A Qualitative Evaluation of Caregiver Support Services Offered at the Atlanta Veterans Affairs Health Care System.
The Department of Veterans Affairs (VA) provides health care to approximately 300,000 patients with dementia. Recognizing the critical role caregivers play in veterans' health, the Cognitive Disorders Specialty Care Education Center of Excellence (COE) at the Atlanta VA Health Care System implemented a suite of caregiver support services, including formal programs and resource linkages. We evaluated the effectiveness of these services and identified caregiver-perceived gaps in them. We conducted 11 semistructured interviews from November 2016 through February 2017 with caregivers of veterans seen in the COE who had participated in support services. After coding transcripts, we established a codebook of 9 major themes and conducted a thematic analysis of all transcripts. Caregivers spoke positively of COE caregiver services that offered information on dementia, social support, an emphasis on caregiver well-being and self-efficacy, and methods for behavioral change. Gaps identified included the need for additional dementia information and practical support in such matters as advanced directives and eligibility for VA benefits. Our findings will inform future improvements to COE caregiver support services, such as an expansion of COE's caregiver educational content and capacity building of existing components such as resource referrals. These results also highlight opportunities for COE to interface with internal and external organizations to enhance existing caregiver services.


Regulation of mouse mammary tumor virus gene expression by glucocorticoid hormones.
Several laboratories have documented that glucocorticoid hormones markedly stimulate the expression of mouse mammary tumor virus genes in a variety of mouse mammary tumor cells and in infected heterologous cells. The effect of the hormone appears to be a rapid and specific augmentation of the synthesis of viral RNA, mediated by interaction with glucocorticoid receptor proteins. The availability of virus-specific reagents and recent developments in the molecular biology of RNA tumor viruses now permit a highly refined analysis of hormonal regulation in this experimental system.


Generation of Parkinson's disease patient-derived human induced pluripotent stem cells line (PNUSCRi001-A) carrying a N227S mutation in GBA gene.
The hiPSC line was generated from peripheral blood mononuclear cells (PBMCs) collected by a male patient with young onset Parkinson's disease, carrying on heterozygous c.680 A > G (N227S) mutation in the GBA gene. The PBMCs was reprogrammed into an induced pluripotent stem cell (iPSC) line (PNUSCRi001-A hiPSCs) using non-integrative sendai virus. The hiPSC line, PNUSCRi001-A displayed a normal karyotype and the Expression of pluripotency markers that is capable of producing derivatives of three germ layers (Ectoderm, Endoderm and Mesoderm).


A review of the new provisions for sanctioning mentally disordered offenders in China, in a broader historical context.
Reforms of the criminal justice system in China in recent years have included the 2012 Code of Criminal Procedure (CCP), which resulted in new disposals for mentally disordered offenders. From a Western perspective, changes in Chinese criminal law are sometimes clichéd as toothless window dressing, but they may represent a genuine step forward in safeguarding human rights. Taking a historical perspective, this paper reveals that in the East, as much as in the West, there is a 'moral tradition' of not punishing mentally disordered offenders who are not considered responsible for their acts. There are clear differences in disposal for those acquitted having been found 'not guilty by reason of insanity'. Whereas Western jurisdictions have offered (criminal) courts the opportunity for commitment in (forensic) mental hospitals from the early 19th Century, in China, disposal has remained, until the recent changes, the responsibility of the administration (mainly the police) or the family of the offender. A few high profile cases brought to light the inadequacy of these arrangements and the general disregard of obvious mental health issues when sentencing offenders. There was lack of clarity regarding who would take responsibility for treatment and issues of future public protection arising from a mental disorder. The 2012 CCP introduces the power of mental health commitment by the judiciary for those found non-responsible for an offense because of a mental disorder. Similar to provisions in Western jurisdictions there remain human rights concerns regarding aspects of 2012 CCP and the role of 'preventive detention' for mentally disordered offenders on indeterminate secure mental health detention. Nevertheless, the shift to judicial decision making in such cases and the possibility of mental health commitment are welcome steps in improving the human rights of this vulnerable population.


Suprachoroidal Space Alterations Following Delivery of Triamcinolone Acetonide: Post-Hoc Analysis of the Phase 1/2 HULK Study of Patients With Diabetic Macular Edema.
To study anatomic changes in the suprachoroidal space (SCS) following suprachoroidal injection of CLS-TA, triamcinolone acetonide injectable suspension. Eyes with diabetic macular edema receiving CLS-TA were imaged serially using anterior segment spectral-domain optical coherence tomography to examine the SCS. At the final imaging session, the SCS was not significantly different in study eyes (n = 14; 8.4 μm) compared to fellow eyes (n = 10; 8.1 μm; P = .698). Two eyes were imaged immediately before and 30 minutes after suprachoroidal injections; in these eyes, mean suprachoroidal width increased significantly following CLS-TA injection, 9.9 μm to 75.1 μm (P < .001), and subsequently returned to 14.9 μm 1 month after the final injection (P = .221). Suprachoroidal CLS-TA injection caused a measurable increase in the SCS, which returned to preinjection levels by 1 month following injection with no apparent lasting impact on SCS anatomy. [Ophthalmic Surg Lasers Imaging Retina. 2018;49:692-697.].


[Length-force relation of rat-carotid artery at different transmural pression. Experiments and models (author's transl)].
The relation between force and extension in longitudinal direction as a function of the internal pressure was examined in isolated carotid arteries of rats using a device for testing the stress-strain relation of fibers (Vibrodyn). In the region of small longitudinal extensions the longitudinal force is decreased by raising the internal pressure. The longitudinal force is increased as a function of the internal pressure in the region of high longitudinal extensions. This behaviour can be explained by a model of the arterial wall which takes into account the spiral structure of the fibers. We could find a good qualitative agreement between this model and our experiments. It can be concluded that the histological structure plays an important role in determining the elastic behaviour of the arterial wall.


Red cell metabolism affects lactate and pyruvate partition across the plasma membrane.
The influence on red cell metabolism of increasing the glucose concentration or the pH of the medium has been examined in order to compare the modification of the lactate/pyruvate ratio inside and outside the cell. In both situations, we evidenced a constancy in the intracellular lactate/pyruvate ratio and an increase in the extracellular one, thus suggesting that the cell efficiently opposes cytoplasmic modifications. In fact an increase of lactate efflux takes place when the intracellular pyruvate decreases. The implications of these changes in the two compartments are discussed on the basis of the available results.


An Approach to Intraoperatively Identify the Coronal Plane Deformities of the Distal Tibia When Treating Tibial Fractures with Intramedullary Nail Fixation: a Retrospective Study.
To develop a new approach to intraoperatively identify the presence of coronal plane deformities (both valgus and varus) when treating tibial fractures with closed reduction and intramedullary nail fixation. A retrospective analysis was conducted by enrolling 33 consecutive patients with tibial fractures who received closed reduction and intramedullary nail fixation from January 2018 to January 2019 at our trauma center. Out of the 33 patients, 23 were males and 10 were females and the average age was 41 years (ranging 22 to 69 years of age). Standard anteroposterior and lateral preoperative radiographs were routinely performed. After intraoperatively inserting the tibial intramedullary nail through the standard entry point, the parallel relationship between the distal horizontal interlocking screw and the tibiotalar joint surface on the anteroposterior fluoroscopy was used to determine the occurrence of valgus or varus deformities of the distal tibial fragment. Radiographic and clinical outcomes were analyzed using the average interval from injury to surgery, the lateral distal tibial angle (LDTA) of the unaffected and affected sides, complications and the Olerud-Molander ankle score. All 33 patients were postoperatively followed for 13 to 25 months (mean 18.7 months). The fractures achieved bone union at an average of 4.3 months (ranging from 3 to 6 months). The total complication rate was 60.6% (20 cases), including four cases that showed deep vein thrombosis, one case showing an infection and delayed union and 15 cases showing slight to moderate anterior knee pain. The postoperative LDTA of the unaffected side measured 87.3° to 89.6 ° (average 88.7° ± 0.8°), and the LDTA of the affected side was 87.5° to 90.4° (average 88.9° ± 1.1°). There was no significant difference between the unaffected and affected sides (t = -1.865, P = 0.068). The intraoperative measurement indicated six cases of valgus angulation and three cases of varus angulation deformities, and all deformities were corrected during surgery. According to the Olerud-Molander ankle score, clinical outcomes demonstrated 22 excellent cases, eight good cases, two fair cases, and one poor case 12 months after surgery. The parallel relationship between the distal horizontal interlocking screw and tibiotalar joint surface on intraoperative anteroposterior films were able to determine the onset of valgus or varus angulations of the distal tibial fragment in the fracture zone after the tibial intramedullary nail was inserted through the standard entry point.


Mother-bingo bonding: screening for gambling addiction in the neonatal intensive care unit.
As the prevalence of gambling increases in society, problem or pathologic gambling in new mothers may interfere with the bonding process as well as with infant and child care. Although neonatal nurses routinely screen for maternal addictions to substances such as alcohol, cocaine, and heroin, they are less likely to be alerted to signs of gambling addiction in new parents. This article presents a case study of a mother with a bingo preoccupation, provides neonatal nurses with an overview of problem and pathologic gambling, presents brief interview questions and a standardized screening tool for assessing gambling problems in parents, and suggests areas for intervention.


Performance of first trimester biochemical markers and mean arterial pressure in prediction of early-onset pre-eclampsia.
To develop a predictive risk model for early-onset pre-eclampsia (EO-PE) using maternal characteristics, combined screening markers, previously reported biomarkers for PE and mean arterial pressure (MAP). This retrospective study was conducted at Oulu University hospital between 2006 and 2010. Maternal serum from first trimester combined screening was further analyzed for alpha fetoprotein (AFP), placental growth factor (PlGF), soluble tumor necrosis factor receptor-1 (sTNFR1), retinol binding protein-4 (RBP4), a disintegrin and metalloprotease-12 (ADAM12), soluble P-selectin (sP-selectin), follistatin like-3 (FSTL3), adiponectin, angiopoietin-2 (Ang-2) and sex hormone binding globulin (SHBG). First, the training sample set with 29 cases of EO-PE and 652 controls was developed to study whether these biomarkers separately or in combination with prior risk (maternal characteristics, first trimester pregnancy associated plasma protein-A (PAPP-A) and free beta human chorionic gonadotrophin (fβ-hCG)) could be used to predict the development of EO-PE. Second, the developed risk models were validated with a test sample set of 42 EO-PE and 141 control subjects. For the test set MAP data was also available. Single marker statistically significant (ANOVA p<0.05) changes between control and EO-PE pregnancies were observed with AFP, RBP4 and sTNFR1 with both training and test sample sets. Based on the test sample set performances, the best detection rate, 47% for a 10% false positive rate, was achieved with PlGF and sTNFR1 added with prior risk and MAP. Based on our results, the best first trimester biomarkers to predict the subsequent EO-PE were AFP, PlGF, RBP4 and sTNFR1. The risk models that performed best for the prediction of EO-PE included prior risk, MAP, sTNFR1 and AFP or PlGF or RBP4.


