Effects of a mixture of phytobiotic-rich herbal extracts on growth performance, blood metabolites, rumen fermentation, and bacterial population of dairy calves.
Forty-eight newborn Holstein dairy calves [40 ± 3.4 (SD) of kg of body weight (BW); 24 females and 24 males] were used in a completely randomized design to investigate the effects of a mixture of phytobiotic-rich herbal extract (Immunofin, IMPE) incorporated into milk on performance, ruminal fermentation, bacterial population, and serum biochemical metabolites during the preweaning period. Calves had free access to calf starter and clean water from d 6 until weaning. The treatments were the control (CON; without additive) and IMPE at 4, 8, and 12 mL/d. The treatments had no significant effect on total dry matter intake, weight gain, and BW at weaning. The incidence of diarrhea was lower in calves fed 8 mL of IMPE/d compared with CON. At weaning, body measurements (except for front leg circumference) were not affected by IMPE treatment. Relative to the CON group, front leg circumference was significantly decreased by IMPE supplementation. Serum IgG concentration was not significantly increased by IMPE supplementation compared with the CON group. Triglyceride concentration decreased in calves receiving 4, 8, and 12 mL/d of IMPE compared with the CON groups. In contrast to the CON group, serum albumin and total serum protein concentrations increased with IMPE supplementation. Calves receiving 4 mL/d of IMPE had a greater abundance of total bacteria, Ruminococcus albus, Ruminococcus flavefaciens, and Fibrobacter succinogenes compared with the other treatments. Molar proportions of acetate increased in calves fed IMPE (at 12 mL/d) compared with calves fed CON. Ruminal N-NH3 concentrations decreased linearly with the increase in IMPE supplementation. The results of the present study suggest that the addition of IMPE to milk may improve some health and immunity conditions, blood metabolite concentrations, and increase the abundance of some cellulolytic bacteria in the rumen of Holstein dairy calves. The use of IMPE may be an alternative to feeding antibiotics at subtherapeutic concentrations to improve calf health and immunity status.


[The influence of patatin-like phospholipase domain-containing protein 3 on palmitic acid-induced hepatocyte apoptosis].
To explore the influence of patatin-like phospholipase domain-containing protein 3 (PNPLA3) on palmitic acid (PA)-induced hepatocyte apoptosis and its mechanism. Human hepatocarcinoma cell line HepG2 cells were transfected with PNPLA3(WT)-pcDNA3.1 (PNPLA3(WT) group) and PNPLA3(I148M)-pcDNA3.1 (PNPLA3(I148M) group) plasmids respectively to overexpress wild type or mutant type PNPLA3, and cells transfected with empty vector pcDNA3.1 (NC group) were set as control group.After 24 h PA incubation, Oil red staining was used to determine lipid deposition, and terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling assay (TUNEL) was used to measure apoptosis.Western blot was used to detect the protein level of endoplasmic reticulum (ER) stress and associated apoptosis.Enzyme-linked immunosorbent assay (ELISA) was used to test lysolecithin (LPC) levels in the cellular supernatant. After 24 h PA incubation, there was no significant difference in lipid deposition among three groups.Compared to NC group, the cell apoptosis rates of PNPLA3(WT) and PNPLA3(I148M) groups were increased by 2 times and 3 times respectively.The levels of ER stress PRKR-like endoplasmic reticulum kinase (PERK) pathway associated proteins, immunoglobulin-binding protein (BIP), p-PERK, p-eIF2α, and ER stress associated apoptosis pathway proteins, CCAAT/enhancer binding homologous protein (CHOP), p53 upregulated modulator of apoptosis (PUMA), Bax, caspase-3 were higher, and were more significant in PNPLA3(I148M) group.The LPC level in the supernatant of PNPLA3(WT) and PNPLA3(I148M) groups were about 5 times and 1.5 times of NC group respectively after PA incubation. PNPLA3 may be involved in palmitic acid-induced apoptosis mediated by hepatocyte ER stress through regulating LPC metabolism.


Invasive fungal rhinosinusitis: what is the appropriate follow-up?
Early detection and aggressive surgical and medical management have been associated with higher overall survival rates among patients with invasive fungal rhinosinusitis (IFS). With improved survival comes the question of how to appropriately manage these patients once disease stability has been achieved. Previous reports suggest follow-up only as long as the patients remain immunocompromised. This study attempts to answer the question of long-term clinical follow-up and suggests a regimen suitable for ensuring minimal posttreatment complications. A retrospective review included all patients diagnosed with IFS between 1988 and 2004. The study group included patients who survived the initial treatment course, with at least 30 days of posttreatment follow-up of their IFS. Patient records were reviewed for significant complications, evidence of chronic sinus disease, the clinical status of their underlying medical comorbidities, and frequency and mode of follow-up. Thirteen patients were included. The average follow-up time was 633 days. Significant complications included one patient with acute bacterial sinusitis with resultant visual loss and one patient with chronic osteomyelitis of the orbit and skull base. Six of 13 patients had persistent chronic bacterial rhinosinusitis with crusting and bone sequestration. All complications were noted to occur after initial IFS eradication was thought to have taken place. Significant complications of IFS can occur after medical remission and recovery of immune competence. Patients with IFS should be followed long term until remucosalization of the sinuses, resolution of crusting, and cessation of bony sequestration has occurred.


EDTA assisted highly selective detection of As(3+) on Au nanoparticle modified glassy carbon electrodes: facile in situ electrochemical characterization of Au nanoparticles.
A facile electrochemical characterization technique of Au nanoparticles (AuNPs) developed by Wang et al. ( Wang, Y.; Laborda, E.; Salter, C.; Crossley, A.; Compton, R. G. Analyst 2012 , 137 , 4693 - 4697 ) was used to track the variation in the particle size and density of AuNPs in situ and to assist in optimizing the conditions of analysis and catalysis. In this method, the combination of total surface area determined by Pb underpotential deposition (UPD) and the amount of Au obtained by anodic stripping of Au in HCl solution was used to evaluate the average diameter of AuNPs and the number of particles on the electrode. The detection of As(3+) in aqueous solution by a AuNP modified glassy carbon electrode (Aunano@GCE) using the electrochemical characterization technique was examined. The AuNPs with a uniform shape and size, deposited onto the GCEs using multiple-scan cyclic voltammetry (MSCV), were suitable for the electrochemical evaluation. The calibration curve for the detection of As(3+) had a dynamic range of 0.1-15.0 μg L(-1) (from 1.30 to 200 nM, y = 0.21x (in μA L μg(-1)) + 0.01 (R(2) = 0.999)) and showed a sensitivity of 0.21 μA L μg(-1) (16.15 μA μM(-1)). A detection limit as low as 0.0025 μg L(-1) (32.5 pM) was achieved. The chelating agent ethylenediaminetetraacetate (EDTA) selectively chelated with the interfering metal ions and effectively inhibited the interfering ions from competing with the ion of interest (As(3+)), in the preconcentration process. The presence of EDTA effectively eliminated interference from several metal ions, especially Cu(2+) and Hg(2+). This method was validated by analyzing the As(3+) content in real water samples.


Increased expression of long non-coding RNA CCEPR is associated with poor prognosis and promotes tumorigenesis in urothelial bladder carcinoma.
Recent emerging evidences have showed that long non-coding RNAs play important regulatory roles in diverse biological processes of tumor development and progression. CCEPR (cervical carcinoma expressed PCNA regulatory lncRNA) is a novel identified lncRNA that acts as a potential biomarker and involves in development and progression of cervical carcinoma. Nevertheless, we know nothing about the clinical significance and molecular mechanism of CCEPR in bladder cancer. In this study, we found that CCEPR was significantly up-regulated in bladder cancer. Furthermore, up-regulated CCEPR expression was positively correlated with advanced TNM stage and higher histological grade. Moreover, further experiments demonstrated that CCEPR promotes cell proliferation and suppresses cell apoptosis in bladder cancer. Mechanistically, we found CCEPR upregulates the expression of PCNA in mRNA and protein level to promote cancer growth. In conclusions, these findings demonstrated that CCEPR plays an important regulatory role in bladder cancer and may serve as a potential diagnostic biomarker and therapeutic target.


The efficacy of a cell-mediated reaction in the disposal of M. leprae in human skin.
The inability of lepromatous leprosy patients to mount a cellular immune response against Mycobacterium leprae antigens is not understood. The extensive intracellular replication of bacilli in the phagocytes and the relative paucity of T lymphocytes in the lesions suggest that these patients might be incapable of generating normal delayed type hypersensitivity responses in their skin. In order to elucidate this problem we evaluated the patient's response to local antigen administration. Our observations suggest that the majority of lepromatous patients can respond normally to intradermal injections of a soluble antigen such as purified protein derivative of tuberculin. The underlying lepromatous lesions do not inhibit mononuclear cell infiltration or differentiation. Moreover, the generation of a cellular immune response in the lesions appears to modify the lepromatous lesion to a lesion resembling the tuberculoid type. This process involves local T cells recruitment, granuloma formation and a reduction in the bacterial load at the antigen responsive site.


