in the top impacted domains; the younger patients were willing to pay more than the older ones; the highest median WTP amounts appear in the lowest income group; the highest median WTP is associated with smaller psoriasis affected health VAS scores. The largest proportion and number of patients (37.3%, n=19) stated preferences for the systemic therapy. The second preferred choice was the thalassotherapy (29.4%, n=15). The utility and reliability of the instrument based on the assessment of WTP stated preferences for 8 domains of HRQOL for evaluation the individual burden of psoriasis were strongly supported.


Production and purification of bacterial membrane vesicles for biotechnology applications: Challenges and opportunities.
Bacterial membrane vesicles (BMVs) are bi-layered nanostructures derived from Gram-negative and Gram-positive bacteria. Among other pathophysiological roles, BMVs are critical messengers in intercellular communication. As a result, BMVs are emerging as a promising technology for the development of numerous therapeutic applications. Despite the remarkable progress in unveiling BMV biology and functions in recent years, their successful isolation and purification have been limited. Several challenges related to vesicle purity, yield, and scalability severely hamper the further development of BMVs for biotechnology and clinical applications. This review focuses on the current technologies and methodologies used in BMV production and purification, such as ultracentrifugation, density-gradient centrifugation, size-exclusion chromatography, ultrafiltration, and precipitation. We also discuss the current challenges related to BMV isolation, large-scale production, storage, and stability that limit their application. More importantly, the present work explains the most recent strategies proposed for overcoming those challenges. Finally, we summarize the ongoing applications of BMVs in the biotechnological field.


A prospective study of definite bites by spiders of the family Sparassidae (huntsmen spiders) with identification to species level.
Spiders of the family Sparassidae occur on most continents in tropical and temperate regions of the world. They are large Araneomorphae (modern spiders) which are often feared. There are few reports of confirmed sparassid bites and some suggest that particular genera (Neosparassus) can cause severe effects. This study investigated the circumstances and clinical effects of bites by Australian sparassid spiders with correlation to taxonomic species level. Cases were recruited prospectively from calls to Australian poison information centres and two emergency departments. Patients were included if there was a clear history of bite and the spider was caught, and were followed up over a week. There were 173 patients with spider injuries and 168 with definite bites by sparassid spiders over a 27 month period. Ninety-five percent of bites were by spiders from four genera: Isopeda (32%), Isopedella (21%), Neosparassus (27%) and Heteropoda (14%); other genera included Delena and Holconia. The seasonal and diurnal distribution of bites differed between genera, although the majority occurred in warmer months in daylight hours. Seventy-six percent of bites occurred because the spider was interfered with (picked up, dressing or picked up an object with the spider) consistent with the behaviour of most of the family requiring provocation to bite. Ninety-five percent of bites occurred on limbs, 82% on distal limbs, again consistent with handling the spider. Pain/discomfort occurred in all cases, and was severe in 27%. The median duration of pain was 5 min which was significantly less than other spiders. Puncture marks (40%) or localised initial bleeding (35%) occurred in 54% of bites, and both occurred significantly more often in all sparassid bites compared to all other spider bites (p<0.0001). Swelling occurred in 16%, itchiness in 14% and redness/red mark in 57% of cases, the latter significantly less common than other spiders (p=0.0002). Systemic effects occurred in 4% of bites and were minor (nausea and/or headache). There were no cases of necrotic ulcers or allergic reactions. There were four cases with features consistent with local infection. Bites by sparassid spiders cause minor effects, characterised by immediate and transient pain, associated with bleeding, puncture marks and local redness. The mechanism of effects appeared to be trauma rather than envenoming. There were no major differences between genera, and Neosparassus spp. did not cause major effects.


Evaluating the perceptions of a required didactic Mental Health First Aid training course among first-year pharmacy students.
Mental Health First Aid (MHFA) training was designed to help participants identify mental health challenges and provide support to those in need. The objective of this study was to evaluate the perceptions of a required first-year didactic MHFA course and to determine if MHFA reduces stigma and increases confidence in first-year pharmacy students' abilities to offer help to those experiencing a mental health challenge or a crisis. Participants were first-year doctor of pharmacy students at a single site in the United States. Methods include the utilization of a survey-based study among a single pharmacy program. Students enrolled in this required MHFA training were asked to voluntarily complete pre- and post-training surveys. A total of 69 first-year pharmacy students participated in the MHFA training. Results demonstrated that students agreed with being able to identify stigmatizing words associated with mental health disorders and that they will work to reduce or avoid using words that can be perceived as stigmatizing related to mental health (P < .001). Students reported increased confidence in their ability to start a conversation with someone who may need mental health help compared to their confidence level before the training (P < .001). Completing MHFA training during the early phases of didactic education can help reduce stigma and increase confidence in abilities to help those experiencing a mental health challenge. Programs should consider implementing MHFA or similar training early in the didactic curriculum.


Secondary depression in severe anxiety disorders: a population-based cohort study in Denmark.
Depression and anxiety disorders are highly comorbid conditions and a worldwide disease burden; however, large-scale studies delineating their association are scarce. In this retrospective study, we aimed to assess the effect of severe anxiety disorders on the risk and course of depression. We did a population-based cohort study with prospectively gathered data in Denmark using data from three Danish population registers: The Danish Civil Registration System, the Danish Psychiatric Central Register, and the Danish National Hospital Registry. We selected the cohort from people born in Denmark between Jan 1, 1955, and Dec 31, 2002, who we followed up from Jan 1, 1994, to Dec 31, 2012. The cohort was restricted to individuals with known parents. First, we investigated the effect of specific anxiety diagnoses on risk of single depressive episodes and recurrent depressive disorder. Second, we investigated the effect of comorbid anxiety on risk of readmission for depression, adjusting for sex, age, calendar year, parental age, place at residence at time of birth, and the interaction of age with sex. We included 3,380,059 individuals in our study cohort. The adjusted incidence rate ratio (IRR) for single depressive episodes was 3·0 (95% CI 2·8-3·1, p<0·0001) and for recurrent depressive disorder was 5·0 (4·8-5·2) in patients with severe anxiety disorders compared with the general population. Compared with control individuals, the offspring of parents with anxiety disorders were more likely to be diagnosed with single depressive episodes (1·9, 1·8-2·0) or recurrent depressive disorder (2·1, 1·9-2·2). Comorbid anxiety increased the readmission rates in both patients with single depressive episodes and patients with recurrent depressive disorder. Severe anxiety constitutes a significant risk factor for depression. Focusing on specific anxiety disorders might help to identify individuals at risk of depression, thereby providing new insights for prevention and treatment. The Lundbeck Foundation Initiative for Integrative Psychiatric Research (iPSYCH).


