The ubiquitously expressed human CYP51 encodes lanosterol 14 alpha-demethylase, a cytochrome P450 whose expression is regulated by oxysterols.
Sterol biosynthesis requires the removal of the 14 alpha-methyl group from lanosterol in animals and fungi and from obtusifoliol in plants. This reaction is catalyzed by a microsomal cytochrome P450, the sterol 14 alpha-demethylase (P450(14DM), which is the only P450 described so far to be expressed in different phyla. A cDNA encoding human P450(14DM) was isolated from a liver cDNA library using a partial rat lanosterol 14 alpha-demethylase cDNA probe. The deduced amino acid sequence is 93% and 38--42% identical to rat and fungal P450(14DM), respectively. Expression of the human CYP51 cDNA in Escherichia coli showed that the cDNA encodes an enzyme having lanosterol 14 alpha-demethylase activity. Northern blot analysis showed that CYP51 mRNA is ubiquitously expressed with highest levels in testis, ovary, adrenal, prostate, liver, kidney, and lung. Many genes involved in cholesterol homeostasis are regulated by cholesterol or its metabolites. In the case of CYP51, cholesterol deprivation led to a 2.6- to 3.8-fold induction of mRNA levels in human adrenocortical H295R cells and this effect was suppressed by the addition of 25-hydroxycholesterol. In human hepatoma HepG2 cells, no effect of cholesterol deprivation was observed; however, the levels of CYP51 mRNA were reduced 4- to 6-fold by the addition of 25-hydroxycholesterol. Thus, like several other genes in the cholesterol biosynthetic pathway, including the 3-hydroxy-3-methylglutaryl coenzyme A (HMG CoA) synthase, HMG CoA reductase, squalene synthase, and farnesyl diphosphate synthase, the expression of the human CYP51 is suppressed by oxysterols.


Treatment of primary Sjögren's syndrome with hydroxychloroquine: a retrospective, open-label study.
To determine whether patients with primary Sjögren's syndrome (SS), diagnosed according to San Diego criteria, had improvement in their laboratory or clinical features during treatment with hydroxychloroquine (6-7 mg/kg/day) for at least two years. The study population included 50 consecutive patients with primary SS who were diagnosed according to San Diego criteria, and in whom hydroxychloroquine was suggested as treatment. This group included 10 patients who were early dropouts (side effects or desire not to take antimalarial drugs) and 40 patients who received drugs for at least two years (range 24-48 months). In a subset of SS patients, values for ESR (westergren) and quantitative immunoglobulins were available for comparison. Improvement with therapy was defined as: (a) > or = 20% improvement in variables of tear flow (Schirmer's test I) or corneal integrity (rose Bengal): (b) > or = 20% salivary function (flow rate); and (c) improvement in at least two of the following measures: physicians assessment of global disease activity by > or = 20%, patient assessment of improvement in pain or fatigue by > or = 20%, and ESR improved by > or = 20 mm/hr. In a retrospective study of SS patients who completed the trial, a significant improvement was noted in ocular symptoms (pain and dryness) in patients (55 and 57%) and improved corneal integrity (rose Bengal straining) in 53% of patients. The Schirmer's test was improved by > or = 2 mm/5 minutes in 50% in patients. Improvement was noted in oral symptoms (pain and dryness) in patients (57 and 60%) and salivary flow rate was increased in 82% of patients. In a subset of SS patients evaluated, the ESR improved by > or = 20 mm/hr in 17/32 patients (53%) and quantitative IgG level by > or = 20% in 8/13 patients (61%). Physician global assessment of overall patient status and patient assessment of overall status indicated improvement in over 62% of patients. In a retrospective study of patients fulfilling San Diego Criteria for SS, we found: (a) sustained improvement of local symptoms (painful eyes, painful mouth) and improvement of systemic manifestations (arthralgias and myalgias) after treatment with hydroxychloroquine 6-7 mg/kg/day over mean three-year follow-up; (b) laboratory analysis showed a significant improvement in their ESR and their quantitative IgG levels; (c) no significant late toxicity was observed in this study cohort. A prospective study of hydroxychloroquine in patients fulfilling San Diego criteria for SS is indicated.


PHF2, a novel PHD finger gene located on human chromosome 9q22.
We have isolated and characterized a novel PHD finger gene, PHF2, which maps to human Chromosome (Chr) 9q22 close to D9S196. Its mouse homolog was also characterized and mapped to the syntenic region on mouse Chr 13. The predicted human and mouse proteins are 98% identical and contain a PHD finger domain, eight possible nuclear localization signals, two potential PEST sequences, and a novel conserved hydrophobic domain. Northern analysis shows widespread expression of PHF2 in adult tissues, while in situ hybridization on mouse embryos reveals staining in the neural tube and dorsal root ganglia significantly above a ubiquitous low level expression signal. From its expression pattern and its chromosomal localization, PHF2 is a candidate gene for hereditary sensory neuropathy type I, HSN1.


Spermiogenesis and spermatozoon ultrastructure of the cranial digenean Troglotrema acutum (Leuckart, 1842).
Ultrastructure of spermiogenesis and the main characters of the mature spermatozoon of Troglotrema acutum are described by means of transmission electron microscopy. Specimens were obtained from the nasolacrimal sinuses of an American mink (Mustela vison). Spermiogenesis in T. acutum follows the general pattern of digeneans. The zone of differentiation is a conical-shaped area bordered by cortical microtubules and delimited at its base by a ring of arched membranes. This area contains 2 centrioles associated with striated rootlets and an intercentriolar body between them. The centrioles develop 2 free flagella that grow ortogonally to the median cytoplasmic process. The posterior flagellar rotation and proximodistal fusion of the free flagella with the median cytoplasmic process originate the spermatozoon. The mature spermatozoon of T. acutum is characterized by the presence of 2 axonemes of different lengths presenting the 9+'1' trepaxonematan pattern, 2 bundles of parallel cortical microtubules, 2 mitochondria, a nucleus, and granules of glycogen. These ultrastructural characters are compared with other digenean species previously studied and the importance of different spermatological features is discussed.


Cloning and expression of murine sister of P-glycoprotein reveals a more discriminating transporter than MDR1/P-glycoprotein.
Sister of P-glycoprotein (SPGP), a novel murine cDNA and member of the ATP-binding cassette superfamily highly homologous to P-glycoprotein (Pgp), was cloned. Moreover, its genomic clone was isolated and localized to chromosome 2 by fluorescence in situ hybridization. SPGP was functionally evaluated relative to MDR1 after subcloning SPGP cDNA into a retroviral bicistronic vector capable of expressing both SPGP and the green fluorescent protein. LLC-PK1 and MDCKII cells were transduced with this retrovirus and SPGP-positive clones were isolated. Drug uptake and efflux was compared in cells ectopically expressing either SPGP or human MDR1. SPGP cells had decreased uptake of taurocholate and vinblastine compared with LLC-PK1 cells. Additional studies revealed that vinblastine efflux was accelerated by SPGP compared with LLC-PK1. Further comparison revealed that although MDR1 easily impaired uptake of vincristine, daunomycin, paclitaxel, and digoxin, SPGP had no effect on uptake of these drugs. However, further studies demonstrated that, like MDR1, SPGP effluxed calcein-acetoxymethyl ester (AM). Unlike MDR1, SPGP was incapable of effluxing rhodamine 123. Although cyclosporine A and reserpine blocked calcein-AM transport by MDR1, these drugs had either minimal or no effect, respectively, on blocking SPGP efflux of calcein-AM. In contrast, ditekiren, a linear hexapeptide, readily and preferentially inhibited SPGP efflux of calcein-AM. Further studies with three structural analogs of ditekiren revealed that one analog inhibited SPGP efflux of calcein-AM, although not as potently as ditekiren. These are the first studies to reveal that SPGP has distinct transport properties compared with MDR1.


[The Expression and Significance of Annexin II in Lung Squamous Cell Cancer and Adenocarcinom.].
Annexin II is a calcium-dependent phosphnolipid-binding protein that plays a role in many cellular functions. It has been proven that annexin II has been involved in many human cancers. The aim of this study is to examine the expression of annexin II in lung cancer tissues, and analyze the correlations between annexin II and clinicopathological parameters. Annexin II expression was examined in 110 lung cancer specimens using immunohistochemistry (SP method). Western Blot was used to detect the expression of annexin II in lung cancer tissues and the corresponding normal lung tissues. Among 110 lung cancer specimens, 67 samples showed positive expression of annexin II (60.9%). It showed membranous staining in lung squamous cell carcinoma. The positive membranous expression and cytoplasmic expression of annexin II in lung adenocarcinoma mainly showed membranous staining. Annexin II expression level was positively correlated with lymphoid node metastasis (P =0.008) and TNM stages (P =0.002). There was expression of annexin II in both lung cancer tissues and the corresponding normal lung tissues, and in lung cancer tissues, the total levels of annexin II proteins were significantly higher than that in the corresponding normal lung tissues (P <0.001). The expression of annexin II positively may correlated with lung squamous cell carcinoma and adenocarcinoma development.


[Methods of diagnosing unstable stenocardia and monitoring its treatment].
The informative value of various functional and laboratory diagnostic methods was reviewed in 150 patients with unstable angina (UA). High informativeness (92%) and sensitivity (83%) of isometric hand grip under computerized electrocardiographic monitoring was demonstrated for the diagnosis and control of treatment in these patients. Isometric loading brought out a dramatically reduced coronary reserve, latent heart failure and electrical instability of the myocardium. The cold test with a small area of skin exposure is of low diagnostic value in UA cases and cannot be recommended for clinical use. Serial myoglobin assays add to the accuracy of diagnosis and have high predictive value.


Effects of a lactoperoxidase system-containing toothpaste on levels of hypothiocyanite and bacteria in saliva.
Lactoperoxidase (LPO)/H2O2/SCN(-)-system-generated hypothiocyanite ions (OSCN-) and hypothiocyanous acid (HOSCN) are inhibitory against a number of oral bacteria, including mutans streptococci. A commercially available toothpaste (Biotene) comprises the complete LPO system. Generation of HOSCN/OSCN- by Biotene was studied in vitro both in sterilized and nonsterilized saliva of 10 healthy subjects. The HOSCN/OSCN- yield ranged from 100 to 300 microM with Biotene, while the salivary levels of HOSCN/OSCN- before the addition of Biotene were 30.1 +/- 25.1 microM. Two in vivo trials were carried out. In the first study, resting saliva was collected from 12 individuals before, immediately after, and 2, 5, 10 and 20 min after brushing with Biotene to evaluate the in vivo generation and decomposition of HOSCN/OSCN-. In the second study, 26 healthy individuals attended a 1-month crossover trial with Biotene and a control toothpaste, Vademecum (no LPO system), both containing F- and xylitol. The salivary counts of total streptococci, mutans streptococci (MS), lactobacilli and the total flora (TF), as well as the peroxidase, thiocyanate ion and HOSCN/OSCN- levels were determined before and after 2 and 4 weeks daily use of the toothpastes. Twice-a-day use of Biotene for 1 month resulted in an elevation of 'resting levels' of HOSCN/OSCN-. No such effect was found with the control toothpaste. No significant changes were found in the salivary levels of total streptococci, MS, lactobacilli or TF after 1-month use of either toothpaste. The results show the capability of the LPO-system-containing toothpaste to elevate the salivary levels of HOSCN/OSCN-, although no bactericidal effect was observed.