Liposomes for entrapping local anesthetics: a liposome electrokinetic chromatographic study.
Bupivacaine is a lipophilic, long-acting, amide class local anesthetic commonly used in clinical practice to provide local anesthesia during surgical procedures. Several cases of accidental overdose with cardiac arrest and death have been reported since bupivacaine was introduced to human use. Recent case reports have suggested that Intralipid (Fresenius Kabi) is an effective therapy for cardiac toxicity from high systemic concentrations of, e.g. bupivacaine, even though the mechanism behind the interaction is not fully clear yet. Our long-term aim is to develop a sensitive, efficient, and non-harmful lipid-based formulation to specifically trap harmful substances in vivo. In this study, the in vitro interaction of local anesthetics (bupivacaine, prilocaine, and lidocaine) with Intralipid or lipid vesicles containing phosphatidylglycerol, phosphatidylcholine, cardiolipin, cholesterol, and N-palmitoyl-D-erythro-sphingosine (ceramide) was determined by liposome electrokinetic chromatography. The interactions were evaluated by calculating the retention factors and distribution constants. Atomic force microscopy measurements were carried out to confirm that the interaction mechanism was solely due to interactions between the analytes and the moving pseudostationary phase and not by interactions with a stationary lipid phase adsorbed to the fused-silica wall. The heterogeneity of the liposomes was also studied by atomic force microscopy. The liposome electrokinetic chromatography results demonstrate that there is higher interaction between the drugs and negatively charged liposome dispersion than with the commercial Intralipid dispersion.


Expression of TM4SF10, a Claudin/EMP/PMP22 family cell junction protein, during mouse kidney development and podocyte differentiation.
Cell junctions in the nephron are highly specialized to perform specific and distinct filtration and reabsorption functions. The mature kidney forms complex cell junctions including slit diaphragms that prevent the passage of serum proteins into the filtrate, and tubule cell junctions that regulate specific paracellular ion reuptake. We have investigated the expression of TM4SF10 (Trans-Membrane tetra(4)-Span Family 10) in mouse kidneys. TM4SF10 is the vertebrate orthologue of Caenorhabditis elegans VAB-9, a tetraspan adherens junction protein in the PMP22/EMP/Claudin family of proteins. We found that TM4SF10 localizes at the basal-most region of podocyte precursors before the capillary loop stage, at some tubule precursors, and at the ureteric bud junction with S-shaped bodies. Overall expression of TM4SF10 peaked at postnatal day 4 and was virtually absent in adult kidneys. The very limited expression of TM4SF10 protein that persisted into adulthood was restricted to a few tubule segments but remained localized to the basal region of lateral membranes. In undifferentiated cultured podocytes, TM4SF10 localized to the perinuclear region and translocated to the cell membrane after Cadherin appearance at cell-cell contacts. TM4SF10 colocalized with ZO1 and p120ctn in undifferentiated confluent podocytes and also colocalized with the tips of actin filaments at cell contacts. Upon differentiation of cultured podocytes, TM4SF10 protein disappeared from cell contacts and expression ceased. These results suggest that TM4SF10 functions during differentiation of podocytes and may participate in the maturation of cell junctions from simple adherens junctions to elaborate slit diaphragms. TM4SF10 may define a new class of Claudin-like proteins that function during junctional development.


Angiotensin II-induced upregulation of MAP kinase phosphatase-3 mRNA levels mediates endothelial cell apoptosis.
Angiotensin II (Ang II) is central to the pathobiology of atherosclerosis. In endothelial cells (EC), Ang II induces apoptosis. The MAP kinase ERK1/2 plays a key role in regulating cell survival. We therefore investigated the effect of Ang II on ERK1/2. Incubation of EC with Ang II led to the dephosphorylation of ERK1/2 (43% of control). To characterize the phosphatase involved, we investigated the effect of Ang II on MAP kinase phosphatase expression. Ang II induced MAP kinase phosphatase-3 (MKP-3) mRNA levels to about 2-fold, whereas MKP-1 expression was not affected. Transfection with a dominant negative MKP-3 construct (dnMKP-3mt) prevented the Ang II-induced ERK1/2 dephosphorylation and apoptosis in EC (p < 0.001). ERK1/2 inactivation has been shown to result in the dephosphorylation and proteasomal degradation of the antiapoptotic protein Bcl-2. Ang II induced the degradation of Bcl-2 wild type, whereas the dephosphorylation-resistant Bcl-2 construct mimicking phosphorylation by ERK1/2 was resistant to Ang II stimulation. These results indicate that Ang II-induced apoptosis signaling in human EC is mediated via MKP-3-dependent dephosphorylation of ERK1/2, which in turn leads to the degradation of Bcl-2.


A double-blind, placebo-controlled trial of tiagabine given three-times daily as add-on therapy for refractory partial seizures. Northern European Tiagabine Study Group.
In a multicentre, double-blind, parallel-group, placebo-controlled trial, a three-times daily regimen of tiagabine was evaluated as add-on therapy in 154 adult patients with refractory partial seizures. A total of 77 patients were randomised to treatment in each arm. Tiagabine HCl was titrated from an initial dose of 12-30 mg/day over 4 weeks. During the 12-week fixed-dose period, there was a significant reduction in the median 4-weekly seizure rate for all partial seizures and simple partial seizures (P < 0.05 in each case). Furthermore, the proportion of patients with a reduction of 50% or more in all partial seizures was higher in the tiagabine group than in the placebo group (14 versus 6%), though the difference did not achieve statistical significance. The difference with respect to simple partial seizures was significant (21 versus 6%, P < 0.01). The percentage of patients achieving an increase of at least 50% in the proportion of days free of all partial seizures was significantly greater in the tiagabine group compared to placebo (14 versus 4%, P<0.01). Tiagabine did not appear to influence the plasma concentrations of other concomitant antiepileptic drugs and was generally well tolerated, with most drug-related adverse events being mild or moderate in severity. The most common adverse events were dizziness, asthenia, headache and somnolence. Adverse event incidence was similar between tiagabine and placebo groups, except for dizziness which was more common with tiagabine (29 versus 10%, P < 0.01). Tiagabine had no significant effects on laboratory tests or vital signs. The present study shows that tiagabine, at a dose of 10 mg administered three-times daily, which is at the lower end of the usual recommended dose range (30-50 mg/day, tiagabine base), is generally well tolerated and demonstrates efficacy for the treatment of refractory partial seizures.


Self-mutilation, substance abuse, and the psychoanalytic approach: four cases.
While self-injury and substance abuse are difficult symptoms for both analyst and patient to cope with, and relapses are frequent, the emphasis does not have to be on managing crisis. The initial ego support and therapeutic boundary setting in these difficult cases must be matched by psychoanalytic exploration. In working with these patients, I find that, through mutual projective identification processes, the analyst and the patient are frequently resurrecting certain aspects of the patient's archaic phantasy life as defined by various self and object representations. Therefore, the continuous analysis of the transference and the countertransference is certainly essential. However, the additional willingness on the part of the analyst and the patient to explore the frequent and mutual interpersonal/intrapsychic acting out is paramount.


Inhibition of phenylalanyl-tRNA synthetase by aromatic guanidines and amidines.
Aromatic guanidines and amidines were investigated for their ability to inhibit phenylalanyl-tRNA synthetase from E. coli B. 2-Phenylacetamidine (1), benzylguanidine (2), and N-benzylbenzamidine (3) are competive inhibitors with respect to phenylalanine, binding nearly as well as the substrate, The remainder of the inhibitors was unexpectedly found to be noncompetitive, indication the presence of a secondary binding site on the enzyme. Inhibition by these compounds appears to be specific for phenylalanyl-tRNA synthetase and requires the presence of a phenyl ring as well as the amidine or guanidine moiety.


Perforator Flaps Based on the Terminal Perforators of the Pectoral Branch of Thoracoacromial Vessels Axis: A Viable Option in the Head and Neck Reconstructions.
The purpose of this article is to develop a new method for elevating the pedicled terminal pectoral perforator flaps (PTPPFs) and to resolve the difficulties and controversies that shroud the anatomy of terminal pectoral perforators from the pectoral branch of the thoracoacromial axis. Anatomical study was conducted to assess the feasibility of perforator flaps based on the terminal perforators of the pectoral branch of the thoracoacromial vessel axis. The knowledge that gleaned from this anatomical study was applied in the clinical scenario. A clinical retrospective study was conducted to evaluate the outcome of PTPPFs for the reconstruction of postexcisional head and neck defects. Thirty cadaveric specimens were examined for the anatomy of the terminal musculocutaneous pectoral branch perforators. Thirty-two cases (22 males, 10 females) of various head and neck postexcision defects were reconstructed with the PTPPFs in a single-stage manner. Pectoralis major muscle with its innervation was spared in all cases. All the secondary defects were closed primarily. The maximum cutaneous paddle size was 156 cm harvested on a single best terminal pectoral branch perforator. The average size of the cutaneous paddle harvested was 73.47 cm. The flap complications noted (9.3%) were trivial, and all the flaps survived well. The average institutional computed score for esthetic and functional recovery at the donor and reconstructed site done by 2 independent observers were 4.56 and 3.78, respectively. The recruitment of the chest skin on the terminal pectoral branch perforators resulted in a long pedicle flap with adequate qualities like color match, texture match, and pliability. This flap shall serve as a primary option for pedicle and free flap head and neck reconstructions.


The detection of circulating immune complexes containing immunoglobulin G.
An immune complex assay using radiolabelled immunospecific antibodies against human IgG and polyethylene glycol precipitation (IgG-PEG assay) is described. The material reactive in this assay was evaluated using aggregated immunoglobulins, immune complexes prepared in vitro, sera of patients with a variety of disorders and normal human serum. Sucrose density gradient ultracentrifugation showed that only large-sized immune complexes (greater than 25 S) were detected. Comparison of the results of the IgG-PEG assay with those of the C1q binding assay showed a highly significant positive correlation (p less than 0.001). It was found that rheumatoid factors do not influence the results of the IgG-PEG assay. The method described in this study detects specifically immune complexes containing IgG and might be extended to the detection of other constituents of circulating immune complexes.