Diffusion-weighted MRI for locally recurrent prostate cancer after external beam radiotherapy.
The objectives of our study were to establish the apparent diffusion coefficients (ADCs) of tumor and nontumor irradiated tissues in patients with suspected postradiation recurrence of prostate cancer and to determine the sensitivity and specificity of a combination of T2-weighted and diffusion-weighted imaging (DWI) for detecting local recurrence. Twenty-four patients with rising prostate-specific antigen levels after having completed radiation therapy 30-130 months earlier (median, 62 months) underwent endorectal T2-weighted imaging and DWI (b = 0, 100, 300, 500, and 800 s/mm(2)) followed by transrectal ultrasound (TRUS)-guided biopsy. Images were scored prospectively as positive for tumor if a region of low signal intensity on T2-weighted imaging within the prostate corresponded with a focally restricted area on the ADC map. A region of interest (ROI) was drawn around the suspicious lesion on a single slice of the ADC map and a corresponding ROI was drawn around presumed nontumor irradiated peripheral zone and central gland tissues on the opposite side of the prostate. The sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) were determined against TRUS-guided biopsy reference standard (octant, n = 17; sextant, n = 5; two samples, n = 1; 12 samples, n = 1). Sixteen of 24 patients (66.7%) had positive histology findings. The median tumor ROI area was 0.37 cm(2) (quartiles, 0.30 and 0.82 cm(2)). The sensitivity, specificity, PPV, and NPV for detecting tumor were 93.8%, 75%, 88.2%, and 85.7%, respectively. A cutoff ADC of 1216 × 10(-6) mm(2)/s could predict tumor with 100% sensitivity and 96% specificity (area under the receiver operating characteristic curve = 0.992). An ADC derived from DWI is a useful adjunct to T2-weighted MRI for detecting local tumor recurrence larger than 0.4 cm(2) within the prostate.


Biocatalytic conversion of poultry processing leftovers: Optimization of hydrolytic conditions and peptide hydrolysate characterization.
Peptide hydrolysate (PH) was produced by deep controllable bioconversion of poultry processing leftovers (broiler necks), by means of a multienzyme composition, containing four commercially available enzyme preparations (Alcalase, Neutrase, Flavourzyme, Protamex). The design of multienzyme composition (MEC) was applied to yield a hydrolysate with adjusted properties, including minimized antigenicity and bitterness. The protein recovery was optimized using Box-Behnken response surface design. The individual and interactive effects of hydrolysis conditions (time, hydromodule and MEC dosage) were studied. The experimental data were analyzed by ANOVA method and a well-predictive, second order polynomial model was developed using multiple regression analysis. Optimal hydrolysis conditions were found to be: hydrolysis time 3 h, hydromodule 2.25 l/kg and dosage of MEC 0.25%. The corresponding predicted value for protein recovery was 75.34%, 2 times higher compared to traditional long-term heating hydrolysis. The PH obtained is a low allergenic product with high antioxidant capacity.


Nerve crush but not displacement-induced stretch of the intra-arachnoidal facial nerve promotes facial palsy after cerebellopontine angle surgery.
Little is known about the reasons for occurrence of facial nerve palsy after removal of cerebellopontine angle tumors. Since the intra-arachnoidal portion of the facial nerve is considered to be so vulnerable that even the slightest tension or pinch may result in ruptured axons, we tested whether a graded stretch or controlled crush would affect the postoperative motor performance of the facial (vibrissal) muscle in rats. Thirty Wistar rats, divided into five groups (one with intact controls and four with facial nerve lesions), were used. Under inhalation anesthesia, the occipital squama was opened, the cerebellum gently retracted to the left, and the intra-arachnoidal segment of the right facial nerve exposed. A mechanical displacement of the brainstem with 1 or 3 mm toward the midline or an electromagnet-controlled crush of the facial nerve with a tweezers at a closure velocity of 50 and 100 mm/s was applied. On the next day, whisking motor performance was determined by video-based motion analysis. Even the larger (with 3 mm) mechanical displacement of the brainstem had no harmful effect: The amplitude of the vibrissal whisks was in the normal range of 50°-60°. On the other hand, even the light nerve crush (50 mm/s) injured the facial nerve and resulted in paralyzed vibrissal muscles (amplitude of 10°-15°). We conclude that, contrary to the generally acknowledged assumptions, it is the nerve crush but not the displacement-induced stretching of the intra-arachnoidal facial trunk that promotes facial palsy after cerebellopontine angle surgery in rats.


Involvement of differential metallothionein expression in free radical sensitivity of RTG-2 and CHSE-214 cells.
The role of metallothionein (MT) in free radical regulation and scavenging was investigated using two fish cell lines, the rainbow trout gonadal (RTG-2) cell line and the chinook salmon embryonic (CHSE-214) cell line. Exposure of RTG-2 cells to H(2)O(2) resulted in upregulation of both MT mRNA and MT protein and was also demonstrated by immunocytochemistry, confirming that MT was regulated by free radicals. We then compared the H(2)O(2) resistance in RTG-2 and CHSE-214 cells following metal treatment with Zn or Cd to induce MT. Comparison of survival of control cells and metal-exposed cells showed that metal treatment, which induced MT, significantly raised the H(2)O(2) tolerance in a dose-dependent manner in RTG-2 cells, while no increased H(2)O(2) resistance was observed in CHSE-214 cells. Transient over-expression of MT in CHSE-214: 59 cells also resulted in a dose-dependent increase in resistance to H(2)O(2) exposure. The raised resistance against H(2)O(2) in metal treated RTG-2 cells as well as transfected CHSE-214: 59 cells strongly demonstrate that MT is involved in the protection against H(2)O(2) and suggest a physiologically important function for MT when cells or whole organisms are exposed to oxidative stress.


Lateral relocation of auxin efflux regulator PIN3 mediates tropism in Arabidopsis.
Long-standing models propose that plant growth responses to light or gravity are mediated by asymmetric distribution of the phytohormone auxin. Physiological studies implicated a specific transport system that relocates auxin laterally, thereby effecting differential growth; however, neither the molecular components of this system nor the cellular mechanism of auxin redistribution on light or gravity perception have been identified. Here, we show that auxin accumulates asymmetrically during differential growth in an efflux-dependent manner. Mutations in the Arabidopsis gene PIN3, a regulator of auxin efflux, alter differential growth. PIN3 is expressed in gravity-sensing tissues, with PIN3 protein accumulating predominantly at the lateral cell surface. PIN3 localizes to the plasma membrane and to vesicles that cycle in an actin-dependent manner. In the root columella, PIN3 is positioned symmetrically at the plasma membrane but rapidly relocalizes laterally on gravity stimulation. Our data indicate that PIN3 is a component of the lateral auxin transport system regulating tropic growth. In addition, actin-dependent relocalization of PIN3 in response to gravity provides a mechanism for redirecting auxin flux to trigger asymmetric growth.


[Inflammatory pathology and rhinosinusal CT scan: the radiologist's report].
Pave the way to a better management of the chronical rhino-sinusitis diseases: Description and classification of the chronical rhino-sinusitis pathology, its medical and surgical treatments in order to help the radiologist in its CT-scan report. Review of the literature and anatomical considerations. Paranasal sinuses CT-scan analysis. The execution and the interpretation of CT-scans, in accordance with the progress made in the past 20 years in the knowledge of the rhino-sinusitis physiology and pathology, are today absolutely necessary for the management of chronical rhino-sinusitis diseases.