Amiodarone-induced thyrotoxicosis: clinical presentation and expanded indications for thyroidectomy.
Amiodarone-induced thyrotoxicosis (AIT) is a rare disorder that is frequently refractory to conventional pharmacologic therapy. An analysis of seven patients who underwent thyroidectomy for control of AIT between 1987 and 1993 was completed to examine the clinical manifestations and the indications for surgical therapy. Five men and two women, 53 to 72 years of age, had AIT after 3 to 55 months of amiodarone treatment (mean, 21 months). The primary manifestations of AIT were ventricular tachycardia (five), exacerbation of chronic pulmonary disease (one), and occult hyperthyroidism (one). Medical therapy included propylthiouracil in doses up to 1200 mg/day in five patients, a beta-receptor antagonist in three, and withdrawal of amiodarone in five. Near-total or total thyroidectomy resulted in resolution of thyrotoxicosis in all patients. Morbidity included pneumonia (one) and cardiac dysrhythmias (two). One patient died of ventricular dysrhythmias 4 months after thyroidectomy. The mean thyroid weight was 50 gm (range, 17 to 216 gm). Microscopic examination showed destructive follicular lesions with fibrosis in all patients. Associated thyroid pathologic condition included multinodular goiter in four patients and a follicular adenoma in one. AIT may be clinically occult or manifested by unusual symptoms requiring a high index of suspicion for diagnosis. Near-total thyroidectomy is safe and effective in producing rapid resolution of AIT and is indicated for the initial treatment of patients who present with a resurgence of life-threatening cardiac arrhythmias and for all patients with AIT refractory to medical therapy.


Circulating Differentially Methylated Amylin DNA as a Biomarker of β-Cell Loss in Type 1 Diabetes.
In type 1 diabetes (T1D), β-cell loss is silent during disease progression. Methylation-sensitive quantitative real-time PCR (qPCR) of β-cell-derived DNA in the blood can serve as a biomarker of β-cell death in T1D. Amylin is highly expressed by β-cells in the islet. Here we examined whether demethylated circulating free amylin DNA (cfDNA) may serve as a biomarker of β-cell death in T1D. β cells showed unique methylation patterns within the amylin coding region that were not observed with other tissues. The design and use of methylation-specific primers yielded a strong signal for demethylated amylin in purified DNA from murine islets when compared with other tissues. Similarly, methylation-specific primers detected high levels of demethylated amylin DNA in human islets and enriched human β-cells. In vivo testing of the primers revealed an increase in demethylated amylin cfDNA in sera of non-obese diabetic (NOD) mice during T1D progression and following the development of hyperglycemia. This increase in amylin cfDNA did not mirror the increase in insulin cfDNA, suggesting that amylin cfDNA may detect β-cell loss in serum samples where insulin cfDNA is undetected. Finally, purified cfDNA from recent onset T1D patients yielded a high signal for demethylated amylin cfDNA when compared with matched healthy controls. These findings support the use of demethylated amylin cfDNA for detection of β-cell-derived DNA. When utilized in conjunction with insulin, this latest assay provides a comprehensive multi-gene approach for the detection of β-cell loss.


Reducing cancer risk with vitamins C, e, and selenium.
Abstract Current research has provided evidence that nearly 90 percent of all cancers may be related to diet, environment, and lifestyle. Of this number, 30 to 40 percent of cancers in men and up to 60 percent of cancers in women may be related to diet and nutrition. The two-stage process in the formation of many cancers, defined as initiation and promotion, is influenced by many dietary components. Vitamins C, E, and the mineral selenium are nutrients that function as antioxidants, reducing potential cancer-causing chemicals in the body. These natural anticarcinogens are thought to alter the cancer process and are currently under study for their cancer prevention properties. The functions, Recommended Dietary Allowances, food sources, research evidence for cancer prevention, and recommendations for supplementation are presented for these three nutrients. Research suggests that the proper and prudent use of nutrients, along with a healthy diet and lifestyle, may offer protection against this devastating disease.


Iron nanoparticle templates for constructing 3D graphene framework with enhanced performance in sodium-ion batteries.
This study examines the synthesis and electrochemical performance of three-dimensional graphene for Li-ion batteries and Na-ion batteries. The in situ formation of iron hydroxide nanoparticles (Fe(OH)x NPs) of various weights on the surface of graphene oxide, followed by thermal treatment at elevated temperature and washing using hydrochloric acid, furnished 3D graphene. The characterization studies confirmed the prevention of graphene layer stacking by over 90% compared with thermal treatment without Fe(OH)x. The electrochemical performance of the 3D graphene was evaluated as a counter electrode for lithium metal and sodium metal in a half-cell configuration. This material showed good performances with a charging capacity of 507 mA h g-1 at 372 mA g-1 in Li-ion batteries and 252 mA h g-1 at 100 mA g-1 in Na-ion batteries, which is 1.4 and 1.9 times higher, respectively, than the graphene prepared without Fe(OH)x templates.


Metabolic depletion decreases the aggregability of erythrocytes.
We studied the influence of metabolic depletion on red blood cell (RBC) aggregability, which is a determinant of blood flow. Heparinized blood was stored at room temperature for 0, 24, and 48 h. RBCs were washed twice and resuspended in Tris-buffer containing 3% dextran 70 (hematocrit 30%). Suspension viscosities were measured at 37 °C and shear rates of 37.6 and 0.1 s(-1), RBC aggregability was analysed by the sedimentation rate, direct microscopic visualization and a Myrenne aggregometer. RBCs in autologous plasma showed an increasing echinocytic shape transformation, which was reversible in buffer. The viscosities of RBC suspensions in buffer remained unchanged at both low (0.1 s(-1)) and high shear rate (37.6 s(-1)), the latter result indicating an unchanged RBC deformability. RBC aggregability decreased: The RBC sedimentation rates were 40.7 ± 5.0, 29.3 ± 13.4, and 13.3 ± 11.2 mm/h (p < 0.001) at 0, 24, and 48 h, respectively, which correlated well with the visual aggregability index and the Myrenne aggregation parameters M and M1. We conclude that metabolic depletion for 48 h leads to RBC swelling and a reversible echinocytic shape transformation. These ATP-depleted, but normally shaped RBCs had a decreased aggregability. In contrast to all other methods used, low shear viscosity was inaccurate and should not be used to test RBC aggregability.


Rat Plasma Oxidation Status After Nigella Sativa L. Botanical Treatment in CCL(4)-Treated Rats.
ABSTRACT Nigella sativa Linn. (family Ranunculaceae), commonly known as black cumin, is native to the Mediterranean area and has been used for thousands of years as a health and beauty aid. The present study investigated the protective effects of Nigella sativa (NS) extract (NSE) and oil (NSO) on CCl(4)-induced nitrosative stress and protein oxidation in rat. CCl(4) (0.8 mg/kg) was used as an aid for induction of nitrosative stress. In vitro antioxidant potential was tested in the presence of 2,4-dinitrophenylhyrdazine (DPPH) as an organic nitrogen radical. Doses of 0.2, 0.3, and 1 mg/kg of the NS extract and oil were administered to CCL(4)-treated rats for 10 days. At the end of treatment, blood was taken from rats under anesthesia and plasma was separated. The concentration of nitric oxide (NO), total antioxidant power (TAP), carbonyl molecules (CM) as measure of protein oxidation (PO), tumor necrosis factor-alpha (TNF-alpha), and total thiol molecules (TTM) were measured in plasma. In vitro evaluation of antioxidant effects of NSE and NSO showed that the highest antioxidant activity (80%) was observed with the concentration of 10 and 20 mg/ml, respectively, that were equal to vitamin E (200 mg/ml). Administration of CCL(4) increased plasma PO, NO, TNF-alpha and decreased TAP and TTM. Both NSE and NSO showed significant protection against CCl(4)-induced changes in biochemical parameters, but not dose-dependently. Doses of 0.3 and 1 mg/kg were more effective than doses of 0.2 mg/kg for both NSE and NSO, but dose of 1 mg/kg was the most effective one. The results indicate the potential of NS in preventing CCL(4)-induced toxic nitrosative stress. It is concluded that NS has marked antioxidant potentials that may be beneficial in alleviating complications of many illnesses related to oxidative/nitrosative stress in humans, but preclinical safety measures should be completed before clinical trials.


Autoimmune Regulator (AIRE) Deficiency Does Not Affect Atherosclerosis and CD4 T Cell Immune Tolerance to Apolipoprotein B.
Atherosclerosis is a chronic, lipid-driven disease of medium sized arteries which causes myocardial infarction and stroke. Recently, an adaptive immune response against the plaque-associated autoantigen Apolipoprotein B100 (ApoB), the structural protein component of low-density lipoprotein, has been implicated in atherogenesis. In healthy individuals, CD4+ T cells responding to ApoB mainly comprised regulatory T cells, which confer immune tolerance and atheroprotection. Mice and patients with atherosclerosis harbor increased numbers of proatherogenic ApoB-reactive T-helper cell subsets. Given the lack of therapies targeting proatherogenic immunity, clarification of the underlying mechanisms is of high clinical relevance. T cells develop in the thymus, where strong autoreactive T cells are eliminated in the process of negative selection. Herein, we investigated whether the transcription factor autoimmune regulator (AIRE), which controls expression of numerous tissue-restricted self-antigens in the thymus, is involved in mediating tolerance to ApoB and whether Aire deficiency might contribute to atherogenesis. Mice deficient for Aire were crossbred to apolipoprotein E-deficient mice to obtain atherosclerosis-prone Aire -/- Apoe -/- mice, which were fed a regular chow diet (CD) or western-type diet (WD). CD4+ T cells responding to the ApoB peptide p6 were analyzed by flow cytometry. We demonstrate that Aire deficiency influences neither generation nor activation of ApoB-reactive T cells and has only minor and overall inconsistent impacts on their phenotype. Furthermore, we show that atherosclerotic plaque size is not affected in Aire -/- Apoe -/- compared to Aire +/+ Apoe -/-, irrespective of diet and gender. In conclusion, our data suggests that AIRE is not involved in regulating thymic expression of ApoB or atherosclerosis. Alternative mechanisms how ApoB-reactive CD4 T cells are selected in the thymus will have to be investigated.


