Substitution of tyrosine residues at the aromatic cluster around the betaA-betaB loop of rubisco small subunit affects the structural stability of the enzyme and the in vivo degradation under stress conditions.
Ribulose 1,5-bisphosphate carboxylase/oxygenase (rubisco) plays a central metabolic role in photosynthetic eukaryotes, and its catabolism is a crucial process for the nutrient economy of higher plants. The rubisco holoenzyme is assembled from eight chloroplast-encoded large subunits and eight nuclear-encoded small subunits. We have identified a cluster of conserved tyrosines at the interface between subunits (comprising Y67, Y68, and Y72 from the betaA-betaB loop of the small subunit and Y226 from the large subunit) that may contribute to holoenzyme stability. To investigate the role of these tyrosines in rubisco structure and in vivo degradation, we have examined site-directed mutants of these residues (Y67A, Y68A, Y72A, and Y226L) in Chlamydomonas reinhardtii. Even if all mutant strains were able to grow photoautotrophically, they exhibited a reduction in rubisco activity and/or the level of expression, especially the Y67A and Y72A mutants. Besides, all mutant rubiscos were inactivated at a lower temperature than the wild type. The kinetics of proteolysis of the mutant enzymes with subtilisin revealed structural alterations, leading to facilitated disassembly (in the cases of Y67A and Y72A) or aggregation propensity (for Y68A and Y226L). When subjected to oxidative stress in vivo through exposure of liquid cultures to hydrogen peroxide, all mutant strains degraded rubisco at a faster rate than the wild type. These results demonstrate that the tyrosine cluster around the betaA-betaB loop of rubisco small subunit plays a stabilizing role by affecting the catalytic activity and the degradation rate of the enzyme in stressed cells.


Expression of human inducible nitric oxide synthase in Escherichia coli.
We have expressed active full-length human inducible nitric oxide synthase (iNOS) in E. coli. Expression required co-expression with calmodulin, a particularly tight-binding cofactor. The extracts also required tetrahydrobiopterin to display activity. Specific activity of the purified recombinant iNOS was similar to iNOS purified from murine macrophages. This result indicates that no special processing events unique to eucaryotic cells are necessary for iNOS activity.


Laboratory and clinical acute effects of active and passive indoor group water-pipe (narghile) smoking.
Indoor group water-pipe tobacco smoking, commonly referred to as water-pipe smoking (WPS), especially in coffee shops, has gained worldwide popularity. We performed a comprehensive laboratory and clinical evaluation of the acute effects of active and passive indoor group WPS. This comparative study evaluated pre- and post-30-min active and passive indoor group WPS. The outcome parameters were carboxyhemoglobin (COHb), nicotine, and cotinine levels; CBC count; and cardiorespiratory parameters. Exhaled breath condensate (EBC) cytokines and endothelial function (using the EndoPat device [Itamar Medical Ltd]) were measured only in active smokers. Statistical methods used were Student t test, Wilcoxon signed rank test, Fisher exact test, analysis of variance, and Newman-Keuls post hoc test where relevant. Sixty-two volunteers aged 24.9±6.2 years were included; 47 were active smokers, and 15 were passive smokers. COHb level increased postactive WPS (active smokers, 2.0%±2.9% vs 17.6%±8.8%; P<.00001); six subjects (12.7%) had a >25% increase, and two subjects (4.2%) had a >40% increase. Plasma nicotine level increased postactive WPS (active smokers, 1.2±4.3 ng/mL vs 18.8±13.9 ng/mL; P<.0001); plasma cotinine and urinary nicotine and cotinine levels also increased significantly. EBC IL-4, IL-5, IL-10, IL-17, and γ-interferon decreased significantly with postactive smoking; endothelial function did not change. WPS was associated with adverse cardiorespiratory changes. In passive smokers, COHb level increased (0.8%±0.25% vs 1.2%±0.8%, respectively, P=.003) as did respiratory rate. One session of active indoor group WPS resulted in significant increases in COHb and serum nicotine levels (eightfold and 18-fold, respectively) and was associated with adverse cardiorespiratory health effects. The minor effects found in passive smokers suggest that they too may be affected adversely by exposure to WPS. The results call for action to limit the continuing global spread of WPS in coffee shops. ClinicalTrials.gov; No.: NCT1237548; URL: www.clinicaltrials.gov.


Patchy haemopoiesis in long-term remission of idiopathic aplastic anaemia.
13 patients with idiopathic aplastic anaemia in remission for more than 2 years were examined to define the haemopoietic status by means of bone marrow scintigraphy, ferrokinetics and bone marrow culture for haemopoietic progenitor cells. Haemoglobin levels reached the normal range in all these patients although mild neutropenia and thrombocytopenia were still observed in 5 patients. Bone marrow scintigrams using indium-111 showed normal distribution in 2, diffuse low accumulation in 3, patchy distribution in 7, and expanded distribution with patchy uptake in 1 patient. The defective haemopoiesis was also confirmed by ferrokinetic and bone marrow culture studies. The patchy haemopoiesis appears to characterize the residual marrow damage in remission of idiopathic aplastic anaemia.


Sol-gel co-assembly of hollow cylindrical inverse opals and inverse opal columns.
A facile approach of fabricating hollow cylindrical inverse opals and inverse opal columns by sol-gel co-assembly method was proposed. Polystyrene (PS) colloidal suspension added with hydrolyzed silicate precursor solution was used to self-assemble composite colloidal crystals which consist of PS colloidal crystal template and infiltrated silica gel in the interstitial of microspheres. Continuous hollow cylindrical composite colloidal crystal films have been produced on capillaries' outside and internal surface. Composite colloidal crystal columns which filling up the interior of a capillary were fabricated by pressure assisted sol-gel co-assembly method. Hollow cylindrical inverse opals and inverse opal columns were obtained after removing PS colloidal crystal from the composite colloidal crystal. Optical properties of the silica hollow cylindrical inverse opals were characterized by transmission spectrum and a stop band was observed. Structure and optical properties of the inverse opal columns were investigated.


Fibrinolytic treatment of intraventricular haemorrhage preceding surgical repair of ruptured aneurysms and arteriovenous malformations.
Previous studies have indicated that intraventricular administration of tissue-type plasminogen activator (TPA) might improve the prognosis of patients with intraventricular haemorrhage (IVH). In aneurysmal IVH, fibrinolytic treatment was always preceded by surgical repair of the aneurysm, since the risk of recurrent haemorrhage from a non-occluded aneurysm was estimated to be high. We reviewed a series of patients with IVH secondary to ruptured aneurysms (n = 4) or arteriovenous malformation (AVM; n = 1) who underwent emergency intraventricular administration of TPA before repair of the bleeding source. Fibrinolysis resulted in rapid decrease of haematoma volume and of ventricular dilatation, and prevented ventricular catheters from becoming obstructed. No intracranial haemorrhages or other complications occurred. The results suggest that the presence of recently ruptured aneurysms or AVM is not necessarily a contraindication for intraventricular administration of TPA. The potentially life saving benefits might outweigh the inherent risks of recurrent haemorrhage in carefully selected patients with massive IVH, in whom ventricular distension, periventricular brain compression, obstruction of CSF flow, and elevated ICP appear to be major determinants for the outcome.


Variation in patient charges for vaccines and well-child care.
Several state and federal programs have attempted to boost immunization rates by reducing or eliminating provider vaccine costs. The relation between patient vaccine and well-child visit charges and vaccine financing systems is unknown. To determine patient charges for vaccines and well-child visits in three states with varying vaccine financing systems and to examine the effects of a short-term reduction in provider vaccine costs. Cross-sectional survey study of a random sample of physicians in three states. A total of 2797 pediatricians and family physicians in North Carolina, Texas, and Massachusetts were surveyed. Current charges to patients for diphtheria-tetanus-pertussis vaccine (DTP), measles-mumps-rubella vaccine, Haemophilus influenzae type b vaccine (Hib), and combined DPT-Hib vaccine for well-child visits; changes in charges over the previous 8 months. Response rate was 62%. Vaccine and well-child visit charges were comparable in North Carolina and Texas. Massachusetts' average charges for well-child visits were higher than in the other states, although vaccine charges were lower; with the use of combined DPT-Hib vaccine, total simulated charges for vaccines and well-child care during the first 6 months of life averaged only 10% less in Massachusetts vs Texas and North Carolina. Neither regional variation in cost of living nor Medicaid reimbursement rates explained this difference. The average cost and composition of charges for well-child care in Massachusetts, a state with universal purchase of vaccines, compared with the other states, warrant further study to explore whether physicians shift costs to other preventive services to compensate for lower allowable immunization charges. If such cost shifting occurs, current federal immunization initiatives that lower or eliminate provider cost may not provide increased access to preventive services.


[Administration of acyclovir (virustatic) to cats].
Herpes felis infections belong to the feline viral rhinotracheitis complex. Acyclovir, a virustatic of the nucleoside-analogue-group, inhibits the in vitro replication of herpesvirus felis. Acyclovir (0.1% in isotonic saline solution) is well tolerated by cats when applied t.i.d. subcutaneously and has a high and long lasting plasma level. The treatment is not too expensive.


Automated large-core needle biopsy of surgically removed breast lesions: comparison of samples obtained with 14-, 16-, and 18-gauge needles.
To determine the needle size that would consistently enable the pathologist to correctly diagnose tissue core biopsy specimens. Fifty-seven surgically removed mass lesions of the breast were sampled with a short-throw automated biopsy gun and an 18-gauge needle first, followed by a 16- and a 14-gauge needle. Samples were evaluated independently by three pathologists, and findings were compared with the final diagnosis made at surgical excision biopsy. The sensitivity for the diagnosis of all 26 malignant lesions was 100%, 92%, and 65% for biopsy samples obtained with 14-, 16-, and 18-gauge needles, respectively. Specificity for malignancy was 100% for all three needle sizes. All of the benign diseases were correctly diagnosed in samples obtained with all three size needles except for two samples that did not survive processing. Biopsy samples obtained with a 14-gauge needle provide the most accurate diagnosis, which correlates with the diagnosis made with the surgical excision biopsy technique.