Heat release rate of enhanced large-scale open oil slick fires with Outdoor Gas Emission Sampling (OGES) system.
The Outdoor Gas Emission Sampling (OGES) system was developed to serve as an economical alternative to expensive industrial gas monitoring equipment. By establishing a sampling plane with four discrete sampling points along the radial direction of the smoke plume, the heat release rate (HRR) was measured for large-scale open oil slick fires. This newfound technique was particularly noteworthy during enhanced burns involving Flame Refluxer™ technology, where it is believed that partial premixing of the fuel and air by the apparatus resulted in a higher HRR than existing flame height correlations would suggest, evident by the HRR calculated using mass burning rate and gas analysis methods, which were in good agreement. Results from OGES show the potential of using point sampling within the plume regime to measure the HRR of fires that exceed the capabilities of conventional hood-based calorimeters, especially when it pertains to large-scale open burns.


Balancing immunity and immunosuppression: vaccinating patients receiving treatment with efalizumab.
Although effective in the treatment of immunodysregulatory diseases such as psoriasis, targeted immunosuppressive agents may confer risks of both enhanced susceptibility to infection and decreased responsiveness to vaccination. In a recent study, Krueger et al. (this issue) investigated these issues by testing the immune response to both a model antigen and a therapeutic vaccination in psoriasis patients during and after treatment with an LFA-1 inhibitor, efalizumab.


Thermal effects of various drill materials during implant site preparation-Ceramic vs. stainless steel drills: A comparative in vitro study in a standardised bovine bone model.
The aim of this study was to evaluate thermal effects of ceramic and metal implant drills during implant site preparation using a standardised bovine model. A total of 320 automated intermittent osteotomies of 10- and 16-mm drilling depths were performed using zirconium dioxide-based and stainless steel drills. Various drill diameters (2.0/ 2.2, 2.8, 3.5, 4.2 mm ∅) and different cooling methods (without/ with external saline irrigation) were investigated at room temperature (21 ± 1°C). Temperature changes were recorded in real time using two custom-built multichannel thermoprobes in 1- and 2-mm distance to the osteotomy site. For comparisons, a linear mixed model was estimated. Comparing thermal effects, significantly lower temperatures could be detected with steel-based drills in various drill diameters, regardless of drilling depth or irrigation method. Recorded temperatures for metal drills of all diameters and drilling depths using external irrigation were below the defined critical temperature threshold of 47°C, whereas ceramic drills of smaller diameters reached or exceeded the harmful temperature threshold at 16-mm drilling depths, regardless of whether irrigation was applied or not. The results of this study suggest that the highest temperature changes were not found at the deepest point of the osteotomy site but were observed at subcortical and deeper layers of bone, depending on drill material, drill diameter, drilling depth and irrigation method. This standardised investigation revealed drill material and geometry to have a substantial impact on heat generation, as well as external irrigation, drilling depth and drill diameter.


Phenotypic and Genetic Features that Differ Between Hereditary and Sporadic Melanoma: Results of a Preliminary Study from a Single Center from Turkey.
Most melanoma patients under our supervision lack characteristic phenotypic features for melanoma. In contrast, history of cancers other than melanoma and early age at onset were common. This observation was in favor of hereditary melanoma. To search for the phenotypic and genetic features that differ between sporadic and hereditary melanomas. In order to reveal phenotypic features, detailed physical exam was conducted to all melanoma patients (N = 43) and for genetic features. CDKN2A and MC1R mutations were detected with Sanger sequencing method. Assignment to hereditary and sporadic groups was done according to the "melanoma cancer syndrome assessment tool". Patients who were diagnosed before the age of 50 were also assigned to the hereditary melanoma group. Thirty-one patients were assigned to the hereditary group and 12 to the sporadic group. Fair eye color was statistically significantly higher in the sporadic group (P = 0.000). CDKN2A was detected in only 1 patient in the hereditary group. MC1R mutations were found in 12 out of 13 (92.3%) in the hereditary group with a score =3 points, 13 out of 18 (72.2%) in the early age at onset group and 5 out of 12 (41.7%) in the sporadic group (P = 0.024). Incidence of CDKN2A mutations in our hereditary group is in accordance with the reported incidences from Mediterranean countries. The difference between the hereditary and sporadic groups in terms of MC1R mutations supports the idea that MC1R genetic testing might help to determine patients with higher risk for hereditary melanoma.


Effect of aging on the microglial response to peripheral nerve injury.
Microglial morphology and immunophenotype have been studied extensively in aging-related neurodegenerative diseases, but to a lesser extent in the normally aged CNS, and little is known about how aging affects the ability of microglia to respond to neuronal injury. The goal of the current study was to determine if aging affects the ability of microglia to divide during the early response to facial nerve axotomy. In addition, we investigated the incidence of microglial cell death during later post-axotomy time points to determine if aging had an effect on microglial turnover. We employed DNA labeling with 3H-thymidine, TUNEL and lectin histochemistry after facial nerve axotomy in young (3 months), middle-aged (15 months), and old (30 months) Fisher344-Brown Norway hybrid rats. Proliferation of microglia in old rats remained significantly higher than in young rats 4 days after injury, suggesting that regulation of microglial proliferation changes with aging. There was no aging-related difference in microglial TUNEL staining at 7, 14 or 21 days post-axotomy. Lectin histochemistry in the unoperated facial nucleus revealed aging-related morphological changes in resting microglia, including hypertrophy of the cytoplasm with dense perinuclear staining. Aging-related differences in activated microglia on the lesioned side were more subtle, although many activated microglia of aged animals continued to exhibit dense perinuclear lectin reactivity. We propose that aging-related changes in morphology in conjunction with a less regulated proliferative response in the aged facial nucleus may be a reflection of microglial senescence.


Trends in the texts of national anthems: A comparative study.
In a recent previous investigation of national anthems, preferred topics and their bias (e.g., towards identity, fighting, or well-being) were identified subjectively (Silaghi-Dumitrescu, 2020). The present report aims to verify whether a more objective, automated, comparison of the texts of national anthems across the world can also reveal systematic trends - and to what extent. To this end, the Tropes and Semantria software packages are employed, revealing preferred topics (e.g., state, feeling, body, time, land, religion, family, fight), how their relative weights differ across continents and cultures, and how the conveyed sentiments vary. For instance, "liberty" is more common in Latin-country anthems while almost absent in Asia, "feelings" are less mentioned in Germanic-language anthems, and the first-person singular "I" is essentially absent African anthems. The sentiment scores of the anthems vary from neutral in Latin and Mediterranean anthems to much more positive in Central and Western Asian, Germanic and Slavic countries.