"Single-cell pattern" of adenocarcinoma cells in effusion cytology: Morphologic challenges of lung cancer.
Lung adenocarcinomas present as tight clusters and three-dimensional balls in effusion specimens. Unlike carcinomas of breast and stomach where singly lying malignant cells are seen in effusion samples, lung adenocarcinomas usually show cohesive morphology. This single-cell pattern may also be confused with reactive mesothelial cells. We studied the frequency of pulmonary adenocarcinoma with single-cell pattern cytomorphology in pleural effusion specimens. All cases reported as either suspicious or positive for malignancy on pleural effusion cytology (PFC) over the past 1 year were retrieved. The clinical details were obtained from requisition forms. Cases with predominant single-cell pattern, clinically suspicious of carcinoma lung were segregated. These were de-stained and immunocytochemistry (ICC) for TTF-1 was performed. Of 103 cases reported as either suspicious or positive for malignancy on PFC, 29 had a predominant single-cell pattern. Of these, 13 (44.8%) were primary lung carcinoma. The rest were metastasis from ovary (5; 17.2%), breast (2; 6.9%), stomach (2; 6.9%), lymphoma (1; 3.5%), and Ewing's sarcoma (1; 3.5%). Five (17.2%) were those with unknown primary. All cases of lung carcinoma were positive for TTF-1 ICC. Single-cell pattern of pulmonary adenocarcinoma is commoner than popularly believed. This pattern may be difficult to differentiate from carcinoma cells of other sites as well as from reactive mesothelial cells. A high degree of suspicion is therefore needed to perform relevant ICC to clinch the correct diagnosis.


Effort required in eligibility screening for clinical trials.
Determining eligibility for a clinical trial (CT) typically requires a lengthy manual review of data for a single evaluation. The cost associated with eligibility screening is typically not compensated through contracts supporting CTs. We used a real-time tracking system that captures CT evaluations and provides information on evaluation outcomes and time spent on each eligibility screening by research staff. Using these data, we describe the effort and costs of eligibility screening overall and per enrolled patient for cancer CTs. The study sample included all completed eligibility assessment (evaluation) records for the 18-month study period. We used generalized multinomial modeling to predict evaluation outcomes and then used the resulting parameter coefficients to estimate the effort associated with each participant, adjusted for probability of being enrolled. From these data, we calculated cost associated with eligibility screening. We found substantial variation in attributed cost by study type and phase. The cost of eligibility screening ranged by study phase from $129.15 to $336.48 per enrolled patient. The estimated annual cost of screening was more than $90,000. This study provides results based on prospectively captured effort to estimate the largely nonreimbursed costs of eligibility screening and suggests that screening can be a significant financial burden to an institution. Centers performing CTs may need to acknowledge the differences in screening costs for different study types when negotiating contracts with funding organizations. Information such as that captured here could support such negotiations to reduce the gap between reimbursed and nonreimbursed costs.


Studies on kinetic parameters and stability of aminoacylase in non-conventional media.
Catalytic properties and conformational stability of aminoacylase (N-acylamino acid amidohydrolase, EC 220.127.116.11) were studied in water-N,N-dimethylformamide (DMF) and water-dioxane solvent mixtures. Beside the prompt inhibition the solvents caused further inactivation during incubations. In the presence of 5% DMF content the inactivation proceeds with a well-measurable rate (t1/2 39 min), while in the case of 20% DMF the enzyme practically lost its starting activity during 50 min incubation (t1/2 13 min). The K(m) value of the enzyme increased about three times with increasing DMF concentrations up to about 2.6 M DMF, while the Vmax value decreased practically to zero in the same concentration range.


Soft tissue radiography of the testes.
A technique of soft tissue radiography of the testes is presented. The normal radiological appearances are described, and also the radiological features seen in 75 patients with testicular lesions. Of these, 18 patients had malignant neoplasms and 57 patients had benign conditions. Testicular tumours appeared as very radio-opaque, quite sharply defined shadows. Benign conditions were less radio-opaque and often had ill-defined margins. A diagnostic accuracy of 88% was achieved in differentiating between testicular tumours and benign conditions.


[Giant mandibular exostosis. Apropos of a case. Description, therapeutic approach. Review of the literature].
The authors present a cases of localised exostosis of the angle and the ascending ramus of the mandible. They stress the technical problems posed by this giant exostosis and they propose an original operative approach together with a complete review of the literature.


Retrograde axoplasmic transport of neurosecretory material. An immunocytochemical and electron-microscopic study of transected axons in normal and colchicine-treated rats.
The axonal transport of neurosecretory material was studied in neurosecretory axons of the supraoptico-posthypophyseal system after in-situ transection of the median eminence. Two hours, 8 h, and 18 h after the lesion, both vasopressin and oxytocin antibodies revealed progressive accumulations of immunoreactive material not only in the proximal but also in the distal stumps of the transected axons. The electron-microscopic examination of these axonal portions revealed that such intense immunopositive labelings could be correlated, in both stumps, to a conspicuous accumulation of neurosecretory granules. It is concluded that, under normal physiological conditions, a significant amount of axoplasmic neurosecretory material is transported in retrograde direction and that such a retrograde transport mainly involves neurosecretory granules.


Novel, human cell line-derived recombinant factor VIII (Human-cl rhFVIII, Nuwiq® ) in children with severe haemophilia A: efficacy, safety and pharmacokinetics.
Nuwiq® (Human-cl rhFVIII) is a new-generation recombinant factor VIII (rFVIII) protein, without chemical modification or fusion to any other protein, produced in a human cell line. This prospective, open-label, multinational phase III study assessed the efficacy and safety of Human-cl rhFVIII in 59 previously treated patients (PTPs) with severe haemophilia A aged 2-12 years (2-5 [N = 29]; 6-12 [N = 30]) during standard prophylaxis (≥50 exposure days and ≥6 months). Efficacy in treating breakthrough bleeds and during surgical prophylaxis was also assessed. An initial pharmacokinetic assessment (N = 13 per age subgroup) demonstrated comparable results with the one-stage and chromogenic assays. Mean (SD) half-life was 11.9 (5.4) and 13.1 (2.6) hours in children aged 2-5 years and 6-12 years respectively (one-stage assay). Prophylactic efficacy, based on mean monthly bleeding rate, was 'excellent' or 'good' in 91.5% of children for all bleeds and in 96.6% of children for spontaneous bleeds. Mean (SD) annualized bleeding rate was 4.12 (5.22) [median 1.9] for all bleeds, 1.50 (3.32) [median 0] for spontaneous bleeds and 2.34 (3.54) [median 1.57] for traumatic bleeds. There were no major, life-threatening bleeds. Efficacy was 'excellent' or 'good' in the treatment of 82.4% of breakthrough bleeds. Overall efficacy during five major surgeries was rated as 'excellent'. There were no FVIII inhibitors or treatment-related serious adverse events. These results in paediatric PTPs indicate that Human-cl rhFVIII is effective for the prevention and treatment of bleeds.


Deletion of the fungus specific protein phosphatase Z1 exaggerates the oxidative stress response in Candida albicans.
Candida albicans is an opportunistic pathogen which is responsible for widespread nosocomial infections. It encompasses a fungus specific serine/threonine protein phosphatase gene, CaPPZ1 that is involved in cation transport, cell wall integrity, oxidative stress response, morphological transition, and virulence according to the phenotypes of the cappz1 deletion mutant. We demonstrated that a short-term treatment with a sublethal concentration of tert-butyl hydroperoxide suppressed the growth of the fungal cells without affecting their viability, both in the cappz1 mutant and in the genetically matching QMY23 control strains. To reveal the gene expression changes behind the above observations we carried out a global transcriptome analysis. We used a pilot DNA microarray hybridization together with extensive RNA sequencing, and confirmed our results by quantitative RT-PCR. Novel functions of the CaPpz1 enzyme and oxidative stress mechanisms have been unraveled. The numbers of genes affected as well as the amplitudes of the transcript level changes indicated that the deletion of the phosphatase sensitized the response of C. albicans to oxidative stress conditions in important physiological functions like membrane transport, cell surface interactions, oxidation-reduction processes, translation and RNA metabolism. We conclude that in the wild type C. albicans CaPPZ1 has a protective role against oxidative damage. We suggest that the specific inhibition of this phosphatase combined with mild oxidative treatment could be a feasible approach to topical antifungal therapy.


Morphological abnormalities in zebrafish cryopreserved sperm.
The aim of the present study was to evaluate the effect of cryopreservation on the morphology of zebrafish sperm (Danio rerio). Sperm from 30 males were collected and divided in two treatments: fresh and cryopreserved semen. The following were measured sperm morphology, motility and membrane integrity. Cryopreservation reduced motility, the number of normal cells and the membrane integrity, as well as increased the percentage of sperm abnormalities. The most frequent types of morphological changes found in cryopreserved semen were macrocephaly, loose head, degenerated head, proximal gout, curled tail and short tail. This study opens the way for further investigations on morphological changes and for a new classification of these changes in fish semen due to cryopreservation.


Sustained clinical response in psoriatic arthritis patients treated with anti-TNF agents: a 5-year open-label observational cohort study.
To investigate the efficacy, toxicity, and drug discontinuation in patients with psoriatic arthritis treated with anti-tumor necrosis factor agents. Sixty-five patients with active disease were included in this open-label study. They had tender or swollen joint count ≥5, Psoriatic Arthritis Severity Index (PASI) score ≥10, and erythrocyte sedimentation rate ≥28 mm Hg/1st hour and/or C-reactive protein ≥10 mg/L. All were refractory to at least 2 disease-modifying antirheumatic drugs. Thirty were treated with infliximab, 25 with etanercept, and 10 with adalimumab. Infliximab (5 mg/kg body weight) was given intravenously at weeks 0, 2, 6, and every 8 weeks thereafter; etanercept was given subcutaneously (25 mg twice a week), while adalimumab was given subcutaneously (40 mg every other week) for a period of 5 years. Data concerning anti-tumor necrosis factor efficacy tolerability, adverse events, and drug discontinuation were recorded. The percentage of patients who achieved the Psoriatic Arthritis Response Criteria (PSARC), the improvement of PASI, the improvement according to the American College of Rheumatology (ACR) criteria, and the disease activity for 28 joint indices score (DAS-28) were recorded. After 5 years, PSARC was 60%, PASI 70 was 66.7%, PASI 90 was 63.3%, while ACR 50 was 56.7% for the patients treated with infliximab. Moreover, PsARC was 64%, PASI 70 and PASI 90 were 68%, while ACR 50 was 56% for those treated with etanercept. Furthermore, in the adalimumab group PsARC was 56%, PASI 70 and PASI 90 were 58% and 50%, respectively, while ACR 50 was 50%. Additionally, DAS-28 scores were significantly improved. Thirteen patients treated with infliximab, 6 with etanercept, and 5 patients with adalimumab were withdrawn. At the end of treatment, the survival of infliximab was 56.7%, for etanercept 76%, and for adalimumab 50%. All drugs were effective, safe, and well-tolerated. The clinical improvement was maintained through the 5 years with satisfying infliximab and adalimumab survival and high etanercept survival.