Expression of adhesion molecules and extracellular matrix proteins in glioblastomas: relation to angiogenesis and spread.
We studied the immunohistochemical expression of inducible adhesion molecules, integrins and extracellular matrix proteins in 10 cases of glioblastoma multiforme in order to investigate their angiogenesis, local invasiveness, poor metastasizing properties and their lack of tumour infiltrating leukocytes. In glioblastomas endothelial proliferations represent the majority of vascular structures; they were positive for endothelial markers (vWF, CD31, VE-cadherin) and negative for macrophage markers (CD68, PAM-1). Immunohistologically, they were subtyped into: 1 solid-glomeruloid ICAM-1, alpha 2 beta 1, alpha 3 beta 1, alpha 5 beta 1 negative; 2 channelled-branching ICAM-1 negative and alpha 2 beta 1, alpha 3 beta 1, alpha 5 beta 1 positive; 3 channelled-telangiectatic ICAM-1, alpha 2 beta 1, alpha 3 beta 1, alpha 5 beta 1 positive. In channelled proliferations, the expression and distribution of tenascin and merosin in the basal membrane was similar to that of normal brain vessels. The expression of all these molecules might indicate different steps of maturation of endothelial proliferations. The majority of endothelial proliferations may be immunohistologically considered as incomplete vascular structures; this might account for the low metastasizing tendency and low recruitment of leukocytes by these tumours. Neoplastic astrocytes were GFAP-1, ICAM-1, VCAM-1, alpha 2 beta 1, alpha 3 beta 1 and alpha 5 beta 1 immunoreactive and alpha 6 beta 4 negative: this allows them to interact with extracellular matrix proteins and might, in part, explain the tendency of glioblastomas to infiltrate locally.


Current Strategies for the Gene Therapy of Autosomal Recessive Congenital Ichthyosis and Other Types of Inherited Ichthyosis.
Mutations in genes such as transglutaminase-1 (TGM1), which are responsible for the formation and normal functioning of a lipid barrier, lead to the development of autosomal recessive congenital ichthyosis (ARCI). ARCIs are characterized by varying degrees of hyperkeratosis and the presence of scales on the body surface since birth. The quality of life of patients is often significantly affected, and in order to alleviate the manifestations of the disease, symptomatic therapy with moisturizers, keratolytics, retinoids and other cosmetic substances is often used to improve the condition of the patients' skin. Graft transplantation is commonly used to correct defects of the eye. However, these approaches offer symptomatic treatment that does not restore the lost protein function or provide a long-term skin barrier. Gene and cell therapies are evolving as promising therapy for ARCIs that can correct the functional activity of altered proteins. However, these approaches are still at an early stage of development. This review discusses current studies of gene and cell therapy approaches for various types of ichthyosis and their further prospects for patient treatment.


Study of Profile of Mucormycosis During the Second Wave of COVID-19 in a Tertiary Care Hospital.
Introduction and aim Mucormycosis is a lethal opportunistic infection caused by filamentous fungi of the family Mucoraceae (black fungus). There has been a sudden increase in the incidence of these cases during the second wave of the COVID-19 pandemic due to the immunocompromised state caused by the disease and its treatment. Early diagnosis and appropriate medical management are essential to reduce disease morbidity and mortality. Through this study, we aim to study the clinical features, risk factors, laboratory investigations, and radiological findings of patients with mucormycosis as well as evaluate the clinical outcomes in each case. Methods and materials This was a prospective study that included only confirmed mucormycosis cases admitted in Tata Main Hospital (TMH) from April 2021 to July 2021. A case of mucormycosis was defined as the one in which clinical and radiological features were consistent with mucormycosis and fungus was demonstrated in the tissue by potassium hydroxide (KOH) mount/culture/histopathological examination (HPE). Data relating to epidemiology, risk factors, clinico-radiological features, and outcomes were analyzed and expressed as a percentage of total cases. Results Of the total 15 cases, three patients (33.3%) had active COVID-19 infection, eight (53.3%) were in the post-COVID-19 state, two (13.4%) had COVID-19 like illness and two (13.4%) patients did not have COVID-19 in the recent past. There was male predominance with the male to female ratio being 2.75:1. The commonest associated co-morbid condition was diabetes mellitus (13 patients, 86.7%). Amongst the myriad manifestations, periorbital swelling was the commonest symptom (11 patients, 73.3%). Among neurological manifestations, involvement of cranial nerves was found in nine (60%) patients with the third cranial nerve being the most commonly affected nerve (eight patients, 53.3%). Cavernous sinus thrombosis (CST) was found in one (6.7%) patient. Diagnostic nasal endoscopy (DNE) revealed eschar at various sites in 13 patients (86.7%). Central retinal artery occlusion (CRAO) was found bilaterally in one patient (6.7%) while two patients (13.3%) had CRAO on the left. Radiologically, the most commonly involved sinuses were maxillary and ethmoidal (eight patients, 53.3%). Bilateral sinus involvement was more common (46.7%) than unilateral sinus involvement. The average length of stay (LOS) was 17.5±7.8 days. The overall mortality was 40%. Five (33.3%) patients developed secondary bacterial infections. All patients received medical therapy with intravenous amphotericin B. In addition, seven (46.7%) patients underwent functional endoscopic sinus surgery (FESS) with debridement of which, five (71.4%) patients survived and made a good recovery. One patient (6.7%) with pulmonary mucormycosis underwent lobectomy. Conclusion New-onset headache, black nasal discharge, periorbital swelling, retro-orbital pain, visual diminution, restriction of eye movements should prompt an immediate search for mucormycosis especially in the background of history of diabetes mellitus in patient with recent or current COVID-19 disease. Radio-imaging with computerized tomography and magnetic resonance imaging are complementary to clinical evaluation in assessing the disease extent and diagnosis of complications. Prompt diagnosis is essential due to the angio-invasive nature of the mucor and requires aggressive anti-fungal therapy and debridement of the devitalized tissue.


Truncated TrkB receptor-induced outgrowth of dendritic filopodia involves the p75 neurotrophin receptor.
The Trk family of receptor tyrosine kinases and the p75 receptor (p75NTR) mediate the effects of neurotrophins on neuronal survival, differentiation and synaptic plasticity. The neurotrophin BDNF and its cognate receptor tyrosine kinase, TrkB.FL, are highly expressed in neurons of the central nervous system. At later stages in postnatal development the truncated TrkB splice variants (TrkB.T1, TrkB.T2) become abundant. However, the signalling and function of these truncated receptors remained largely elusive. We show that overexpression of TrkB.T1 in hippocampal neurons induces the formation of dendritic filopodia, which are known precursors of synaptic spines. The induction of filopodia by TrkB.T1 occurs independently of neurotrophin binding and of kinase activity of endogenous TrkB.FL. Coexpression of a p75NTR lacking an intracellular domain inhibits the TrkB.T1-induced effect in a dominant negative manner. Steric hindrance of extracellular p75NTR interactions with a specific antibody, or absence of p75NTR with an intact extracellular domain also inhibit this TrkB.T1-induced effect. We thus propose a novel signalling pathway initiated by neurotrophin-independent extracellular or intramembrane interaction of TrkB.T1 with the p75NTR receptor, which modulates dendritic growth via p75NTR signalling cascades.


[Principles and clinical applications of diffusion weighted echo planar MR imaging].
The ultrafast capability of echo planar imaging(EPI) made diffusion-weighted MRI (DWI) measurements practical, without the need of head fixation and without the need to exclude patients unable to hold still. The DW hyperintensity and reduced apparent diffusion coefficient(ADC) were observed in all hyperacute strokes, initially at 2.5 hours. In contrast to decreased ADC within 10 days and DW hyperintensity within 30 days, ADC was increased without DW hyperintensity in chronic stage. This suggests that DWI can discriminate between acute and chronic strokes. Furthermore, DWEPI and T2-weighted EPI were also be useful to detect and distinguish acute hemorrhagic stroke from nonhemorrhagic stroke. This review highlighted the promising technique of DWEPI to examine stroke patients in routine clinical practice.


Thrombin generation in vitro and in vivo, and disturbed tissue factor regulation in patients with acute pancreatitis.
Being a central link between inflammation and coagulation, tissue factor (TF) and its inhibitor (TFPI) might be associated with the severity of acute pancreatitis (AP) and the development of organ failure (OF). The study comprises 9 severe AP patients with OF and 24 reference patients (11 mild AP and 13 severe AP without OF). Plasma samples were collected on admission. TF-induced thrombin generation in plasma samples was studied using the thrombogram method. In vivo thrombin generation was estimated by prothrombin fragment F1+2. Free and total TFPI levels were measured. To evaluate coagulation status the activated partial thromboplastin time, prothrombin time, platelet count, D-dimer, fibrinogen, antithrombin (AT) 3 and protein C (PC) were determined. There was no significant difference in F1+2 levels between the patient groups. Patients with severe AP tended to show low platelet counts, PC and AT3 levels, and high D-dimer levels. In 11 patients the standard TF stimulation did not trigger thrombin generation in the thrombogram. All deaths occurred in these patients. Free TFPI levels and free/total TFPI ratios were significantly higher in these patients and in non-survivors. Failure of TF-initiated thrombin generation in the thrombogram assay explained by high levels of circulating free TFPI may be associated with OF and mortality in AP. and IAP.


The added value of apparent diffusion coefficient to cerebral blood volume in the preoperative grading of diffuse gliomas.
In cerebral gliomas, rCBV correlates with tumor grade and histologic findings of vascular proliferation. Moreover, ADC assesses water diffusivity and is inversely correlated with tumor grade. In the present work, we have studied whether combined rCBV and ADC values improve the diagnostic accuracy of MR imaging in the preoperative grading of gliomas. One hundred sixty-two patients with histopathologically confirmed diffuse gliomas underwent DWI and DSC. Mean rCBV and ADC values were compared among the tumor groups with the Student t test or ANOVA. ROC analysis was used to determine rCBV and ADC threshold values for glioma grading. rCBV had significantly different values between grade II and IV gliomas and between grade III and IV tumors, but there were no significant differences between grade II and III gliomas (P > .05). Grade II and III tumors also did not differ when astrocytomas, oligodendrogliomas, and oligoastrocytomas were considered separately. ADC values were significantly different for all 3 grades. The ADC threshold value of 1.185 × 10(-3) mm(2)/s and the rCBV cutoff value of 1.74 could be used with high sensitivity in the characterization of high-grade gliomas. The area under the ROC curve for the maximum rCBV and minimum ADC was 0.72 and 0.75, respectively. The combination of rCBV and ADC values increased the area under the ROC curve to 0.83. ADC measurements are better than rCBV values for distinguishing the grades of gliomas. The combination of minimum ADC and maximum rCBV improves the diagnostic accuracy of glioma grading.


Heavy metals induce expression of the TPA-inducible sequence (TIS) genes.
We previously cloned a set of primary response genes, which we call TIS (TPA-Inducible Sequence) genes, from a cDNA library prepared from Swiss 3T3 cells treated with tetradecanoyl phorbol acetate (TPA) and cycloheximide. TPA, polypeptide growth factors, and serum induce TIS gene expression in 3T3 cells. We now report that cadmium and zinc elevate mRNA levels for the TIS genes, including TIS28 (c-fos), in Swiss 3T3 cells. The time-course of TIS gene mRNA accumulation after metal exposure is delayed in comparison to the accumulation of TIS gene mRNA after treatment with TPA and growth factors. Cadmium induction of the TIS gene message accumulation is blocked by actinomycin D. Moreover, cadmium treatment does not significantly stabilize TIS gene messages. TIS gene induction by metal is a primary response; TIS8, which encodes a zinc-finger transcription factor, and TIS28 (c-fos) can be induced in the presence of cadmium and cycloheximide, an inhibitor of protein synthesis. Down-regulation of protein kinase C does not attenuate TIS gene induction by heavy metals.