Improvement of glutathione production by metabolic engineering the sulfate assimilation pathway of Saccharomyces cerevisiae.
Glutathione (GSH) is a valuable tri-peptide that is widely used in the pharmaceutical, food, and cosmetic industries. Glutathione is produced industrially by fermentation using Saccharomyces cerevisiae. In this study, we demonstrated that engineering in sulfate assimilation metabolism can significantly improve GSH production. The intracellular GSH content of MET14 and MET16 over-expressing strains increased up to 1.2 and 1.4-fold higher than that of the parental strain, respectively, whereas those of APA1 and MET3 over-expressing strains decreased. Especially, in the MET16 over-expressing strain, the volumetric GSH concentration was up to 1.7-fold higher than that of the parental strain as a result of the synergetic effect of the increases in the cell concentration and the intracellular GSH content. Additionally, combinatorial mutant strains that had been engineered to contain both the sulfur and the GSH synthetic metabolism synergistically increased the GSH production. External addition of cysteine to S. cerevisiae is well known as a way to increase the intracellular GSH content; however, it results a decrease in cell growth. This study showed that the engineering of sulfur metabolism in S. cerevisiae proves more valuable than addition of cysteine as a way to boost GSH production due to the increases in both the intracellular GSH content and the cell growth.


A theoretical analysis of the relative influences of peak BMD, age-related bone loss and menopause on the development of osteoporosis.
Factors that determine a post-menopausal woman's bone mineral density (BMD) include her mass at the time of skeletal maturity (peak BMD), menopause and the rate of loss she experiences as she ages. Understanding the relative influence of each of these factors may help identify important preventive treatments and provide new ways to identify women at risk for osteoporosis. In this analysis we utilize a computer model of the bone remodeling process to predict the relative influences of peak BMD, menopause and age-related bone loss on the development of osteoporosis. The delay in the onset of osteoporosis (defined as BMD <2.5 SD from the young adult mean) caused by modifying peak BMD, age-related bone loss or the age at menopause is quantified. A 10% increase in peak BMD is predicted to delay the development of osteoporosis by 13 years, while a 10% change in the age at menopause or the rate of non-menopausal bone loss is predicted to delay osteoporosis by approximately 2 years, suggesting that peak BMD may be the single most important factor in the development of osteoporosis.


Risk of imported malaria infections in Zanzibar: a cross-sectional study.
Zanzibar has made substantial progress in malaria control with vector control, improved diagnosis, and artemisinin-based combination therapy. Parasite prevalence in the population has remained around 1% but imported infections from mainland Tanzania contribute to sustained local transmission. Understanding travel patterns between mainland Tanzania and Zanzibar, and the risk of malaria infection, may help to control malaria importation to Zanzibar. A rolling cross-sectional survey linked to routine reactive case detection of malaria was carried out in Zanzibar between May 2017 and October 2018. Households of patients diagnosed with malaria at health facilities were surveyed and household members were tested for malaria using rapid diagnostic tests and a sub-sample by quantitative PCR (qPCR). Interviews elicited a detailed travel history of all household members who had travelled within the past two months, including trips within and outside of Zanzibar. We estimated the association of malaria infection with travel destinations in pre-defined malaria endemicity categories, trip duration, and other co-variates using logistic regression. Of 17,891 survey participants, 1177 (7%) reported a recent trip, of which 769 (65%) visited mainland Tanzania. Among travellers to mainland Tanzania with travel destination details and a qPCR result available, 241/378 (64%) reported traveling to districts with a 'high' malaria endemicity and for 12% the highest endemicity category was 'moderate'. Travelers to the mainland were more likely to be infected with malaria parasites (29%, 108/378) than those traveling within Zanzibar (8%, 16/206) or to other countries (6%, 2/17). Among travellers to mainland Tanzania, those visiting highly endemic districts had a higher odds of being qPCR-positive than those who travelled only to districts where malaria-endemicity was classified as low or very low (adjusted odd ratio = 7.0, 95% confidence interval: 1.9-25.5). Among travellers to the mainland, 110/378 (29%) never or only sometimes used a mosquito net during their travel. Strategies to reduce malaria importation to Zanzibar may benefit from identifying population groups traveling to highly endemic areas in mainland Tanzania. Targeted interventions to prevent and clear infections in these groups may be more feasible than attempting to screen and treat all travellers upon arrival in Zanzibar.


Identification of a novel 17,000-dalton parathyroid hormone-like adenylate cyclase-stimulating protein from a tumor associated with humoral hypercalcemia of malignancy.
Humoral hypercalcemia of malignancy is a common paraneoplastic syndrome which is characterized by hypercalcemia resulting from secretion by tumors of a circulating bone-resorbing factor. Evidence suggests that in many instances this factor is an adenylate cyclase-stimulating protein which shares features with, but is distinct from, parathyroid hormone (PTH). The current report describes the purification to homogeneity from a humoral hypercalcemia of malignancy-associated tumor of a novel, basic, highly potent PTH-like adenylate cyclase-stimulating protein. This factor differs from previously described PTH-like factors with respect to size, amino acid composition, and specific activity.


Seronegative rheumatoid arthritis: a rare cause of effusive-constrictive pericarditis requiring pericardiectomy.
Effusive-constrictive pericarditis is a rare condition. In this report, we describe a case of effusive-constrictive pericarditis caused by seronegative rheumatoid arthritis which was successfully treated with surgical pericardiectomy.


The treatment of carcinoma in situ of the uterine cervix in the Mersey Regional Cancer Registry Area with some comments on incidence.
One thousand and twenty-four patients were registered with carcinoma in situ of the cervix at the Mersey Regional Cancer Registry from 1962 to 1973. The number of new cases reached a peak in the over-35s during 1968-70, but later in the under-35s. Fifty-five per cent of patients were treated initially by hysterectomy and 39% by cone biopsy or amputation. In the former group two patients developed invasive recurrence and one patient in situ recurrence. In the latter group one patient developed invasive recurrence and 15 patients developed in situ recurrence. This evidence is in agreement with other studies and supports the view that cone biopsy is an effective treatment for c. in situ of the cervix.


Association of ST-elevation and non-ST-elevation presentation on ECG with transmurality and size of myocardial infarction as assessed by contrast-enhanced magnetic resonance imaging.
It is the general perception, that ST-elevation myocardial infarction is associated with transmural ischemia while Non-ST elevation myocardial infarction is found in non-transmural subendocardial ischemia. This association, however, derives primarily from post mortem studies. A total of 220 patients with acute myocardial infarction (MI) who had PCI on admission and contrast-enhanced cardiac magnetic resonance imaging (CMR) within one week were included into the study. Size and transmural extent of MI was quantified by CMR and correlated with the ECG on admission. Based on the ECG findings, 57% were classified as STEMI and 43% as NSTEMI. CMR infarct size was significantly larger in STEMI than NSTEMI (23.2 vs. 14.2 LV%, p<0.001). As assessed by CMR, STEMI patients were transmural in 63% as compared to 27% of patients with NSTEMI (p<0.001). In a multivariable logistic regression model, total infarct size was significantly associated with presence of STEMI (OR: 1.045, 95% CI [1.014-1.077], p=0.004) whereas the number of transmural segments did not significantly add further information for a STEMI/NSTEMI classification (p=0.054, change of c-index from 0.69 to 0.70). The electrocardiographic STEMI/NSTEMI classification does rather characterize the total size of MI than the transmural extent as assessed by CMR.