Fatal intracranial extension of spinal hemorrhage after lumbar puncture.
Lumbar puncture (LP) is sometimes performed in addition to computed tomography (CT) to exclude hemorrhage prior to anticoagulation for suspected cerebral ischemia. The LP may be complicated by spinal hemorrhage during subsequent anticoagulation. We report such a case with an eventually fatal outcome due to intracranial extension of blood. On balance, the use of LP in addition to CT in this setting appears to be unwise.


[LASER TECHNOLOGIES IN LAPAROSCOPIC HERNIOLOGY].
Objective - to analyze the expediency of using a universal laser coagulator "Lika-surgeon" with a wavelength of 1470 nm as an alternative to an electrocoagulation while performing dissection for TAPP. The work is based on the analysis of 88 TAPP patients, using a universal laser "Lika-surgeon" with a wavelength of 1470 nm at a radiation power of 10-12.5 W and 102 patients operated according to the trivial standard TAPP procedure, where an electrocoagulation hook with a coagulation and cutting power of 35 W is used as a dissection instrument. Evaluation considered the criteria of operating time, duration of hospital stay, severity of pain syndrome, morphological assessment of the prevalence of laser and electrical effects on the tissues in the affected area.. All surgical interventions were performed in the Department of General and Vascular Surgery of the PNCO "Kyiv City Hospital №1, Shalimov's National Institute Of Surgery And Transplantation National Academy Of Medical Sciences Of Ukraine and Republican Clinical Hospital Of Batumi in 2020. The cutting mode in an electrosurgical apparatus is accompanied by tissue wrinkling and smoke in the operating field. The laser device is devoid of these disadvantages. At the same time, laser coagulation requires practical skills of the operator to select the optimal distance between the optical fiber and the object. The operating time and the duration of hospital stay in patients of the first group are significantly shorter. The severity of pain in the early postoperative period was comparable in both groups. In the late postoperative period, the pain syndrome was more severe in the second group - it accompanied 6.1% of patients in the first group and 15.2% in the second group. Analysis of the morphological material showed significantly less tissue trauma in the affected zone in the first group - the coagulation zone is 372.91 µm versus 835.88 µm in the second group. A laser coagulator with a wavelength of 1470 nm has a number of significant advantages over an electrosurgical coagulator: more efficient cutting and coagulation modes; optical clarity of the operating field; minimal thermal damage of the underlying tissues (the depth of tissue damage in the first case is 166.36 microns, in the second 835.88 microns); significant reduction of operating tine and the duration of hospital stay; significant reduction of pain in the postoperative period. This opens up new perspectives for its use in laparoscopic surgery of inguinal hernia.


Leadership 99. Interview by Terese Hudson.
Cost control, customer service and collaboration among health care sectors rank as top concerns with panelists in H&HN's annual Leadership Report. Efforts to improve community health are a priority, too, but are often frustrated by financial and other constraints. The 16 panelists represent managed care, physicians, and hospitals and health systems.


The results of radial artery Y-graft for complete arterial revascularization.
Harvesting of multiple arterial grafts is commonly associated with prolonged operating times and increased trauma in complete arterial coronary artery bypass grafting (CABG). Using sequential grafting techniques, CABG is possible with only two arterial grafts in multi-vessel coronary artery disease (CAD). However, sequential grafting may not be convenient for all circumstances and sometimes surgical technique may be challenging. We present our experience in the use of radial artery (RA) Y-graft on a routine basis. Between January 1996 and November 2001, 127 patients (aged 63+/-8 years) with the diagnosis of multi-vessel disease underwent complete arterial revascularization using left internal mammarian artery (LIMA) and RA. Left ventricular ejection fraction ranged from 23 to 65% (mean 51+/-11%). Triple-vessel disease was present in 73.2% of patients. We used the division technique of RA during harvesting and formation of one or more composite Y-grafts of the RA itself to allow end-side rather than sequential anastomoses without any significant decrease the usable conduit length. The results of this technique were compared with the data of patients (n=109) who underwent completely arterial CABG with the use of the multiple arterial grafts in the same period. LIMA was anastomosed to the left anterior descending coronary artery (LAD) system in all patients. Two to four (mean 2.8+/-0.6) anastomoses were performed with RA Y-graft per patient. Proximal end of the radial graft was anastomosed to LIMA (60.6%) or aorta (39.4%). Mean operating time was 185 (45 min; bypass time, 68+/-23 min; and cross-clamp time, 49+/-17 min). Perioperative intraaortic balloon pump was necessary in five patients (3.9%). There was no operative mortality or morbidity. During the follow-up period of 2-30 months, none of the patients had any complication. Postoperative coronary angiography in 54 patients (42.5%) documented excellent early patency rates (LIMA 100%, and RA 98.1%). We believe that keeping our technique in their armamentarium will be useful for cardiac surgeons as an alternative method during complete arterial revascularization. This approach allows for complete arterial revascularization in multi-vessel CAD using only single IMA and RA grafts with excellent early results.


The CC homozygosis of the -174G>C IL-6 polymorphism predicts a lower efficacy of rituximab therapy in rheumatoid arthritis.
Identification of genetic biomarkers of response to biologics in rheumatoid arthritis (RA) is a relevant issue. Being IL-6 a key cytokine for B cell survival, the interleukin-6 (IL-6) -174G>C and the IL-6 receptor (IL-6R) D358A gene polymorphisms were investigated in 158 RA patients treated with rituximab (RTX). One hundred and twenty-eight (81.0%) were RF positive and 126 (79.7%) were anti-CCP positive. Response to therapy was evaluated at the end of the sixth month after the first RTX infusion, by using both the EULAR and the ACR criteria. The possible relationship with IL-6 serum levels was also studied. By univariate analysis, lack of response by the EULAR criteria was more prevalent in RA patients with the IL-6 -174 CC genotypes (39.1%), than in the GC/GG patients (18.5%) (OR 2.83; 95%CI=1.10-7.27; p=0.031). A good response was noticed in only one patient (4.3%) with the IL-6 -174 CC genotype, while it was present in 24.4% of GG/GC cases (p=0.06). By stepwise multivariate analysis (including RA duration, baseline DAS28, baseline HAQ, RF status, anti-CCP status and IL-6 genotype as covariates), the IL-6 -174CC genotype was selected as an independent predictor of no response to RTX by both EULAR and ACR≥50 criteria, while the IL-6R polymorphism resulted as not associated. No definite association between gene polymorphisms and IL-6 serum levels was noticed. Present results suggest a possible role for IL-6 genotyping to better plan treatment with RTX in RA, and larger studies are worthwhile.


New evidence for the intermediate position of relaxed accommodation.
Experimental evidence is presented that the focus of the eye tends to return passively to an individually characteristic intermediate resting position or dark-focus whenever (1) the stimulus to accommodation is degraded or (2) when the quality of the image is independent of focus. Based on measurements of the dark-focus with a laser optometer, it is possible to predict the magnitude of night, empty field, and instrument myopia on an individual basis. The role of the intermediate dark-focus as a factor in accommodation also provides an explanation for the paradoxical variation of visual acuity with observation distance. Applications to night myopia and night driving are described, and implications for clinical practice are discussed.


GABAA receptor but not muscarinic receptor density was decreased in the brain of patients with Parkinson's disease.
The activity of glutamic acid decarboxylase (GAD) and choline acetyltransferase (ChAT) as presynaptic markers of gamma-aminobutyric acid (GABA)- and acetylcholine (ACh)-containing neurons, and the binding of [3H]muscimol and [3H]quinuclidinyl benzilate ([3H]QNB) as postsynaptic ones were measured in autopsied samples of the caudate nucleus, putamen, pallidum, substantia nigra and the cerebral cortex from L-dopa-treated patients with Stage V (terminally bedridden) patients with Parkinson's Disease (PD). In PD, GAD activities were significantly reduced in the caudate nucleus and substantia nigra relative to normal controls, but were normal when the values from protracted terminal illness (PTI) cases were used as the controls. ChAT activities were reduced in all regions studied. These reductions in GAD and ChAT activities were not accompanied by a concomitant increase in the density of GABAA or muscarinic receptors. GABAA receptor densities were significantly decreased in both the cortical and subcortical brain regions, while muscarinic receptor densities remained unchanged. We suggest that the decreased density of GABAA receptor in PD brains reflects degeneration of neurons on which the receptor is localized, i.e., degeneration of ascending monoaminergic neurons including nigral dopamine (DA) neurons.


The "new" immigration and the politics of cultural diversity in the United States and Japan.
"Certain parallels between the recent experience of Japan and that of the United States with immigration from Third World countries are increasingly evident. In this discussion, I shall focus on these key similarities rather than the obvious differences between the two countries, in terms of culture, economy, political system, and 'immigration profile'." Aspects considered include the structural nature of the demand for foreign labor, the composition of immigrant stocks and flows, public tolerance, and contradictions in government policies.


Towards Structure-Guided Development of Pain Therapeutics Targeting Voltage-Gated Sodium Channels.
Voltage-gated sodium (NaV) channels are critical molecular determinants of action potential generation and propagation in excitable cells. Normal NaV channel function disruption can affect physiological neuronal signaling and lead to increased sensitivity to pain, congenital indifference to pain, uncoordinated movement, seizures, or paralysis. Human genetic studies have identified human NaV1.7 (hNaV1.7), hNaV1.8, and hNaV1.9 channel subtypes as crucial players in pain signaling. The premise that subtype selective NaV inhibitors can reduce pain has been reinforced through intensive target validation and therapeutic development efforts. However, an ideal therapeutic has yet to emerge. This review is focused on recent progress, current challenges, and future opportunities to develop NaV channel targeting small molecules and peptides as non-addictive therapeutics to treat pain.


Direct Detection of Dilute Solid Chemicals with Responsive Lateral Organic Diodes.
Organic field-effect transistors (OFETs) have emerged as promising sensors targeting chemical analytes in vapors and liquids. However, the direct detection of solid chemicals by OFETs has not been achieved. Here for the first time, we describe the direct detection of solid chemical analytes by organic electronics. An organic diode structure based on a horizontal side-by-side p-n junction was adopted and shown to be superior to OFETs for this purpose. The diodes showed more than 40% current decrease upon exposure to 1 ppm melamine powders. The estimated detection limit to melamine can potentially reach the ppb range. This is the first demonstration of an electronic signal from an interaction between a solid and an organic p-n junction directly, which suggests that our lateral organic diodes are excellent platforms for the development of future sensors when direct detection of solid chemicals is needed. The approach developed here is general and can be extended to chemical sensors targeting various analytes, opening unprecedented opportunities for the development of low-cost and high-performance solid chemical sensors.