State-of-the-art surgical management of testicular tumors.
Testicular cancer is the most common malignancy in men aged 20-35 years and accounts for approximately 1% of all male malignancies. Testicular cancer has a propensity to spread via the lymphatic system to the retroperitoneal lymph nodes, and retroperitoneal lymph node dissection remains an essential component in the cure of these patients. This review summarizes the basic principles of surgical management of germ cell tumors.


Depression among southern rural African American women: a factor analysis of the Beck Depression Inventory-II.
This is the first reported study involving a factor analysis of the Beck Depression Inventory-II, which was administered to a sample of southern rural African American women. To determine the factor structure of the Beck Depression Inventory-II using data collected from southern rural African American women. Using a correlational, descriptive design, 206 southern rural African American women were invited to participate in a face-to-face interview that occurred in a variety of community-based settings. The factor analysis of the Beck Depression Inventory-II resulted in a two-factor solution. Symptoms such as pessimism and worthlessness loaded high on the first factor (cognitive). The second factor explained somatic-affective symptoms of depression, with factor loadings high on tiredness and fatigue and loss of energy. The application of the Beck Depression Inventory-II among African American people would generate needed information about how depressive symptoms may be expressed among them. Knowledge gained from this study promises to be useful for developing appropriate research studies and population-specific treatment approaches for this group of women.


Factors influencing growth of rumen, liver, and other organs in kids weaned from milk replacers to solid foods.
From pooled data of many feeding trials with kids fed various types of solid foods from 37 days of age and slaughtered thereafter, relationships between such independent variables as age, cumulative intake of solid food, weight gain, and/or final body weight and such dependent variables as weights and such dependent variables as weights of stomach-compartments, heart, liver, kidneys, and spleen were analyzed. Growth of forestomachs, and especially rumen mucosa, relative to body weight was stimulated only in feeding periods of solid foods. Gain, intake of solid food, and final body weight were the best predictors for growth of forestomach-compartments, liver, and heart, respectively. Correlations were large between ruminoreticulum weight and liver weight and among weights of forestomach-compartments. Second degree polynomials were fitted to predict the curvilinear relationships between solid-food intake and either ruminoreticulum weight or rumen mucosal weight, and a linear equation was fitted to predict the relationship between solid-food intake and liver weight. Feeding of roughage increased rumen muscular and omasal weights. Growth of the rumen under stimulation of solid-food intake may be regulated by the same biological factors contributing to increased growth rate of the whole animal. In this situation, growth of the rumen may be correlated closely with growth of the liver.


Publication Bias in Gastrointestinal Oncology Trials Performed over the Past Decade.
Randomized controlled trials (RCTs) are the gold standard for evidence-based practice, but their development and implementation is resource intensive. We aimed to describe modern RCTs in gastrointestinal (GI) cancer and identify predictors of successful accrual and publication. ClinicalTrials.gov was queried for phase III GI cancer RCTs opened between 2010 and 2019 and divided into two cohorts: past and recruiting. Past trials were analyzed for predictors of successful accrual and the subset with ≥3 years follow-up were analyzed for predictors of publication. Univariate and multivariable (MVA) logistic regression were used to identify covariates associated with complete accrual and publication status. A total of 533 GI RCTs were opened from 2010 to 2019, 244 of which are still recruiting. In the "past" trials cohort (235/533) MVA, Asian continent of enrollment was a predictor for successful accrual, whereas trials with prolonged enrollment (duration longer than median of 960 days) trended to failed accrual. Predictors for publication on MVA included international enrollment and accrual completion. Sponsorship was not associated with accrual or publication. Notably, 33% of past trials remain unpublished, and 60% of trials that were closed early remain unpublished. Accrual rate and the primary continent of enrollment drive both trial completion and publication in GI oncology. Accrual must be streamlined to enhance the impact of RCTs on clinical management. A large portion of trials remain unpublished, underscoring the need to encourage dissemination of all trials to, at a minimum, inform future trial design. Two-thirds of gastrointestinal (GI) oncology phase III randomized controlled trials successfully accrue; however, one third of these trials are unpublished and more than half of trials that close early are unpublished. The strongest predictors for publication are successful accrual and international collaborations. Initiatives to optimize the trial enrollment process need to be explored to maximize the potential for trials to engender progress in clinical practice. Moreover, this study identified a significant publication bias in the realm of GI oncology, and the field should promote reporting of all trials in order to better inform future trial questions and design.


Discovery and development of anticancer aptamers.
Aptamers, also termed as decoys or "chemical antibodies," represent an emerging class of therapeutics. They are short DNA or RNA oligonucleotides or peptides that assume a specific and stable three-dimensional shape in vivo, thereby providing specific tight binding to protein targets. In some cases and as opposed to antisense oligonucleotides, effects can be mediated against extracellular targets, thereby preventing a need for intracellular transportation. The first aptamer approved for use in man is a RNA-based molecule (Macugen, pegaptanib) that is administered locally (intravitreally) to treat age-related macular degeneration by targeting vascular endothelial growth factor. The most advanced aptamer in the cancer setting is AS1411, formerly known as AGRO100, which is being administered systemically in clinical trials. AS1411 is a 26-mer unmodified guanosine-rich oligonucleotide, which induces growth inhibition in vitro, and has shown activity against human tumor xenografts in vivo. The mechanism underlying its antiproliferative effects in cancer cells seems to involve initial binding to cell surface nucleolin and internalization, leading to an inhibition of DNA replication. In contrast to other unmodified oligonucleotides, AS1411 is relatively stable in serum-containing medium, probably as a result of the formation of dimers and a quartet structure. In a dose escalation phase I study in patients with advanced solid tumors, doses up to 10 mg/kg/d (using a four or seven continuous infusion regime) have been studied. Promising signs of activity have been reported (multiple cases of stable disease and one near complete response in a patient with renal cancer) in the absence of any significant adverse effects. Further trials are ongoing in renal and non-small cell lung cancers. In preclinical studies, additional aptamers have been described against several cancer targets, such as tenascin-C, the transcription factor signal transducer and activator of transcription 3, and antiapoptotic and Ku proteins.


KIR genotyping in the selected population in Andhra Pradesh (India).
The population is not always homogeneous in relation to the representation and functioning of genes. Therefore, the presence of allogenicity is a universal phenomenon. The profound variability is noticed among the members of the human population with reference to the resistance against infections and late onset of diseases. In this line, a few sets of alleles which come under the domain of immune function namely KIRs (Killer immunoglobulin-like receptor genes) and HLA-I have been chosen to report in the population of Puttaparthi (India). The genotyping of the population is the current ongoing focus of our team wherein the distribution of the following alleles has been taken up in the mixed ethnic groups of Puttaparthi as a prelude to earmark them as genotypic markers in future studies relating to susceptible diseases. The PCR protocols for the identified immune related genes viz., KIR- 2DL1, 2DL2, 2DL3, 2DL4, 2DL5, 3DL1, 3DL2, 3DL3, 3DS1, 2DS1, 2DS2, 2DS3, 2DS4, 2DS5, 2DP1, 3DP1; HLA- C1 and HLA-C2 have been standardized. In the present study, except KIR 2DL2, the other non-framework inhibitory KIR genes were represented at higher percentage and ranged from 57% to 80% in the chosen population which would suggest its higher survival adaptation. Interestingly, the majority of activating KIR genes were least represented and varied between 5% to 32.5% which is also in compliance with the survival adaptation of the chosen population. The carrier gene frequencies of KIRs were compared with the other populations' viz., Chinese Mongolian, Chinese Han, Greek and Brazilian data. The expected heterozygosity of KIR alleles and their rank in gene diversity among the population of Puttaparthi were also discussed.


Acceleration rate of mitral inflow E wave: a novel transmitral doppler index for assessing diastolic function.
We performed comprehensive transmitral and pulmonary venous Doppler echocardiographic studies to devise a novel index of diastolic function. This is the first study to assess the utility of the acceleration rate (AR) of the E wave of mitral inflow as a primary diagnostic modality for assessing diastolic function. Study group consisted of 84 patients (53 + 11 years) with left ventricle (LV) diastolic dysfunction and 34 healthy people (35 ± 9 years) as control group, who were referred for clinically indicated two-dimensional transthoracic echocardiogram (TTE) during 2012 and 2013 to Imam Hospital. Normal controls were defined as patients without clinical evidence of cardiac disease and had normal TTE. LV diastolic function was determined according to standardized protocol of American Society of Echocardiography (ASE). As our new parameter, AR of E wave of mitral inflow was also measured in all patients. It was represented by the slope of the line between onset of E wave and peak of it. Correlation between AR of E wave and LV diastolic function grade was measured using the Spearman correlation coefficient. Receiver operating characteristic (ROC) curve was used to determine the sensitivity and specificity of AR of E wave in diagnosing LV diastolic dysfunction in randomly selected two-thirds of population then its derived cutoff was evaluated in rest of the population. The institutional review board of the hospital approved the study protocol. All participants gave written informed consent. This investigation was in accordance with the Declaration of Helsinki. The mean value of AR was 1010 ± 420 cm/s(2) in patients whereas the mean value for the normal controls was 701 ± 210 cm/s(2). There was a strong and graded relation between AR of E wave of mitral inflow and LV diastolic function grade (Spearman P ≤0.0001, rs =0.69). ROC curve analysis revealed that AR of E wave of mitral inflow =750 cm/s(2) predicted moderate or severe LV diastolic dysfunction with 89 % sensitivity and 89 % specificity (area under curve [AUC] = 0.903, P <0.0001). Application of this cutoff on test group showed 96 % sensitivity and 77 % specificity with AUC = 0.932 and P <0.0001. AR of E wave of mitral inflow could be used for assessment of diastolic function, especially moderate or severe diastolic dysfunction. However, before its clinical application, external validation should be considered.