Some relations between dissolution rates and physical parameters of a drug in aqueous micellar solutions of a non-ionic surfactant.
Dissolution rates of salicylic acid from a constant surface area into a series of aqueous micellar polysorbate 20 solutions at pH 1-0 to 4-0 have been measured using two different methods; a stirred beaker and a rotating disc technique. The micellar molecular weight of polysorbate 20 has been obtained from light scattering and differential refractometry data and used with other independently determined physical data to calculate diffusion coefficients of the diffusing species. Linear multiple regression analysis was used to assess the dependence of drug dissolution rate on the diffusion coefficient and the viscosity of the dissolution medium.


Efficient degradation of the refractory organic pollutant by underwater bubbling pulsed discharge plasma: performance, degradation pathway, and toxicity prediction.
It is essential to develop an efficient technology for the elimination of refractory contaminants due to their high toxicity. In this study, a novel underwater bubbling pulsed discharge plasma (UBPDP) system was proposed for the degradation of Orange II (OII). The degradation performance experiments showed that by enhancing the peak voltage and pulse frequency, the degradation efficiency of OII increased gradually. The removal efficiencies under different air flow rates were close. Reducing OII concentration and solution conductivity could promote the elimination of OII. Compared with neutral and alkaline conditions, acidic condition was more beneficial to OII degradation. The active species including ·OH, ·O2-, 1O2, and hydrated electrons were all involved in OII degradation. The concentrations of O3 and H2O2 in OII solution were lower than those in deionized water. During discharge, the solution pH increased while conductivity decreased. The variation of UV-vis spectra with treatment time indicated the effective decomposition of OII. Possible degradation pathways were speculated based on LC-MS. The toxicity of intermediate products was predicted by the Toxicity Estimation Software Tool. Coexisting constituents including Cl-, SO42-, HCO3-, and humic acid had a negative effect on OII removal. Finally, the comparison with other technology depicted the advantage of the UBPDP system.


Long-term respiratory functional results after pneumonectomy.
The aim of this study is to evaluate the long-term respiratory outcome of patients who underwent pneumonectomy for non-small cell lung cancer (NSCLC), analysing functional tests. Twenty-seven consecutive patients who were candidates for pneumonectomy performed spirometry before and at least 24 months after surgery in the same laboratory. Diffusion of carbon monoxide and the most common dynamic and static lung volumes were evaluated in percentage of predicted and compared. A significant inverse correlation was observed between the preoperative FEV1 (%) and FVC (%) and their postoperative loss, respectively r=-641 (p<0.0001) and r=-789 (p<0.0001). Also the correlation between the RV/TLC ratio and the FEV1 loss confirmed a better postoperative outcome in patients with major airway obstruction (p=0.02). To investigate these data, the series were divided into two groups: group A included BPCO patients with a FEV1 lower than 80%, the others were considered group B. Group B showed a significant major postoperative FEV1 (%) and FVC (%) impairment, 31% versus 12%, p=0.005, and FVC (%) loss, 37% versus 16% (p=0.02), meanwhile group A showed a significant major postoperative RV (%) reduction, 43% versus 17%, p=0.03. Despite being significantly higher preoperatively in BPCO patients, the RV% becomes similar between the two groups in the postoperative. In our experience patients with major preoperative airway obstruction who underwent pneumonectomy had lower impairment in FEV1% at almost one year after surgery than those with normal respiratory function. The resection of a certain amount of non-functional parenchyma associated with the mediastinal shift, with an improvement of the chest cavity for the remaining lung, could give a reduction volume effect in BPCO/emphysematous patients.


The proposal of architecture for chemical splitting to optimize QSAR models for aquatic toxicity.
One of the challenges in the field of quantitative structure-activity relationship (QSAR) analysis is the correct classification of a chemical compound to an appropriate model for the prediction of activity. Thus, in previous studies, compounds have been divided into distinct groups according to their mode of action or chemical class. In the current study, theoretical molecular descriptors were used to divide 568 organic substances into subsets with toxicity measured for the 96-h lethal median concentration for the Fathead minnow (Pimephales promelas). Simple constitutional descriptors such as the number of aliphatic and aromatic rings and a quantum chemical descriptor, maximum bond order of a carbon atom divide compounds into nine subsets. For each subset of compounds the automatic forward selection of descriptors was applied to construct QSAR models. Significant correlations were achieved for each subset of chemicals and all models were validated with the leave-one-out internal validation procedure (R(2)(cv) approximately 0.80). The results encourage to consider this alternative way for the prediction of toxicity using QSAR subset models without direct reference to the mechanism of toxic action or the traditional chemical classification.


Role of the microenvironment in tumor growth and in refractoriness/resistance to anti-angiogenic therapies.
Angiogenesis is critical for growth of many tumor types and the development of anti-angiogenic agents opened a new era in cancer therapy. However, similar to other anti-cancer therapies, inherent/acquired resistance to anti-angiogenic drugs may occur in cancer patients leading to disease recurrence. Recent studies in several experimental models suggest that both tumor and non-tumor (stromal) cell types may be involved in the reduced responsiveness to the treatments. The current review focuses on the role of stromal cells in tumor growth and in refractoriness to anti-VEGF treatment.


Chitosan/cyclodextrin nanoparticles can efficiently transfect the airway epithelium in vitro.
The main goal of the present study was to investigate the potential of a new generation of hybrid polysaccharide nanocarriers, composed of chitosan (CS) and anionic cyclodextrins (CDs), for gene delivery to the airway epithelium. More specifically, these nanocarriers were investigated with regard to their ability to enter epithelial cells and promote gene expression in the Calu-3 cell culture model. In the search for the most suitable nanocarrier composition for gene delivery, the effect of CS molecular weight (Mw) on the nanocarriers characteristics and their ability to transfect cells was investigated. Thus, hybrid CS/CD nanoparticles were prepared with two different CS Mw, medium (110 kDa) and low (10 kDa), and loaded with pSEAP (plasmid DNA model that encodes the expression of secreted alkaline phosphatase). The resulting nanoparticles presented an adequate size range (100-200 nm, depending on CS Mw), a positive surface charge (+22 to +35 mV) and very high DNA association efficiency values (>90%). Cellular uptake studies showed that the nanoparticles were effectively internalized by the cells, providing a good indication of their potential as gene carriers. The transfection efficiency of the different formulations, measured by the concentration of secreted gene product (SEAP), indicated that all the nanoparticles were able to elicit a significantly higher response than the naked DNA (control), the transfection efficiency being more important for low MwCS nanoparticles than for those composed of medium MwCS. Overall, this report is the first evidence of the potential of a new generation of safe polysaccharide nanocarriers for gene delivery to the airway epithelium.