Activation of the ATX/LPA/LPARs axis induces a fibrotic response in skeletal muscle.
Several common chronic diseases, muscular dystrophies (MDs), and aging lead to progressive fibrous connective tissue (fibrosis) accumulation in skeletal muscle. Cumulative past evidence points to the role of signaling lipids such as lysophosphatidic acid (LPA) and its receptors (LPARs) in different models of fibrosis. However, the potential contribution of these molecules to the fibrotic process in skeletal muscle has not been explored. Here, we show the expression of ATX/LPA/LPARs axis components in skeletal muscle, which suggests their potential relevance for the biology of this tissue. We investigated if the skeletal muscle responds to the stimulus of intramuscular (IM) LPA injections, finding an early induction of the pro-fibrotic factor connective tissue growth factor/Cellular Communication Network factor 2 (CCN2) and extracellular matrix (ECM) proteins. Also, we found that LPA induces an increase in the number of fibro/adipogenic progenitors (FAPs), which are the primary cellular source of myofibroblasts. These effects were for the most part prevented by the inhibitor Ki16425, which inhibits the LPA receptors LPA1 and LPA3, as well as in the LPA1-KO mice. We also evaluated the in vivo activation of extracellular signal-regulated kinases (ERK 1/2), AKT, c-Jun N-terminal kinase (JNK), and Yes-asocciated protein 1 (YAP) in response to LPA. Our results show that LPA induces ERK 1/2 phosphorylation in WT muscle, but not in LPA1-KO mice. Treatment with the ERK 1/2 inhibitor U0126 prevented the induction of fibronectin in response to LPA, suggesting that this pathway is involved in LPA-induced fibrosis. Altogether, these results demonstrate that ATX/LPA/LPARs constitute a pro-fibrotic axis and suggest a possible role in muscular diseases.


Bacterial and fungal contaminants in caprine and ovine cheese: A meta-analysis assessment.
The dairy industry is of great importance to the European economy contributing towards € 8.7 billion of the total trade surplus. Caprine and ovine milk amount to 3.1% of the 152 million tonnes of milk produced in Europe, 95% of which is transformed into dairy products such as cheese. This cheese is mostly produced in small holdings from untreated milk, making it a high-risk dairy product for human consumption. A total of 49 foodborne disease outbreaks caused by dairy products were registered in 2017 in Europe. Therefore, these products remain a serious health risk. This meta-analysis examined 30 studies assessing bacterial or fungal contamination of caprine or ovine milk cheeses. The significantly contaminating microbes were found to be Acremonium spp. (19%), Aspergillus spp. (23%), Bacillus spp. (2%), Brucella spp. (34%), Enterobactericae spp. (36%), Enterococcus spp. (28%), Escherichia spp. (15%), Fusarium spp. (21%), Geotrichum spp. (22%), Listeria spp. (11%), Mucor spp. (15%), Penicillium spp. (25%), Phoma spp. (20%), Rhizopus spp. (15%), Salmonella spp. (3%), Scopulariopsis spp. (19%) and Staphylococcus spp. (25%) in caprine and ovine cheese, indicating a variety of food pathogens as well as spoilers. Raw milk is nutritious hence prone to contamination. However, since traditional cheese is often made from untreated milk, it is important to educate cheesemakers of key safety measures and good manufacturing practice allowing for the safe production of these food items.


The G to A transformation of rs4702 polymorphism in 3'UTR of FURIN reduced the risk of radiotherapy-induced cognitive impairment in glioma patients.
The G allele of rs4702 polymorphism has been reported to reduce the production of mature BDNF and FURIN, both of which were closely associated with cognitive functions. Real-time PCR, ELISA and luciferase assay were performed to explore the interactions between miR-338-3p, FURIN and BDNF. T-RFLP was used to assess the intestinal flora in the stool samples of glioma patients after radiotherapy. We grouped the 106 glioma patients recruited according to the rs4702 polymorphism. The results showed no obvious correlation between rs4702 polymorphism and the expression of miR-338-3p. However, rs4702-A was associated with increased expression of FURIN and BDNF in the serum and PBMC of glioma patients after radiotherapy. Besides, the study found that rs4702-A was remarkably associated with increased enterotype I and decreased enterotype III in the stool of glioma patients after radiotherapy. Rs4702-A was also proved to be closely associated with increased MMSE, role functioning and social functioning at three months after radiotherapy. Furthermore, miR-338-3p repressed the expression of FURIN-G. Compared with G allele, the presence of A allele of rs4702 polymorphism in FURIN could obstruct the suppressive effect of miR-338-3p upon the expression of FURIN and BDNF in intestinal flora. Therefore, the carriers of A allele will be challenged with less risk of radiotherapy-induced cognitive impairment.


Overnight S100B in Parkinson's Disease: A glimpse into sleep-related neuroinflammation.
Calcium-binding protein B (S100B), a primary product of astrocytes, is a proposed marker of Parkinson's Disease (PD) pathophysiology, diagnosis and progression. However, it has also been implicated in sleep disruption, which is very common in PD. To explore the relationship between S100B, disease severity, sleep symptoms and polysomnography (PSG) findings, overnight changes in serum S100B levels were investigated for the first time in PD. 17 fully treated, non-demented, moderately advanced PD patients underwent PSG and clinical assessment of sleep symptoms. Serum S100B samples were collected immediately before and after the PSG. Results are shown as median [interquartile range]. Night and morning S100B levels were similar, but uncorrelated (rs=-0.277, p=0.28). Morning S100B levels, as opposed to night levels, positively correlated with the Unified Parkinson's Disease rating scale (UPDRS) subsections I and II (rs=0.547, p=0.023; rs=0.542, p=0.025). Compared to those with overnight S100B reduction, patients with overnight S100B elevation had higher H&Y scores (2.5 [0.87] vs. 2 [0.25], p=0.035) and worse total Pittsburgh Sleep Quality Index (PSQI) and Parkinson's Disease Sleep Scores (10 [3.2] vs. 8 [4.5], p=0.037; 92.9 [39] vs. 131.4 [28], p=0.034). Correlation between morning S100B levels and total UPDRS score was strengthened after controlling for total PSQI score (rs=0.531, p=0.034; partial rs=0.699, p=0.004, respectively). Overnight S100B variation and morning S100B were associated with PD severity and perceived sleep disruption. S100B is proposed as a putative biomarker for sleep-related neuroinflammation in PD. Noradrenergic-astrocytic dysfunction is hypothesized as a possible mechanism underlying these findings.


Evidences for salbutamol metabolism by respiratory and liver cell lines.
This study aimed to investigate the enantiomeric biotransformation of salbutamol in the human respiratory and liver cells. The cells from the different cell growth cycles were treated with various concentrations of salbutamol sulfate. Salbutamol and its metabolites were analyzed using chiral liquid chromatography and mass spectrometry. There were no metabolites of salbutamol found in the extracellular medium, intracellular, and cell lysate of respiratory cell lines. The S/R ratios of salbutamol were found to be 0.99-1.10 in all cell lines, cell cycles, and salbutamol concentrations in this study. Salbutamol metabolites were found only in intracellular HepG2 cells. The S/R ratios of the salbutamol inside the liver cells were 10 times greater than the S/R ratios of the salbutamol in the liver extracellular medium (0.99-1.10). It is important to note that the S/R ratios of salbutamol in liver cell lysate enzyme were 0.99-1.10 whereas the S/R salbutamol metabolites inside the liver cell were around 1.91-2.14. Both salbutamol and sulfate conjugation metabolites were detected in MS chromatograms with an m/z of 239.2 and 317.6, respectively. Hence, the delivery of salbutamol directly to the respiratory system is a right target that can avoid first-pass metabolism.


Plasma protein binding of the reversible type A MAO inhibitor cimoxatone (MD 780515).
Binding of a new selective reversible type A MAO inhibitor cimoxatone (MD 780515) to plasma proteins was studied in vitro by equilibrium dialysis. Binding to 580 microM human serum albumin (HSA) and to total plasma proteins was 93-96% and independent of cimoxatone concentration (0.15-207 microM). The drug was mainly bound to HSA with two binding sites and a moderate association constant (K = 2.9 X 10(4) M-1). Free fatty acids did not modify cimoxatone binding to HSA. Cimoxatone was also moderately bound to isolated lipoprotein fractions; alpha 1-acid glycoprotein and gamma-globulins did not play an important role in the binding of cimoxatone. MD 770222, the O-demethyl metabolite, appeared to be bound to HSA at the same binding sites as cimoxatone. However, no interaction occurred between the two compounds for 580 microM HSA. L-Tryptophan, bilirubin, the benzodiazepines flunitrazepam and oxazepam, imipramine and aspirin, did not displace cimoxatone from its binding sites. On the other hand, warfarin and phenylbutazone decreased cimoxatone binding to 29 microM HSA but no interaction occurred with 580 microM HSA.


Some amino sugars structurally related to 6-deoxymannojirimycin precursors prepared from methyl 6-deoxy-2,3-O-isopropylidene-alpha-D-lyxo-hexofuranosid-5-ulose and methyl 2,3-O-isopropylidene-alpha-D-lyxo-pentodialdo-1,4-furanoside.
Methyl (5S)-5-C-amino-5-cyano-5-deoxy-2,3-O-isopropylidene-alpha-D-lyxofuranoside has been synthesised from methyl 2,3-O-isopropylidene-alpha-D-lyxo-pentodialdo-1,4-furanoside, applying the Strecker synthesis. Analogously, methyl (5S) and (5R)-5-C-amino-5-cyano-5,6-dideoxy-2,3-O-isopropylidene-alpha-D-lyxo-hexofuranosides were prepared from methyl 6-deoxy-2,3-O-isopropylidene-alpha-D-lyxo-hexofuranosid-5-ulose. The 5-S configuration was unambiguously determined by single-crystal X-ray diffraction analysis of corresponding N-acetyl derivatives. Conformations of five-membered rings are discussed. The conversion of N-acetylated amino nitriles to N-acetylamino acid ethyl ester and amide, respectively, is also described.