[Measurement of clinical pain intensity: quantitative sensory testing].
Pain is a private internal event, the degree of which can not be evaluated directly by others. The measurement of the nociceptive pain threshold may apply to the evaluation of the degree of the pain. The instruments using mechanical and electrical stimulation are available for the clinical application. Thermotest allows the testing of quantitative evaluation of thermal thresholds such as heat, cold, and heat and cold pain sensation. A machine for the thermotest using radiant heat and contact heat is available in Japan. There is an instrument using puncturing stimulus and pressure stimulus for a mechanical stimulation. A puncturing stimulus is suitable for the clinical use with a handy instrument such as von Frey filaments and needle type algometry. In clinical practice, the pressure algometry is usually applied over a bony surface or over the muscle. The method seems to be well suited for the evaluation of pain in musculoskeletal disorders. The electrical stimulation test provides a quantitative measure of pain tolerance to a transcutaneous stimulation, neuroselective for large and small myelinated and unmyelinated nerve fibers. There are characteristics including advantages and faults in each method, and the method of measurement corresponding to the purpose and application is important.


Field testing of decision coaching with a decision aid for parents facing extreme prematurity.
The objective of this study is to assess and modify an existing decision aid and field-test decision coaching with the modified aid during consultations with parents facing potential delivery at 23 to 24 weeks gestation. International Patient Decision Aid Standards instrument (IPDASi) scoring deficits, multi-stakeholder group feedback and α-testing guided modifications. Feasibility/acceptability were assessed. The Decisional Conflict Scale was used to measure participants' decisional conflict before (T1) and immediately after (T2) the consultation. IPDASi assessment of the existing aid (score 11/35) indicated it required updated data, more information and a palliative care description. Following modification, IPDASi score increased to 26/35. Twenty subjects (12 pregnancies) participated in field-testing; 15 completed all questionnaires. Most participants (89%) would definitely recommend this form of consultation. Decisional conflict scores decreased (P<0.001) between T1 (52±25) and T2 (10±16). Field testing demonstrated that consultations using the aid with decision coaching were feasible, reduced decisional conflict and may facilitate shared decision-making.


Health status assessment in a hypertension section of an internal medicine clinic.
The authors obtained health status instrument information with the SF-36 and COOP charts distributed in random order (along with selected items from the Hypertension Technology of Patient Experience [TyPE] tool) to patients attending the hypertension section of an internal medicine clinic. The goal was to examine and compare potential associations of clinical, demographic, and/or treatment variables with SF-36/COOP health status scale variables in a nonuniversity urban clinic. One hundred fifty-eight pairs of health status instruments were returned (62% response rate). One hundred (64%) had a diagnosis of hypertension, 81 (51%) were males, 94 (60%) were older than 65 years, and 122 (78%) were white. Clinical, demographic, and treatment measurements were studied using regression analysis; the estimated regressions accounted for 4-32% of the variation in the COOP scales and 8-19% in the SF-36 scales. The number of coexisting diseases, gender, and diagnosis of hypertension were the most frequent significant variables associated with health status scale outcomes for each health status instrument. For most COOP and some SF-36 scales, there was a significant hypertension by gender interaction indicating that women with a hypertension diagnosis report better health status than women seen for other conditions; hypertension diagnosis had little effect on men's reported health status for most scales. Further study is necessary to confirm these results, yet the regression models developed in this study suggest that health status as assessed by these instruments is affected by multiple and not always obvious factors.


A Spatiotemporal and Multisensory Approach to Designing Wearable Clinical ICU Alarms.
In health care, auditory alarms are an important aspect of an informatics system that monitors patients and alerts clinicians attending to multiple concurrent tasks. However, the volume, design, and pervasiveness of existing Intensive Care Unit (ICU) alarms can make it difficult to quickly distinguish their meaning and importance. In this study, we evaluated the effectiveness of two design approaches not yet explored in a smartwatch-based alarm system designed for ICU use: (1) using audiovisual spatial colocalization and (2) adding haptic (i.e., touch) information. We compared the performance of 30 study participants using ICU smartwatch alarms containing auditory icons in two implementations of the audio modality: colocalized with the visual cue on the smartwatch's low-quality speaker versus delivered from a higher quality speaker located two feet away from participants (like a stationary alarm bay situated near patients in the ICU). Additionally, we compared participant performance using alarms with two sensory modalities (visual and audio) against alarms with three sensory modalities (adding haptic cues). Participants were 10.1% (0.24s) faster at responding to alarms when auditory information was delivered from the smartwatch instead of the higher quality external speaker. Meanwhile, adding haptic information to alarms improved response times to alarms by 12.2% (0.23s) and response times on their primary task by 10.3% (0.08s). Participants rated learnability and ease of use higher for alarms with haptic information. These small but statistically significant improvements demonstrate that audiovisual colocalization and multisensory alarm design can improve user response times.


The psychotherapy of genetics.
The evolution of genomic science and its effect on medicine and health care offer opportunities for family therapists to participate in the comprehensive care of patients and families with genetic disorders. This article provides an overview of what we now know about the psychological and interpersonal experience of patients and families facing some of these illnesses. Case examples illustrate the process of decision-making about testing and treatment, and the importance of understanding developmental issues and transgenerational family dynamics in any related psychotherapy. Challenging emotional issues include managing anger, ambivalence, and guilt; challenging interpersonal issues include dealing with differing coping and communication styles, decisions about disclosure and secrets, and conflict resolution. Family-oriented interventions include individual, couple, and family therapy, and psychoeducational groups. Recommendations are made for family therapists to participate as part of the genetic healthcare team.


Exploring the Steric and Electronic Factors Governing the Regio- and Enantioselectivity of the Pd-Catalyzed Decarboxylative Generation and Allylation of 2-Azaallyl Anions.
The impact of the steric and electronic factors in both the para-substituted benzaldimine and 2,2-diarylglycine components on the regioselectivity and enantioselectivity of the palladium-catalyzed decarboxylative allylation of allyl 2,2-diarylglycinate aryl imines was explored. These studies revealed that using 2,2-di(2-methoxyphenyl)glycine as the amino acid linchpin allowed for the exclusive synthesis of the desired homoallylic benzophenone imine regioisomers, independent of the nature of the imine moiety, in typically high yields. The resulting enantiomeric ratios, however, are slightly decreased in comparison to the transformations involving the corresponding allyl 2,2-diphenylglycinate imines, but this is more than balanced out by the increases in yield and regioselectivity. Overall, these studies suggest a general strategy for the highly regioselective functionalization of 2-azaallyl anions.


Virological and immunological impact of tuberculosis on human immunodeficiency virus type 1 disease.
Unlike other opportunistic infections associated with human immunodeficiency virus (HIV) type 1, tuberculosis (TB) occurs throughout the course of HIV-1 infection, and, as a chronic infection, its impact on viral activity is sustained. In dually infected subjects, HIV-1 load and heterogeneity are increased both locally and systemically during active TB. Studies over the past decade have indicated that Mycobacterium tuberculosis (MTB) infection supports HIV-1 replication and dissemination through the dysregulation of host cytokines, chemokines, and their receptors. Furthermore, concentrations of HIV-1 inhibitory chemokines are limited during TB and at sites of MTB infection. Cumulatively, these data indicate that TB provides a milieu of continuous cellular activation and irregularities in cytokine and chemokine circuits that are permissive of viral replication and expansion in situ. I address new research that has identified the basis for the augmentation of HIV-1 replication during TB and discuss potential immunotherapies to contain viral expansion during TB.


Clinical applications of L-glutamine: past, present, and future.
This review will attempt to summarize recent clinical data on glutamine's use. It will present the concept of glutamine as a "drug" or "nutraceutical," given in addition to standard nutrition support. Key references will be discussed, and clinical recommendations with regard to patients who may benefit and dosing are also provided. Glutamine, traditionally considered a nonessential amino acid, now is considered "conditionally essential" after critical illness, stress, and injury. States of illness or injury can lead to a significant decrease in plasma levels of glutamine, and when this decrease is severe, it has been correlated with increased mortality. Laboratory data have demonstrated numerous benefits of glutamine in experimental models of critical illness, cancer, and cardiac injury. The mechanism of these protective effects includes attenuated proinflammatory cytokine expression, improved gut barrier function, enhanced ability to mount a stress response, improved immune cell function, and decreased mortality. Over the last 10 years, clinical trials of glutamine supplementation in critical illness, surgical stress, and cancer have shown benefit with regard to mortality, length of stay, and infectious morbidity. However, data demonstrating a lack of benefit with glutamine supplementation in some patients have been presented as well. It appears that dose and route of administration clearly influence the benefit observed from glutamine administration, with high-dose parenteral glutamine demonstrating an advantage over low-dose enteral glutamine. High-dose or parenteral (> 0.25 to 0.30 g/kg/day IV or >or=30 g/day enterally) glutamine appears to demonstrate the greatest potential for benefit in hospitalized patients. No evidence of harm has been observed in studies conducted to date; thus, further clinical trials using glutamine as a pharmacologic supplement to standard nutrition are warranted.


CADASIL: what component of the vessel wall is really a target for Notch 3 gene mutations?
Cerebral Autosomal Dominant Arteriopathy with Subcortical Infarcts and Leukoencephalopathy (CADASIL) is a hereditary cerebrovascular disease leading to cognitive decline, dementia and recurrent strokes. The underlying angiopathy of the small vessels is characterized by basophilic degeneration of the media, Notch 3 protein accumulation in vessel wall and a unique type of ultrastructural deposits located nearby the basal lamina. In some cases of CADASIL, morphological changes similar to those observed in panarteritis nodosa (PAN) were found. PAN-like changes manifested as fibrinoid necrosis of the tunica media and perivascular inflammatory infiltrates were found in arteries not only in the central nervous system but also in internal organs. Presence of PAN-like changes indicates that some autoimmunological mechanisms can participate in the CADASIL process. Although vascular smooth muscle cells seem to be a primary target of the pathogenic process triggered by mutations in Notch 3 gene they are probably not the only target. This article gives a brief overview on the morphologic spectrum of the vascular pathological changes in CADASIL and discusses some of the relevant mechanisms that lead from Notch 3 mutations to ischemic infarcts.