Basophils Trigger Fibroblast Activation in Cardiac Allograft Fibrosis Development.
Fibrosis is a major component of chronic cardiac allograft rejection. Although several cell types are able to produce collagen, resident (donor-derived) fibroblasts are mainly responsible for excessive production of extracellular matrix proteins. It is currently unclear which cells regulate production of connective tissue elements in allograft fibrosis and how basophils, as potential producers of profibrotic cytokines, are involved this process. We studied this question in a fully MHC-mismatched model of heart transplantation with transient depletion of CD4(+) T cells to largely prevent acute rejection. The model is characterized by myocardial infiltration of leukocytes and development of interstitial fibrosis and allograft vasculopathy. Using depletion of basophils, IL-4-deficient recipients and IL-4 receptor-deficient grafts, we showed that basophils and IL-4 play crucial roles in activation of fibroblasts and development of fibrotic organ remodeling. In the absence of CD4(+) T cells, basophils are the predominant source of IL-4 in the graft and contribute to expansion of myofibroblasts, interstitial deposition of collagen and development of allograft vasculopathy. Our results indicated that basophils trigger the production of various connective tissue elements by myofibroblasts. Basophil-derived IL-4 may be an attractive target for treatment of chronic allograft rejection.


Transcriptional activity of isolated maize chloroplasts.
Chloroplasts and etioplasts, isolated from light- or dark-grown Zea mays plants, respectively, can incorporate labeled UTP into RNA in a reaction stimulated by light or ATP. This in organello RNA synthesis proceeded at a linear rate for up to 2 h. When expressed per unit protein, plastids from dark-grown plants incorporated more UTP than those from light-grown plants, and the highest rate of UTP incorporation was found in plastids from light-stimulated leaves (grown previously in the dark). The in organello newly synthesized RNA was heterodispersed, with most transcripts smaller than 14 S. Specific transcripts were detected in organelles from both dark- and light-grown plants that contain sequences that are homologous to the mRNAs for the rbcL gene (coding for the large subunit of ribulose bisphosphate carboxylase (LS-RuBPCase] and for the psbA gene (32-kDa thylakoid membrane protein). Qualitatively, the newly synthesized in organello transcripts were similar from the dark and light organelles.


Effect of isotypic and allotypic variations of MHC class II molecules on staphylococcal enterotoxin presentation to murine T cells.
Staphylococcal enterotoxins (SE) are known to be potent T cell activators, stimulating +/- proliferation and lymphokine production. These toxins have recently have been termed "superantigens" because of their ability to bind directly to class II molecules forming a ligand that interacts with particular V beta gene elements within the TCR complex. This interaction between SE and MHC class II molecules plays a central role in toxin-induced mitogenesis. In the present study we have examined the effect of polymorphism on the ability of MHC class II molecules to bind and present SE. Through the use of H-2 congenic mouse strains, it was possible to look directly at haplotype differences within the MHC and their effect on SE presentation to a panel of responsive V beta-bearing T cells. The results demonstrate that toxin presentation by class II-bearing accessory cells to murine T cells is greatly affected by polymorphisms within the H-2 complex. Toxin-pulsed accessory cells obtained from mice of an H-2k and H-2u haplotype were found to be less efficient in activating a variety of T cell clones and hybridomas. However, one T cell clone responded similarly to the enterotoxins presented on all H-2 haplotypes, suggesting that differences in responses of T cells are not simply a function of the degree of binding of these toxins to various class II molecules. Neutralization analysis with monoclonal anti-class II antibodies demonstrates that both I-A and I-E molecules play a significant role in SEA and SEB presentation to murine T cells. These results suggest that the differential activation of T cells by a particular enterotoxin may reflect a difference in recognition of an SE:class II ligand by a surface T cell receptor complex.


Dissociable contribution of 5-HT1A and 5-HT2A receptors in the medial prefrontal cortex to different aspects of executive control such as impulsivity and compulsive perseveration in rats.
Serotonin (5-HT) receptors are increasingly recognized as major targets for cognitive enhancement in schizophrenia. Several lines of evidence suggest a pathophysiological role for glutamate NMDA receptors in the prefrontal cortex in schizophrenia and associated disorders in attention and executive functioning. We investigated how the interactions between 5-HT1A and 5-HT2A and glutamate NMDA receptor mechanisms in the medial prefrontal cortex (mPFC) contribute to the control of different aspects of attentional performance. Rats were trained on a five-choice serial reaction time (5-CSRT) task, which provides indices of attentional functioning (percentage of correct responses), executive control (measured by anticipatory and perseverative responses), and speed. The competitive NMDA receptor antagonist CPP (50 ng/side) was infused directly into the mPFC 5 min after infusion of either 8-OH-DPAT (30 and 100 ng/side) or M100907 (100 and 300 ng/side) into the same brain area. Impairments in attentional functioning induced by CPP were completely abolished by both doses of 8-OH-DPAT or M100907. In addition, M100907 abolished the CPP-induced anticipatory responding but had no effects on perseverative over-responding, while 8-OH-DPAT reduced the perseverative over-responding but had no effects on anticipatory responding induced by CPP. The selective 5-HT(1A) receptor antagonist WAY100635 (30 ng/side) antagonized the effects of 8-OH-DPAT (100 ng/side). 8-OH-DPAT at 30 ng/side reduced the latency of correct responses in controls and CPP-injected rats and lowered the percentage of omissions in CPP-injected rats. The data show that 5-HT1A and 5-HT2A receptors in the mPFC exert opposing actions on attentional functioning and demonstrate a dissociable contribution of 5-HT1A and 5-HT2A receptors in the mPFC to different aspects of executive control such as impulsivity and compulsive perseveration.


Therapeutic Hypothermia in STEMI.
In this review article we tried to find an answer to the question, should local coronary hypothermia be a part of the early reperfusion strategy in patients with STEMI to prevent reperfusion injury, no-reflow phenomenon, and to reduce the infarct size and mortality. Hypothermia can save cardiomyocytes if achieved in a timely fashion before reperfusion. Intracoronary hypothermia can be adjunct to PCI by lessening ischemia/reperfusion injury on cardiomyocytes and reduction in infarct size. Reperfusion induced Calcium overload, generation of ROS and subsequent activation of Mitochondrial permeability transition pore (MPT) are major contributors to reperfusion injury. Hypothermia reduces calcium loading of the cell and maintains cellular energy and tissue level glucose which can scavenger ROS. Hypothermia reduces MPT activation and thus reduces infarct size. Systemic cooling trials failed to reduce infarct size, perhaps because the target temperature was not reached fast enough, and it was associated with systemic side effects. The need for rapid induction of hypothermia to <35 °C with the ethical concern of delaying reperfusion while cooling the patient and the inconsistency of endovascular cooling results lead to a belief that endovascular cooling may exceed the acceptable level of invasiveness in the context of other novels cardioprotective, regenerative and reperfusion therapies. Clinical trials showed the safety and feasibility of novel intracoronary hypothermia with rapid induction and maintenance of hypothermia using routine PCI equipment ahead of reperfusion. Two phases of cooling were applied without significant delay in the door to balloon time. Cooling of the coronary artery leads to cooling of its dependant myocardium without affecting adjacent myocardium. Heat transfer occurred by heat conduction during the occlusion phase and heat convention during the reperfusion phase. Fine-tuning of saline temperature and infusion rate helped to improve the protocol. The best duration of hypothermia before and after reperfusion is not known and needs further investigation. A balance between the undoubted cardioprotective effects of hypothermia with iatrogenic prolongation of ischemia time needs to be established. A reduction in infarct size was observed but needs to be validated with large randomized trials. Furthermore, it might be possible to augment the cardioprotective effects of intracoronary hypothermia by combination with other cardioprotective approaches such as antioxidant drugs and afterload reducing agents.