Total Synthesis of the Proposed Structure of Marineosin A.
A total synthesis of a proposed structure of marineosin A has been achieved. The key steps involve Lewis acid catalyzed spirocyclizaton, ring-closing metathesis, Paal-Knorr pyrrole synthesis, and a Vilsmeier-Haack type reaction with Tf2O.


Long-term results of Heller myotomy without an antireflux procedure in achalasic patients.
Both open and laparoscopic myotomies have been used in the treatment of achalasia. Postoperative gastro-oesophageal reflux is among the commonly reported side effects of myotomy. The addition of an antireflux procedure to the standard surgical approach has given rise to controversy. The objective of our study was to determine whether or not an antireflux procedure should be used in addition to Heller myotomy. Over the period from 1980 to 1990, 94 patients (mean age: 47.9 years) with achalasia underwent Heller myotomy calibrated by intraoperative oesophageal manometry without fundoplication. In 1999-2000, all patients filled in a clinical questionnaire: all underwent radiographic oesophageal imaging, oesophageal manometry, ambulatory 24-h oesophageal pH monitoring, and oesophagogastroduodenoscopy, when necessary. Ten healthy age-matched subjects were compared in the manometric and radiological studies. Myotomy improved the clinical profiles and instrumental data results in all patients. Gastro-oesophageal reflux was present in 10 patients (10.6%); none of these 10 subjects presented oesophagitis. Heller open myotomy yields good long-term results. Intraoperative manometric calibration reduces the side effects of myotomy, such as gastro-oesophageal reflux. The addition of fundoplication is not justified in all patients.


Prevalence of Mental Health problems in sentenced men in prisons from Andalucía (Spain).
To estimate the prevalence of different mental health problems in men serving prison sentences in Andalusia. Descriptive, cross-sectional study of a random sample of 472 men interned in two prisons located in Andalusia. We collected socio-demographic and general criminal and penitentiary data, and we identified mental health problems with two validated instruments for epidemiological research in mental health: the SCID-I interview to diagnose Axis 1 disorders of the DSM-IV and the self-applied questionnaire IPDE to estimate personality disorders. We analyzed the data (proportions and confidence intervals) with the SPSS-18 statistical package. 82.6% of the sample had a history of having suffered some type of mental health problem throughout their life (prevalence-life) and 25.8 have suffered from them in the past month (month prevalence). The most common disorders of the Axis I (DSM-IV) are related to abuse of and dependence on psychoactive substances (prevalence life of 65.9% and month prevalence of 6.6%), with an important but less frequent presence of affective (31.4%-9.3%), anxiety (30.9%-10, 4%) and psychotic disorders (9.5%-3, 4%). As regards personality disorders, the estimated probable prevalence lies between the 56.6% ("5" cutoff point) and the 79.9 ("4" cut-off point). The male inmate population in prisons in Andalucía shows a high prevalence of mental health problems, similar to that found in other Spanish and international prisons, but their care needs should take into account the different pathologies that they present.


[Study of breast carcinoma with non-inflammatory skin involvement (t4b)].
In this study, we evaluated the clinical presentation of patients with T4b breast cancer, and analyzed the impact of non- inflammatory skin involvement on long-term survival. We treated 1 228 cases of breast cancer patients during 2004 to 2010. The T4b breast cancers were 52 cases out of these 1,228 patients. The primary treatment methods of these T4b patients were chemotherapy (67.3%), surgical therapy (26.9%), endocrine therapy (5.8%), and radiation therapy (0%). The probability that surgical resection was performed over the duration of treatment was 88. 5%. The mean time to progression (TTP) was 3.9 years. The disease specific mean survival time (MST) of T4b breast cancer patients was 5.9 years. There was no difference among the surgical and non-surgical therapy groups in the long-term survival period, but the surgical therapy group was much better than non-surgical therapy group in terms of non-recurrent survival ratio. In viewing from the control of bleeding of the patients, the surgical therapy was effective and improved the quality of life during a time of the therapy.


SPATA18 Expression Predicts Favorable Clinical Outcome in Colorectal Cancer.
Dysregulation of mitochondrial quality control has been reported to be associated with cancer and degenerative diseases. SPATA18 (spermatogenesis-associated 18, also known as Mieap) encodes a p53-inducible protein that can induce lysosome-like organelles within mitochondria that eliminate oxidized mitochondrial proteins and has tumor suppressor functions in mitochondrial quality control. In the present study, 268 primary colorectal cancers (CRCs) were evaluated immunohistochemically for SPATA18 expression to assess its predictive utility and its association with cellular proliferation activity. Furthermore, the association with p53 immunoreactivity, a surrogate marker for TP53 mutation, was analyzed. Non-neoplastic colonic mucosa showed cytoplasmic SPATA18 expression. Seventy-two percent of the lesions (193/268) displayed high SPATA18 expression in the cytoplasm of CRC cells. Univariate analyses revealed significant associations between SPATA18 expression and tumor size (p < 0.0001), histological differentiation (p = 0.0017), and lymph node metastasis (p = 0.00039). The log-rank test revealed that patients with SPATA18-high CRCs had significantly better survival than SPATA18-low patients (p < 0.0001). Multivariate Cox hazards regression analysis identified tubular-forming histology (hazard ratio [HR] = 0.25), age < 70 years (HR = 0.50), and SPATA18-high (HR = 0.55) as potential favorable factors. Lymph node metastasis (HR = 1.98) and peritoneal metastasis (HR = 5.45) were cited as potential independent risk factors. Cellular proliferation activity was significantly higher in SPATA18-high tumors. However, no significant correlation was detected between SPATA18 expression and p53 immunoreactivity or KRAS/BRAF mutation status. On the basis of our observations, SPATA18 immunohistochemistry can be used in the prognostication of CRC patients.


[BDNF measurement in stress-related mood disorders: a review of clinical studies].
The study of brain-derived neurotrophic factor (BDNF) in mood disorders and other psychiatric disorders is increasing. Of particular interest is whether or not BDNF can be a marker for psychiatric disorders. The aim of this study was to review the published findings on BDNF in stress-related mood disorders. Searches were conducted of the PubMed and Psikiyatri Dizini databases to access international and national publications, respectively. Database searches were conducted with the keywords BDNF, and stress and mood disorders. BDNF plays a role in the physiopathology of stress-related changes and is supposedly related to a decrease in the number and size of neurons. Additionally, it has been suggested that serum levels of BDNF are lower in depressive patients than in the healthy controls. Nonetheless, BDNF levels increase after antidepressant treatment, along with symptom recovery, to those seen in healthy control subjects. This increase cannot be achieved in patients that do not respond to antidepressant treatment. On the other hand, in patients with stress-related non-depressive psychiatric disorders, serum BDNF is also low. To date, even though consistent findings of reduced BDNF levels have been reported, the specificity of these findings is doubtful. Serum BDNF might be considered a marker for stress-related mood disorders.


Effects of Immediate and Delayed Cementations for CAD/CAM Resin Block after Alumina Air Abrasion on Adhesion to Newly Developed Resin Cement.
The purpose of this study was to evaluate the effect of one week of Computer-aided design/Computer-aided manufacturing (CAD/CAM) crown storage on the μTBS between resin cement and CAD/CAM resin composite blocks. The micro-tensile bond strength (μTBS) test groups were divided into 4 conditions. There are two types of CAD/CAM resin composite blocks, namely A block and P block (KATANA Avencia Block and KATANA Avencia P Block, Kuraray Noritake Dental, Tokyo, Japan) and two types of resin cements. Additionally, there are two curing methods (light cure and chemical cure) prior to the μTBS test-Immediate: cementation was performed immediately; Delay: cementation was conducted after one week of storage in air under laboratory conditions. The effect of Immediate and Delayed cementations were evaluated by a μTBS test, surface roughness measurements, light intensity measurements, water sorption measurements and Scanning electron microscope/Energy dispersive X-ray spectrometry (SEM/EDS) analysis. From the results of the μTBS test, we found that Delayed cementation showed significantly lower bond strength than that of Immediate cementation for both resin cements and both curing methods using A block. There was no significant difference between the two types of resin cements or two curing methods. Furthermore, water sorption of A block was significantly higher than that of P block. Within the limitations of this study, alumina air abrasion of CAD/CAM resin composite restorations should be performed immediately before bonding at the chairside to minimize the effect of humidity on bonding.


High Glucose Induces Down-Regulated GRIM-19 Expression to Activate STAT3 Signaling and Promote Cell Proliferation in Cell Culture.
Recent studies indicated that Gene Associated with Retinoid-IFN-Induced Mortality 19 (GRIM-19), a newly discovered mitochondria-related protein, can regulate mitochondrial function and modulate cell viability possibly via interacting with STAT3 signal. In the present study we sought to test: 1) whether GRIM-19 is involved in high glucose (HG) induced altered cell metabolism in both cancer and cardiac cells, 2) whether GRIM-19/STAT3 signaling pathway plays a role in HG induced biological effects, especially whether AMPK activity could be involved. Our data showed that HG enhanced cell proliferation of both HeLa and H9C2 cells, which was closely associated with down-regulated GRIM-19 expression and increased phosphorylated STAT3 level. We showed that GRIM-19 knock-down alone in normal glucose cultured cells can also result in an increase in phosphorylated STAT3 level and enhanced proliferation capability, whereas GRIM-19 over-expression can abolished HG induced STAT3 activation and enhanced cell proliferation. Importantly, both down-regulated or over-expression of GRIM-19 increased lactate production in both HeLa and H9C2 cells. The activated STAT3 was responsible for increased cell proliferation as either AG-490, an inhibitor of JAK2, or siRNA targeting STAT3 can attenuate cell proliferation increased by HG. In addition, HG increased lactate acid levels in HeLa cells, which was also observed when GRIM-19 was genetically manipulated. However, HG did not affect the lactate levels in H9C2 cells. Of note, over-expression of GRIM-19 and silencing of STAT3 both increased lactate production in H9C2 cells. As expected, HG resulted in significant decreases in phosphorylated AMPKα levels in H9C2 cells, but not in HeLa cells. Interestingy, activation of AMPKα by metformin was associated with a reversal of the suppressed GRIM-19 expression in H9C2 cells, the fold of changes in GRIM-19 expression by metformin were much less in HeLa cells. Metformin did not affect the phosphorylated STAT3 lelvels, however, decreased its levels in H9C2, especially in the setting of HG culture. Not like HG alone which resulted in no changes in lactate acid in H9C2 cells, metformin can increase lactate acid levels in H9C2 cells. Increased lactate induced by metformin was also observed in HeLa cells.