The natural product mensacarcin induces mitochondrial toxicity and apoptosis in melanoma cells.
Mensacarcin is a highly oxygenated polyketide that was first isolated from soil-dwelling Streptomyces bacteria. It exhibits potent cytostatic properties (mean of 50% growth inhibition = 0.2 μm) in almost all cell lines of the National Cancer Institute (NCI)-60 cell line screen and relatively selective cytotoxicity against melanoma cells. Moreover, its low COMPARE correlations with known standard antitumor agents indicate a unique mechanism of action. Effective therapies for managing melanoma are limited, so we sought to investigate mensacarcin's unique cytostatic and cytotoxic effects and its mode of action. By assessing morphological and biochemical features, we demonstrated that mensacarcin activates caspase-3/7-dependent apoptotic pathways and induces cell death in melanoma cells. Upon mensacarcin exposure, SK-Mel-28 and SK-Mel-5 melanoma cells, which have the BRAFV600E mutation associated with drug resistance, showed characteristic chromatin condensation as well as distinct poly(ADP-ribose)polymerase-1 cleavage. Flow cytometry identified a large population of apoptotic melanoma cells, and single-cell electrophoresis indicated that mensacarcin causes genetic instability, a hallmark of early apoptosis. To visualize mensacarcin's subcellular localization, we synthesized a fluorescent mensacarcin probe that retained activity. The natural product probe was localized to mitochondria within 20 min of treatment. Live-cell bioenergetic flux analysis confirmed that mensacarcin disturbs energy production and mitochondrial function rapidly. The subcellular localization of the fluorescently labeled mensacarcin together with its unusual metabolic effects in melanoma cells provide evidence that mensacarcin targets mitochondria. Mensacarcin's unique mode of action suggests that it may be a useful probe for examining energy metabolism, particularly in BRAF-mutant melanoma, and represent a promising lead for the development of new anticancer drugs.


Vitamin A deficiency alters the expression of mucin genes by the rat ocular surface epithelium.
To study effects of depletion of retinoic acid on expression of the mucins ASGP (rMuc4), rMuc5AC, and rMuc1, by the corneal and conjunctival epithelia of the rat. Nineteen-day-old Sprague-Dawley male rats were fed a casein-based vitamin A- deficient diet or casein-based diet with vitamin A as control. Rats from both groups were killed at 1, 3, 5, 13, 15, 18, and 20 weeks after initiation of feeding. Expression of the three mucin genes by the ocular surface epithelium was assayed by reverse transcription-polymerase chain reaction (RT-PCR) and in situ hybridization. In vitamin A-deficient rats, ASGP mRNA was not detected by RT-PCR after 15 weeks of feeding. rMuc5AC mRNA was detected by RT-PCR at 15 weeks, but by 18 and 20 weeks was no longer detectable. By in situ hybridization, ASGP mRNA was localized in the entire ocular surface epithelium after 1 week of feeding, was diminished but detectable above background by 13 weeks, and was not detectable at 20 weeks. rMuc5AC mRNA was detected in the goblet cells of vitamin A- deficient rats by in situ hybridization at 13 weeks, but was lost by 20 weeks, as were identifiable goblet cells. rMuc1 mRNA were detected by RT-PCR through all time points of 1 to 20 weeks in both vitamin A-deficient and control rats, indicating no significant change in rMuc1 mRNA expression with vitamin A deficiency. Both the membrane-spanning mucin ASGP (rMuc4) and the secretory mucin rMuc5AC are directly or indirectly regulated by vitamin A in the ocular surface epithelium, whereas the membrane-spanning mucin rMuc1 is not.


What factors influence patient preferences regarding cardiopulmonary resuscitation?
The aims of this study were to investigate the impact of medical and non-medical factors on the cardiopulmonary resuscitation (CPR) preferences of patients, to determine which of them are the most important to patients when considering CPR, and to compare the views of older (> or = 70 years) and younger (< 70 years) patients. We interviewed 180 patients, 86 of whom were aged 70 years or older. 'I do not want to be a burden on my family' was the most important factor for older patients, and they were more ready to leave the decision to the doctor than were the younger patients. 'I want to retain my capacity to think clearly' was most important to younger patients. In general, younger patients gave higher ratings in favour of CPR than older patients. Increased age, drug abuse, dementia, pain, poor functional status and a low likelihood of success were associated with a lower preference rating for CPR in both age groups.


Differential location of NKT and MAIT cells within lymphoid tissue.
Natural Killer T (NKT) cells and Mucosal-Associated Invariant T (MAIT) cells are innate-like T cells that express semi-invariant αβ T cell receptors (TCRs) through which they recognise CD1d and MR1 molecules, respectively, in complex with specific ligands. These cells play important roles in health and disease in many organs, but their precise intra-organ location is not well established. Here, using CD1d and MR1 tetramer staining techniques, we describe the precise location of NKT and MAIT cells in lymphoid and peripheral organs. Within the thymus, NKT cells were concentrated in the medullary side of the corticomedullary junction. In spleen and lymph nodes, NKT cells were mainly localised within T cell zones, although following in vivo activation with the potent NKT-cell ligand α-GalCer, they expanded throughout the spleen. MAIT cells were clearly detectable in Vα19 TCR transgenic mice and were rare but detectable in lymphoid tissue of non-transgenic mice. In contrast to NKT cells, MAIT cells were more closely associated with the B cell zone and red pulp of the spleen. Accordingly, we have provided an extensive analysis of the in situ localisation of NKT and MAIT cells and suggest differences between the intra-organ location of these two cell types.


Experiences of people with multiple sclerosis participating in a social cognitive behavior change physical activity intervention.
Understanding the experiences of people with MS taking part in physical activity interventions is critical to inform future interventions. The aim was to gain insight into the experiences of people with MS taking part in a behavior change group physical activity intervention with a novel social cognitive theory component. A qualitative semi-structured interview format was utilized. Questions focussed on expectations, views and beliefs at being involved in the study, beliefs on physical activity, and subjective evaluation of the trial content and delivery. Seventeen people were interviewed and data were analyzed using thematic analysis. Three themes were generated: psychological and social factors, intervention processes, and MS identity. The acceptance of MS as an identity acted as an initial barrier to exercise, with a more positive, exercise-enabling identity post-intervention. Psychological factors such as self-efficacy and anxiety, as well as social factors such as social support, were found to play an important role in how participants experienced the program. Similarly, intervention processes included support for group-based activities and structure of exercise classes, and were also inter-linked to the other themes. It appears that group-based exercise interventions are acceptable and feasible for people with MS. The qualitative findings support previously reported quantitative findings that the Step it Up intervention is effective at promoting physical activity and improving psychological outcomes.


Saccharomyces cerevisiae goes through distinct metabolic phases during its replicative lifespan.
A comprehensive description of the phenotypic changes during cellular aging is key towards unraveling its causal forces. Previously, we mapped age-related changes in the proteome and transcriptome (Janssens et al., 2015). Here, employing the same experimental procedure and model-based inference, we generate a comprehensive account of metabolic changes during the replicative life of Saccharomyces cerevisiae. With age, we found decreasing metabolite levels, decreasing growth and substrate uptake rates accompanied by a switch from aerobic fermentation to respiration, with glycerol and acetate production. The identified metabolic fluxes revealed an increase in redox cofactor turnover, likely to combat increased production of reactive oxygen species. The metabolic changes are possibly a result of the age-associated decrease in surface area per cell volume. With metabolism being an important factor of the cellular phenotype, this work complements our recent mapping of the transcriptomic and proteomic changes towards a holistic description of the cellular phenotype during aging.


Real-time polymerase chain reaction quantification of free DNA in serum of patients with polyps and colorectal cancers.
Colorectal cancer (CRC) is one of the most frequent causes of cancer related deaths worldwide. Recently, the use of cell-free DNA as diagnostic tools to identify cancer has been investigated. The aim of this work was to assess whether circulating DNA could be considered a useful marker for detection of early stage CRC and polyps. A total of 118 patients with CRC were included in the study along with 49 patients with colorectal polyps and 26 control subjects. Cell-free DNA was quantified using a real-time TaqMan-polymerase chain reaction assay. Non-parametric tests (Mann-Whitney test and Spearman correlation) were utilized for statistical analysis. Serum DNA concentrations were significantly higher in CRC patients and patients with polyps (median value 105.0 ng/mL and 40.0 ng/mL) compared with controls (median value 14.0 ng/mL; p<0.05). Although carcinoembryonic antigen was above the cut-off in only 13/66 (19.7%) patients with early stage CRC, serum free DNA showed values above the threshold identified using receiver operator characteristic (ROC) curve analysis in 53/66 (80.3%) patients. Our data confirm that serum DNA concentrations are significantly increased in CRC patients with early stage disease and in patients with polyps. This marker might be useful for identifying high-risk individuals.


Plant microRNAs: Biogenesis, Homeostasis, and Degradation.
MicroRNAs (miRNAs), a class of endogenous, tiny, non-coding RNAs, are master regulators of gene expression among most eukaryotes. Intracellular miRNA abundance is regulated under multiple levels of control including transcription, processing, RNA modification, RNA-induced silencing complex (RISC) assembly, miRNA-target interaction, and turnover. In this review, we summarize our current understanding of the molecular components and mechanisms that influence miRNA biogenesis, homeostasis, and degradation in plants. We also make comparisons with findings from other organisms where necessary.