Extracellular matrix regulates glomerular epithelial cell survival and proliferation.
Glomerular epithelial cell (GEC) injury and apoptosis may contribute to sclerosis in glomerulonephritis. The present study addresses signals that regulate survival of GEC in culture and in the acute puromycin aminonucleoside nephrosis (PAN) model of GEC injury in vivo. Compared with GEC on plastic substratum, adhesion to collagen increased activation of focal adhesion kinase (FAK), c-Src, and ERK and facilitated survival (prevented apoptosis). GEC on plastic exhibited increased caspase-8 and -9 activities, increased expression of the proapoptotic protein, Bax, and decreased the antiapoptotic protein, Bcl-XL, compared with collagen. Stable expression of constitutively active mutants of FAK (CD2-FAK) or MEK (R4F-MEK) activated the ERK pathway and supplanted the requirement of collagen for survival. In contrast, expression of a Ras mutant that activates phosphatidylinositol 3-kinase but blocks ERK activation or pharmacological inhibition of the ERK pathway decreased survival on collagen. Glomeruli isolated from rats with PAN revealed increased beta1-integrin expression, along with increased activation of FAK, c-Src, and ERK, compared with controls. EGF receptor activation was undetectable in PAN. Therefore, adhesion to collagen, resulting in activation of FAK and the Ras-ERK pathway, supports GEC survival. Analogous signals for GEC survival are activated in PAN.


A primary care-based interdisciplinary team approach to the treatment of chronic pain utilizing a pragmatic clinical trials framework.
Chronic pain affects at least 116 million adults in the USA and exacts a tremendous cost in suffering and lost productivity. While health systems offer specialized pain services, the primary care setting is where most patients seek and receive care for pain. Primary care-based treatment of chronic pain by interdisciplinary teams (including behavioral specialists, nurse case managers, physical therapists, and pharmacists) is one of the most effective approaches for improving outcomes and managing costs. To ensure robust integration of such services into sustainable health-care programs, evaluations must be conducted by researchers well versed in the methodologies of clinical trials, mixed methods and implementation research, bioinformatics, health services, and cost-effectiveness. Recent national health policy changes, in addition to the increasing recognition of the high prevalence and cost of chronic pain conditions, present a unique opportunity to shift the care paradigm for patients with chronic pain.


Tissue changes in enlarged sternal bursas of turkeys.
Enlarged sternal bursas evident in turkeys beyond 12 weeks of age were examined grossly and microscopically in 169 turkeys 3 to 26 weeks old. Grossly, the enlarged sternal bursas were sacs of varied size and wall thickness containing fluid in amounts from a fraction of a milliliter up to 70 ml. The fluid was usually the consistency of serum but was sometimes viscid, and in infected bursas it would be purulent. Its color varied from amber to red to dark brown depending on the amounts of and the changes in the blood pigments. The synovial membranes were smooth and glistening and of variable thicknesses. Strands or cords of the synovial membrane of variable thickness and number extended from side to side of the sac. Microscopically, the synovial membrane was focally thickened in the small bursas and generally thickened in the larger bursas. The thickening was the result of an increase in the very cellular fibrous tissue and in the number and size of blood vessels. The lining cells were flat fibrocytes one cell thick in the smaller bursas, and round stratified cells in the larger bursas. In infected bursas, mononuclear and heterophilic infiltration occurred. Hemorrhaging occurred quite often, the result of trauma to the highly vascular synovial membrane.


Identification of the disulfide bonds in the recombinant somatomedin B domain of human vitronectin.
The NH(2)-terminal somatomedin B (SMB) domain (residues 1-44) of human vitronectin contains eight Cys residues organized into four disulfide bonds and is required for the binding of type 1 plasminogen activator inhibitor (PAI-1). In the present study, we map the four disulfide bonds in recombinant SMB (rSMB) and evaluate their functional importance. Active rSMB was purified from transformed Escherichia coli by immunoaffinity chromatography using a monoclonal antibody that recognizes a conformational epitope in SMB (monoclonal antibody 153). Plasmon surface resonance (BIAcore) and competitive enzyme-linked immunosorbent assays demonstrate that the purified rSMB domain and intact urea-activated vitronectin have similar PAI-1 binding activities. The individual disulfide linkages present in active rSMB were investigated by CNBr cleavage, partial reduction and S-alkylation, mass spectrometry, and protein sequencing. Two pairs of disulfide bonds at the NH(2)-terminal portion of active rSMB were identified as Cys(5)-Cys(9) and Cys(19)-Cys(21). Selective reduction/S-alkylation of these two disulfide linkages caused the complete loss of PAI-1 binding activity. The other two pairs of disulfide bonds in the COOH-terminal portion of rSMB were identified as Cys(25)-Cys(31) and Cys(32)-Cys(39) by protease-generated peptide mapping of partially reduced and S-alkylated rSMB. These results suggest a linear uncrossed pattern for the disulfide bond topology of rSMB that is distinct from the crossed pattern present in most small disulfide bond-rich proteins.


Anticoagulation vs Antiplatelet Treatment in Patients with Carotid and Vertebral Artery Dissection: A Study of 370 Patients and Literature Review.
Dissection of the carotid and vertebral arteries is an important cause of stroke in young patients. The objective of this study is to compare antithrombotic treatments in patients with carotid and vertebral dissections. Three hundred seventy patients with carotid and vertebral artery dissections were included. Univariate and multivariate analyses were conducted to analyze the association between treatment and new or recurrent events and clinical outcome. Mean follow-up was 24.3 months. In patients with spontaneous dissection, 55% received antiplatelets, 29.4% anticoagulation, and 12.6% combined treatment. New or recurrent ischemic and hemorrhagic events occurred in 9.6% of patients on antiplatelets, 10.4% on anticoagulation, and 13.3% on combined treatment. For traumatic dissection, 58.3% received antiplatelets, 26.9% anticoagulation, and 10.2% combined treatment. New or recurrent ischemic and hemorrhagic events occurred in 6.9% on antiplatelets, 11.1% on anticoagulation, and 20% on combined treatment. In patients with intracranial dissection, 63.1% were started on antiplatelets, 19.7% on anticoagulation, and 14.5% on combined treatment. Ischemic and hemorrhagic events occurred in 8.5% on antiplatelet treatment, 15.4% on anticoagulation, and 18.2% on combined treatment. In patients with extracranial dissection, 54.4% were on antiplatelets, 28.9% on anticoagulation, and 11.2% on combined treatment. Ischemic and hemorrhagic events occurred in 10.1% on antiplatelet treatment, 9.3% on anticoagulation, and 13.8% on combined treatment. The association between antithrombotic treatment and ischemic/hemorrhagic events and clinical outcome was not significant for all subtypes of dissection. The rate of new or recurrent events is similar with antiplatelet and anticoagulation treatment in treating intracranial and extracranial carotid and vertebral artery dissection.


[Palliative therapy in cancer. 6. Quality of life and BRM therapy in cancer-patients].
Almost all of anti-cancer drugs developed, injured not only the tumor cells but also the normal cells. For this reason, immunotherapy which injures specifically the tumor cells, has been developed. However, the clinical studies conducted in the past 20 years tend to indicate that even though the immunotherapy does not produce any serious side effect, the anti-tumor effect are not totally satisfactory in spite of marked prolongation of survivals. In recent years, various fields of cancer therapy have seen the development and application of treatment modalities which take into account the quality of the patients. Several patient's self-assessment questionnaires are widely used practical at the aims to assess what happens in the cancer-patients and to improve the cancer treatment. The measurements of quality of life have been performed on both immunochemotherapy group and chemotherapy group. The results showed the remarkable improvement of QOL in the immunochemotherapy group. The biotechnological advances have been made in the last 10 years. Various cytokines which participate in the immunological response between cancer and host. Many of these biologicals that will be called as biological response modifiers (BRM) are cell products of lymphocytes or macrophage. Systemic administration of exogenous cytokines to act against a tumor at a distant site may not be as successful as more localized therapy in situation. In most circumstances, cytokines are produced transiently at a local site acting in an autocrine or paracrine manner. Cytokines, whether naturally produced, or administered therapeutically, are rapidly clear from the circulation, both as a result of finding to often ubiquitous cell surface receptors, and by active excretion. So, the local injections of TNF and LAK cells are effective to decrease tumor size without the side effects. Finally, the serum level of IL-2 and TNF-alpha were determined for 24 healthy volunteers and 42 patients with advanced or terminal cancer, and the relationship between these levels and the QOL was investigated. In the cancer-patients who showed a higher serum level of IL-2 than the healthy volunteers, it was elucidated that there was an inverse correlation between the serum IL-2 level and the QOL. This finding cannot be explained at present, but it is surmised that, for this subpopulation of cancer-patients, the serum IL-2 level can probably serve as an objective biochemical index for use in evaluating the QOL.