Testing for genetic structure in different urban Argentinian populations.
Fifteen autosomal short tandem repeat (STR) markers (D3S1358, HUMTH01, D21S11, D18S51, PENTA E, D5S818, D13S317, D7S820, D16S539, CSF1PO, PENTA D, HUMvWA, D8S1179, HUMTPOX, FGA) were analyzed in 1734 individuals living in urban areas of cities from six different Argentinian provinces (Buenos Aires, Neuquén, Tucumán, La Pampa, San Luis, Santa Cruz) in order to determine if a common urban database could be used in Argentina for forensic purposes. Frequencies estimates, Hardy-Weinberg equilibrium (HWE), and other parameters of forensic interest were computed. Comparisons between the six populations, and with published data from one Native American population from Argentina and other urban populations from Argentina and Europe were also performed. Our results reveal evidences for population structure, both when testing for genetic differentiation and when comparing frequencies distributions between different pairs of populations. Therefore, caution should be taken when using a common pooled database with general forensic purposes in Argentina.


Retrospective construction of the judgement of free choice.
The problem of free will lies at the heart of modern scientific studies of consciousness. Some authors propose that actions are unconsciously initiated and awareness of intention is referred retrospectively to the action after it has been performed [e.g. Aarts, H., Custers, R., & Wegner, D. M. (2005). On the inference of personal authorship: Enhancing experienced agency by priming effect information. Consciousness & Cognition, 14, 439-458]. This contrasts with the common impression that our intentions cause those actions. By combining a stop signal paradigm and an intentional action paradigm we show that participants sometimes indicate to have intentionally initiated an action while reaction time data strongly suggest that they in fact failed to stop the action. In a second experiment we demonstrate that the number of trials in which participants misattributed their awareness of intention varied with the intentional involvement during action planning. Our data support the retrospective account of intentional action. Furthermore, we introduce an experimental approach that objectifies introspective judgments of awareness of intention.


Lipoprotein analyses in varying degrees of glucose tolerance. Comparison between non-insulin-dependent diabetic, impaired glucose tolerant, and control populations.
Atherosclerosis is the major cause of death in diabetic patients. Lipoproteins and lipids are frequently altered in non-insulin-dependent diabetes. These lipoprotein alterations are of interest because of their possible role in the origin of the accelerated atherosclerosis found in diabetes. Because of the link between lipoproteins and diabetes, serum lipids and lipoproteins were measured in 215 middle-aged patients (107 female, 108 male) with varying degrees of glucose tolerance: control subjects, subjects with impaired glucose tolerance (IGT), and patients with non-insulin-dependent diabetes mellitus (NIDDM). In male subjects, levels of fasting total triglycerides were significantly greater in those with NIDDM compared with control subjects. In female subjects, fasting total cholesterol levels were significantly greater in NIDDM compared with IGT. Both high-density lipoprotein (HDL) cholesterol and HDL2 cholesterol values were significantly lower in both sexes with NIDDM compared with control subjects. Low-density lipoprotein (LDL) cholesterol levels were elevated in the male subjects with IGT. No differences in HDL cholesterol or its subfractions were seen in both sexes with IGT compared with control subjects. Bivariate analyses showed that the reduced HDL cholesterol and HDL subfraction levels were most closely associated with both total triglycerides and weight. This study shows that reduced HDL cholesterol and HDL2 cholesterol levels occur in NIDDM, whereas persons with "impaired glucose tolerance" do not have the dramatic alterations in HDL levels.


Off-label use of ceftiofur in one-day chicks triggers a short-term increase of ESBL-producing E. coli in the gut.
This trial was designed to evaluate the off-label use of ceftiofur with Marek's vaccine in one-day-old broiler chicks, a prophylactic treatment that has been done in some commercial hatcheries, on the emergence of extended-spectrum beta-lactamase producing Escherichia coli (ESBL-E. coli). A total of 168 chicks (Cobb500®) were used in a completely randomized design. Birds were assigned to two treatments (Marek's vaccine plus saline vs Marek's vaccine plus ceftiofur) and six repetitions, with 14 animals each. Cloacal swabs were collected from 1 to 14 days post-hatch. The majority (86%; p<0.0001) of the ESBL-producing isolates harboring blaCTX-M and blaSHV genes originated from animals receiving the antimicrobial. None of the isolates were positive for plasmid-mediated AmpC betalactamase genes (blaACC, blaCMY-2, blaDHA, blaFOX, blaMOX and blaMIR). These findings indicate that the off-label use of ceftiofur with Marek's vaccine is associated with the short-term increase in ESBL-producing Escherichia coli in the gut of chicks.


Cloning of cDNA for regenectin, a humoral C-type lectin of Periplaneta americana, and expression of the regenectin gene during leg regeneration.
We isolated cDNA for regenectin, a C-type lectin of the American cockroach (Periplaneta americana), and analysed expression of the regenectin gene in the regenerating legs. Regenectin was found to be a member of the Periplaneta lectin-related protein family. We found that the regenectin gene was expressed specifically in the epidermal cells of the newly formed regenerating legs. Together with our previous results, these results suggest that regenectin is synthesized by epidermal cells, secreted into the regenerating leg saccule, and assembles around myoblasts to form leg muscle fibers in situ.


Maternal attitudes in pregnancy predict drinking initiation in adolescence.
The aim of the present study was to examine whether maternal attitudes to pregnancy and infant caregiving predict drinking initiation in adolescence. Data were used from the Mater University Study of Pregnancy, an Australian longitudinal study of mothers and their children from pregnancy to when the children were 14 years. Logistic regression analyses examined whether maternal attitudes to pregnancy and infant caregiving at birth and 6 months contributed to the prediction of drinking initiation at age 14, after controlling for a range of confounding covariates. Mother's negative feelings about being pregnant and not planning/wanting the pregnancy predicted adolescent alcohol initiation at the 14 year follow up. Negative maternal attitudes to infant caregiving at birth and 6 months did not predict adolescent alcohol initiation. Maternal attitudes in pregnancy are important and there is a need for effective perinatal interventions.


N-acetylglucosamine suppresses osteoclastogenesis in part through the promotion of O-GlcNAcylation.
Osteoclasts are the only cells in an organism capable of resorbing bone. These cells differentiate from monocyte/macrophage lineage cells upon stimulation by receptor activator of NF-κB ligand (RANKL). On the other hand, osteoclastogenesis is reportedly suppressed by glucose via the downregulation of NF-κB activity through suppression of reactive oxygen species generation. To examine whether other sugars might also affect osteoclast development, we compared the effects of monomeric sugars (glucose, galactose, N-acetylglucosamine (GlcNAc), and N-acetylgalactosamine (GalNAc)) on the osteoclastogenesis of murine RAW264 cells. Our results demonstrated that, in addition to glucose, both GlcNAc and GalNAc, which each have little effect on the generation of reactive oxygen species, suppress osteoclastogenesis. We hypothesized that GlcNAc might affect osteoclastogenesis through the upregulation of O-GlcNAcylation and showed that GlcNAc increases global O-GlcNAcylation, thereby suppressing the RANKL-dependent phosphorylation of NF-κB p65. Furthermore, an inhibitor of N-acetyl-β-D-glucosaminidase, O-(2-acetamido-2-deoxy-D-glucopyranosylidene) amino N-phenylcarbamate (PUGNAc), which also increases O-GlcNAcylation, suppressed the osteoclastogenesis of RAW264 cells and that of human peripheral blood mononuclear cells. Together, these data suggest that GlcNAc suppresses osteoclast differentiation in part through the promotion of O-GlcNAcylation.


Phase I study of vinorelbine and carboplatin in advanced non-small cell lung cancer.
Thirty-one patients with previously untreated advanced non-small cell lung cancer were included in a Phase I study to determine the optimal dose of Carboplatin (CBDCA) which preserves the best Navelbine (NVB) dose-intensity. NVB was administered at a 30-mg/m2 fixed-dose on days 1-8/q 3 weeks, whereas CBDCA doses were planned to be escalated from 275 to 400 mg/m2 on day 1/q 3 weeks for six successive groups of patients. The toxicity limiting dose of CBDCA in the combination was 350 mg/m2 on day 1/q 3 weeks because of repetitive Grade IV neutropenia, and the optimal dose of CBDCA was 325 mg/m2 on day 1/q3 weeks, offering a 86.4% NVB and a 92.6% CBDCA relative dose-intensity for the first 9 weeks. Responses were observed at each step. This study demonstrates the feasibility and the efficacy of the NVB-CBDCA combination. It suggests that dose-intensity calculation can be helpful to determine the recommended dose for Phase II studies of new drug combinations.


A tentative mechanism for inferior punctate keratopathy.
We analyzed the blinking motions of ten patients with inferior punctate keratopathy and ten normal subjects by means of time-lapse motion pictures. We also determined the frequency of blinking during one-minute intervals for each individual. Four types of blinking were distinguished: complete, incomplete, twitch, and forced blink. The average blinking frequency was about the same for both groups: 16 to 17 blinks per minute. The relative proportion of complete blinks, however, was much lower in patients with inferior punctate keratopathy than in normal subjects. The impaired blinking motion may be responsible for the punctate staining of the inferior corneal epithelium by producing exposure and inadequacies in the tear film. For this reason, these patients may benefit from blink training, application of tear substitutes, or soft contact lenses more than from the customary prescription of topical cortisone eyedrops.


Mitigating Cd and bacterial wilt stress in tomato plants through trico-synthesized silicon nanoparticles and Trichoderma metabolites.
There has been a growing apprehension in recent years about the harmful effects of environmental pollutants on agricultural output, encompassing both living organisms and non-living factors that cause stress. In this study, the soil application of bulk silicon (Si), silicon nanoparticles (SiNPs) and Trichoderma metabolites (TM) were investigated alone or in combination for the management of an important abiotic stress i.e. Cd toxicity and biotic stress i.e. bacterial wilt (BW) in tomato plants. SiNPs were synthesized by Trichoderma and confirmed through XRD, FTIR, and Ranman spectrum analysis. Results showed that Si, SiNPs and TM were all effective treatments. The combine treatment of SiNPs and TM followed by SiNPs alone were superior over other treatments in mitigating Cd toxicity and reducing BW disease on tomato plants. The soil application of these treatments reduced the Cd toxicity by enhancing Cd-tolerance index, decreasing bioavailability of soil Cd, reducing Cd contents and translocation in plants, improving gaseous exchange, photosynthesis, and increasing the antioxidant enzyme activities and their transcriptions. These treatments significantly suppressed BW pathogen leading to the significant decrease in disease index and severity on plants. In vitro evaluation and scanning electron microscopic (SEM) analysis revealed that SiNPs and TM significantly disrupted the cellular morphology of BW pathogen Ralstonia solanacearum. Findings of this study proposes the possible use of SiNPs and TM in mitigating the Cd and BW stress in tomato plants and possibly in other crops.