Simple Scoring System for Predicting TACE Unsuitable among Intermediate-Stage Hepatocellular Carcinoma Patients in the Multiple Systemic Treatment Era.
With the development of systemic treatment methods for unresectable hepatocellular carcinoma (uHCC), the concept of unsuitable for transcatheter arterial chemoembolization (TACE) has become important. This study aimed to establish a simple predictive scoring system for determining TACE unsuitable status. From 1998 to 2015, 196 patients with intermediate-stage uHCC with Child-Pugh A (score 5:6 = 108:88) and given TACE as the initial treatment were enrolled. At the baseline, tumor burden (Milan criteria-out, up-to-7 in/out, and up-to-11 in/out: 0-2 points) and modified albumin-bilirubin grade 1/2a or 2b (0-1 point) were added to determine the score for TACE unsuitable (CITRUS-MICAN score; low <2 and high ≥2). In addition, a previously reported tumor marker (TM) score, in which alpha-fetoprotein (AFP) was ≥100 ng/mL, fucosylated AFP ≥10%, and des-gamma-carboxy prothrombin ≥100 mAU/mL (each 1 point) (total 0, 1, or ≥2 points), was used for additionally evaluating tumor malignancy potential. Prognosis was retrospectively evaluated based on those scores. Median survival time (MST) was better for low compared to high CITRUS-MICAN score (42.0 vs. 26.4 months) (p = 0.002). A 2-step evaluation using the combination of CITRUS-MICAN and TM scores showed an MST of 43.2 months for low CITRUS-MICAN/TM score 0/1 (rank-A) and 39.6 months for low CITRUS-MICAN/TM score ≥2 (rank-B2), while it was 46.8 months for high CITRUS-MICAN/TM score 0 (rank-B1), 28.8 months for high CITRUS-MICAN/TM score 1 (rank-B2), and 22.8 months for high CITRUS-MICAN/TM score ≥2 (rank-C). For rank-A cases (n = 51), MST was 43.2 months, while it was 46.8 months for rank-B1 (n = 12), 31.2 months for rank-B2 (n = 82), and 22.8 months for rank-C (n = 51) (p = 0.001). The results showed that rank-C indicates absolute TACE unsuitable status. For rank-A patients, good prognosis with TACE can be expected, while TACE refractoriness status during the clinical course should be carefully evaluated so as to anticipate the appropriate timing for switching to systemic treatment in rank-B1 and -B2 patients.


DIFFUSE CHOROIDAL HEMANGIOMA MASQUERADING AS CENTRAL SEROUS CHORIORETINOPATHY TREATED WITH ORAL PROPRANOLOL.
To describe a case of diffuse choroidal hemangioma with exudative retinal detachment treated successfully with oral propranolol. Single interventional case report of a 14-year-old boy, known case of Sturge-Weber syndrome, presented with decreased vision in the right eye since 2 months. Presenting vision was finger-counting at 3 m. Examination revealed a neurosensory detachment at the macula clinically appearing like central serous chorioretinopathy. Fundus fluorescein angiogram, B-scan ultrasound, and optical coherence tomography were consistent with an underlying diffuse choroidal hemangioma with secondary exudative macular detachment. The patient could not afford radiotherapy and thus was treated with oral propranolol. Serial clinical examinations and optical coherence tomographies showed gradual resolution of the subretinal fluid. Vision at the last visit 4 months after treatment was 6/9 best corrected. Choroidal hemangioma can be mistaken clinically to be central serous chorioretinopathy. Oral propranolol is an effective and economical treatment option in patients not affording other standard modalities of treatment.


Carditis After COVID-19 Vaccination With a Messenger RNA Vaccine and an Inactivated Virus Vaccine : A Case-Control Study.
Case reports of carditis after BNT162b2 vaccination are accruing worldwide. To examine the association of BNT162b2 and CoronaVac (Sinovac) vaccination with carditis. Case-control study with hospital control participants. Territory-wide, public health care database with linkage to population-based vaccination records in Hong Kong. Inpatients aged 12 years or older first diagnosed with carditis were selected as case patients. All other hospitalized patients without carditis were treated as control participants. Ten control participants were randomly matched with each case patient by age, sex, and admission date. Vaccination with BNT162b2 or CoronaVac. Incident diagnosis of carditis based on the International Classification of Diseases, Ninth Revision, and elevated troponin levels. A total of 160 case patients and 1533 control participants were included. Incidence of carditis per 100 000 doses of CoronaVac and BNT162b2 administered was estimated to be 0.31 (95% CI, 0.13 to 0.66) and 0.57 (CI, 0.36 to 0.90), respectively. Multivariable analyses showed that recipients of the BNT162b2 vaccine had higher odds of carditis (adjusted odds ratio [OR], 3.57 [CI, 1.93 to 6.60]) than unvaccinated persons. Stratified by sex, the OR was 4.68 (CI, 2.25 to 9.71) for males and 2.22 (CI, 0.57 to 8.69) for females receiving the BNT162b2 vaccine. The ORs for adults and adolescents receiving the BNT162b2 vaccine were 2.41 (CI, 1.18 to 4.90) and 13.79 (CI, 2.86 to 110.38), respectively. Subanalysis showed an OR of 9.29 (CI, 3.94 to 21.91) for myocarditis and 1.06 (CI, 0.35 to 3.22) for pericarditis associated with BNT162b2. The risk was mainly seen after the second dose of BNT162b2 rather than the first. No association between CoronaVac and carditis with a magnitude similar to that for BNT162b2 was seen. Limited sample size, absence of electrocardiography and other clinical investigative data, and unrecorded overseas vaccination exposure. Despite a low absolute risk, there is an increased risk for carditis associated with BNT162b2 vaccination. This elevated risk should be weighed against the benefits of vaccination. Health and Medical Research Fund.


General practitioners' knowledge of and attitudes to inflammatory bowel disease.
Inflammatory bowel disease (IBD) is a chronic disease requiring long-term management. General practitioners (GPs) are often the first point of contact for initial symptoms and flares. Thus we assessed GPs' attitudes to and knowledge of IBD. A state-wide postal survey of GPs was performed collecting demographic details, practice and attitudes in IBD-specific management and knowledge. Of 1800 GPs surveyed in South Australia, 409 responded; 58% were male, 80% Australian trained and 73% practised in metropolitan areas. Most GPs (92%) reported seeing zero to five IBD patients per month. Overall, 37% of the GPs reported being generally 'uncomfortable' with IBD management. Specifically, they were only somewhat comfortable in providing/using maintenance therapy, steroid therapy or unspecified therapy for an acute flare. They were uncomfortable with the use of immunomodulators and biologicals (71 and 91% respectively). No GP reported never referring, referring sometimes (12%), often (34%) or always (55%). Most (87%) GPs rated their communication with private specialists positively; while only 32% were satisfied with support from public hospitals. Of concern, most (70%) monitored patients on immunosuppression on a case-by-case basis rather than by protocol. In multivariable analyses, GPs' IBD-specific knowledge did not influence comfort with overall management, nor did knowledge influence GP comfort with any particular therapy. Individual GPs care for few IBD patients and have variable attitudes in their practice. Whether improvement can realistically be achieved given individual GP's paucity of patients is questionable. These data support the provision of better support and specific action plans for IBD patients.


Risk factors for breast cancer in Iranian women: a hospital-based case-control study in tabriz, iran.
Breast cancer is the most common cancer in women worldwide and the leading cause of cancer death among women. In Asian countries such as Iran, the incidence of breast cancer is increasing. The present study aimed to assess the risk factors for breast cancer of women in Tabriz, Iran. A hospital-based case-control study was undertaken to identify breast cancer risk factors. The study consisted of 140 cases confirmed via histopathological analysis and 280 group-matched controls without any malignancy. Data were analyzed using descriptive and inferential statistical methods via the SPSS software version 18. In a multivariate analysis, educational level (odds ratio [OR], 4.7; 95% confidence interval [CI], 2.11-10.83), menopausal status (OR, 2.54; 95% CI, 1.41-4.59), a high-fat diet (OR, 2.76; 95% CI, 1.51-5.04), abortion (OR, 2.13; 95% CI, 1.20-3.79), passive smoking (OR, 2.76; 95% CI, 1.51-5.04), oral contraceptive use (OR, 3.18; 95% CI, 1.80-5.59), stress (OR, 3.05; 95% CI, 1.74-5.36), and migration (OR, 3.09; 95% CI, 1.39-6.90) were factors associated with a significantly increased risk of breast cancer. Breastfeeding (OR, 0.39; 95% CI, 0.16-0.97) and a diet containing sufficient fruit and vegetables (OR, 0.22; 95% CI, 0.12-0.39) had protective roles against breast cancer. The study revealed that the risk factors for breast cancer among women in the Tabriz area of Iran are related to the lifestyle. Therefore, the provision of education to change unhealthy lifestyle choices and periodic check-ups for early breast cancer detection are recommended.


Antibiotic Profile of Pathogenic Bacteria Isolated from Postsurgical Site Infections in Public Hospitals in Northern Jordan.
Surgical site infections are hospital-acquired, vary from one hospital to another, and can cause significant postoperative morbidity, mortality, and prolonged hospital stay. The main aim of the study was to identify the bacterial pathogens associated with surgical site infections and their antibiotic susceptibility in a public hospital in northern Jordan. Postsurgical wound samples were collected and processed in a microbiology laboratory using standard microbiological techniques. Antibiotic susceptibility tests were performed using 13 antibiotics covering the gram-positive and gram-negative bacteria using the disc diffusion test. The bacterial species that were identified in this study include Escherichia coli 8 (29%), Pseudomonas aeruginosa 3 (11%), Proteus mirabilis 1 (3.5%), Klebsiella pneumoniae 4 (14%), Salmonella enterica 2 (7%), Staphylococcus aureus 8 (29%), Staphylococcus epidermidis 1 (3.5%), and Streptococcus pyogenes 1 (4%). The antibiotic profiles of these bacteria showed high resistance. The MAR indices showed that 17 of 28 bacteria isolated were above 0.2 indicating high resistance. Resistant bacteria are becoming more dominant in wound infections with a high prevalence of multidrug resistant isolates. Hospital disinfection and treatment protocols regarding the use of antibiotics should be practiced vigorously and monitored regularly to combat resistance.


Karyotypic evolution: cytogenetics follow-up study in childhood acute lymphoblastic leukemia.
Forty seven children affected with acute lymphoblastic leukemia (ALL) were cytogenetically investigated at diagnosis and all through different stages of the disease (remission and relapse). A clonal karyotypic abnormality was found in 32% at diagnosis (mainly comprised of cALLa+). A hyperdiploid mode with chromosome counts ranging from 47-58, was found to be most prominent among cALLa+ patients. The most common numerical aberrations were gain of chromosomes 2, 5, and 21. The structural aberrations at diagnosis were found to be del(9)(p22), inv(9)(p11q13) and del(19)(p12). None of the children showed ph+ chromosome. A good prognosis was found in cALLa+ children with an abnormal karyotype at diagnosis and of these children, those who showed karyotypic instability, had a significantly longer first remission time. The karyotypic evolution through remission(s) and relapse(s) revealed the occurrence of structural alterations, including changes in chromosomes 3, 6, 9, 21 and 22. However, irrespective of the karyotypic clonal nature at diagnosis, chromosome 9 was the most commonly involved chromosome through the course of disease.