Relations among aging, antioxidant status, and cataract.
Light and oxygen are necessary for the function of the eye. However, when present in excess or in uncontrolled circumstances, they appear to be related, probably causally, to the development of cataract. Compromises of function of the lens and retina with aging are exacerbated by depleted or diminished primary antioxidant reserves, antioxidant enzyme capabilities, and diminished secondary defenses such as proteases. Smoking appears to provide an additional oxidative challenge associated with depletion of antioxidants as well as with enhanced risk for cataract formation. Poor education and lower socioeconomic status are associated with poorer nutriture and are also significantly related to increased risk for these debilities. Optimizing nutriture, including diets rich in fruit and vegetables, may provide the least costly and most practicable means to delay cataract.


Outcome of acute respiratory distress syndrome patients treated with extracorporeal membrane oxygenation and brought to a referral center.
Patients with severe acute respiratory distress syndrome (ARDS) are candidates for extracorporeal membrane oxygenation (ECMO) therapy. The evaluation of organ severity is difficult in patients considered for cannulation in a distant hospital. This study was designed to identify early factors associated with hospital mortality in ARDS patients treated with ECMO and retrieved from referring hospitals. Data from 85 consecutive ARDS patients equipped with ECMO by our mobile team and consequently admitted to our ICU were prospectively collected and analyzed. The main ARDS etiologies were community-acquired bacterial pneumonia (35%), influenza pneumonia (23%) (with 12 patients having been treated during the first half of the study period), and nosocomial pneumonia (14%). The median (interquartile range) time between contact from the referring hospital and patient cannulation was 3 (1-4) h. ECMO was venovenous in 77 (91%) patients. No complications occurred during transport by our mobile unit. Forty-eight patients died at the hospital (56%). Based on a multivariate logistic regression, a score including age, SOFA score, and a diagnosis of influenza pneumonia was constructed. The probability of hospital mortality following ECMO initiation was 40% in the 0-2 score class (n = 58) and 93% in the 3-4 score class (n = 27). Patients with an influenza pneumonia diagnosis and a SOFA score before ECMO of less than 12 had a mortality rate of 22%. Age, SOFA score, and a diagnosis of influenza may be used to accurately evaluate the risk of death in ARDS patients considered for retrieval under ECMO from distant hospitals.


The effects of interval feedback on the self-efficacy of netball umpires.
The present study used quantitative and qualitative methods to explore the relationship between interval feedback and self-efficacy toward umpiring netball games. Grade "A "level umpires (n = 7) provided feedback to umpires (n = 40) under two conditions; 1) interval feedback given at the end of one tournament game (after 14 minutes) and again at the end of a second consecutive game (after 28 minutes), and 2) feedback at the end of the game (after 28 minutes). Umpires in both conditions completed an Umpiring Self-Efficacy Questionnaire (USEQ) which was a 14-item measure designed to assess factors relevant to netball umpire performance. Participants completed the USEQ immediately before game one, during the interval, and after a second game. Umpires also completed a feedback questionnaire which enabled them to reflect on the feedback received. A repeated measures factorial (time x feedback condition) ANOVA indicated no significant interaction effect (F = 0.05, p > .05), and no main effect for condition (F = 0.06, p > .05) or time (F = 1.61, p > .05) for changes in self-efficacy. Although there were no significant effects, qualitative data alluded to aspects of feedback perceived to enhance umpire self-efficacy, thus identifying ways in which feedback might have a more consistent effect. Practical implications of the study in relation to verbal interval feedback are discussed. Key pointsInterval feedback may detract from umpires processing of information during a game.Interval feedback can enhance self-efficacy.Findings show that personal and situational factors interacted to determine the outcome of feedback.Feedback should be tailored for the needs of the individual and take into account situational factors, to enhance the possibility of accruing positive outcomes.


Application of anticytokeratin antibodies to colon carcinomas.
Mouse monoclonal anticytokeratin antibodies (BA 16; BA 17; CO 8.2; C18.2; C 10; LE 41) were used for examination of 5 patients with large bowel carcinoma ("left colon") using the indirect immunoperoxidase technique. For each case cryostat sections were examined from three localizations: primary tumour of large bowel, mucosa in the vicinity of the tumour, and mucosa of distant part from the primary tumour. The antibodies BA 16 and BA 17 produced strongly positive results. Antibody C 18.2 seems to be less suitable for detection of cytokeratin expression in this part of large bowel. The obtained results indicate that no significant differences were found in expression of cytokeratins in the three localizations by histologically well or moderately differentiated adenocarcinoma.


Mechanisms of passive potassium influx in corn mitochondria.
Corn mitochondria in 100 millimolar KCl show accelerated passive swelling upon addition of uncoupler. This unusual response has been compared with swelling produced by valinomycin, tripropyltin, and nigericin. It is concluded that the driving force for swelling lies with the chloride gradient and a high P(Cl):P(K) ratio, the chloride influx creating a negative membrane potential. The action of uncoupler is to facilitate K(+) influx via the endogenous H(+)/K(+) antiporter. The antiporter is active over the pH range 6 to 8, is not sensitive to Mg(2+) concentration, and is not inactivated by aging. It is not clear why corn mitochondria show this exceptional activity of the H(+)/K(+) antiporter in K(+) influx. It is speculated that during isolation the antiporter may be exposed or activated, and that it contributes to cyclic K(+) transport and high State 4 respiration rates.


Effect of Cuttlebone on Healing of Indomethacin-Induced Acute Gastric Mucosal Lesions in Rats.
The continuing use of nonsteroidal anti-inflammatory drugs (NSAIDs) usually increases the side effects such as peptic ulcer and acute gastric lesions in the gastrointestinal tract. Cuttlebone (CB), isolated from Sepiella maindroni de Rochebrune, was reported to have antioxidant activities, but its role in the treatment of indomethacin-induced gastric lesions has not yet been confirmed. In this research, we investigate the protective effect of cuttlebone on indomethacin-related ulcers in rats and possible mechanisms. Here, gastric ulcers were induced by oral administration of indomethacin, and then the rats were treated with omeprazole (4 mg/kg) or different doses (750, 1500, and 3000 mg/kg of body weight) of cuttlebone. We evaluated lesion index, inflammation score, and a series of oxidant/antioxidant parameters. The data demonstrated that cuttlebone could protect against gastric ulcers induced by indomethacin in a dose-dependent manner (positive correlation). Also, these effects were associated with attenuating the expression of malonaldehyde (MDA) and increasing the levels of some protective ingredients like epidermal growth factor (EGF), prostaglandin E2 (PGE2), and superoxide dismutase (SOD). Thus, considering its ability to protect indomethacin-induced acute gastric mucosal lesions and the underlying mechanisms, CB might be a potential candidate for treating gastric damage caused by NSAIDs.


Tissue-specific expression of the nonneuronal promoter of the aromatic L-amino acid decarboxylase gene is regulated by hepatocyte nuclear factor 1.
The rat aromatic l-amino acid decarboxylase (AADC) gene contains alternative promoters which direct expression of neuronal and nonneuronal mRNAs that differ only in their 5'-untranslated regions (UTRs). We have analyzed the expression of the nonneuronal promoter of the rat AADC gene in the kidney epithelial cell line LLC-PK1 and in cells which do not express the nonneuronal form of AADC by transient transfection. These studies revealed that the first 1.1 kilobases of the nonneuronal promoter, including the nonneuronal-specific 5'-UTR (Exon 1), contains sufficient information to direct tissue-specific expression. Serial deletions of this promoter localized the cis-active element to a region between -52 and -28 base pairs upstream of the nonneuronal transcription start site. An A/T-rich sequence, within this region which we have termed KL-1, was found to bind a kidney and liver-specific factor by DNase footprint analysis and was capable of directing tissue-specific expression from a heterologous promoter. Moreover, when the KL-1 sequence was mutated in the context of the entire promoter sequence, all transcriptional activity was abolished. DNA sequence comparison revealed that the KL-1 fragment is highly homologous to the binding site for hepatocyte nuclear factor-1 (HNF-1). Mobility shift studies utilizing an antibody to HNF-1 demonstrated binding of HNF-1 to the KL-1 fragment and cotransfection of HNF-1 cDNA into cells which do not express the nonneuronal form of AADC resulted in activation of transfected AADC nonneuronal promoter constructs. These results strongly suggest that the transcription factor which regulates the tissue-specific expression of the nonneuronal form of AADC mRNA is HNF-1.


Faecal elastase-1 and fat-soluble vitamin profiles in patients with cystic fibrosis in Western Norway.
Exocrine pancreatic insufficiency is a major clinical manifestation of cystic fibrosis (CF). Almost nine of ten patients develop signs and symptoms of maldigestion and malabsorption, which often deteriorates nutritional status and therefore worsens the prognosis. Human faecal elastase-1 (FE-1) has shown promising results to assess exocrine pancreatic insufficiency, and this test has been used at Haukeland University Hospital since 1996. To evaluate FE-1 values and fat-soluble vitamin profiles in patients with CF and to correlate exocrine pancreatic function as measured as FE-1 to fat-soluble vitamin profiles. Moreover, we wanted to assess if there are differences between fat-soluble vitamin profiles in patients with impaired versus patent exocrine pancreatic function, and thirdly, if fat-soluble vitamin deficiency at diagnosis is effectively treated by supplementation. Consecutive analyses (N = 212) of fat-soluble vitamin profiles and 35 analyses of FE-1 were investigated in 35 patients with CF. In 17 out of 35 patients fat-soluble vitamin profiles were also assessed at diagnosis. Results Mean value of FE-1 for all CF patients was 256.9 microg/g faeces (median 24.1 microg/g faeces). CF patients considered to have maldigestion (N=24) showed a mean value of 19.9 microg/g faeces (median 18.7 microg/g faeces), those without pancreas affection had a mean value of 773.9 microg/g faeces (median 728.9 microg/g faeces, p < 0.01). There was no difference in fat-soluble vitamin profiles among patients with or without exocrine pancreatic insufficiency while on appropriate supplementation. Median value for vitamin E in patients with exocrine pancreatic insufficiency at diagnosis was low (3.6 mg/L). Supplementation of pancreatic enzymes and vitamins normalised profiles in this group at follow-up. There was no significant correlation between exocrine pancreatic function as measured as FE-1 and fat-soluble vitamin profiles, neither in patients with impaired nor in those with patent pancreatic function. Severe degree of exocrine pancreatic insufficiency is common in patients with cystic fibrosis. There was no correlation of faecal elastase-1 levels to fat-soluble vitamin status. Fat-soluble vitamins (A, D, E) given in appropriate dosages combined with pancreatic enzymes ensured normal profiles in our patients with CF and malabsorption. Officially recommended supplementation of vitamin A and D in Norway during infancy and childhood may explain why so few patients had vitamin deficiencies at diagnosis.