Efficacy of indacaterol as a single therapy versus salmeterol/fluticasone therapy in patients with milder chronic obstructive pulmonary disease.
In non-exacerbation chronic obstructive pulmonary disease (COPD) with mild lung function impairment, single bronchodilator therapy might be as effective as combined inhaled corticosteroid/bronchodilator therapy, whereas the risk of pneumonia associated with the latter would be practically absent. We performed an analysis of a recent study evaluating the efficacy and safety of inhaled indacaterol versus inhaled salmeterol/fluticasone in COPD patients. Both therapies were found to exert comparable effects on lung function, symptom severity and health status.


Cost-Effectiveness of iGlarLixi Versus iDegLira in Type 2 Diabetes Mellitus Inadequately Controlled by GLP-1 Receptor Agonists and Oral Antihyperglycemic Therapy.
The fixed-ratio combinations (FRCs) of glucagon-like peptide 1 receptor agonists (GLP-1 RAs) and basal insulin, insulin glargine 100 U/mL plus lixisenatide (iGlarLixi), and insulin degludec plus liraglutide (iDegLira), have demonstrated safety and efficacy in patients with type 2 diabetes mellitus (T2DM) inadequately controlled on GLP-1 RAs. However, a comparative cost-effectiveness analysis between these FRCs from a UK Health Service perspective has not been conducted. The IQVIA Core Diabetes Model was used to estimate lifetime costs and outcomes in patients with T2DM receiving iGlarLixi (based on the LixiLan-G trial) versus iDegLira (based on relative treatment effects from an indirect treatment comparison using data from DUAL III). Utilities, medical costs, and costs of diabetes-related complications were derived from literature. Model outputs included costs and quality-adjusted life years (QALYs). Incremental cost-effectiveness ratios were calculated with a local willingness-to-pay threshold of £20,000 per QALY. Extensive scenario, one-way sensitivity, and probabilistic sensitivity analyses were conducted to evaluate the robustness of the model. iGlarLixi was less costly (iGlarLixi, £30,011; iDegLira, £40,742), owing to lower acquisition costs, and similar in terms of QALYs gained (iGlarLixi, 8.437; iDegLira, 8.422). Extensive scenario and sensitivity analyses supported the base case findings. In patients with T2DM and inadequate glycemic control despite GLP-1 RAs, use of iGlarLixi was associated with substantial cost savings and comparable utility outcomes. iGlarLixi can be considered as cost-effective versus iDegLira from the UK Health Service perspective.


Immunohistochemical localization of chondroitin sulphate and dermatan sulphate proteoglycans in tumour tissues.
Immunohistochemical localization of chondroitin sulphate and dermatan sulphate proteoglycans (PGs) was observed in 70 tumour tissues, using monoclonal antibodies 9A-2 and 3B-3 raised against core molecules obtained from chondroitin sulphate PG by chondroitinase ABC-treatment. They recognize a stub of delta Di-4S and delta Di-6S binding to core protein via a linkage tetrasaccharide, respectively. The antibody 6B6 raised against dermatan sulphate PG obtained from an ovarian fibroma capsule in our laboratory was also used. The interstitial fibrous elements, so-called 'specific stroma' within the cancer cell nests contained chondroitin 4-sulphate PG as revealed with 9A-2, whereas the surrounding connective tissue and the preexisting fibrous connective tissue involved in the tumour growth consisted of dermatan sulphate PG with a considerable amount of chondroitin 4-sulphate PG. Chondroitin 6-sulphate PG as revealed with 3B-3 was located in the connective tissue proliferating from blood vessels and muscle tissue in association with the invasive growth of tumour cells. Chondroitin 6-sulphate PG was also observed in the basement membrane components of some tumours. In non-epithelial tumours (fibrogenic, chondrogenic, osteogenic and neurogenic tumours), chondroitin 4-sulphate was in fibrous portions. When collagenization and hyalinization progressed, dermatan sulphate PG was observed to increase in quantity.


Pediatric otosyphilis-An unusual cause of conductive hearing loss.
We present the case of a 5 year old female with a unilateral conductive hearing loss which had a relapsing and remitting course over a 3 year period. An initial noncontrast CT temporal bone study was unremarkable and a diagnosis of otitis media was made in the first instance. However, a second CT temporal bone study performed 3 years later demonstrated bilateral demineralisation of the ossicles and abnormal lucency affecting both the otic capsules. A diagnosis of otosyphilis was proposed on the basis of the imaging features and a prior medical history of previously treated congenital syphilis. With the benefit of hindsight, early pericochlear lucency was identified on the initial CT temporal bone study. There has been a steady rise of syphilis cases since the millennium with resurgence in many high income countries. Otosyphilis has a highly variable clinical presentation and there is limited data to establish the pattern of hearing loss in pediatric patients with a background of congenital otosyphilis. Temporal bone and otic capsule demineralisation carries a broad differential diagnosis including osteogenesis imperfecta, otosclerosis, Paget's disease and radiation related changes. Otosyphilis is a rare but potentially treatable cause of deafness and a high index of suspicion is required to make the diagnosis. In conjunction with a positive syphilis serology, a noncontrast temporal bone CT can aid the diagnosis and expedite the treatment.


Aflatoxin M1 in milk powders: processing, homogeneity and stability testing of certified reference materials.
As part of the certification campaign of three candidate reference materials for the determination of aflatoxin M1 (AfM1) in whole milk powders, homogeneity, short- and long-term stability tests of naturally contaminated milk powders have been performed. The homogeneity of two AfM1-contaminated milk powders was studied by taking samples at regular intervals of the filling sequences and analysing in triplicate for their AfM1 contents by liquid chromatography with fluorescence detection (LC-FLD) using random stratified sampling schemes. The homogeneity testing of an AfM1 'blank' milk powder material was performed by determining the nitrogen content because AfM1 levels were below the limit of detection of the most sensitive determination method. The short-term stability of AfM1-contaminated milk powders was evaluated at three different storage temperatures (4, 18 and 40 degrees C). After storage times of 0, 1, 2 and 4 weeks, samples were investigated using LC-FLD. The long-term stability study comprised of measurements after 0, 6, 12 and 18 months after storage at -20 and 4 degrees C. Analyses were done by LC-FLD. Based on the homogeneity tests, the materials were sufficiently homogenous to serve as certified reference materials. Corresponding uncertainty contributions of 0.23-0.89% were calculated for the homogeneity. The stability measurements showed no significant trends for both short- and long-term stability studies. The long-term stability uncertainties of the AfM1-contaminated milk powders were 7.4 and 6.3%, respectively, for a shelf-life of 6 years and storage at -20 degrees C. Supplementary stability monitoring schemes over a long period of several years are currently ongoing.