Immunohistochemical stains in Mohs surgery: a review.
During Mohs surgery, there are instances in which residual tumor cells may be difficult to detect, thereby increasing the risk of incomplete excision and tumor recurrence. It is possible to employ immunohistochemical techniques as an adjunct to routine hematoxylin and eosin staining to aid in ensuring negative margins. To review the literature regarding the use of immunostains in Mohs surgery. Various immunostains have proved useful in detecting tumor cells in various malignancies, including melanoma, basal cell carcinoma, squamous cell carcinoma, dermatofibrosarcoma protuberans, extramammary Paget's disease, primary cutaneous mucinous carcinoma, granular cell tumor, and trichilemmal carcinoma. In this article, we review immunohistochemical stains that have been employed in Mohs micrographic surgery and evaluate their utility in enhancing detection of residual tumors with respect to tumor type, particularly in situations in which detection of residual tumor may be difficult.


Decreased cytochrome c oxidase subunit VIIa in aged rat heart mitochondria: immunocytochemistry.
Aging decreases oxidative phosphorylation through cytochrome oxidase (COX) in cardiac interfibrillar mitochondria (IFM) in 24-month old (aged) rats compared to 6-month old adult Fischer 344 rats, whereas subsarcolemmal mitochondria (SSM) located beneath the plasma membrane remain unaffected. Immunoelectron microscopy (IEM) reveals in aged rats a 25% reduction in cardiac COX subunit VIIa in cardiac IFM, but not in SSM. In contrast, the content of subunit IV remains unchanged in both SSM and IFM, irrespective of age. These subunits are localized mainly on cristae membranes. In contrast, semi-quantitative immunoblotting, which detects denatured protein, indicates that the content of COX VIIa is similar in IFM and SSM from both aged and adult hearts. IEM provides a sensitive method for precise localizing and quantifying specific mitochondrial proteins. The lack of immunoreaction of COX VIIa subunit by IEM in aged IFM is not explained by a reduction in protein, but rather by a masking phenomenon or by an in situ change in protein structure affecting COX activity.


Curcumin protects rat myocardium against isoproterenol-induced ischemic injury: attenuation of ventricular dysfunction through increased expression of Hsp27 along with strengthening antioxidant defense system.
This investigation examines the role of heat shock protein (Hsp) 27 and its modulation by curcumin in isoproterenol-induced myocardial ischemic injury in rats. Evidence from hemodynamic functions and oxidative stress parameters were also included in the study. The animals were divided into control, isoproterenol, and curcumin 100, 200, and 400 mg/kg treatment groups. Curcumin was administered orally for 15 days to all the treated groups. On 13th and 14th day, isoproterenol (85 mg/kg, s.c.) was injected to curcumin-treated and isoproterenol group. On day 15, hemodynamic parameters were recorded. Thereafter, animals were sacrificed and hearts were kept for biochemical and Western blot analysis. We found dose-dependent increase in the expression of Hsp27 with drastic fall at highest dose. Hemodynamically, the lower 2 doses also restored the cardiac function as evident by improved contractile functions, decreased left ventricular end-diastolic pressure, restored arterial pressures, and heart rate. In addition, there was an increase in then level of superoxide dismutase, catalase, reduced glutathione, and decreased production of thiobarbituric acid reactive substances and leakage of cardiac necroenzyme creatine kinase-MB isoenzyme and lactate dehydrogenase in curcumin 100 and 200 mg/kg group as compared with isoproterenol. However, at a dose of 400 mg/kg, there was ineffectual protection against isoproterenol-induced myocardial damage. Our results suggested 200 mg/mg as the most optimum therapeutic dose showing improved cardiac function due to stabilization of cytoskeleton structure which in turn is attributed to Hsp27 expression along with fortified antioxidant defense system.


Modern management of a patent foramen ovale.
A patent foramen ovale (PFO) has been associated with medical conditions such as cryptogenic stroke, migraine with aura, and decompression illness. Whether closure of the PFO has clinical benefit has been suggested from registry studies, but not yet confirmed in multiple randomized trials. Methods of diagnosis of a PFO and a summary of the current evidence for treatment is presented and discussed as a guide to patient-centred decision-making.


Low Frequency Transcranial Alternating Current Stimulation Accelerates Sleep Onset Process.
The aim of this study is to find a kind of low frequency oscillation transcranial alternating current stimulation, which is directly applied to the scalp epidermal, to stimulate the cerebral cortex with a large spatial range of electric field oscillation across the brain hemisphere, and then trigger the start of the Top-Down processing of sleep homeostasis, in the daytime nap. Thirty healthy subjects, to take naps, underwent an intervention of electrical stimulation at 5 Hz, applied to the dorsal lateral prefrontal cortex. The subjects in the experiments were strictly controlled, and opened their eyes when stimulation was transmitted. Subsequently, after 15 min transcranial alternating current stimulation, subjects entered the experimental procedure of sleep. Electroencephalograph was taken at baseline and during sleep. Behavioral indicators were also added to the experiment. We found that the total power of Electroencephalograph activity in the theta band, as well as low-frequency power at 1-7 Hz, was significantly entrained and increased, and that alpha activity was attenuated faster and spindle activity active earlier. Even more, the transition from awake to Non-rapid eye movement stages occurs earlier. Alertness also decreased when the subjects woke up after brief sleep. The intervention of low frequency brain rhythmic transcranial alternating current stimulation may induce accelerated effect on sleep onset process, thereby possibly alleviating the problems related to sleep disorders such as difficulty to reach the real sleep state quickly after lying down.


[Rabbit fever--a neck tumor].
Twenty-two-year-old previously healthy male is referred to the ENT clinic with a neck tumour. The patient was an animal caretaker. Ultrasonic examination showed a lymph gland conglomerate on the left side of the neck. Primary serologic examination was negative and the lymph tumour was removed. The biopsy showed necrotic lymphadenitis. Three weeks after the first consultation the patient tests positive for F. tularensis. The patient was treated with ciprofloxacin 750 mg x 2 by the oral route for two weeks. Seven weeks after the onset of treatment, the titer for F. tularensis lowers.


Can the hexamethylhydrazinium dication [Me(3)N-NMe(3)](2+) be prepared?
The long sought hexamethylhydrazinium(2+) dication, Me(3)N-NMe(3)(2+), calculationally unstable towards "coulombic explosion" because of formal positive charges on adjacent N atoms, can be synthesized and isolated as a CHB(11)Cl(11)(-) carborane salt.


Isopropanol protein precipitation for the analysis of plasma free metanephrines by liquid chromatography-tandem mass spectrometry.
High-performance liquid chromatography-tandem mass spectrometric (LC-MS/MS)1 analysis of plasma free metanephrines is the most diagnostically sensitive and specific screening test for the diagnosis of pheochromocytoma. We sought to develop an in-house method for this expensive test We used off-line isopropanol protein precipitation of plasma to remove interfering substances before LC-MS/MS analysis. We compared the extraction efficiency and limits of quantification of protein precipitation to those of previously reported solid-phase techniques. The new method had limits of quantification of 0.09 nmol/L and 0.17 nmol/L for metanephrine and normetanephrine, respectively. Method comparison with a previously described solid-phase extraction method revealed Deming regression slopes of 0.904 and 0.994, intercepts of 0.007 and 0.023, and SEs of the residuals (S(y/x)) of 0.071 and 0.284 for metanephrine and normetanephrine, respectively. Extraction efficiency of isopropanol protein precipitation was 66% for metanephrine and 35% for normetanephrine, results that were superior to the efficiencies of 4% and 1% for our adapted solid-phase extraction method. No ion suppression was observed at the retention times for metanephrine and normetanephrine. Isopropanol protein precipitation is a novel and effective off-line sample preparation method for metanephrines that offers a less expensive alternative to on-line solid-phase extraction for low-volume testing and requires a sample volume of only 200 microL. The mass spectrometric analysis time is equivalent to that of solid-phase techniques.


Analysis of digitalized ECG signals based on artificial intelligence and spectral analysis methods specialized in ARVC.
Arrhythmogenic right ventricular cardiomyopathy (ARVC) is an inherited heart muscle disease that appears between the second and forth decade of a patient's life, being responsible for 20% of sudden cardiac deaths before the age of 35. The effective and punctual diagnosis of this disease based on electrocardiograms (ECGs) could have a vital role in reducing premature cardiovascular mortality. In our analysis, we first outline the digitalization process of paper-based ECG signals enhanced by a spatial filter aiming to eliminate dark regions in the dataset's images that do not correspond to ECG waveform, producing undesirable noise. Next, we propose the utilization of a low-complexity convolutional neural network for the detection of an arrhythmogenic heart disease, that has not been studied through the usage of deep learning methodology to date, achieving high classification accuracy, namely 99.98% training and 98.6% testing accuracy, on a disease the major identification criterion of which are infinitesimal millivolt variations in the ECG's morphology, in contrast with other arrhythmogenic abnormalities. Finally, by performing spectral analysis we investigate significant differentiations in the field of frequencies between normal ECGs and ECGs corresponding to patients suffering from ARVC. In 16 out of the 18 frequencies where we encounter statistically significant differentiations, the normal ECGs are characterized by greater normalized amplitudes compared to the abnormal ones. The overall research carried out in this article highlights the importance of integrating mathematical methods into the examination and effective diagnosis of various diseases, aiming to a substantial contribution to their successful treatment.


Nicotine-induced neuroplasticity in striatum is subregion-specific and reversed by motor training on the rotarod.
Nicotine is recognized as one of the most addictive drugs, which in part could be attributed to progressive neuroadaptations and rewiring of dorsal striatal circuits. Since motor-skill learning produces neuroplasticity in the same circuits, we postulate that rotarod training could be sufficient to block nicotine-induced rewiring and thereby prevent long-lasting impairments of neuronal functioning. To test this hypothesis, Wistar rats were subjected to 15 days of treatment with either nicotine (0.36 mg/kg) or vehicle. After treatment, a subset of animals was trained on the rotarod. Ex vivo electrophysiology was performed 1 week after the nicotine treatment period and after up to 3 months of withdrawal to define neurophysiological transformations in circuits of the striatum and amygdala. Our data demonstrate that nicotine alters striatal neurotransmission in a distinct temporal and spatial sequence, where acute transformations are initiated in dorsomedial striatum (DMS) and nucleus accumbens (nAc) core. Following 3 months of withdrawal, synaptic plasticity in the form of endocannabinoid-mediated long-term depression (eCB-LTD) is impaired in the dorsolateral striatum (DLS), and neurotransmission is altered in DLS, nAc shell, and the central nucleus of the amygdala (CeA). Training on the rotarod, performed after nicotine treatment, blocks neurophysiological transformations in striatal subregions, and prevents nicotine-induced impairment of eCB-LTD. These datasets suggest that nicotine-induced rewiring of striatal circuits can be extinguished by other behaviors that induce neuroplasticity. It remains to be determined if motor-skill training could be used to prevent escalating patterns of drug use in experienced users or facilitate the recovery from addiction.