Experimental and simulation studies of multivariable adaptive optimization of continuous bioreactors using bilevel forgetting factors.
A multivariable on-line adaptive optimization algorithm using a bilevel forgetting factor method was developed and applied to a continuous baker's yeast culture in simulation and experimental studies to maximize the cellular productivity by manipulating the dilution rate and the temperature. The algorithm showed a good optimization speed and a good adaptability and reoptimization capability. The algorithm was able to stably maintain the process around the optimum point for an extended period of time. Two cases were investigated: an unconstrained and a constrained optimization. In the constrained optimization the ethanol concentration was used as an index for the baking quality of yeast cells. An equality constraint with a quadratic penalty was imposed on the ethanol concentration to keep its level close to a hypothetical "optimum" value. The developed algorithm was experimentally applied to a baker's yeast culture to demonstrate its validity. Only unconstrained optimization was carried out experimentally. A set of tuning parameter values was suggested after evaluating the results from several experimental runs. With those tuning parameter values the optimization took 50-90 h. At the attained steady state the dilution rate was 0.310 h(-1) the temperature 32.8 degrees C, and the cellular productivity 1.50 g/L/h.


Immunological consequences of stress-related proteins--cytosolic tryparedoxin peroxidase and chaperonin TCP20--identified in splenic amastigotes of Leishmania donovani as Th1 stimulatory, in experimental visceral leishmaniasis.
In earlier studies, proteomic characterization of splenic amastigote fractions from clinical isolates of Leishmania donovani, exhibiting significant cellular responses in cured Leishmania subjects, led to the identification of cytosolic tryparedoxin peroxidase (LdcTryP) and chaperonin-TCP20 (LdTCP20) as Th1-stimulatory proteins. Both the proteins, particularly LdTCP20 for the first time, were successfully cloned, overexpressed, purified and were found to be localized in the cytosol of purified splenic amastigotes. When evaluated against lymphocytes of cured Leishmania-infected hamsters, the purified recombinant proteins (rLdcTryP and rLdTCP20) induced their proliferations as well as nitric oxide production. Similarly, these proteins also generated Th1-type cytokines (IFN-γ/IL-12) from stimulated PBMCs of cured/endemic Leishmania patients. Further, vaccination with rLdcTryP elicited noticeable delayed-type hypersensitivity response and offered considerably good prophylactic efficacy (~78% inhibition) against L. donovani challenge in hamsters, which was well supported by the increased mRNA expression of Th1 and Th2 cytokines. However, animals vaccinated with rLdTCP20 exhibited comparatively lesser prophylactic efficacy (~55%) with inferior immunological response. The results indicate the potentiality of rLdcTryP protein, between the two, as a suitable anti-leishmanial vaccine. Since, rLdTCP20 is also an important target, for optimization, further attempts towards determination of immunodominant regions for designing fusion peptides may be taken up.


[The dependence of diet macronutrient composition and energy intake from human phase of the weekly cycle – weekdays/weekends].
When planning your diet regime is desirable to focus on the natural feeding behavior of the person. However, many aspects of eating behavior are not studied. The aim of this work was to study the changes of diet calorie and nutrient composition depending on the body mass index (BMI), time of day, week cycle phase: weekends/weekdays, and the availability of rational installation in nutrition. The study involved 67 women aged 29 to 49 years with a BMI of 18.3 to 34.2 kg/m2. Macronutrient composition and energy value of the diet were evaluated by analyzing the food diaries, which were filled by all respondents continuously for 8 workdays and 4 weekends. The results of this study showed no significant correlation between BMI and calorie intake, while a positive correlation was observed between BMI and fat component of the diet (r=0.362, p≤0.05). It was also found that the restriction of food intake in the evening (after 8 pm) did not lead to a decrease in total daily energy value of the diet, and was characterized by the consumption in the evening about 31% of the total fat intake. Diet energy value was significantly higher on weekends than during the week and was 2376±394 against 1940±402 kcal (p<0.05). According to the data obtained, compliance «restriction of supply in the evening» does not lead to a significant reduction in daily caloric intake.


Sea urchin spine injuries of the hand.
Sea urchin spine injuries are common. They usually cause local pain and swelling that subsides. Chronic granulation is rare. We report two cases of sea urchin granulomata involving finger metacarpophalangeal joints. Both resolved following surgery.


Linearization of excitatory synaptic integration at no extra cost.
In many theories of neural computation, linearly summed synaptic activation is a pervasive assumption for the computations performed by individual neurons. Indeed, for certain nominally optimal models, linear summation is required. However, the biophysical mechanisms needed to produce linear summation may add to the energy-cost of neural processing. Thus, the benefits provided by linear summation may be outweighed by the energy-costs. Using voltage-gated conductances in a relatively simple neuron model, this paper quantifies the cost of linearizing dendritically localized synaptic activation. Different combinations of voltage-gated conductances were examined, and many are found to produce linearization; here, four of these models are presented. Comparing the energy-costs to a purely passive model, reveals minimal or even no additional costs in some cases.


Nebulized budesonide inhalation suspension compared with cromolyn sodium nebulizer solution for asthma in young children: results of a randomized outcomes trial.
The availability of antiinflammatory asthma medications for infants and young children has been limited. The objective of this study was to compare effects of nebulized budesonide inhalation suspension and cromolyn sodium nebulizer solution on asthma-related health outcomes in young children with asthma. We conducted a randomized, parallel-group, 52-week, open-label study in 36 US clinical sites. Patients included 335 children who were 2 to 6 years of age and had persistent asthma that had been treated with at least 1 long-term control medication; 287 children (86%) completed the study. Patients received budesonide inhalation suspension, 0.5 mg daily (n = 168), or cromolyn sodium nebulizer solution, 20 mg 4 times daily (n = 167), for 8 weeks, followed by dose titration at the investigator's discretion. The main outcome measure was the rate of asthma exacerbations over 52 weeks. Secondary measures included times to first asthma exacerbation and first use of additional asthma therapy, asthma symptom scores, rescue medication use, and health care resource use. The budesonide group had a mean (median) asthma exacerbation rate of 1.23 (0.99) per year compared with 2.41 (1.85) for the cromolyn group, significantly longer times to first exacerbation and first use of additional long-term asthma medication, greater improvements in asthma symptom scores, reduced use of rescue medication, and fewer urgent care visits. Both treatments were well tolerated. Budesonide inhalation suspension was more effective than nebulized cromolyn sodium in young children with persistent asthma. Both treatments were well tolerated with similar adverse event profiles.


Immunodetection of PrPSc using Western and Slot blotting techniques.
Prion infectivity is often linked to presence of the protease-resistant isoform of prion protein (PrP), PrP(res); therefore, it is of highest interest to have convenient methods for rapid detection of PrP(res) in the research laboratory. For detection of PrP(res) in model systems to confirm infectivity, there are several methods that can be applied. This chapter focuses on detection of PrP(res) by proteinase K digestion followed by Western blot, which is the only method that is both quantitative and qualitative. For large-scale screening of PrP(res) content in samples, the dot blot method offers a great advantage for detecting PrP(res), and this method is also thoroughly described in this chapter.


How 'sensible' is the UK Sensible Drinking message? Preliminary findings amongst newly matriculated female university students in Scotland.
Current levels and patterns of alcohol drinking continue to cause concern, particularly amongst young females. Effective interventions remain elusive. The aim of this study was to investigate the levels of knowledge relating to UK 'Sensible Drinking' guidelines, definitions of binge drinking and attitude to drink labelling initiatives amongst female school leavers enrolling at university. Non-standardized questionnaires were administered and completed by researchers during the process of matriculation (response rate 94%; n = 180). The survey revealed that recommended daily guidelines for women were not recalled by 54% of participants; 52% could not quote the unit content of their favoured alcoholic drink, whereas only 14% reported the use of the UK unit system to guide drinking. Personal quantitative definitions of binge drinking varied by a factor of 18. A rewording of the UK Sensible Drinking message to one recognizing the needs of different population groups and their particular language of consumption measure is timely.


Present and future activities of the IAEA on internal dosimetry: lessons learned from international intercomparisons.
The International Atomic Energy Agency (IAEA) conducts safety activities to support the assessment of occupational exposure due to intakes of radionuclides; a comprehensive set of safety documents will soon be completed. In recent years, extensive improvements in measurement techniques, phantoms and computational tools have been made. Thus, it is important for laboratories involved in internal dosimetry to undergo performance testing procedures to demonstrate the correctness of the methods applied and also to determine the consistency of their results with those obtained by other laboratories. Several intercomparisons were organised, and they revealed significant differences among laboratories in their approaches, methods and assumptions, and consequently in their results. This paper presents the current and future IAEA activities in support of assessment of occupational exposure due to intakes of radionuclides in the IAEA Member States, as well as the lessons learned from several intercomparison exercises in the last 5 years.


Assessment of knowledge and factors that may predict willingness to volunteerism: a pilot study of community-directed distributors in Anambra state.
Malaria as a leading cause of death in many developing countries requires urgent interventions. In order to improve access to healthcare, trained volunteers are used to distribute health commodities. The present study aims at determining knowledge and factors that may predict willingness to volunteerism in a developing country. This was a cross-sectional descriptive study carried out in 2014 among 284 community-directed distributors in three rural communities in Anambra, Southeast Nigeria using pretested semi-structured questionnaires. The age range of volunteers was 21-79 years. Most (71.8%) are females and are married (83.1). Only 5.6% of the volunteers did not have any formal education. The predominant occupation is trading (52.5). Most volunteers (78.5%) could define the term volunteerism. Less than half (40.1%) knew the resources that could be volunteered. Most (67.3%) felt that volunteerism is most needed in church activities. Many respondents (58.8%) had volunteered for one or more programs previously. The most common challenge faced was interference with other income generating activities (66.5%). Retired males were more likely to volunteer than retired females (P ≤ 0.01). However, females are more likely to volunteer if the main reason of volunteering is to help people (P ≤ 0.01). The more educated ones believe that volunteerism will help them to be selected for other community programs. Most respondents had volunteered for other programs and the motivating factors included the satisfaction derived from helping others and the hope of being used for other community programs.