Parathyroids: the forgotten glands.
Symptomatic calcium disorders--most notably, hypocalcemia--are common problems in preterm and certain term infants. Many factors predispose susceptible neonates to hypo- or hypercalcemia, but in many cases, the root of the problem is altered function of the parathyroid glands. Parathyroid gland dysfunction may affect calcium homeostasis and alter physiologic functioning, resulting in significant clinical symptoms. A review of how the parathyroid glands and parathyroid hormone affect calcium balance and of the problems that result from altered function promotes a better appreciation of the important role played by these sometimes "forgotten" glands in maintaining normal neonatal physiologic functioning.


Repeated Intravenous Treatment with Recombinant Tissue-Type Plasminogen Activator in Patients with Acute Ischemic Stroke.
Increased use of systemic thrombolysis, demographic changes, and higher chances of surviving first-ever strokes all lead to an increasing number of patients with recurrent stroke. However, data on repeated thrombolysis are limited. Here, we report on the safety and clinical effects of repeated intravenous recombinant tissue-type plasminogen activator (rt-PA) treatment in a large consecutive cohort of stroke patients. We identified all stroke patients who received repeated thrombolysis. We determined safety and 3-month clinical outcome after the first and second thrombolysis. All patients received follow-up brain imaging. Good clinical outcome was defined as a modified Rankin Scale of 0-2 or recovery to the prestroke status. In total, 24 patients were included (i.e. 1.5% of all stroke patients treated with rt-PA at our center who survived the first treatment; male 45.8%; median age at first event: 74.5 years). No allergic or anaphylactic reactions were recorded after the first time of treatment, but oral angioedema developed once during the second treatment. No symptomatic intracerebral hemorrhage was observed. Clinical outcome was good in 75.0% after the first, but in only 41.7% after the second treatment (p = 0.021). Repeated thrombolysis was not associated with a higher rate of complications. However, the clinical outcome appears to be less satisfactory than after the first treatment.


CMAHP promotes metastasis by reducing ubiquitination of Snail and inducing angiogenesis via GM-CSF overexpression in gastric cancer.
Pseudogenes are generally considered "junk" DNA or "genomic fossils" generated during the evolution process that lack biological activity. However, accumulating reports indicate that pseudogenes have biological functions critical for cancer development. Experiments from the current study showed marked overexpression of the cytidine monophospho-N-acetylneuraminic acid hydroxylase pseudogene (CMAHP) in gastric cancer, which was associated with poor overall survival. However, the mechanisms underlying the activity of CMAHP in tumor development are largely unknown. Gene Set Enrichment Analysis (GSEA) revealed that CMAHP-correlated genes are significantly involved in epithelial-mesenchymal transition (EMT) and angiogenesis. Functional studies further confirmed that CMAHP mediates metastasis and angiogenesis in vitro and in vivo. Furthermore, CMAHP promoted cancer cell migration, invasion, and metastasis through Snail overexpression, which decreased ubiquitination mediated by NF-κB signaling. Angiogenesis is known to be induced by granulocyte-macrophage colony-stimulating factor (GM-CSF) stimulation. CMAHP increased GM-CSF transactivation via promoting direct binding of c-Jun to the -1981/-1975 region of the GM-CSF promoter. Notably, CMAHP interacts with Histone H1.4 promoting histone acetylation to enhance c-Jun and RelA (p65) expression. Our collective findings provide novel evidence that CMAHP contributes to tumor progression and modulates metastasis and angiogenesis in gastric cancer.


Animal models of self-injurious behaviour: an overview.
Self-injurious behaviour is highly prevalent in neurodevelopmental disorders. Interestingly, it is not restricted to any individual diagnostic group. Rather, it is exhibited in various forms across patient groups with distinct genetic defects and classifications of disorders. This suggests that there may be shared neuropathology that confers vulnerability. Convergent evidence from clinical pharmacotherapy, brain imaging studies, postmortem neurochemical analyses, and animal models indicates that dopaminergic insufficiency is a key culprit. This chapter provides an overview of studies in which animal models have been used to investigate the biochemical basis of self-injury, and highlights the convergence in findings between these models and expression of self-injury in humans.


Comparison of the subjective satisfaction of the donor site morbidity: Free radial forearm flap versus anterolateral thigh flap for reconstruction in tongue cancer patients.
The purpose of the study was to compare the differences of the subjective satisfaction of the donor site morbidity between the free radial forearm flap (FRFF) and anterolateral thigh flap (ALTF) for tongue reconstruction. One hundred and nineteen patients underwent FRFF or ALTF reconstruction were retrospectively evaluated by a standardized self-established donor site morbidity questionnaire which included 5 domains, sensibility, movement disabilities, cosmetics, social activities and general impacts on the quality of life. The Cronbach's coefficient alpha of the questionnaire was 0.707. The exploratory factor analysis revealed that the 5 items of the questionnaire might load onto two distinct subscales. Patients with ALTF had higher scores in the sensibility, cosmetics and the composite score (P < 0.05). No significant differences were found in the movement disabilities, social activities and general impacts on the quality of life between the two groups (P > 0.05). ALTF has the advantage of better results of donor site morbidity, such as sensibility and cosmetics, over FRFF.


Glycolysis and pyruvate oxidation in cardiac hypertrophy--why so unbalanced?
Cardiac hypertrophy, induced by chronic pressure or volume overload, is associated with abnormalities in energy metabolism as well as characteristic increases in muscle mass and alterations in the structure of the heart. Hypertrophied hearts display increased rates of glycolysis and overall glucose utilization, but rates of pyruvate oxidation do not rise in step with rates of pyruvate generation. Glycolysis and glucose oxidation, therefore, become markedly less 'coupled' in hypertrophied hearts than in non-hypertrophied hearts. Because the pyruvate dehydrogenase complex (PDC) contributes so powerfully to the control of glucose oxidation, we set out to test the hypothesis that the function of PDC is impaired in cardiac hypertrophy. In this review we describe evidence indicating that the alterations in glucose metabolism in hypertrophied hearts cannot be explained simply by changes in PDC expression or control. Additional mechanisms that may lead to an altered balance of pyruvate metabolism in cardiac hypertrophy are discussed, with commentaries on possible changes in pyruvate transport, NADH shuttles, lactate dehydrogenase, and amino acid metabolism.


The development of an interdepartmental audit as part of a physics quality assurance programme for external beam therapy.
A cost-effective audit system has been developed that will both detect systematic error in data and procedures and evaluate the quality assurance programme provided by a physics department for radiotherapy. The audit has been developed for external beam radiotherapy and assesses one modality and one treatment machine per year. The audit is carried out on an interdepartmental basis and can be undertaken by two physicists from each department in one working day. The method of assessing the quality assurance programme and the schedule of measurements are described. The process is illustrated using the results of trial audits between the medical physics departments at Coventry and Leicester.


[Adrenergic control of myocardial contractile function in rats with incorporated 137Cs].
It was shown on isolated myocardium preparations that internal gamma irradiation of rats with 137Cs (1.6 MBq/kg) decreased the contractile function of ventricles and had no essential effect on the auricle contractility. The inotropic response of myocardium to activation of alpha-adrenergic receptors decreased, and of beta-adrenergic receptors decreased.


Induction and purification by three-phase partitioning of aryl alcohol oxidase (AAO) from Pleurotus ostreatus.
Aryl alcohol oxidase (AAO) is an extracellular flavoenzyme involved in lignin degradation by white rot fungi. Screening of lignolytic and AAO activity from twenty different fungal species were carried out. Among them, seven species showed lignolytic activity and three of them (Pleurotus ostreatus, Pleurotus eous, and Pleurotus platypus) were found to be AAO positive. Maximal AAO activity was observed in batch cultures of P. ostreatus and was found to be induced by aromatic amino acids and aryl alcohols up to a level of 289 U/l. Purification of AAO was carried out by three-phase partitioning (TPP). The 67 kDa enzyme was purified up to 10.19-fold by TPP with an overall recovery of 10.95%. Optimum pH and temperature for P. ostreatus AAO activity was found to be around 6 and 40 °C, respectively. From the LB plot, K (m) value of AAO for oxidizing veratryl alcohol was determined to be 0.6 mM. Results of the study indicate that P. ostreatus is the best producers of AAO, and they could be employed as promising fungal species for biotechnological applications.


Metabolic response to elevated CO2 levels in Pinus pinaster Aiton needles in an ontogenetic and genotypic-dependent way.
Global climate changes involve elevated atmospheric [CO2], fostering the carbon allocation to tree sink tissues, partitioning it into metabolic pathways. We use metabolomics analysis in adult and juvenile needles of four Pinus pinaster genotypes exposed to two levels of growth [CO2]: ambient (400 μmol mol-1) and enriched (800 μmol mol-1), to know if the metabolic responses are genotype-dependent and vary according to the stage of needle ontogeny. The eCO2-induced changes in the needle metabolomes are more significant in secondary metabolism pathways and especially meaningful in juvenile needles. The heteroblasty has important consequences in the expression of the metabolome, and on the plasticity to CO2, determining the level of specific metabolite accumulation, showing an interdependence between adult and juvenile needles. The P. pinaster needle metabolomes also show clear quantitative differences linked to genotype, as well as regarding the metabolic response to eCO2, showing both, common and genotype-specific biochemical responses. Thus, the changes in flavonol levels are mainly genotype-independent, while those in terpenoid and free fatty acids are mainly genotype-dependent, ratifying the importance of genotype to determine the metabolic response to eCO2. To understand the adaptation mechanisms that tree species can develop to cope with eCO2 it is necessary to know the genetically distinct responses within a species to recognize the CO2-induced changes from the divergent approaches, what can facilitate knowing also the possible interrelation of the physiological and metabolic responses. That could explain the controversial effects of eCO2 on the carbon-based metabolite in conifers, at the inter- and intra-specific level.