Impact of Nanoparticles on Brain Health: An Up to Date Overview.
Nanoparticles are zero-dimensional nanomaterials and, based on their nature, they can be categorized into organic, inorganic, and composites nanoparticles. Due to their unique physical and chemical properties, nanoparticles are extensively used in a variety of fields, including medicine, pharmaceutics, and food industry. Although they have the potential to improve the diagnosis and treatment of brain diseases, it is fundamentally important to develop standardized toxicological studies, which can prevent the induction of neurotoxic effects. The focus of this review is to emphasize both the beneficial and negative effects of nanoparticles on brain health.


Manipulation of malic enzyme in Saccharomyces cerevisiae for increasing NADPH production capacity aerobically in different cellular compartments.
The yeast Saccharomyces cerevisiae is an attractive cell factory, but in many cases there are constraints related with balancing the formation and consumption of redox cofactors. In this work, we studied the effect of having an additional source of NADPH in the cell. In order to do this, two strains were engineered by overexpression of malic enzyme. In one of them, malic enzyme was overexpressed as its wild-type mitochondrial form, and in the other strain a short form lacking the mitochondrial targeting sequence was overexpressed. The recombinant strains were analyzed in aerobic batch and continuous cultivations, and the basic growth characteristics were generally not affected to a great extent, even though pleiotropic effects of the manipulations could be seen by the altered in vitro activities of selected enzymes of the central metabolism. Moreover, the decreased pentose-phosphate pathway flux and the ratios of redox cofactors showed that a net transhydrogenase effect was obtained, which can be directed to the cytosol or the mitochondria. This may find application in redirecting fluxes for improving specific biotechnological applications.


The miRNA targetome of coronary artery disease is perturbed by functional polymorphisms identified and prioritized by in-depth bioinformatics analyses exploiting genome-wide association studies.
In recent years, genome-wide association studies (GWAS) have made great progress in elucidating the genetic influence on complex traits. An overwhelming number of GWAS signals resides in regulatory elements, therefore most post-GWAS studies focused only on transcriptional regulatory variants. However, recent findings have expanded the spectrum of trait/disease-associated regulatory variants beyond transcriptional level and highlighted the importance of post-transcriptional variants like those in miRNA targetome. The present work integrated genome-wide association data of coronary artery disease (CAD) with population-specific linkage disequilibrium structures from 1000 Genomes Project to map disease associations to miRNA targetome. Moreover, we performed a variety of functional prediction analyses to prioritize disease-associated variants (DAVs) influencing miRNA targetome and in-silico analyses to get insights into their functional significance. In conclusion, although the role of miRNA targetome variations in the development of CAD still has to be fully elucidated, we provided a systematic bioinformatics approach to the miRNA targetome variations in CAD. The results of this study will be valuable for researchers interested in the identification of CAD GWAS signals that may implicate polymorphic miRNA targeting.


Salt sensitivity in wheat : a case for specific ion toxicity.
Two selected lines of bread wheat, Triticum aestivum L., differing in their relative salt resistance, were grown in isosmotic solutions of different ionic compositions to investigate sensitivity to specific ions. Growth rates and ion accumulation were determined. The salt composition of the various solutions had little effect on the growth of the salt-resistant line, but significantly affected that of the salt-sensitive line. Specifically, solutions containing high Na(+) concentrations were more toxic than those containing high Cl(-) concentrations or high concentrations of nutrient ions. There were few differences in ion accumulation between lines in a given treatment, although the sensitive line tended to accumulate more Na(+) than the tolerant line in the salt treatments with high Na(+) concentrations. The overall results provide evidence that there is a definite specific ion effect which is related to salt sensitivity in wheat. It is suggested that superior compartmentation of toxic ions, principally Na(+), may be a mechanism of salt resistance in this case.


Neurotrophin 3 rescues neuronal precursors from apoptosis and promotes neuronal differentiation in the embryonic metanephric kidney.
We analyzed the developmental regulation and role of the neurotrophins during metanephric kidney morphogenesis. RNase protection assay revealed the presence of nerve growth factor, neurotrophin 3 (NT-3), and brain-derived neurotrophic factor mRNAs and the regulation of their expression during embryonic development of rat metanephros. NT-3 induced differentiation (neurite outgrowth) and survival (inhibition of apoptosis) of the neuronal precursors in cultured nephrogenic mesenchymes and neuronal differentiation in cultured whole kidneys, whereas NT-4/5, brain-derived neurotrophic factor, and nerve growth factor were without effect. The neurotrophins did not trigger tubular differentiation of isolated nephrogenic cells, which underwent apoptosis when cultured with or without the neurotrophins. NT-3 is thus an inducer of differentiation and a survival factor for renal neuronal cells, but none of the neurotrophins is a morphogen in kidney tubule induction.


Crowter v Secretary of State for Health and Social Care [2021] EWHC 2536: Discrimination, Disability, and Access to Abortion.
In a recent case before the High Court of England and Wales, Crowter v Secretary of State for Health and Social Care, three claimants challenged the disability ground for abortion contained in section 1(1)(d) of the Abortion Act 1967. It was argued on behalf of the claimants that this provision was incompatible with a number of rights protected by the European Convention on Human Rights. Among the arguments made was a claim that this provision perpetuates discriminatory attitudes and negative stereotypes towards disabled people. While the case was rightly unsuccessful, as restricting or removing section 1(1)(d) would only force pregnant people to continue pregnancies in difficult circumstances, the claim around discrimination carries some weight. The High Court rejected this line of argument, missing an opportunity to consider the ways in which the Abortion Act is both inadequate in relation to access to abortion and perpetuates stigma towards disabled people. However, the reform required to address this must take place in Parliament and cannot be done by the courts.


Lung torsion after tracheoesophageal fistula repair in an infant.
Lung torsion is a very rare event that has been reported in only 9 cases in the pediatric literature but has not yet been reported in Korean infants. We present a case of lung torsion after tracheoesophageal fistula repair in an infant. Bloody secretion from the endotracheal tube and chest radiographs and computed tomographic scan results indicated lung torsion. Emergency exploration indicated 180° torsion of the right upper lobe (RUL) and right middle lobe (RML). After detorsion of both lobes, some improvement in the RUL color was observed, but the color change in the RML could not be determined. Although viability of the RML could not be proven, pexy was performed for both the lobes. Despite reoperation, clinical signs and symptoms did not improve. The bronchoscopy revealed a patent airway in the RUL but not in the RML. Finally, the RML was surgically removed. The patient was discharged on the 42nd day after birth.


In Vitro and In Vivo Antiamebic Activity of Iron-Targeting Polypyridine Compounds against Enteric Protozoan Parasite Entamoeba histolytica.
The infectious protozoan parasite Entamoeba histolytica is responsible for amebiasis causing colitis and liver abscesses, which is an epidemic in developing countries. To develop a drug discovery strategy targeting the iron source required for the proliferation of E. histolytica, an untapped chemical group consisting of low-molecular-weight compounds with metal-binding affinity was investigated. Electrochemically neutral polypyridine compounds, PHN-R2, that showed specific Fe(II)-binding affinity and growth inhibitory ability against E. histolytica were identified. Furthermore, the iron-dependent IC50 values of PHN-R2 and the spectrometric analytical data of their iron complexes clarified the relationship between the antiamebic activity and the iron-targeting specificity. Notably, when PHN-H2 was administrated to E. histolytica-infected hamsters as an animal model of amebiasis, it exhibited a prominent therapeutic efficacy to completely cure liver abscesses without serious side effects. Deciphering the antiamebic activity of iron-targeting compounds in vitro and in vivo provides valuable insights into the development of a next-generation drug against amebiasis.


[German Total Ankle Replacement Register of the German Foot and Ankle Society (D. A. F.) - presentation of design and reliability of the data as well as first results].
Even though arthroplasty of the ankle joint is considered to be an established procedure, only about 1,300 endoprostheses are implanted in Germany annually. Arthrodeses of the ankle joint are performed almost three times more often. This may be due to the availability of the procedure - more than twice as many providers perform arthrodesis - as well as the postulated high frequency of revision procedures of arthroplasties in the literature. In those publications, however, there is often no clear differentiation between revision surgery with exchange of components, subsequent interventions due to complications and subsequent surgery not associated with complications. The German Orthopaedic Foot and Ankle Association's (D. A. F.) registry for total ankle replacement collects data pertaining to perioperative complications as well as cause, nature and extent of the subsequent interventions, and postoperative patient satisfaction. The D. A. F.'s total ankle replacement register is a nation-wide, voluntary registry. After giving written informed consent, the patients can be added to the database by participating providers. Data are collected during hospital stay for surgical treatment, during routine follow-up inspections and in the context of revision surgery. The information can be submitted in paper-based or online formats. The survey instruments are available as minimum data sets or scientific questionnaires which include patient-reported outcome measures (PROMs). The pseudonymous clinical data are collected and evaluated at the Institute for Evaluative Research in Medicine, University of Bern/Switzerland (IEFM). The patient-related data remain on the register's module server in North Rhine-Westphalia, Germany. The registry's methodology as well as the results of the revisions and patient satisfaction for 115 patients with a two year follow-up period are presented. Statistical analyses are performed with SAS™ (Version 9.4, SAS Institute, Inc., Cary, NC, USA). About 2½ years after the register was launched there are 621 datasets on primary implantations, 1,427 on follow-ups and 121 records on re-operation available. 49 % of the patients received their implants due to post-traumatic osteoarthritis, 27 % because of a primary osteoarthritis and 15 % of patients suffered from a rheumatic disease. More than 90 % of the primary interventions proceeded without complications. Subsequent interventions were recorded for 84 patients, which corresponds to a rate of 13.5 % with respect to the primary implantations. It should be noted that these secondary procedures also include two-stage procedures not due to a complication. "True revisions" are interventions with exchange of components due to mechanical complications and/or infection and were present in 7.6 % of patients. 415 of the patients commented on their satisfaction with the operative result during the last follow-up: 89.9 % of patients evaluate their outcome as excellent or good, 9.4 % as moderate and only 0.7 % (3 patients) as poor. In these three cases a component loosening or symptomatic USG osteoarthritis was present. Two-year follow-up data using the American Orthopedic Foot and Ankle Society Ankle and Hindfoot Scale (AOFAS-AHS) are already available for 115 patients. The median AOFAS-AHS score increased from 33 points preoperatively to more than 80 points three to six months postoperatively. This increase remained nearly constant over the entire two-year follow-up period. Covering less than 10 % of the approximately 240 providers in Germany and approximately 12 % of the annually implanted total ankle-replacements, the D. A. F.-register is still far from being seen as a national registry. Nevertheless, geographical coverage and inclusion of "high-" (more than 100 total ankle replacements a year) and "low-volume surgeons" (less than 5 total ankle replacements a year) make the register representative for Germany. The registry data show that the number of subsequent interventions and in particular the "true revision" procedures are markedly lower than the 20 % often postulated in the literature. In addition, a high level of patient satisfaction over the short and medium term is recorded. From the perspective of the authors, these results indicate that total ankle arthroplasty - given a correct indication and appropriate selection of patients - is not inferior to an ankle arthrodesis concerning patients' satisfaction and function. First valid survival rates can be expected about 10 years after the register's start.