Classification algorithm for high-dimensional protein markers in time-course data.
Identification of biomarkers is an emerging area in oncology. In this article, we develop an efficient statistical procedure for the classification of protein markers according to their effect on cancer progression. A high-dimensional time-course dataset of protein markers for 80 patients motivates us for developing the model. The threshold value is formulated as a level of a marker having maximum impact on cancer progression. The classification algorithm technique for high-dimensional time-course data is developed and the algorithm is validated by comparing random components using both proportional hazard and accelerated failure time frailty models. The study elucidates the application of two separate joint modeling techniques using auto regressive-type model and mixed effect model for time-course data and proportional hazard model for survival data with proper utilization of Bayesian methodology. Also, a prognostic score is developed on the basis of few selected genes with application on patients. This study facilitates to identify relevant biomarkers from a set of markers.


Physical, psychological, and menopause-related symptoms and minor psychiatric disorders in a community-based sample of Brazilian premenopausal, perimenopausal, and postmenopausal women.
The aim of this study was to identify the prevalence of physical, psychological, and menopause-related symptoms and their association with minor psychiatric disorders in premenopausal, perimenopausal, and postmenopausal women. This was a nested cross-sectional study. Demographic characteristics, education, and climacteric symptoms were investigated. The 20-item Self-Reporting Questionnaire was used to screen for minor psychiatric disorders, with a score of 8 or higher indicating positive screening. We studied 324 Brazilian women aged 36 to 62 years (86 premenopausal women, 156 perimenopausal women, and 82 postmenopausal women). Mean (SD) age was 44.8 (3.6), 46.3 (4.6), and 53.3 (3.8) years, respectively (P < 0.001); 52.4% had 8 years or less of schooling (whereas 22.8% had 4 years or less). Forty-five (28.8%) perimenopausal women and 32 (39%) postmenopausal women were users of hormone therapy; 15 (17.4%) and 21 (13.5%) premenopausal and perimenopausal participants, respectively, were users of oral contraceptives. Hormone therapy and oral contraceptive users were excluded from the analysis of symptom prevalence. Hot flashes, night sweats, and vaginal dryness were more prevalent among perimenopausal women (P < 0.001). Fatigue was the most frequent complaint in all groups (61%, 81%, and 88% in premenopausal, perimenopausal, and postmenopausal women, respectively). The variables most frequently associated with positive findings during the screening for minor psychiatric disorders were very low education level and memory loss and irritability. Classic vasomotor complaints were weakly associated with nonpsychotic disease. In turn, perimenopausal women, but not postmenopausal women, were at greater risk of minor psychiatric disorders. Low education level, memory loss, irritability, and the menopausal transition represent risk factors for positive findings in a screening for minor psychiatric disorders.


Lung diseases due to environmental exposures to erionite and asbestos in Turkey.
Asbestos deposits have been used locally by the rural inhabitants of Central and Southeastern Anatolia for domestic purposes for many years. Mineralogical analysis revealed that tremolite is the most prominent asbestos type found in the region. There is in addition another mineral fiber found particularly in three villages located in the Cappadocian region of Central Anatolia (zeolite villages). This is a non-asbestos mineral, which has been identified as the fibrous zeolite, erionite. This fiber is present in the volcanic tuffs, which are used as building stone. Hence, exposure to erionite fibers is always possible in the houses, annexes, and streets of the villages. It has been demonstrated that both asbestos and erionite cause a variety of benign and malignant chest diseases. Among the diseases, calcified pleural plaques (CPP) are the most frequently seen and may be used as an indicator of mineral fiber exposure. Asbestos and erionite exposure are the main causes of malignant mesotheliomas in Turkey. In zeolite villages malignant mesothelioma is responsible for more than 50% of the total deaths. A recent study showed that simian virus 40 is not a cofactor in the pathogenesis of environmental malignant mesothelioma in Turkey. An additional recent genetic-epidemiological study showed that there are some families, which are genetically predisposed to mesothelioma.


Sequential inoculation of flocculent Torulaspora delbrueckii with Saccharomyces cerevisiae increases color density of Pinot Noir wines.
Pinot noir grapes require careful management in the winery to prevent loss of color density and promote aging stability. Winemaking with flocculent yeast has been shown to increase color density, which is desirable to consumers. This research explored interspecies sequential inoculation and co-flocculation of commercial yeast on Pinot noir wine color. Sedimentation rates of six non-Saccharomyces species and two Saccharomyces cerevisiae strains were assayed individually and in combination. The most flocculent pairings, Torulaspora delbrueckii BIODIVA with S. cerevisiae RC212 or VL3, were used to ferment 20 L Pinot noir must. Sequential fermentations produced wines with greater color density at 420 + 520 nm, confirmed by sensory panel. Total and monomeric anthocyanin concentrations were decreased in sequentially fermented wines, despite being the main source of red wine color. BIODIVA adsorbed more anthocyanins than S. cerevisiae, indicating a greater number of cell wall mannoproteins in flocculent yeast, that could then result in a later release of anthocyanins and enhance copigment formation in red wines.


Robert Allen Gardner (1930-2021).
Memorializes Robert Allen Gardner (1930-2021). Gardner and his late wife and research partner, Beatrix "Trixie" Gardner, are best known for their ground-breaking cross-fostering project with the chimpanzee Washoe, the first nonhuman to learn a human language (American Sign Language). Between 1972 and 198, Allen and Trixie replicated their success with four additional infant chimpanzees. Allen cofounded and was a psychology fellow of the Center for Advanced Studies at the University of Nevada in Reno, and served as director from 1990 to 1993. (PsycInfo Database Record (c) 2022 APA, all rights reserved).


Hypoglycemia pathogenesis in children with dumping syndrome.
Three children with severe hypoglycemic reactions secondary to dumping syndrome were studied to discern the mechanism by which hypoglycemia occurred. Symptoms in patient 1 developed after fundoplication, generalized autonomic dysfunction occurred in patient 2, and dumping syndrome developed in patient 3 after malplacement of a feeding gastrostomy tube. Average blood glucose levels studied during and after two to seven meals in each child were 375 +/- 97 mg/dL (mean +/- SD) 30 minutes postprandially and 35 +/- 10 mg/dL greater than 120 minutes later. Swings in glucose values were proportional to the volume of meals. Insulin and glucagon levels were followed during a single meal challenge test in each patient; the average glucose concentration increased to 356 +/- 59 mg/dL 30 minutes postprandially and decreased to 32 +/- 11 mg/dL at 150 +/- 30 minutes. Hormonal analyses indicated (1) inappropriate early release of glucagon (300 pg/mL at 15 minutes) in patient 1, (2) exuberant early release of insulin (maximum 190 +/- 15 microU/mL) resulting in rapid decrease in glucose concentration in all patients, (3) development and/or persistence of hypoglycemia after the decline in circulating insulin to undetectable levels, and (4) inadequate glucagon response to hypoglycemia resulting in sustained hypoglycemia. These data indicate that gross disturbances of the insulin-glucagon axis attend childhood dumping syndrome.


Water Permeability during Tomato Fruit Development in Normal and rin Nonripening Mutant.
This work tested one aspect of the relations between membrane permeability and fruit ripening. Membrane permeability was measured as [(3)H]water efflux rate from preloaded fruit pericarp disks. Different stages of fruit development were compared between two tomato (Lycopersicon esculentum Mill) strains: the normal Rutgers and the isogenic nonripening rin strain. The first significant increase in permeability was measured in Rutgers tissue at 110% of development, after fruit ripening had already begun as indicated by ethylene and CO(2) evolution and lycopene synthesis. The rin did not show any increase in tissue permeability during fruit development or maturation.Our results do not support the idea that the first event of the ripening process is an increase in membrane permeability. Nevertheless, the nonripening mutant fails to show the normal increase in permeability.


Eosinophilic granuloma masquerading as aggressive periodontitis.
Eosinophilic granuloma (EG) is the mildest and mainly localized form of the clinicopathologic spectrum of Langerhans' cell histiocytosis. It is a destructive osseous lesion characterized by a vast number of eosinophils and histiocytes. The etiology remains unknown. In this paper, a case of EG is presented that was initially diagnosed and treated as aggressive periodontitis (AP). Despite treatment procedures, the EG continued to expand very quickly, destroying the lingual cortical bone and the neighboring soft tissues and exhibiting periosteal reaction. Diagnosis of EG was established on the basis of histopathologic and immunohistochemical evaluation. Moreover, certain manifestations in the skeletal and respiratory system were observed. Surgical curettage of the lesions was effective; however, corticosteroids and low-dose radiation were used as adjunctive therapy. The rapid progress of eosinophilic granuloma, the diagnostic problems, and the consequences of late diagnosis and treatment are discussed.


Effect of modified diabetic splenocytes on mice injected with multiple low-dose streptozotocin.
This work reports the effects of a previous injection of mitomycin-modified splenocytes from multiple-low dose streptozotocin-treated mice (mld-sz) on autoimmune diabetes produced by mld-sz. Our work shows that a previous inoculation of modified mononuclear splenocytes from mld-sz mice prevents alterations in glycemia, in insulin secretion (IS) pattern from isolated perifused islets, and in mass of pancreatic islets. Immunohistochemistry showed an alteration in the number of beta, but not of alpha or delta cells. While a mononuclear intra-islet infiltration was observed in mld-sz mice, a predominantly polar or peri-islet infiltration was seen in vaccinated mice. Islet-associated mononuclear cells from mld-sz mice produced diabetes and induced a diminished IS when transferred to normal receptors. Those cells from previously vaccinated mld-sz mice had no effect when injected into normal receptors. In addition, they also inhibited the damage induced in normal receptors by the islet-associated mononuclear cells from mld-sz animals. Cellular death was also prevented by previous vaccination. Our results suggest that vaccination with modified splenocytes from mld-sz mice is capable of shifting the islet cells infiltration pattern from an aggressive one toward a protective one and thus preventing the beta cell destruction observed in mld-sz mice.


Frictional rheology of a confined adsorbed polymer layer.
The sliding dynamics of a confined adsorbed polymer layer is investigated at the nanoscale. A combined mechanical and physical approach is used to model the rheology and structure of the adsorbed layer. The confinement at short distances governs the nanotribological behavior of the polymer layer formed close to the surface. It appears that the Amontons' proportionality between frictional and normal stresses does not hold here: the higher the contact pressure, the lower the friction. Besides, the sliding stress is strongly dependent on the velocity: it increases with the sliding velocity. Using a model based on the kinetics of formation and rupture of adhesive bonds between the two shearing surfaces theoretically accounts for the behavior of this system. This approach allows us to correlate the frictional properties to the molecular organization on the surfaces.