Incorporating a peer-teaching, virtual, educational activity to teach newly approved medications in a senior seminar capstone course.
Peer teaching and serious gaming are effective pedagogical approaches to actively engage learners, foster collaboration, and promote student accountability for self-directed learning. Literature describing how students learn about and are assessed on knowledge of newly approved medications is lacking. The objective of this project is to describe the design and implementation of a peer-taught virtual educational gaming activity, assess impact on knowledge of newly approved medications, and describe student perceptions of the activity. Pharmacy students in the fourth professional year developed and delivered a topic discussion focusing on newly approved medications to peers utilizing a virtual escape room gaming format. The activity was delivered via Zoom, Version 4.6.9 (Zoom Video Communications, Inc). A pre- and post-activity knowledge-based assessment and perception-based questionnaire were administered to participants. Observations regarding teamwork and communication were collected by facilitators. Sixteen students participated in this activity. Mean scores on the knowledge quiz were higher post-activity compared to pre-activity (8.1 vs. 6.3). The activity was perceived positively by participants with 100% of respondents agreeing or strongly agreeing that they "enjoyed the escape room theme for the activity and would recommend it to a friend." Zoom was perceived an effective platform to deliver this activity. All facilitators observed students working effectively as a team to complete the activity. Virtual gaming developed and facilitated by peer teachers may be an effective method for delivering educational content and positively influencing student learning.


Spotty calcification as a marker of accelerated progression of coronary atherosclerosis: insights from serial intravascular ultrasound.
The purpose of this study was to determine atheroma progression in patients with spotty calcification. Although extensively calcified atherosclerotic lesions have been proposed to be clinically quiescent, the presence of spotty calcification within plaque has been reported to be associated with an increased incidence of ischemic cardiovascular events. The relationship between spotty calcification and disease progression has not been investigated. A total of 1,347 stable patients with angiographic coronary artery disease underwent serial evaluation of atheroma burden with intravascular ultrasound imaging. Patients with spotty calcification were identified based on the presence of lesions (1 to 4 mm in length) containing an arc of calcification of <90°. Clinical characteristics and disease progression were compared between patients with spotty calcification (n = 922) and those with no calcification (n = 425). Patients with spotty calcification were older (age 56 years vs. 54 years; p = 0.001), more likely to be male (68% vs. 54%; p = 0.01), and have a history of diabetes mellitus (30% vs. 24%; p = 0.01) and myocardial infarction (28% vs. 20%; p = 0.004), and have lower on-treatment high-density lipoprotein cholesterol levels (48 ± 16 mg/dl vs. 51 ± 17 mg/dl; p = 0.001). Patients with spotty calcification demonstrated a greater percent atheroma volume (PAV) (36.0 ± 7.6% vs. 29.0 ± 8.5%; p < 0.001) and total atheroma volume (174.6 ± 71.9 mm(3) vs. 133.9 ± 64.9 mm(3); p < 0.001). On serial evaluation, spotty calcification was associated with greater progression of PAV (+0.43 ± 0.07% vs. +0.02 ± 0.11%; p = 0.002). Although intensive low-density lipoprotein cholesterol and blood pressure lowering therapy slowed disease progression, these efficacies were attenuated in patients with spotty calcification. The presence of spotty calcification is associated with more extensive and diffuse coronary atherosclerosis and accelerated disease progression despite use of medical therapies.


Prevalence of human papillomaviruses in lung carcinomas: a study of 218 cases.
High-risk human papillomaviruses (HPV) are largely implicated in the carcinogenesis of cervical carcinomas. Their role in bronchopulmonary carcinomas is still unclear. In the present study, we have explored 218 fresh frozen lung tumours for the presence of HPV with the Roche line blot assay and for the expression of mRNAs encoding E6 oncoprotein in HPV positive tumours. Only four samples were positive for HPV detection, one poorly differentiated squamous cell carcinoma and three large cell carcinomas. E6 mRNA was undetectable in these four samples. Our data confirm the low prevalence of HPV in lung carcinomas in Western European countries and do not plead in favour of a carcinogenic role for HPV in these carcinomas.


Effect of Hepatic Perfusion on Microwave Ablation Zones in an Ex Vivo Porcine Liver Model.
To compare the size of ablation zones derived from nonperfused ex vivo livers with ablation zones created using an ex vivo perfused porcine liver model. Six fresh porcine livers were used to evaluate microwave ablation (MWA). Perfused (n = 3) and nonperfused (n = 3) livers were warmed to 37°C by oxygenated, O-positive human blood reconstituted with Ringer solution, using an organ perfusion circuit. During MWA, perfusion was discontinued in the nonperfused group and maintained in the perfused group. After MWA (140 watts × 2 min at 2.45 GHz) with the Acculis MTA System (AngioDynamics, Latham, New York), ablation zones were bisected sagittally. Sections were stained with nicotinamide adenine dinucleotide (NADH) and hematoxylin-eosin to assess viability of cells in ablation and marginal zones. Comparison of 22 MWA zones (9 in perfused group, 13 in nonperfused group) was performed. Ablation zones demonstrated a central "white" and peripheral "red" zone. Cells in the white zone were nonviable with no NADH staining. The red zone showed progressive NADH staining toward the periphery, suggesting incomplete cell death. White and red zones of the perfused group were significantly smaller compared with the nonperfused group (short axis, 17.8 mm ± 2.7 vs 21.1 mm ± 3.2, P = .003; long axis, 40.69 mm ± 3.9 vs 39.63 mm ± 5.2, P = .44; intermediate zone,1.33 mm ± 0.04 vs 2.7 mm ± 0.14, P < .0001; mean ± SD). MWA algorithms provided by this manufacturer are based on nonperfused organ data, which overestimate ablation zone size. Data from perfused liver models may be required for more accurate dosimetry guidelines.


The needs and hopes for an AIDS vaccine.
This article reviews the many difficulties met in the development of an effective vaccine against human immunodeficiency virus (HIV), including the considerable antigenic variability of the virus, its intracellular mode of transmission, its mucosal port of entry, and the persistent nature of the infection. Progresses in the development of prototype vaccines in animal models are discussed together with the results of initial clinical trials in human volunteers.


Ubiquitin in the peroxisomal protein import pathway.
PEX5 is the shuttling receptor for newly synthesized peroxisomal matrix proteins. Alone, or with the help of an adaptor protein, this receptor binds peroxisomal matrix proteins in the cytosol and transports them to the peroxisomal membrane docking/translocation module (DTM). The interaction between cargo-loaded PEX5 and the DTM ultimately results in its insertion into the DTM with the concomitant translocation of the cargo protein across the organelle membrane. PEX5 is not consumed in this event; rather it is dislocated back into the cytosol so that it can promote additional rounds of protein transportation. Remarkably, the data collected in recent years indicate that dislocation is preceded by monoubiquitination of PEX5 at a conserved cysteine residue. This mandatory modification is not the only type of ubiquitination occurring at the DTM. Indeed, several findings suggest that defective receptors jamming the DTM are polyubiquitinated and targeted to the proteasome for degradation.


Bioefficacy and persistence of acephate in mungbean Vigna radiata (L.) Wilczek.
The bioefficacy, persistence and metabolism of foliar application of acephate 75 SP at the respective recommended and double doses of 1500 and 3000 g a.i. ha(-1) were studied on kharif mungbean crop. Acephate gave a very effective control of the pod borer, Helicoverpa armigera (Hubner) at the tested doses on mungbean crop throughout the cropping season, besides recording lower percent pod damage and higher grain yield. The recommended dose of acephate also recorded higher net returns and thus indicating its superiority. Persistence of acephate in mungbean leaves and soil was studied following applications of acephate @ 1500 and 3000 g a.i. ha(-1) at 30 days after sowing. Residues of acephate in mungbean leaves and soil were estimated using gas liquid chromatograph (GLC) and confirmed on gas chromatograph-mass spectrometry (GC-MS). Half-life periods for acephate in mungbean leaves were observed to be 2.98 and 3.27 days at single and double the application rates, respectively. Residues of acephate dissipated below its limit of quantification (LOQ) of 0.05 mg kg(-1) after 20 and 25 days at single and double the application dosage, respectively. Similarly, half-life periods for acephate in mungbean soil were observed to be 1.86 and 1.94 days at single and double the application rates, respectively. Residues of acephate dissipated below its LOQ of 0.05 mg kg(-1) after 10 and 15 days at single and double the application dosage, respectively.


Effects of in vitro growth culture duration and prematuration culture on maturational and developmental competences of bovine oocytes derived from early antral follicles.
Bovine ovaries offer a large pool of oocytes that could be used for in vitro production of embryos of genetically valuable animals. The effects of in vitro growth (IVG) culture duration (10, 12, and 14 days) on the viability and growth of bovine oocytes derived from early antral follicles (0.5-1 mm diameter) in this study. In addition, the effect of pre-IVM culture with phosphodiesterase inhibitor (3-isobutyl-1-methylxanthine) on nuclear maturation of IVG oocytes was also evaluated. In experiment 1, oocyte viability observed after 10 or 12 days of IVG culture was greater (P < 0.05) than that observed after 14 days of culture. Oocyte diameters and proportions of oocytes at metaphase II stage were greater (P < 0.05) when 12 or 14 days of IVG culture where used when compared with 10 days culture. In addition, the proportion of oocytes at metaphase II stage was greater (P < 0.05) when pre-IVM culture was performed for oocytes derived from 12 and 14 days of IVG culture. When 12 and 14 days of IVG culture followed by pre-IVM culture were compared in experiment 2, cumulus cell membrane integrity was greater (P < 0.05) after 12 days. Blastocyst production rate for oocytes obtained after 12 days of IVG culture (24.5%) was greater (P < 0.05) than for oocytes obtained after 14 days (9.9%). In conclusion, 12 days IVG followed by pre-IVM culture was considered the optimal processing system for bovine oocytes derived from early antral follicles when oocyte viability, diameter, maturation, and development competences were considered.


Raptor/mTORC1 loss in adipocytes causes progressive lipodystrophy and fatty liver disease.
Normal adipose tissue growth and function is critical to maintaining metabolic homeostasis and its excess (e.g. obesity) or absence (e.g. lipodystrophy) is associated with severe metabolic disease. The goal of this study was to understand the mechanisms maintaining healthy adipose tissue growth and function. Adipose tissue senses and responds to systemic changes in growth factor and nutrient availability; in cells mTORC1 regulates metabolism in response to growth factors and nutrients. Thus, mTORC1 is poised to be a critical intracellular regulator of adipocyte metabolism. Here, we investigate the role of mTORC1 in mature adipocytes by generating and characterizing mice in which the Adiponectin-Cre driver is used to delete floxed alleles of Raptor, which encodes an essential regulatory subunit of mTORC1. Raptor (Adipoq-cre) mice have normal white adipose tissue (WAT) mass for the first few weeks of life, but soon thereafter develop lipodystrophy associated with hepatomegaly, hepatic steatosis, and insulin intolerance. Raptor (Adipoq-cre) mice are also resistant to becoming obese when consuming a high fat diet (HFD). Resistance to obesity does not appear to be due to increased energy expenditure, but rather from failed adipose tissue expansion resulting in severe hepatomegaly associated with hyperphagia and defective dietary lipid absorption. Deleting Raptor in WAT also decreases C/EBPα expression and the expression of its downstream target adiponectin, providing one possible mechanism of mTORC1 function in WAT. mTORC1 activity in mature adipocytes is essential for maintaining normal adipose tissue growth and its selective loss in mature adipocytes leads to a progressive lipodystrophy disorder and systemic metabolic disease that shares many of the hallmarks of human congenital generalized lipodystrophy.