Motives that cancer patients in oncological care have for consulting a psychologist--an empirical study.
Most people with cancer are able to deal with the mental turmoil with support from ordinary health care, family and friends. However, by themselves or by attentive health-care workers some patients are referred to specialists within the psychosocial field, foremost social workers and psychologists. This paper deals with patients' motives for seeing a psychologist. The case books for all patients who had met with the psychologist at a department of oncology in Sweden during a 10-year period were read through and categorised according to what the patients wanted to talk about. The most commonly found motives were in sliding order: coping with anxiety and worries caused by the disease; dealing with relational problems in life outside the disease; dealing with relational problems actualised by the disease; living with a malignant disease-despair in a new life situation; and finally dealing with a particular problem. Patients seeing a psychologist in oncology do not just ask for help to come to terms with anxiety and worries. More than a third of the patients wanted to talk about distressing relationships, which were not connected to, or only remotely connected to the cancer disease. Patients have different needs and competence in different psychological treatment perspectives is therefore important. The found diversity of motives bears impact on the external validity of screening instruments for distress and randomised controlled intervention studies.


Vaccinated Patients Admitted in ICU with Severe Pneumonia Due to SARS-CoV-2: A Multicenter Pilot Study.
Background: The aim of this study was to analyze the percentage of patients admitted to the ICU having received the vaccine against COVID-19, to describe the clinical profile of vaccinated patients admitted to the ICU, and to assess the humoral immune response to vaccination. Methods: In this multicenter prospective descriptive cohort study, consecutive critically ill patients with confirmed SARS-CoV-2 pneumonia who received at least one dose of the SARS-CoV-2 vaccine were included. The time of study was from 1 July to 10 August of 2021. Results: Of the 94 consecutive patients from seven Andalusian ICUs admitted during the time of study, 50 (53.2%) received at least one dose of anti SARS-CoV-2 vaccine. No patient was admitted having previously had SARS-CoV-2 infection. The B.1.617.2 (Delta) variant was the most frequently identified, in 80.76% of cases. Patients with a complete vaccination with non-optimal antibody levels were immunocompromised. Fifteen patients were admitted to the ICU with Acute Respiratory Distress Syndrome (ARDS) without having completed their vaccination; the clinical profile was younger and with less comorbidities compared to patients with full vaccination. There were no differences in severity of ARDS. Conclusions: Most of the patients who were admitted to the ICU having received a dose of the vaccine were not optimally vaccinated; fully vaccinated patients who did not obtain optimal serum antibody levels were patients considered immunocompromised.


Extrachromosomal chromatin: novel target for bleomycin cleavage in cells and solid tumors.
The preference of bleomycin, a DNA strand scission antitumor agent, to damage extrachromosomal (episomal) DNA was investigated. These episomes contain transcriptional promoters, replication origins, and oncogenes from MMTV, BPV, and v-Ha-ras and confer a neoplastic phenotype to a mouse fibroblast cell line. We found that bleomycin induces dose-dependent single- and double-stranded cleavage of intracellular episomes as measured by topological forms conversion. Bleomycin scission of episomes occurs within 1 min, and upon drug removal, damaged episomes are as rapidly repaired. By expressing the episomal and genomic damage as breaks per nucleotide, bleomycin has a 30-50-fold cleavage preference for episomal chromatin compared to genomic DNA. The episomes have preferred regions of the bleomycin-induced damage, particularly within the MMTV LTR and BPV origin of replication. Also, it is possible to assess bleomycin action on episomes in solid tumors in mice. Single intravenous injections of BLM into tumor-bearing mice result in single- and double-stranded cleavage of episomes that are dose related and occur within 1 min. Specific double-stranded breaks occur in the same regulatory regions of episomes in solid tumors and in cultured cells. Finally, we observe that damage to the episomal drug target occurs at therapeutic doses in mice.


Synergistic Mechanisms Underlie the Peroxide and Coagent Improvement of Natural-Rubber-Toughened Poly(3-hydroxybutyrate-co-3-hydroxyvalerate) Mechanical Performance.
Poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) is a promising bio-based and biodegradable thermoplastic with restricted industrial applications due to its brittleness and poor processability. Natural rubber (NR) has been used as a toughening agent, but further physical improvements are desired. In this study, rubber toughening efficiency was significantly improved through the synergistic use of a trifunctional acrylic coagent and an organic peroxide during reactive extrusion of PHBV and NR. The rheological, crystallization, thermal, morphological, and mechanical properties of PHBV/NR blends with 15% rubber loading were characterized. The peroxide and coagent synergistically crosslinked the rubber phase and grafted PHBV onto rubber backbones, leading to enhanced rubber modulus and cohesive strength as well as improved PHBV⁻rubber compatibility and blend homogeneity. Simultaneously, the peroxide⁻coagent treatment decreased PHBV crystallinity and crystal size and depressed peroxy-radical-caused PHBV degradation. The new PHBV/NR blends had a broader processing window, 75% better toughness (based on the notched impact strength data), and 100% better ductility (based on the tensile elongation data) than pristine PHBV. This new rubber-toughened PHBV material has balanced mechanical performance comparable to that of conventional thermoplastics and is suitable for a wide range of plastic applications.


Ten Years of National Seasonal Surveillance for Severe Complications of Influenza in Australian Children.
Severe complications of influenza in children are uncommon but may result in admission to hospital or an intensive care unit (ICU) and death. Active prospective surveillance using the Australian Paediatric Surveillance Unit with monthly reporting by pediatricians of national demographic and clinical data on children with <15 years of age hospitalized with severe complications of laboratory-confirmed influenza during ten influenza seasons 2008-2017. Of 722 children notified, 613 had laboratory-confirmed influenza and at least one severe complication. Most (60%) were <5 years of age; 10% were <6 months, hence ineligible for vaccination. Almost half of all cases were admitted to ICU and 30 died. Most children were previously healthy: 40.3% had at least one underlying medical condition. Sixty-five different severe complications were reported; pneumonia was the most common, occurring in over half of all cases. Influenza A accounted for 68.6% hospitalizations; however, influenza B was more often associated with acute renal failure (P = 0.014), rhabdomyolysis (P = 0.019), myocarditis (P = 0.015), pericarditis (P = 0.013), and cardiomyopathy (P = 0.035). Children who died were more likely to be older (5-14 years), have underlying medical conditions, be admitted to ICU, and have encephalitis, acute renal failure, or myocarditis. Only 36.1% of all children reported received antiviral medications, and 8.5% were known to be vaccinated for seasonal influenza. Severe influenza complications cause morbidity and mortality in children, which may increase if coinfection with COVID-19 occurs in the 2020 season and beyond. Increased vaccination rates, even in healthy children, early diagnosis and timely antiviral treatment are needed to reduce severe complications and death.


The use of bioelectric dressings in skin graft harvest sites: a prospective case series.
Despite advances in wound care treatments for the management of acute and chronic wounds, there remains an unmet need for interventions that accelerate epithelialization. Many authors in the past have advocated the use of electric currents to accelerate wound healing. Novel wound dressings with inherent electric activity are emerging, and studies of these specific modalities are lacking. The principal aim of this study is to evaluate the impact of a bioelectric dressing on acute wound healing. Thirteen patients who underwent skin grafting were enrolled. One half of all skin graft donor sites were treated with the bioelectric dressing and semi-occlusive dressing (SOD) and the other half using solely a SOD. Epithelialization was rated by a blinded burn surgeon attending. Participants also provided a self-assessment of their scar appearance. At week 1 postprocedure, average epithelialization of 71.8% was noted on the bioelectric dressing-treated side, compared with 46.9% on the SOD side, representing an average 34.62% faster wound healing (P = .015). At 1 month, patients rated the bioelectric dressing-treated half as superior in terms of scar color (P = .198), stiffness (P = .088), thickness (P = .038), and overall quality (P = .028). These early data show promise in terms of faster healing, improved scarring, and improved patient subjective outcome with the use of the bioelectric dressing on acute wounds. With fulfillment of an extended study population, the authors hope to provide a solid foundation for extrapolating their data beyond skin graft donor sites to all areas of wound care.


Detection of single-nucleotide mutations including substitutions and deletions by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry.
Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOFMS) was applied to the detection of mutations involving single nucleotides. For detection of a single-nucleotide deletion, a normally 44 bp region of the L1CAM gene was amplified in a 50 microL solution, and measurement was carried out on the DNA sample after phenol extraction and ethanol precipitation. A molecular mass decrease of 300 Da corresponding to a single nucleotide was identified in the amplified product of patient DNA. For detection of a substitution, an amplified product from a 50 bp region of the human beta-globin gene was cleaved with restriction endonucleases HaeIII and Bsp12861. Measurement of a mixture of digested fragments, or restriction fragment mass mapping, clearly identified a heterozygous G/C mutation in the molecular ion signals for both sense and antisense single-stranded DNAs. The results indicate that MALTI-TOFMS is feasible for genetic diagnosis of point mutations.


A case report of an inflammatory myofibroblastic tumor of the neck: A focus on the computed tomography and magnetic resonance imaging findings.
Inflammatory myofibroblastic tumors (IMTs) of the neck are rare, with only a few patients reported in the literature. The present study discusses the clinical manifestations, radiographic characteristics and management of these tumors, with a focus on imaging modalities. A case of IMT of the neck is presented and the associated literature is reviewed. In total, seven patients in seven English-language studies, including the present case, and one patient in one Chinese-language study were found. On CT scans, all tumors appeared as soft-tissue densities. Upon magnetic resonance imaging (MRI), all tumors displayed a heterogeneous hypointense-isointense signal on T1-weighted sequences and an isointense-hyperintense signal on T2-weighted sequences. All tumors showed enhancement on enhanced CT and MR images. The imaging features of the neck IMTs can be summarized as follows: i) When enhanced, the tumor displays enhancement on CT and MR images; ii) MRI is superior to CT scans in the differential diagnosis of this disease; iii) in general, the lesion displays a hypointense-isointense signal on T1-weighted sequences and an isointense-hyperintense signal on T2-weighted sequences; iv) due to the fibrous tissue in the tumor, delayed enhancement may be observed on gadolinium-enhanced MR images; and v) due to its benign or intermediate features, the tumor is usually a well-defined mass.