Primaquine for preventing relapse in people with Plasmodium vivax malaria treated with chloroquine.
Plasmodium vivax infections are an important contributor to the malaria burden worldwide. The World Health Organization recommends a 14-day course of primaquine (0.25 mg/kg/day, giving an adult dose of 15 mg/day) to eradicate the liver stage of the parasite and prevent relapse of the disease. Many people find a 14-day primaquine regimen difficult to complete, and there is a potential risk of haemolytic anaemia in people with glucose-6-phosphate-dehydrogenase enzyme (G6PD) deficiency. This review evaluates primaquine in P. vivax, particularly alternatives to the standard 14-day course. To compare alternative primaquine regimens to the recommended 14-day regimen for preventing relapses (radical cure) in people with P. vivax malaria treated for blood stage infection with chloroquine. We also summarize trials comparing primaquine to no primaquine that led to the recommendation for the 14-day regimen. We searched the Cochrane Infectious Diseases Group's Specialized Register, CENTRAL (The Cochrane Library), MEDLINE, EMBASE and LILACS up to 8 October 2013. We checked conference proceedings, trial registries and reference lists and contacted researchers and pharmaceutical companies for eligible studies. Randomized controlled trials (RCTs) and quasi-RCTs comparing various primaquine dosing regimens with the standard primaquine regimen (15 mg/day for 14 days), or with no primaquine, in people with vivax malaria treated for blood stage infection with chloroquine. We independently assessed trial eligibility, trial quality, and extracted data. We calculated risk ratios (RR) with 95% confidence intervals (CI) for dichotomous data, and used the random-effects model in meta-analyses if there was significant heterogeneity. We assessed the overall quality of the evidence using the GRADE approach. We included 15 trials (two cluster-RCTs) of 4377 adult and child participants. Most trials excluded people with G6PD deficiency. Trials compared various regimens of primaquine with the standard primaquine regimen, or with placebo or no treatment. All trials treated blood stage infection with chloroquine. Alternative primaquine regimens compared to 14-day primaquineRelapse rates were higher over six months with the five-day primaquine regimen than the standard 14-day regimen (RR 10.05, 95% CI 2.82 to 35.86; two trials, 186 participants, moderate quality evidence). Similarly, relapse over six months was higher with three days of primaquine than the standard 14-day regimen (RR 3.18, 95% CI 2.1 to 4.81; two trials, 262 participants, moderate quality evidence; six months follow-up); and with primaquine for seven days followed up over two months, compared to 14-day primaquine (RR 2.24, 95% CI 1.24 to 4.03; one trial, 126 participants, low quality evidence).Relapse with once-weekly supervised primaquine for eight weeks was little different over nine months follow-up compared to 14-day self-administered primaquine in one small study (RR 2.97, 95% CI 0.34 to 25.87; one trial, 129 participants, very low quality evidence). Primaquine regimens compared to no primaquineThe number of people that relapsed was similar between people given five days of primaquine or given placebo or no primaquine (four trials, 2213 participants, high quality evidence; follow-up six to 15 months); but lower with 14 days of primaquine (RR 0.6; 95% CI 0.48 to 0.75; ten trials, 1740 participants, high quality evidence; follow-up seven weeks to 15 months).No serious adverse events were reported. Treatment-limiting adverse events were rare and non-serious adverse events were mild and transient. Trial authors reported that people tolerated the drugs.We did not find trials comparing higher dose primaquine regimens (0.5 mg/kg/day or more) for five days or more with the 14-day regimen. The analysis confirms the current World Health Organization recommendation for 14-day primaquine (15 mg/day) to prevent relapse of vivax malaria. Shorter primaquine regimens at the same daily dose are associated with higher relapse rates. The comparative effects with weekly primaquine are promising, but require further trials to establish equivalence or non-inferiority compared to the 14-day regimen in high malaria transmission settings.


Metformin attenuates high glucose-induced injury in islet microvascular endothelial cells.
As one of the most frequently prescribed antidiabetic drugs, metformin can lower glucose levels, improve insulin resistance manage body weight. However, the effect of metformin on islet microcirculation remains unclear. In the present study, to explore the effect of metformin on islet endothelial cells and investigated the underlying mechanism, we assessed the effects of metformin on islet endothelial cell survival, proliferation, oxidative stress and apoptosis. Our results suggest that metformin stimulates the proliferation of pancreatic islet endothelial cells and inhibits the apoptosis and oxidative stress caused by high glucose levels. By activating farnesoid X receptor (FXR), metformin increases the expression of vascular endothelial growth factor-A (VEGF-A) and endothelial nitric oxide synthase (eNOS), improves the production of nitric oxide (NO) and decreases the production of ROS. After the inhibition of FXR or VEGF-A, all of the effects disappeared. Thus, metformin appears to regulate islet microvascular endothelial cell (IMEC) proliferation, apoptosis and oxidative stress by activating the FXR/VEGF-A/eNOS pathway. These findings provide a new mechanism underlying the islet-protective effect of metformin.


The intracellular nature of Salmonella infection during the early stages of mouse typhoid.
In mouse enteric fever (typhoid) infection with S. typhimurium, the bacteria appear to grow intracellularly, and at least during the early phase of infection they are in splenic PMNs rather than macrophages. Inflammation caused by salmonella infection and other infections, such as MHV, results in inflammatory responses that enhance resistance to salmonella infection. At least in the case of MHV, this effect is most pronounced on the rate of salmonella growth. Since the effects of the Ity locus on salmonella growth rate are readily seen during the first few days of infection, when salmonella are primarily within PMNs, the Ity locus is able to mediate its effect on salmonella pathogenesis in PMNs in vivo. Whether or not macrophages play a predominant role in salmonella pathogenesis later in infection is not yet known.


Mutagenic consequences of cytosine alterations site-specifically embedded in the human genome.
Cytosine residues in CpG dinucleotides often undergo various types of modification, such as methylation, deamination, and halogenation. These types of modifications can be pro-mutagenic and can contribute to the formation of mutational hotspots in cells. To analyze mutations induced by DNA modifications in the human genome, we recently developed a system for tracing DNA adducts in targeted mutagenesis (TATAM). In this system, a modified/damaged base is site-specifically introduced into intron 4 of thymidine kinase genes in human lymphoblastoid cells. To further the understanding of the mutagenesis of cytosine modification, we directly introduced different types of altered cytosine residues into the genome and investigated their genomic consequences using the TATAM system. In the genome, the pairing of thymine and 5-bromouracil with guanine, resulting from the deamination of 5-methylcytosine and 5-bromocytosine, respectively, was highly pro-mutagenic compared with the pairing of uracil with guanine, resulting from the deamination of cytosine residues. The deamination of 5-methylcytosine and 5-bromocytosine rather than that of normal cytosine dramatically enhances the mutagenic potential in the human genome.


Study on the Role of the Common House Fly, Musca domestica, in the Spread of ORF Virus (Poxviridae) DNA under Laboratory Conditions.
ORF virus (Poxviridae) is the causative agent of contagious ecthyma (soremouth), a disease primarily affecting sheep and goats worldwide, but also humans exposed to disease-ridden animals. Pathogens are shed with scabs, and infection mainly occurs by direct contact. Although the disease is relatively benign and self-limiting, the morbidity rate is high in livestock with subsequent significant financial and economic impact. The aim of the study was to experimentally investigate the potential for the housefly, Musca domestica, to act as a mechanical vector of the virus. Homogenate of crusted scabs from ORFV-positive sheep (Italy, Apulia) were used to infect laboratory-reared flies. Flies walking on viral mixture and flies inoculated on their wings were individually placed in Falcon tubes and the ORFV DNA was searched by PCR on tube walls; flies were fed on the same homogenized crusts and their crop and spots (vomit and feces) molecularly examined for ORF DNA at 2, 4, and 6 h. All of the flies (100%) used in the experiments were able to pick up and transmit the viral genome to contact surfaces; 60% were found ORF virus (DNA)-positive in both spots and crop. These results suggest that M. domestica could play a role as potential mechanical vector and/or reservoir in the epidemiology of the ORF virus infection. Thus, house fly management should be considered in the measures to control the disease in ovine-caprine farms.


Unraveling the structure and exciton coupling for multichromophoric merocyanine dye molecules.
The relative orientation of chromophores is a key factor in determining the relationship between the structure and the functionality in molecular multichromophore ensembles. In the case of structurally flexible molecular systems in solution, the task to clarify the relevant effects of accessible chromophore orientations with spectroscopic observations is very demanding. In this study, we address this issue by investigating a series of differently connected multichromophoric systems composed of highly dipolar merocyanine dyes that are systematically varied in their substitution pattern and the number of chromophores attached to a bridging benzene ring. Combining electro-optical absorption (EOA) and UV/Vis spectroscopy with density functional theory (DFT) as well as exciton theory discloses conformational preferences and rationalizes the optical properties of the interacting chromophores. Our findings suggest for all multichromophoric systems there is a relative orientation of the chromophores which compensates for the individual dipole moments of the merocyanine dyes by pointing preferably in opposing directions. These orientations furthermore rationalize the observed spectral properties by partly excitonically-coupled subunits.


Mutants of the tumour suppressor p53 L1 loop as second-site suppressors for restoring DNA binding to oncogenic p53 mutations: structural and biochemical insights.
To assess the potential of mutations from the L1 loop of the tumour suppressor p53 as second-site suppressors, the effect of H115N and S116M on the p53 'hot spot' mutations has been investigated using the double-mutant approach. The effects of these two mutants on the p53 hot spots in terms of thermal stability and DNA binding were evaluated. The results show that: (i) the p53 mutants H115N and S116M are thermally more stable than wild-type p53; (ii) H115N but not S116M is capable of rescuing the DNA binding of one of the most frequent p53 mutants in cancer, R248Q, as shown by binding of R248Q/H115N to gadd45 (the promoter of a gene involved in cell-cycle arrest); (iii) the double mutant R248Q/H115N is more stable than wild-type p53; (iv) the effect of H115N as a second-site suppressor to restore DNA-binding activity is specific to R248Q, but not to R248W; (v) molecular-dynamics simulations indicate that R248Q/H115N has a conformation similar to wild-type p53, which is distinct from that of R248Q. These findings could be exploited in designing strategies for cancer therapy to identify molecules that could mimic the effect of H115N in restoring function to oncogenic p53 mutants.