Low-frequency dielectric properties of lysozyme as a function of hydration and pH of lyophilisation.
Dielectric measurements have been made on lysozyme-compressed powders as a function of hydration and of the pH at which the samples were lyophilised. A dielectric dispersion previously described in the literature and known as the alpha-dispersion is found to be strongly influenced by the lyophilisation pH, and reaches a maximum magnitude at around pH 11. A dielectric loss of the form of the alpha-dispersion is observed in polylysine hydrobromide and the sodium salt of polyglutamic acid, but not for polyglycine. These results can be understood in terms of proton transfer between the ionisable side-groups, as well as by the controlling influence of counter-ions in moderating the energetics of the proton transfer processes. Another, weaker and hitherto unreported, dielectric loss process has been found for lysozyme, polyglycine, polylysine and poly(glutamic acid). This is referred to as the alpha 2-dispersion and its characteristics can be understood in terms of vibrational motions of the polypeptide backbones and the plasticising action of bound water.


The gait disorder in primary orthostatic tremor.
To uncover possible impairments of walking and dynamic postural stability in patients with primary orthostatic tremor (OT). Spatiotemporal gait characteristics were quantified in 18 patients with primary OT (mean age 70.5 ± 5.9 years, 10 females) and 18 age-matched healthy controls. One-third of patients reported disease-related fall events. Walking performance was assessed on a pressure-sensitive carpet under seven conditions: walking at preferred, slow, and maximal speed, with head reclination or eyes closed, and while performing a cognitive or motor dual-task paradigm. Patients exhibited a significant gait impairment characterized by a broadened base of support (p = 0.018) with increased spatiotemporal gait variability (p = 0.010). Walking speed was moderately reduced (p = 0.026) with shortened stride length (p = 0.001) and increased periods of double support (p = 0.001). Gait dysfunction became more pronounced during slow walking (p < 0.001); this was not present during fast walking. Walking with eyes closed aggravated gait disability as did walking during cognitive dual task (p < 0.001). OT is associated with a specific gait disorder with a staggering wide-based walking pattern indicative of a sensory and/or a cerebellar ataxic gait. The aggravation of gait instability during visual withdrawal and the normalization of walking with faster speeds further suggest a proprioceptive or vestibulo-cerebellar deficit as the primary source of gait disturbance in OT. In addition, the gait decline during cognitive dual task may imply cognitive processing deficits. In the end, OT is presumably a complex network disorder resulting in a specific spino-cerebello-frontocortical gait disorder that goes beyond mere tremor networks.


Protocol for a randomised pilot study of a novel Parent-Child Interaction Therapy (PCIT) 're-implementation' intervention.
Despite a number of clinicians having been trained in Parent-Child Interaction Therapy (PCIT) in Aotearoa/New Zealand, few are regularly delivering the treatment, with barriers to use including a lack of suitable equipment and lack of professional support. This pragmatic, parallel-arm, randomised, controlled pilot trial includes PCIT-trained clinicians who are not delivering, or only rarely utilising, this effective treatment. The study aims to assess the feasibility, acceptability and cultural responsivity of study methods and intervention components and to collect variance data on the proposed future primary outcome variable, in preparation for a future, larger trial. The trial will compare a novel 're-implementation' intervention with a refresher training and problem-solving control. Intervention components have been systematically developed to address barriers and facilitators to clinician use of PCIT using implementation theory, and a draft logic model with hypothesised mechanisms of action, derived from a series of preliminary studies. The intervention includes complimentary access to necessary equipment for PCIT implementation (audio-visual equipment, a 'pop-up' time-out space, toys), a mobile senior PCIT co-worker and an optional weekly PCIT consultation group, for a 6-month period. Outcomes will include the feasibility of recruitment and trial procedures; acceptability of the intervention package and data collection methods to clinicians; and clinician adoption of PCIT. Relatively little research attention has been directed at interventions to resurrect stalled implementation efforts. Results from this pragmatic pilot RCT will refine and shape knowledge relating to what it might take to embed the ongoing delivery of PCIT in community settings, providing more children and families with access to this effective treatment. ANZCTR, ACTRN12622001022752, registered on July 21, 2022.


Skin temperature measurement in individuals with spinal cord injury during and after exercise: Systematic review.
Athletes with spinal cord injury (SCI) have difficulties in maintaining thermal homeostasis during exercise due to their lower sweat capacity and skin vasodilation. Skin temperature (Tsk) assessment, as opposed to core temperature, has become more widely accepted due to its non-invasive nature. The aims of this systematic review was to collate research studies that measured Tsk of individuals with SCI during or after exercise, study their Tsk response, taking into account the method employed, the environmental and exercise conditions, and to identify the different cooling strategies and their effect during exercise. Pubmed, Web of Science and Scopus databases were searched to identify the articles published since year 2000. Two reviewers working independently extracted data and assessed the quality of the articles included. If they disagreed, a third reviewer was consulted. ROBINS-I scale was used to assess the quality of the articles, and the review has been conducted in agreement with PRISMA guidelines. Twenty studies were included in this review. 84% of them presented moderate, serious or critical risk of bias. The entire of the studies assessed Tsk during exercise, but only seven studies measured it during rest or after exercise. Eighteen studies used contact thermometry to assess Tsk and the two remaining studies employed non-contact techniques. Seven studies were conducted in warm conditions (>31.5°C) and the remaining studies in moderate conditions (10°C to 26.6°C). According to cooling strategies, ice vests and water spray are effective in reducing Tsk and decreasing the risk of heat stroke. The 90% of the studies applied contact thermometry and due to their effect in the data assessed, it is necessary more research into the SCI population using infrared thermography due to its differences in characteristics, methodology, and applications. The methodological differences among studies make difficult to perform a meta-analysis.


Concentration statistics for mixing-controlled reactive transport in random heterogeneous media.
Uncertainty in the distribution of hydraulic parameters leads to uncertainty in flow and reactive transport. Traditional stochastic analysis of solute transport in heterogeneous media has focused on the ensemble mean of conservative-tracer concentration. Studies in the past years have shown that the mean concentration often is associated with a high variance. Because the range of possible concentration values is bounded, a high variance implies high probability weights on the extreme values. In certain cases of mixing-controlled reactive transport, concentrations of conservative tracers, denoted mixing ratios, can be mapped to those of constituents that react with each other upon mixing. This facilitates mapping entire statistical distributions from mixing ratios to reactive-constituent concentrations. In perturbative approximations, only the mean and variance of the mixing-ratio distribution are used. We demonstrate that the second-order perturbative approximation leads to erroneous or even physically impossible estimates of mean reactive-constituent concentrations when the variance of the mixing ratio is high and the relationship between the mixing ratio and the reactive-constituent concentrations strongly deviates from a quadratic function. The latter might be the case in biokinetic reactions or in equilibrium reactions with small equilibrium constant in comparison to the range of reactive-constituent concentrations. When only the mean and variance of the mixing ratio is known, we recommend assuming a distribution that meets the known bounds of the mixing ratio, such as the beta distribution, and mapping the assumed distribution of the mixing ratio to the distributions of the reactive constituents.


Successful Treatment of Persistent Hiccups in an Advanced Palliative Cancer Patient With Gabapentin: A Case Report.
Hiccups may appear to be a common normal phenomenon that does not warrant treatment in the general population. However, severe and persistent hiccups can become annoying and distressing and may decrease the quality of life, especially in cancer patients. The management of hiccups remains challenging. Despite trying many pharmacological and non-pharmacological methods, there is no clear evidence to support the management guidelines. In our case, we successfully treated persistent hiccups of more than four days duration in a patient with acute myeloblastic leukemia with gabapentin.


Teaching NeuroImages: Comatose patient with bilateral thalamic infarct due to internal carotid artery occlusion.
An 88-year-old woman with a medical history of diabetes, hypertension, and atrial fibrillation presented to the emergency room after being found unresponsive with a NIH Stroke Scale score of 23 and Glasgow Coma Scale score of 3. She was unresponsive to painful stimuli. Noncontrast CT demonstrated bilateral thalamic infarcts (figure 1A).


Case report of paroxysmal atrioventricular block and ventricular arrest in a young pregnant woman: What is the mechanism?
A 30-year-old female patient with 12 weeks of gestation was admitted to our hospital due to dizziness and amaurosis fugax. Moreover, 24 h Holter monitoring showed paroxysmal atrioventricular block (P-AVB) and ventricular arrest. The heart block in the patient was likely a vagally mediated heart block based on the "vagal score." She was not given a pacemaker, and the symptoms and AV nodal conduction were improved following the delivery.


Appropriate treatment of malaria? Use of antimalarial drugs for children's fevers in district medical units, drug shops and homes in eastern Uganda.
To evaluate the quality of pharmaceutical care of malaria for children in eastern Uganda prescribed at government health units and drug shops, and administered by caretakers at home; and to assess its appropriateness in relation to national treatment guidelines, which recommend chloroquine over 3 days. We followed 463 children under 5 years whose caretakers attended two drug shops and two government health units to seek treatment for fever. The children were examined and the caretakers interviewed on the day of enrollment in the study (day 0), and in their homes on days 3 and 7. Data was collected on drug use prior to attending the shop or health unit, the treatment provided at these study sites, and the administration of drugs at home over the following 3 days. Before attending the study sites, 72% of children had already been given some biomedical drugs, and 40% had received the recommended drug, chloroquine. Health workers prescribed chloroquine for 94% of the children, but only 34% of the recommended doses followed guidelines. Two-thirds of the children were prescribed an injection of chloroquine. By day 3, according to caretaker reports, about 38% of the children had received chloroquine in compliance with the instructions given by the health workers and drug shop attendants. Only 28% of the children had received chloroquine at the optimal dose of 20-30 mg/kg recommended by national policy. The methods were useful for examining adherence of both caretakers and health care providers to national guidelines and the extent to which caretakers were compliant with providers' prescriptions. Chloroquine and antipyretics were the drugs of choice for fever in these areas of rural eastern Uganda. But children did not receive the recommended dosage of chloroquine because of lack of compliance on the parts of providers as well as users of health care.