Recent advances on bisphenol-A and endocrine disruptor effects on human prostate cancer.
Endocrine disrupting chemicals (EDCs) are man-made substances widespread in the environment that include, among many others, bisphenol A (BPA), organochlorinated pesticides and hormone derivatives detectable in meat from animals raised in concentrated animal feeding operations. Increasing evidence indicates that EDCs have a negative impact on human health as well as on male and female fertility. They may also be associated with some endocrine diseases and increased incidence of breast and prostate cancer. This review aims to summarize available data on the (potential) impact of some common EDCs, focusing particularly on BPA, prostate cancer and their mechanisms of action. These compounds interfere with normal hormone signal pathway transduction, resulting in prolonged exposure of receptors to stimuli or interference with cellular hormone signaling in target cells. Understanding the effects of BPA and other EDCs as well as their molecular mechanism(s) may be useful in sensitizing the scientific community and the manufacturing industry to the importance of finding alternatives to their indiscriminate use.


Markers of melanocytic tumour progression.
Melanoma progression markers can be defined as molecules with a preferential expression in one or a few stages of melanocytic tumour development. These molecules include growth factors, growth factors receptors, adhesion molecules, proteases and related components. Immunohistochemical studies suggest that some of these molecules are useful as prognostic markers in melanoma patients. In cutaneous melanocytic lesions, the distribution of E-cadherin, a member of a family of cell adhesion molecules that mediate cell-cell interactions by means of Ca2+ dependent, homophilic interactions, appears to be complex. Although a decrease of E-cadherin would be expected with invasive tumour growth in advanced primary melanoma and eventually in metastasis, surprisingly an increase is found, whereas alpha- and beta-catenin, (cytoplasmic) molecules functionally associated with E-cadherin, are detected in all benign and malignant lesions. A possible interpretation includes a difference in the morphogenesis and function of melanocytic cells, compared with epithelial cells. Further research is needed to clarify the role of E-cadherin/catenin during melanoma progression.


Upregulated long non-coding RNA LINC00152 expression is associated with progression and poor prognosis of tongue squamous cell carcinoma.
Altered expression of long non-coding RNAs (lncRNAs) associated with human carcinogenesis and might be used as diagnosis and prognosis biomarkers. However, the expression of lncRNAs in tongue squamous cell carcinoma (TSCC) and their relevance on the diagnosis, progression and prognosis of TSCC have not been thoroughly elucidated. To discover novel TSCC-related lncRNAs, we analyzed the lncRNA expression patterns in two sets of previously published TSCC gene expression profile data (GSE30784 and GSE9844), and found that long intergenic non-coding RNA 152 (LINC00152) was significantly upregulated in TSCC samples. We then detected LINC00152 expression in two other cohorts of TSCC samples. Quantitative Real time PCR (qRT-PCR) results indicated that LINC00152 was highly expressed in 15 primary TSCC biopsies when compared with 14 adjacent non-tumor tongue squamous cell epithelium samples. The expression of LINC00152 was also measured in 182 paraffin-embedded human TSCC tissues by in situ hybridization, increased expression of LINC00152 was significantly correlated with TSCC progression, such as T stage (p = 0.009), N stage (p = 0.036), TNM stage (p = 0.017), and associated with relapse (p < 0.001), and invasion (p < 0.001). Kaplan-Meier analysis demonstrated that increased LINC00152 expression contributed to both poor overall survival (p = 0.006) and disease-free survival (p = 0.007) of TSCC patients. These findings suggest that LINC00152 might serve as a potential biomarker for early detection and prognosis prediction of TSCC.


Pulse Pressure, Instead of Brachium-Ankle Pulse Wave Velocity, is Associated with Reduced Kidney Function in a Chinese Han Population.
In this study, we aim to investigate the association between renal function and arterial stiffness in a Chinese Han population, and further to discuss the effects of smoking on renal function. We collected the data of the brachium-ankle pulse wave velocity (baPWV), blood pressure, blood chemistry and smoking status. Then, the multiple linear regression was done to explore the relationship between estimated glomerular filtration (eGFR) and baPWV. Further, the parameters were compared among the four groups divided according to the quartiles of baPWV. Finally, the baPWV, eGFR and albuminuria values were compared between smokers and non-smokers. baPWV is associated with eGFR in the correlation analysis and univariate linear regression model. After adjustment, the pulse pressure (PP) instead of baPWV showed a significant association with eGFR. Nevertheless, the eGFR values differed among the four baPWV groups; the baPWV values were significantly higher in the subjects at the CKD (eGFR<60 mL/min/1.73 m2) and the early CKD stage (eGFR60-80 mL/min/1.73 m2). The baPWV values and the ratio of proteinuria were significantly increased in smokers. PP but not baPWV is a predictor of declined renal function. Smokers have worse arterial stiffness and worse renal function.


Selective biotransformation of taxol to 6 alpha-hydroxytaxol by human cytochrome P450 2C8.
The principal taxol biotransformation reaction of humans and of human hepatic in vitro preparations is 6 alpha-hydroxylation of the taxane ring, but a separate, minor hydroxylation pathway (metabolite B formation) also exists. Taxol metabolism was studied using membrane fractions from Hep G2 cells infected with recombinant vaccinia viruses that contain complementary DNAs encoding several human cytochrome P450 enzymes. Only P450 2C8 formed detectable 6 alpha-hydroxytaxol. Metabolite B formation was catalyzed by complementary DNA-expressed 3A3 and 3A4, but not by 3A5. Each P450 3A preparation catalyzed felodipine oxidation. The apparent Km and Vmax values for taxol 6 alpha-hydroxylation were 5.4 +/- 1.0 microM and 30 +/- 1.5 nmol/min/nmol P450, respectively, for complementary DNA-expressed P450 2C8; the values were 4.0 +/- 1.0 microM and 0.87 +/- 0.06 nmol/min/mg protein, respectively, for human hepatic microsomes. The inhibition of 6 alpha-hydroxytaxol formation by quercetin was competitive with an apparent Ki of 1.3 or 1.1 microM with 2C8 or hepatic microsomes, respectively; retinoic acid was inhibitory, showing an apparent Ki of 27 microM with hepatic microsomes; inhibition by tolbutamide was complex, weak, and unlikely to be clinically relevant. The correlation between hepatic 2C8 protein content and 6 alpha-hydroxytaxol formation was high (r2 = 0.82), while the correlation with 2C9 content was low (r2 = 0.38). These data show that human biotransformation routes of taxol result from catalysis by specific enzymes of two P450 families and that taxol 6 alpha-hydroxylation is a useful indicator of P450 2C8 activity in human hepatic microsomes.


Neuroanatomical digital image processing in CT-guided stereotactic operations.
In order to utilize computed tomography (CT) for functional neurosurgical procedures, a number of problems must be resolved. One such problem is the development of an appropriate technique for digital processing of neuroanatomical images. Our previous work concerned a numerical reproduction of a well-known stereotactic atlas and its three-dimensional manipulation. In order to efficiently utilize this information with CT images we propose a technique for data reduction that allows the reconstruction of anatomical structures from the atlas. The logic of implementing these data in CT-assisted stereotactic surgery is described.


Spatiotemporal correlation between phase singularities and wavebreaks during ventricular fibrillation.
Phase Singularity and Wavebreak. Phase maps and the detection of phase singularities (PSs) have become a well-developed method for characterizing the organization of ventricular fibrillation (VF). How precisely PS colocalizes with wavebreak (WB) during VF, however, is unknown. We performed optical mapping of 27 episodes of VF in nine Langendorff-perfused rabbit hearts. A WB is a point where the activation wavefront and the repolarization waveback meet. A PS is a site where its phase is ambiguous and its neighboring pixels exhibit a continuous phase progression from -pi to +pi. The correlation coefficient between the number of WBs and PSs was 0.78 +/- 0.09 for each heart and 0.81 for all VF episodes (P < 0.001), indicating a significant temporal correlation. We then superimposed the WBs and PSs for every 100 frames of each episode. These maps showed a high degree of spatial colocalization. To quantify spatial colocalization, the spatial shifts between the cumulative maps of WBs and PSs in corresponding frames were calculated by automatic alignment to obtain maximum overlap between these two maps. The spatial shifts were 0.04 +/- 0.31 mm on the x-axis and 0.06 +/- 0.27 mm on the y-axis over a 20 x 20 mm2 mapped field, indicating highly significant spatial correlation. Phase mapping provides a convenient and robust approach to quantitatively describe wave propagation and organization during VF. The close spatiotemporal correlation between PSs and WBs establishes that PSs are a valid alternate representation of WB during VF and further validated the use of phase mapping in the study of VF dynamics.


Chronic pain in later life women: issues and challenges from the research literature.
Because research on chronic pain in older adults comes from many disciplines, existing information about geriatric pain is widely scattered. In order to establish a foundation from which to construct future research and interventions for older women and men, we developed a comprehensive, multidisciplinary database of the chronic pain and aging literature published between 1990 and 1998. We searched 10 electronic databases that index scientific journals and found 302 articles that focused on chronic pain in later life. Given the disproportionately high number of women in many of the study samples, gender comparisons were not always statistically feasible. Because a limited number of studies reported sex differences in older adults, it is difficult to determine whether older women's experiences with chronic pain are unique and require special attention from health care providers or whether the causes, treatments, and consequences of chronic pain should be considered universal to the older-population as a whole.