Distribution and time course of protein extravasation in the rat spinal cord after contusive injury.
We have previously characterized a graded, spinal cord contusive injury in the rat. We have now used this reproducible model to examine vascular permeability to horseradish peroxidase (HRP) after injury. The relationship between severity of injury and distribution of protein extravasation was evaluated at 3 h after injury. After mild injury, tracer was primarily confined to central gray matter and the ventral part of the dorsal columns. After moderate injury, protein extravasation was similar to that observed after mild injury, with the exception that the central hemorrhage included pericentral while matter and occasionally extended to the pial surface. After severe injury, reaction product (RP) was more densely distributed within the central cord and peripheral white matter. The axial extent of tracer at sites proximal and distal to the impact site increased with severity of injury. At 2.0 cm from the injury, no leakage of tracer was noted after mild injury. In contrast, after moderate and severe injury limited microvascular leakage of HRP was noted. Furthermore, after severe injury, in addition to local sites of microvascular leakage, intense RP was present in the dorsal columns up to at least 2.0 cm from the injury. The time course for re-establishment of the blood-spinal cord barrier to protein was evaluated from 3 h to 14 days after moderate injury. At 3 h to 1 day, protein leakage was maximal and coincided with sites of extravasated blood components, although was consistently more extensive. By 7 days, despite resolution of the initial hemorrhage, there remained scattered evidence for protein extravasation at the injured site and at sites along the axis of the cord. The blood-spinal cord barrier to HRP was reestablished by 14 days after injury.


Neuropilin-2 expression in cancer.
Neuropilin-2 is a coreceptor for vascular endothelial growth factor family members. Blockade of neuropilin-2 is able to suppress lymphogenous metastasis in preclinical models. The aim of this study was to validate a protocol for the evaluation of neuropilin-2 protein expression in situ, by comparison with in-situ hybridization, western blotting, and mRNA expression levels. Immunohistochemistry was performed on normal human tissues, and whole sections for 79 primary non-small-cell lung carcinomas, 65 primary breast carcinomas, 79 primary colorectal cancers, and 52 metastases. Neuropilin-2 expression was observed in lymphatic and blood vessels from all normal and malignant tissues examined. In addition, 32% of primary non-small-cell lung carcinomas, 15% of primary breast carcinomas and 22% of primary colorectal cancers showed tumour cell expression. Fifty-five primary and nine secondary malignant melanomas were also examined for neuropilin-2 expression by in-situ hybridization. All showed vascular expression, and 85% of primary malignant melanomas showed tumour cell expression. In the majority of lung, breast and colorectal cancers, the effects of anti-neuropilin-2 are likely to be restricted to the vasculature. These results will assist in pharmacokinetic evaluations, tolerability assessments and the choice of setting to evaluate the activity of anti-neuropilin-2 therapies.


Enzymatic (R)-phenylacetylcarbinol production in a benzaldehyde emulsion system with Candida utilis cells.
Recent progress in enzymatic (R)-phenylacetylcarbinol (PAC) production has established the need for low cost and efficient biocatalyst preparation. Pyruvate decarboxylase (PDC) added in the form of Candida utilis cells showed higher stability towards benzaldehyde and temperature in comparison with partially purified preparations. In the presence of 50 mM benzaldehyde and at 4 degrees C, a half-life of 228 h was estimated for PDC added as C. utilis cells, in comparison with 24 h for the partially purified preparation. Increasing the temperature from 4 to 21 degrees C for PAC production with C. utilis cells resulted in similar final PAC levels of 39 and 43 g l(-1) (258 and 289 mM), respectively, from initial 300 mM benzaldehyde and 364 mM pyruvate. The overall volumetric productivity was enhanced 2.8-fold, which reflected the 60% shorter reaction time at the higher temperature. Enantiomeric excess values of 98 and 94% for R-PAC were obtained at 4 and 21 degrees C, respectively, and benzyl alcohol (a potential by-product from benzaldehyde) was not formed.


Performance of Echocardiographic Algorithms for Assessment of High Aortic Bioprosthetic Valve Gradients.
Bioprosthetic aortic valve dysfunction (BAVD) is a challenging diagnosis. Commonly used algorithms to classify high-gradient BAVD are the 2009 American Society of Echocardiography, 2014 Blauwet-Miller, and 2016 European Association of Cardiovascular Imaging algorithms. We sought (1) to evaluate the accuracy of existing algorithms against objectively proven BAVD and (2) to propose an improved algorithm. This was a retrospective study of 266 patients with objectively proven BAVD (pathology of explanted valves, four-dimensional computed tomography prior to transcatheter valve-in-valve replacement, or therapeutically confirmed bioprosthetic thrombosis) who were treated. Of those, 191 had obstruction, 48 had regurgitation, 15 had mixed stenosis and regurgitation, and 12 had patient-prosthesis mismatch (PPM). Normal controls were matched 1:1 (age, prosthesis size, and type), of which 43 had high gradients (PPM in 30, high flow in nine, and normal prosthesis in nine). Algorithm assignment was based on the echocardiogram leading to BAVD diagnosis and the predischarge "fingerprint" echocardiogram after surgical or transcatheter aortic valve replacement. A novel algorithm (Mayo Clinic algorithm) incorporating valve appearance in addition to Doppler parameters was developed to improve observed deficiencies. The accuracy of existing algorithms was suboptimal (2009 American Society of Echocardiography, 62%; 2014 Blauwet-Miller, 62%; 2016 European Association of Cardiovascular Imaging, 57%). The most common overdiagnosis was PPM (22%-29% of patients and controls with high gradients). The novel Mayo Clinic algorithm correctly identified the mechanism in 256 of 307 patients and controls (83%). Recognition of regurgitation was substantially improved (42 of 47 patients, 89%), and the number of PPM misdiagnoses was significantly reduced (five patients). Currently recommended algorithms misclassify a significant number of BAVD patients. The accuracy was improved by a newly proposed algorithm.


Characterization of two carotenoid gene promoters in the cyanobacterium Synechocystis sp. PCC 6803.
The in vivo promoter activity of the 5'-region of the genes encoding the carotenogenic enzymes phytoene desaturase (crtP) and phytoene synthase (crtB) from the cyanobacterium Synechocystis sp. PCC 6803 has been studied by deletion mapping, primer extension, Northern blot, and using the chloramphenicol acetyltransferase (cat) reporter gene. crtP and crtB are closely linked in the Synechocystis genome, but it is shown that both genes are independently transcribed. The level of transcription in both cases is very low. Their expression is affected in a similar way by light intensity: very little or no expression is observed in the dark compared with expression under illumination conditions. High light intensities induce a transient increase in transcription initiation as detected with the cat probe. DNA fragments containing 81 base pairs upstream of the crtP putative transcription initiation site or 192 base pairs upstream of the crtB putative transcription initiation site are enough to confer light sensitive expression to a reporter gene.


Cutaneous abscess due to Eubacterium lentum in injection drug user: a case report and review of the literature.
We described the first case, to the best of our knowledge, of cutaneous abscess due to Eubacterium lentum in a parenteral drug user, after complete fracture of the right femor. The case underlines the importance of carefully performed microbiological tests, due to the peculiar cultural needs of the micro-organism.