Adjuvant! Online is overoptimistic in predicting survival of Asian breast cancer patients.
Adjuvant! Online is a free web-based tool which predicts 10-year breast cancer outcomes and the efficacy of adjuvant therapy in patients with breast cancer. As its prognostic performance has only been validated in high income Caucasian populations, we validated the model in a middle income Asian setting. Within the University Malaya Hospital-Based Breast Cancer Registry, all 631 women who were surgically treated for invasive non-metastatic breast cancer between 1993 and 2000 were identified. The discriminative performance of Adjuvant! Online was tested using receiver operating characteristic (ROC) analysis. Calibration of the model was evaluated by comparing predicted 10-year overall survival with observed 10-year survival. Adjuvant! Online was fairly capable in discriminating between good and poor survivors, as attested by the area under ROC curve of 0.73 (95% Confidence Interval: 0.69-0.77). Overall, Adjuvant! Online predicted 10 year survival (70.3%) was significantly higher than the observed 10 year survival (63.6%, difference of 6.7%; 95% CI: 3.0-10.4%). The model was especially overoptimistic in women under 40 years and in women of Malay ethnicity, where survival was overestimated by approximately 20% (95% CI: 9.8-29.8%) and 15% (95% CI: 5.3-24.5%) respectively. Even though Adjuvant! Online is capable of discriminating between good and poor survivors, it systematically overestimates survival. These findings suggest that the model requires adaptation prior to use in Asian settings.


Efficient Decellularization of the Full-Thickness Rat-Derived Abdominal Wall to Produce Acellular Biologic Scaffolds for Tissue Reconstruction: Promising Evidence Acquired from In Vitro Results.
Functional restoration of abdominal wall defects represents one of the fundamental challenges of reconstructive surgery. Synthetic grafts or crosslinked animal-derived biological grafts are characterized by significant adverse reactions, which are mostly observed after their implantation. The aim of this study was to evaluate the efficacy of the decellularization protocol to produce a completely acellular full-thickness abdominal wall scaffold. Full-thickness abdominal wall samples were harvested from Wistar rats and submitted to a three-cycle decellularization process. Histological, biochemical, and DNA quantification analyses were applied to evaluate the effect of the decellularization protocol. Mechanical testing and immunogenicity assessment were also performed. Histological, biochemical, and DNA analysis results showed efficient decellularization of the abdominal wall samples after the third cycle. Decellularized abdominal wall scaffolds were characterized by good biochemical and mechanical properties. The data presented herein confirm the effective production of a rat-derived full-thickness abdominal wall scaffold. Expanding this approach will allow the exploitation of the capacity of the proposed decellularization protocol in producing acellular abdominal wall scaffolds from larger animal models or human cadaveric donors. In this way, the utility of biological scaffolds with preserved in vivo remodeling properties may be one step closer to its application in clinical studies.


Inosine Induces Stemness Features in CAR T cells and Enhances Potency.
Adenosine (Ado) mediates immune suppression in the tumor microenvironment and exhausted CD8+ CAR T cells mediate Ado-induced immunosuppression through CD39/73-dependent Ado production. Knockout of CD39, CD73 or A2aR had modest effects on exhausted CAR T cells, whereas overexpression of Ado deaminase (ADA), which metabolizes Ado to inosine (INO), induced stemness features and potently enhanced functionality. Similarly, and to a greater extent, exposure of CAR T cells to INO augmented CAR T cell function and induced hallmark features of T cell stemness. INO induced a profound metabolic reprogramming, diminishing glycolysis and increasing oxidative phosphorylation, glutaminolysis and polyamine synthesis, and modulated the epigenome toward greater stemness. Clinical scale manufacturing using INO generated enhanced potency CAR T cell products meeting criteria for clinical dosing. These data identify INO as a potent modulator of T cell metabolism and epigenetic stemness programming and deliver a new enhanced potency platform for immune cell manufacturing.


Associations of Antibodies Targeting Periodontal Pathogens With Subclinical Coronary, Carotid, and Peripheral Arterial Atherosclerosis in Rheumatoid Arthritis.
Both periodontal disease and cardiovascular disease (CVD) are overrepresented in rheumatoid arthritis (RA). This study was undertaken to investigate the contribution of periodontal pathogens to CVD in RA. RA patients underwent assessments of coronary artery calcification (CAC), carotid intima-media thickness and plaque, and ankle-brachial index via computed tomography, ultrasound, and Doppler ultrasound, respectively. Sera were assayed for antibodies targeting Porphyromonas gingivalis (Pg), Aggregatibacter actinomycetemcomitans serotype B (Aa), and Aa-derived leukotoxin A (LtxA). Associations of antibodies against these periodontal pathogens with measures of atherosclerosis were explored using generalized linear models. Among 197 RA patients, anti-Pg was detected in 72 patients (37%), anti-Aa in 41 patients (21%), and anti-LtxA in 84 patients (43%). Adjusting for relevant confounders and reported tooth loss, the mean CAC score was 90% higher in those with anti-Aa and/or anti-LtxA compared with those without either antibody (19 units versus 10 units; P = 0.033). The adjusted odds of CAC ≥100 units were 2.23-fold higher in those with anti-Aa and/or anti-LtxA compared with those without either antibody (P = 0.040). Anti-Aa and/or anti-LtxA seropositivity was significantly associated with all other assessed measures of atherosclerosis except carotid plaque. Anti-Pg was not associated with any measure of atherosclerosis. Higher swollen joint count was associated with CAC exclusively in the group with anti-Aa and/or anti-LtxA. Immunoreactivity against Aa and/or its major virulence factor LtxA was associated with atherosclerosis in multiple vascular beds of RA patients and amplified the effect of swollen joints on coronary atherosclerosis, suggesting a role for treatment/prevention of periodontal disease in the prevention of CVD in RA.


The natural course of bridging osteophyte formation on MRI-A pictorial illustration.
Diffuse skeletal hyperostosis is a common spinal disorder, but its pathophysiology is mostly unclear. The disorder can lead to a variety of symptoms, but many patients remain relatively asymptomatic. We present a case demonstrating the development of bridging osteophytes on a series of magnetic resonance images. An elderly person's spine was scanned repeatedly due to non-specific back pain during the last 4 years and the consecutive images revealed the formation of a bony bridge in the lumbar spine. Extensive bone marrow edema was seen during the formation of the osteophyte, suggestive of an ongoing inflammatory process. This case underlines that the inflammatory reaction in diffuse skeletal hyperostosis can be intense and prolonged, and its role might be worth studying further.


Assessment of the reproducibility and repeatability of a method of grading macular subretinal fluid using optical coherence tomography.
To devise a quantative method for the measurement of the extent of macular subretinal fluid using optical coherence tomography (OCT), and to assess the interobserver and intraobserver agreement for this grading system. Observational cohort series. Patients were a cohort who underwent retinal detachment surgery over an 18-month period. All patients had OCT scan at 6 weeks after surgery. The scans were graded by two independent observers experienced in OCT interpretation and each grader was masked to the others findings. Observer 1 then regraded the scans on a later day masked to his previous findings. The interobserver and intraobserver agreement was assessed using weighted Kappa (Kw) statistics. In all, 116 patients were analysed. Both the intraobserver and interobserver agreement was very high, with Kw being 0.9631 and 0.9070, respectively. The grading system for assessment of the extent of macular subretinal fluid using OCT appears to have very good reproducibility and repeatability. We propose that this grading system would be clinically useful when applied to pathologies visible on OCT scan of the macula.


Evaluation of abduction range of motion and avoidance of inferior scapular impingement in a reverse shoulder model.
The purpose of this study was to determine the effects of prosthetic design and surgical technique of reverse shoulder implants on total abduction range of motion and impingement on the inferior scapular neck. Custom implants in three glenosphere diameters (30, 36, and 42 mm), with 3 different centers of rotation offsets (0, +5, and +10 mm), were placed into a Sawbones scapula (Pacific Research Laboratories, Vashon, WA) in 3 different positions: superior, center, and inferior glenoid. Humeral sockets were manufactured with a 130 degrees , 150 degrees , and 170 degrees neck-shaft angle. Four independent factors (glenosphere diameter, center of rotation offset, glenosphere position on the glenoid, and humeral neck-shaft angle) were compared with the 2 dependent factors of range of motion and inferior scapular impingement. Center of rotation offset had the largest effect on range of motion, followed by glenosphere position. Neck-shaft angle had the largest effect on inferior scapular impingement, followed by glenosphere position. This information may be useful to the surgeon when deciding on the appropriate reverse implant.


G-actin diffusion is insufficient to achieve F-actin assembly in fast-treadmilling protrusions.
To generate forces that drive migration of a eukaryotic cell, arrays of actin filaments (F-actin) are assembled at the cell's leading membrane edge. To maintain cell propulsion and respond to dynamic external cues, actin filaments must be disassembled to regenerate the actin monomers (G-actin), and transport of G-actin from sites of disassembly back to the leading edge completes the treadmilling cycle and limits the flux of F-actin assembly. Whether or not molecular diffusion is sufficient for G-actin transport has been a long-standing topic of debate, in part because the dynamic nature of cell motility and migration hinders the estimation of transport parameters. In this work, we applied an experimental system in which cells adopt an approximately constant and symmetrical shape; they cannot migrate but exhibit fast, steady treadmilling in the thin region protruding from the cell. Using fluorescence recovery after photobleaching, we quantified the relative concentrations and corresponding fluxes of F- and G-actin in this system. In conjunction with mathematical modeling, constrained by measured features of each region of interest, this approach revealed that diffusion alone cannot account for the transport of G-actin to the leading edge. Although G-actin diffusion and vectorial transport might vary with position in the protruding region, good agreement with the fluorescence recovery after photobleaching measurements was achieved by a model with constant G-actin diffusivity ∼2 μm2/s and anterograde G-actin velocity less than 1 μm/s.


Use of the radial maze in studies of phencyclidine and other drugs of abuse.
Effects of drugs known to disrupt performance in an 8-arm radial maze are reported in terms of changes caused in the pattern of arm entry. Phencyclidine (PCP) and N-allyl-N-normetazocine (SKF-10,047) alter the pattern of arm entry in a way which distinguishes their actions from those of scopolamine and certain serotonergic agonists. The apparent rank order of potencies for causing this effect is (+)SKF-10,047 greater than PCP greater than (-)SKF-10,047. Results of previous radial maze studies evaluating the interactions of clonidine and verapamil with PCP are summarized. Data are reported which indicate that the ability of verapamil to potentiate PCP's behavioral effects stems from an alteration of the pharmacokinetics of PCP; when verapamil (20 mg/kg, IP) was administered 15 minutes before [3H]PCP (40 microCi/kg, IP), brain levels of tritium were increased by 154 to 225 percent. Finally, possible advantages of using a 4-arm radial maze in studies of PCP and related drugs are discussed.