Site-specific sensitization in a murine model of allergic rhinitis: role of the upper airway in lower airways disease.
Allergic rhinitis (AR) is the most common atopic disease with strong links to asthma. We have developed a murine model of AR to study nasal, bronchial, and systemic immune response to local allergen stimulation. The purpose of this study was to develop and characterize a murine model of AR. Six- to 8-week-old BALB/c mice were sensitized by means of intranasal (local) application of ovalbumin (OVA) or systemic intraperitoneal injection. They were then challenged with intranasal OVA, and allergic response was assessed. Intranasal particle deposition was found to be exclusively in the nares. All sensitized animals showed increased levels of OVA-specific serum IgE and IgG after challenge, although the timing to maximal response varied with the route and dose of allergen used. Histology of the upper and lower airways showed marked eosinophilic infiltration, and analysis of bronchoalveolar lavage fluid showed increased IL-5 and PMN infiltrates after challenge. Using exclusive local sensitization and challenge of mouse nares, we were able to demonstrate inflammatory changes in both the upper and lower airways, even though distribution of allergen particles appeared to be only in the nares of these animals. This provides further evidence for the importance of the upper airway in lower airways disease. We have shown that the route of administration greatly affects the characteristics of the subsequent immune responses.


[A case of penile revascularization by Hauri's method for arteriogenic impotence].
In a 35 year old arteriogenic impotent patient without a history of hypertension, arteriosclerotic disease, or diabetes mellitus, the corpus cavernosum of the penis was revascularized using Hauri's method. Before surgery, erection after the intra-cavernous injection of 20 micrograms of prostaglandin E1 was very weak. In a color ultrasonography the peak systolic velocity of the cavernous arteries was recorded as being only 18 cm/sec. Furthermore, no artery except the right dorsal artery was evident even with a digital subtraction angiography. Accordingly he was diagnosed as having arteriogenic impotence, and we carried out the corpus cavernosum revascularization using Hauri's method under microscopic magnification. The dorsal artery and the deep dorsal vein were anastomosed side-to-side, and the hypogastric artery and dorsal artery were anastomosed end-to-side. After the revascularization surgery, the peak systolic velocity of cavernous arteries returned to normal (53 cm/sec), and the penis showed complete erection after an intracavernous injection of 20 micrograms of prostaglandin E1. Before surgery this patient had no experienced sexual intercourse, but he could achieve full sexual intercourse 2 weeks after the surgery. His erectile ability has been maintained for 4 months since the surgery. This is the 1st case of arteriogenic impotence treated using Hauri's method in Japan.


Quantitative detection of the nosZ gene, encoding nitrous oxide reductase, and comparison of the abundances of 16S rRNA, narG, nirK, and nosZ genes in soils.
Nitrous oxide (N2O) is an important greenhouse gas in the troposphere controlling ozone concentration in the stratosphere through nitric oxide production. In order to quantify bacteria capable of N2O reduction, we developed a SYBR green quantitative real-time PCR assay targeting the nosZ gene encoding the catalytic subunit of the nitrous oxide reductase. Two independent sets of nosZ primers flanking the nosZ fragment previously used in diversity studies were designed and tested (K. Kloos, A. Mergel, C. Rösch, and H. Bothe, Aust. J. Plant Physiol. 28:991-998, 2001). The utility of these real-time PCR assays was demonstrated by quantifying the nosZ gene present in six different soils. Detection limits were between 10(1) and 10(2) target molecules per reaction for all assays. Sequence analysis of 128 cloned quantitative PCR products confirmed the specificity of the designed primers. The abundance of nosZ genes ranged from 10(5) to 10(7) target copies g(-1) of dry soil, whereas genes for 16S rRNA were found at 10(8) to 10(9) target copies g(-1) of dry soil. The abundance of narG and nirK genes was within the upper and lower limits of the 16S rRNA and nosZ gene copy numbers. The two sets of nosZ primers gave similar gene copy numbers for all tested soils. The maximum abundance of nosZ and nirK relative to 16S rRNA was 5 to 6%, confirming the low proportion of denitrifiers to total bacteria in soils.


Introduction to the special section on clinical applications of analogue behavioral observation.
This Special Section addresses methods, validity, and utility of analogue behavioral observation. Separate contributions to the Special Section cover analogue behavioral observation of marital interaction, child behavior problems, parent-child interaction, and adult social functioning. Additional articles address psychometric foundations of analogue behavioral observations, general issues, and future directions in the development and evaluation of this assessment method. Many published studies were reviewed in detail and issues of validity, clinical assessment utility, and sources of variance in obtained measures are addressed.


[Two patients with osteochondroma of the cervical spine].
In two patients, both women of 32 years old, who had a local tumour in the neck firmly attached to the spine, radiologic imaging and biopsy led to the diagnosis of osteochondroma. Both patients recovered, but one had an episode of atrophy of the M. trapezius and the M. rhomboideus. If a solid mass is palpated in the neck a malignant process and cervical exostosis should be considered in the differential diagnosis. Exostoses usually are benign symptomless tumours, diagnosed in the second or third decade of life. They may compress spinal nerves and in rare cases cause severe complications by compression of vital structures such as myelum and arteries. An exostosis should be resected only if it is symptomatic or if a malignant process is suspected. In case of doubt surgical intervention is indicated with special attention to surrounding structures such as nerves and blood vessels.


Capsid assembly and involved function analysis of twelve core protein mutants of duck hepatitis B virus.
The roles of different regions of the duck hepatitis B virus (DHBV) core protein on viral capsid assembly and related functions were examined. Twelve deletion and insertion mutations which covered 80% of the DHBV C open reading frame were constructed and expressed in Escherichia coli. The N-terminal region (amino acids 3 to 66) of DHBV core protein was important for its tertiary structure and function in E. coli. The expressed core mutants without this region apparently inhibited E. coli growth. The results of transmission electron microscopy of E. coli thin sections, capsid agarose gel, and sucrose gradient sedimentation demonstrated that a few DHBV core mutants with insertion in the N terminus and deletion in the C terminus retained the ability to form core-like particles in E. coli. However, other mutations in most of N-terminal and central regions strongly inhibited the self-assembly ability of DHBV core protein in E. coli. In addition, the mutant with a C-terminal region deletion (amino acids 181 to 228) lost most of the nucleic acid-binding activity of the DHBV core protein.


[Future perspectives of bone marrow and stem cell activation for autologous transplantation].
Autologous bone marrow transplantation has gained an increasing role in modern oncology. The use of circulating progenitors from the peripheral blood as autografts might have some important advantages, since leukaphereses are better tolerable than a BM harvest, could be done immediately following chemotherapy, are possible in patients with previous irradiation to the pelvis and might be less contaminated with malignant progenitor cells. We have shown that monitoring transplants with the relatively simple in vitro assay for myeloid progenitor cells is predictive for hematopoietic recovery in animal models as well as in the clinical setting. Moreover, stem cells harvested during repeated leukapheresis were able to repopulate permanently the irradiated BM. Using recombinant human granulocyte/macrophage colony stimulating factor, large quantities of progenitor cells can be mobilized into the peripheral blood and collected for autotransplantation, in particular if used directly following chemotherapy. Cell separation studies showed that a bone marrow/blood barrier retains some subpopulations of progenitor cells which might lead to the preferential circulation of normal stem cells following chemotherapy. Methods are described to distinguish normal and leukemic progenitor cells which will help in the future to assay autografts both for their repopulating ability and their contamination with tumor cells.


Pelvic floor ultrasound: when, why, and how?
Pelvic floor disorders are a significant medical issue, reportedly affecting nearly one in four women in the United States. Nonetheless, until the last decade, there has been relatively limited imaging research into this highly prevalent disorder. The three major imaging modalities utilized to assess pelvic floor function are ultrasound, MRI and fluoroscopy. Pelvic floor ultrasound is a rapidly emerging technique which takes advantage of the widespread availability of ultrasound, the non-invasive and relatively inexpensive approach and the incorporation of real-time imaging and software advances which permit 3-D volume imaging. Pelvic floor ultrasound provides the opportunity to optimize patient counseling and enhance pre-operative planning by providing an anatomic and functional roadmap for the referring clinician. We recommend the consideration of pelvic floor ultrasound, as described here, as an addition to the imaging armamentarium available to physicians and surgeons serving this patient population.


Galactolipid Synthesis in Vicia faba Leaves: IV. Site(s) of Fatty Acid Incorporation into the Major Glycerolipids.
The fatty acids of the major glycerolipids of Vicia faba leaves were analyzed immediately following (14)CO(2) feeding. The leaves were fractionated into chloroplast and cytoplasmic fractions and the location of radioactivity in the fatty acids determined. The results indicate that the major site of incorporation of fatty acids is in the phospholipids. Phosphatidylcholine contained the highest level of radioactivity in the cytoplasmic fraction, whereas phosphatidylglycerol contained radioactivity in both the chloroplast and cytoplasmic fractions. The galactolipids contained very little radioactivity in comparison, this radioactivity being confined to high speed centrifugal fractions believed to contain the envelopes of the chloroplast. Our results suggest that phosphatidylcholine is a major site of incorporation of fatty acids (mainly in oleic acid) in the cytoplasm, whereas phosphatidylglycerol is also a site of incorporation involving both oleic and palmitic acids, inside and outside the chloroplast.


Self-mutilation among male patients with alcohol dependency: the role of dissociation.
The aim of this study was to investigate the relationship of self-mutilative behavior with dissociative experiences among men who are alcohol dependent. Participants were 176 inpatients consecutively admitted to an alcohol dependency treatment center. Dissociative Experiences Scale, Beck Depression Inventory, State-Trait Anxiety Inventory, Symptom Checklist-Revised, and Michigan Alcoholism Screening Test were conducted to all participants. A sizable proportion of patients (29.0%) reported self-mutilation (SM). Childhood abuse, younger age, early onset of alcoholism, and dissociative taxon membership predicted SM. The overall severity of clinical condition and the frequency of suicide attempts among those who reported SM were higher than those of the remaining patients. The Dissociative Experiences Scale-Taxon item "auditory verbal hallucinations" and the Symptom Checklist-Revised dimension "hostility" were predictors of SM. There is a complex relationship between dissociation, alcohol use, and SM. Increased awareness among clinicians on this relationship may increase the effectiveness of treatment interventions.


Organosolv ethanol lignin from hybrid poplar as a radical scavenger: relationship between lignin structure, extraction conditions, and antioxidant activity.
Twenty-one organosolv ethanol lignin samples were prepared from hybrid poplar (Populus nigra xP. maximowiczii) under varied conditions with an experimental matrix designed using response surface methodology (RSM). The lignin preparations were evaluated as potential antioxidants. Results indicated that the lignins with more phenolic hydroxyl groups, less aliphatic hydroxyl groups, low molecular weight, and narrow polydispersity showed high antioxidant activity. Processing conditions affected the functional groups and molecular weight of the extracted organosolv ethanol lignins, and consequently influenced the antioxidant activity of the lignins. In general, the lignins prepared at elevated temperature, longer reaction time, increased catalyst, and diluted ethanol showed high antioxidant activity. Regression models were developed to enable the quantitative prediction of lignin characteristics and antioxidant activity based on the processing conditions.