Elicitation of motion sickness by head movements in the microgravity phase of parabolic flight maneuvers.
During parabolic flight maneuvers in a Boeing KC-135 aircraft 44 college students were tested for motion sickness susceptibility. These subjects were categorized as 1) insusceptible, 2) moderately susceptible, or 3) highly susceptible to motion sickness during exposure to varying gravitoinertial force levels. After categorization, they were tested in the microgravity phase of parabolic flight to see how three types of head movements affected their baseline susceptibility. The head movements evaluated included side-to-side swivel, shoulder-to-shoulder roll, and front-up head and trunk movements; each type of head movement was used on a separate test day for eyes-open and eyes-covered conditions. Ten cycles of head movements were made in each parabola until a motion sickness endpoint, nausea, was reached or 40 parabolas had been completed. All types of head movements significantly increased susceptibility for subjects in all categories; eyes-open conditions were always more stressful than eyes-closed for each kind of head movement. These findings show unequivocally that natural head movements in microgravity can elicit symptoms of motion sickness. They suggest that head movements play an important etiological role in space motion sickness. In ground based studies where head movements are necessary to elicit symptoms, they are also necessary to elicit adaptation. We describe the use of paced and incremented head movement schedules as a possible way of partially alleviating space motion sickness.


Transactivation of the GATA-1 promoter by a myb-ets-containing mouse retrovirus is mediated by CACCC elements.
The myb-ets-containing ME26 virus causes erythroleukemia in mice by a novel mechanism involving the inappropriate activation of erythroid-specific genes in hematopoietic precursor cells. We have previously shown that the ME26 viral protein can transactivate the GATA-1 promoter in transient transactivation assays carried out in mouse fibroblasts. The mouse GATA-1 promoter, whose activity is regulated by the GATA-1 protein itself, contains a double GATA consensus sequence at its 5' end and two CACCC elements at its 3' end, both of which are crucial for promoter activity in erythroid cells, as well as a nonconsensus GATA sequence and several putative c-myb and c-ets binding sites. In order to determine which sequences in the GATA-1 promoter are crucial for activation by the ME26 viral protein, we made deletions of the promoter, cloned them into a luciferase expression vector and tested their activity in mouse fibroblasts, which do not express GATA-1. Our results indicate that sequences in the 3' end of the GATA-1 promoter, which include two CACCC elements, are essential for transactivation by ME26 virus, while other upstream sites contribute to full activation by the virus. Mutation of the CACCC sites abolishes ME26 viral transactivation. The interaction of cell extracts containing ME26 viral protein and the GATA-1 promoter fragment containing the two CACCC elements was examined by electrophoretic mobility shift analysis (EMSA) and the results showed no direct interaction between the two. However, we could detect the ubiquitous transcription factor Sp1 bound to this sequence. These data demonstrate that the CACCC element is necessary for GATA-1 promoter transactivation by ME26 virus and that the viral protein may indirectly transactivate the promoter by binding to Sp1.


Vascularity of the lateral calcaneal flap: a cadaveric injection study.
To describe the arterial blood supply of the subcutaneous tissues of the lateral hindfoot and define the relationships between these arteries and the lateral extensile incision used for open reduction and internal fixation of calcaneal fractures. Human cadaveric lower extremity specimens, doubly injected with India ink and latex, were used to demonstrate the location of the arteries of the subcutaneous tissues of the lateral hindfoot. Anatomy laboratory. Twenty-four randomly obtained, cadaveric elderly lower extremity specimens. India ink and then latex were injected into the superficial femoral artery at the level of the inguinal crease after cleansing of the arterial system. Transtibial amputation specimens were manually debrided of the skin and chemically debrided of subcutaneous tissues with sodium hypochlorite to demonstrate the arterial supply to the soft tissues of the lateral hindfoot. MEAN OUTCOME MEASUREMENTS: The location of the three major arteries was determined relative to the lateral malleolus. The proximity of these vessels to the typical extensile lateral incision was determined radiographically with vascular clips applied along each artery and skin staples placed along the path of the typical skin incision. Three arteries, the lateral calcaneal artery, the lateral malleolar artery, and the lateral tarsal artery, were consistently found along the lateral aspect of the hindfoot. The lateral calcaneal artery appeared to be responsible for the majority of the blood supply to the corner of the flap and, because of its proximity to the vertical portion of the typical incision, it appeared most likely to be injured from inaccurate placement of the incision. The development of wound complications following open reduction and internal fixation of the calcaneus is multifactorial. Disruption of the blood supply to the surgically created flap may play a larger role in the development of wound complications than previously thought. An understanding of the local vascular anatomy may decrease the rate of wound complications during the operative treatment of intraarticular calcaneal fractures.


A critical examination of Danish norms for the Strengths and Difficulties Questionnaire (SDQ).
The objective was to describe observed differences between the official Danish Strengths and Difficulties Questionnaire (SDQ)-norms and data from a sample based on the Danish National Birth Cohort (DNBC) including children born between 1996 and 2003. We compared the risk classification, cut-off bandings and the group sizes between the Danish norms and the values found in our sample at ages 7, 11 and 18 years. Two sets of norms are used in Denmark: Arnfred's norms, based on a sample from one single Danish municipality and Niclasen's norms, based on multiple Danish cohorts, including the DNBC. Inconsistencies were found between banding scores in the two existing norms and the banding scores identified in our sample from DNBC: discrepancies included banding scores for several of the problem scales for children and preadolescents. For adolescents, we found less apparent inconsistencies between Arnfred's sample and the DNBC. Results demonstrate that the existing SDQ norms do not apply well to a large-scale cohort sample in Denmark. The usefulness of the SDQ as a screening instrument for mental health problems depends on appropriate norms. We therefore urge that the current Danish SDQ norms are used with caution, and preferably they should be revised.


Proteomic study of calpeptin-induced differentiation on calpain-interacting proteins of C2C12 myoblast.
Studies on skeletal muscle cell specification and development have demonstrated in the past that calpains interact with various transcriptional factors in regulating the cellular function. It has therefore, been assumed that transcriptional factors like myogenin, MyoD, Myf5, and MRF4 that are active during the myogenic differentiation might be affected and degraded by calpains. Therefore, to examine the biochemical adaptations of myoblasts during myocyte formation and muscle development comprehensively, the current study was designed to identify the effect of calpeptin (calpain inhibitors) on protein expression during differentiation of C2C12 mouse myoblast. Cells were proliferated to near 80% confluence under Dulbecco's modified eagle medium and differentiated further in 2% HS with 50 μM calpeptin. Incubated cells were collected at 0, 12, and 72 h and later the cell proteins were focused onto pH 4-7 IEF strip, followed by 12.5% SDS-PAGE. Obtained spots on the gels were compared and matched using commercial 2-DE analysis software and matched spots were identified by MALDI-ToF and/or Q-Tof systems. Conclusively, cell differentiation was observed to be active from 12 to 72 h however, calpeptin affected the differentiation process and cut down the rate of fusion by approximately 50%. Out of 41 proteins identified, 12 proteins were found to be upregulated where as 29 proteins were downregulated.


Frequent prosthesis refitting to prevent implant exposure in patients with retinoblastoma.
To examine the effects of prosthesis refitting on porous orbital implant exposure in 100 patients who underwent enucleation for retinoblastoma. Parameters analyzed included patient's age at enucleation, gender, implant type, frequency of prosthesis adjustment, and use of chemotherapy or radiation. The main outcome measures consisted of the condition of the fornices, condition of the implant (including conjunctival thinning), and condition and functioning ability of the prosthesis. Increased frequency of visits with the ocularist and number of prosthesis adjustments and refits significantly improved the condition of the implant, fornices, and prosthesis. The patient's age at enucleation, gender, and use of chemotherapy or radiation did not have a significant effect on any of the outcome variables. The condition of the implant was significantly better for porous polyethylene than hydroxyapatite implants (P = .024). No implant exposures were observed. Frequent adjustments and refits by the ocularist are significantly associated with a reduced rate of conjunctival thinning and complete avoidance of implant exposure in patients undergoing enucleation for retinoblastoma. These findings are particularly significant for this population, which historically has demonstrated a high rate of implant exposure.


Sublingual administration of testosterone-hydroxypropyl-beta-cyclodextrin inclusion complex simulates episodic androgen release in hypogonadal men.
In search of a more physiological testosterone (T) replacement therapy for hypogonadal states, we evaluated an inclusion complex of T with 2-hydroxypropyl-beta-cyclodextrin (HPBCD). HPBCD enhances T solubility and absorption, but HPBCD is not absorbed. Five hypogonadal men (mean age, 32.4 +/- 2.3 yr) with serum T levels below the normal range were treated in two separate experimental phases with either a 2.5- or 5.0-mg tablet of sublingual (SL) T-HPBCD three times daily for 7 days. Acute pharmacodynamic changes were monitored at baseline and 10, 20, and 40 min and 1, 1.5, 2, 3, 4, and 8 h after administration of the first dose. At the 5-mg dose, a maximal concentration (Cmax) of T (85.4 +/- 11.0 nmol/L) was achieved in 20 min (63 +/- 24-fold increase), followed by a rapid decline to below the normal range (less than 12 nmol/L) at 2 h, with an estimated half-life of decline of 1.87 +/- 0.19 h. The dihydrotestosterone (DHT) Cmax (4.1 +/- 0.5 nmol/L) occurred at 32 +/- 5 min (8.9 +/- 1.3-fold increase) and declined to below the normal range (less than 1.2 nmol/L) after 3 h. The integrated 8 h value for the ratio of T/DHT was 10.0 +/- 1.1, which fell within the normal range. The increment in androstenedione paralleled that in T, and the Cmax (6.8 +/- 0.9 nmol/L) was reached in 24 +/- 4 min (2.3 +/- 0.6-fold increase). Compared to baseline, the Cmax was significantly greater for T (P less than 0.005), DHT (P less than 0.0005), and androstenedione (P less than 0.005). Both estradiol (E2) and estrone (E1) remained in the normal range (less than 200 pmol/L), although the Cmax for E1 was significantly greater than baseline (P less than 0.05). Serum LH levels were suppressed (19.0 +/- 2.6%) at 2 h (P less than 0.05), without a significant change in FSH. During 7 days of treatment, there was no cumulative increase in basal T, DHT, and E2 levels or further decline in LH or FSH levels. There was no change in sex hormone-binding globulin levels. Similar results were observed with the 2.5-mg dose, suggesting that the capacity of SL absorption may be limited to a certain dose of T-HPBCD. The fluctuations in T after SL administration of T-HPBCD resemble endogenous episodic secretion. We conclude that T, complexed with HPBCD, is rapidly absorbed by the SL route and quickly metabolized without sustained elevations of DHT or E2.