Congenital high airway obstruction syndrome and airway reconstruction: an evolving paradigm.
To refine the classic definition of, and provide a working definition for, congenital high airway obstruction syndrome (CHAOS) and to discuss the various aspects of long-term airway reconstruction, including the range of laryngeal anomalies and the various techniques for reconstruction. Retrospective chart review. Four children (age range, 2-8 years) with CHAOS who presented to a single tertiary care children's hospital for pediatric airway reconstruction between 1995 and 2000. To date, CHAOS remains poorly described in the otolaryngologic literature. We propose the following working definition for pediatric cases of CHAOS: any neonate who needs a surgical airway within 1 hour of birth owing to high upper airway (ie, glottic, subglottic, or upper tracheal) obstruction and who cannot be tracheally intubated other than through a persistent tracheoesophageal fistula. Therefore, CHAOS has 3 possible presentations: (1) complete laryngeal atresia without an esophageal fistula, (2) complete laryngeal atresia with a tracheoesophageal fistula, and (3) near-complete high upper airway obstruction. Management of the airway, particularly in regard to long-term reconstruction, in children with CHAOS is complex and challenging.


A daytime, abbreviated cardio-respiratory sleep study (CPT 95807-52) to acclimate insomnia patients with sleep disordered breathing to positive airway pressure (PAP-NAP).
To assess the impact of a daytime sleep medical procedure--the PAP-NAP--on adherence to positive airway pressure (PAP) therapy among insomnia patients with sleep disordered breathing (SDB) METHODS: The PAP-NAP is based on Current Procedural Terminology (CPT) codes and combines psychological and physiological treatments into one procedure, which increases contact time between SDB patients and polysomnography technologists to enhance PAP therapy adherence. Using a Sleep Dynamic Therapy framework, explicating SDB as a mind-body disorder, the PAP-NAP includes mask and pressure desensitization, emotion-focused therapy to overcome aversive emotional reactions, mental imagery to divert patient attention from mask or pressure sensations, and physiological exposure to PAP therapy during a 100-minute nap period. Patients treated with the PAP-NAP test (n = 39) were compared to an historical control group (n = 60) of insomnia patients with SDB who did not receive the test. All 99 insomnia patients were diagnosed with SDB (mean AHI 26.5 +/- 26.3, mean RDI 49.0 +/- 24.9), and all reported a history of psychiatric disorders or symptoms as well as resistance to PAP therapy. Among 39 patients completing the PAP-NAP, 90% completed overnight titrations, compared with 63% in the historical control group; 85% of the nap-tested group filled PAP therapy prescriptions for home use compared with 35% of controls; and 67% of the nap-tested group maintained regular use of PAP therapy compared with 23% of the control group. Using standards from the field of sleep medicine, the nap-tested group demonstrated objective adherence of 49% to 56% compared to 12% to 17% among controls. All studies were reimbursed using CPT 95807-52. In this pilot study, the PAP-NAP functioned as a brief, useful, reimbursable procedure to encourage adherence in insomnia patients with SDB in comparison to an historical control group that did not undergo the procedure.


Naturally Occurring β-Nicotinamide Adenine Dinucleotide-Independent Avibacterium paragallinarum Isolate in Peru.
The β-nicotinamide adenine dinucleotide (NAD) requirement has been considered to be essential for the isolation of the causal agent of infectious coryza, Avibacterium paragallinarum. Nevertheless, NAD-independent reports from South Africa and Mexico dismissed this paradigm. It is now accepted that both NAD-dependent and NAD-independent agents are able to cause infectious coryza and thus belong to the species A. paragallinarum. Here, we report for the first time in Peru a NAD-independent isolate from broiler chickens with typical signs of infectious coryza that have received a trivalent inactivated vaccine against infectious coryza. The isolate was identified based on its morphology, biochemical and serologic tests, and PCR results. Partial 16S rRNA gene sequence analysis confirmed the isolate as A. paragallinarum. There have been no cases of NAD-independent A. paragallinarum isolates reported in South America. Increasing reports around the world highlight not only the need to reconsider the in vitro nutritional requirements of this species for its correct isolation but also the cross-protection conferred by commercial infectious coryza vaccines against NAD-independent isolates.


Renal brush-border Na+-H+ exchange activity in the aging rat.
Amiloride-sensitive Na+-H+ exchange activity in brush-border membrane vesicles isolated from male rat proximal tubules was decreased in the senescent rat (24 mo) compared with the young adult (6 mo). There was no significant loss in Na+-H+ exchange activity in the kidneys of animals between 6 and 18 mo of age. Amiloride-insensitive Na+ uptake and the rate of pH gradient dissipation were not altered during aging. The decrease in sodium-dependent phosphate transport preceded the decline in Na+-H+ exchange activity by at least 6 mo. Sodium-dependent glucose transport was not significantly altered during aging. Thus various renal plasma membrane transport functions were affected differently in the aging rat. The decrease in Na+-H+ exchange activity during aging contrasted with the increase in exchange activity reported previously in acute ablation models of chronic renal failure.


Population pharmacokinetics and dosing considerations for the use of daptomycin in adult patients with haematological malignancies.
To assess the population pharmacokinetics (popPK) of daptomycin at the conventional dose of 6 mg/kg/day in a cohort of oncohaematological patients. Patients underwent serial blood sampling on day 3 of therapy (before dosing and at 0, 0.5, 1, 2, 3, 5, 7, 9 and 12 h after dosing) to assess the pharmacokinetic profile of daptomycin. PopPK and Monte Carlo simulation were performed to define the probability of target attainment (PTA) with 6, 8, 10 and 12 mg/kg/day of the pharmacokinetic/pharmacodynamic target of AUC 24 /MIC >1081. Thirty patients were recruited. A two-compartment open model with first-order intravenous input and first-order elimination was developed. Estimated creatinine clearance (CL CR ), serum albumin concentration (Alb) and presence of AML were covariates included in the final model. Monte Carlo simulation showed that the conventional 6 mg/kg/day dose resulted in optimal PTAs (≥80%) in the presence of pathogens with an MIC up to 0.5 mg/L only in patients with CL CR 50-100 mL/min/1.73 m 2 , Alb 26-45 g/L and a haematological diagnosis other than AML. Conversely, higher dosages, up to 12 mg/kg/day, were needed to achieve this goal in the presence of pathogens with an MIC of 0.25-0.5 mg/L in all of the other tested scenarios. In patients with CL CR 101-150 mL/min/1.73 m 2 and Alb 15-25 g/L, suboptimal PTAs (<60%) were predicted even with 12 mg/kg/day dosing . Our study provides a strong rationale for considering daptomycin dosages of ≥ 8 mg/kg/day in several clinical scenarios for oncohaematological patients. In some of these scenarios therapeutic drug monitoring could be a useful adjunct for optimized care.


Four copper(II) complexes with potentially tetradentate tripodal ligands.
The four title Cu(II) compounds are chloro[(2-furylmethyl)bis(2-pyridylmethyl)amine-N,N',N"]copper(II) perchlorate, [CuCl(C(17)H(17)N(3)O)]ClO(4), (I), chloro{2-[bis(2-pyridylmethyl)amino]ethanolato-N,N',N",O}copper(II) hemi[tetrachlorocopper(II)], [CuCl(C(14)H(17)N(3)O)][CuCl(4)](1/2), (II), chloro[(2-morpholinoethyl)bis(2-pyridylmethyl)amine-N,N',N",N"']copper(II) perchlorate, [CuCl(C(18)H(24)N(4)O)]ClO(4), (III), and chloro[(2-piperidinylethyl)bis(2-pyridylmethyl)amine-N,N',N",N"']copper(II) hexafluorophosphate, [CuCl(C(19)H(26)N(4))]PF(6), (IV). They have tripodal potentially tetradentate ligands. In (I), the O atom of the furan moiety weakly coordinates to the Cu atom at a distance of 2.750 (3) A.


[Changes of cytokines of splenocytes in mice immunized by mix recombinant BCG-EmII/3 and BCG-Em14-3-3 vaccine of Em].
To investigate the changes of cytokines of splenocytes in mice immunized by mix recombinant BCG-EmII/3 and BCG-Em14-3-3 vaccine of Echinococcus multilocularis(Em) and challenged by Em protoscoleces. BALB/c mice were vaccinated subcutaneously and intranasally by the vaccine respectively. In the eighth week of vaccination they were Challenged by Em protoscoleces.In the eighteenth week of infection they were killed for spleen. After splenocytes were separated,their culture was stimulated by EmAg or ConA and their supernatants were gathered. The levels of IL-2, IFN-gamma, TNF-alpha and IL-4 were measured by ELISA kit. Blank vector, BCG and PBS were served as control. In the groups of vaccine immunization, the levels of IFN-gamma and TNF-alpha increased obviously but the level of IL-4 decreased. The level of TNF-alpha in subcutaneous group was higher than that in intranasal group. Th1 response was induced in mice immunized by mix recombinant BCG-EmII/3 and BCG-Em14-3-3 vaccine of Em against the challenge by Em protoscoleces. Subcutaneous injection with this vaccine may be a good way of vaccination.


Competitive rational inhibitor design to 4-maleylaceto-acetate isomerase.
Tyrosinemia type I is the result of genetic disorder in fomaryl acetoacetase gene that leads to 4-fumaryl acetoacetate accumulation. The current treatment for tyrosinemia type I is nitisinone that inhibits 4-hydroxyphenyl pyruvic dioxygenase in competitive manner. In the present study, we have designed two theoretical chemicals, which could inhibit the direct enzyme responsible for fumarylacetoacetate formation. Subset 2_p.0.5 from Zinc database was screened by PyRx software using a Lamarckian genetic algorithm as the scoring function for docking. Top nine successive hits were selected for further pharmacological analysis and finally the new designed ligands RD6-2 (3Z)- 1,3-Butadiene-1,1,2,4-tetrol and RD-7-1 ((Z)-3-[4-Hydroxy-1-(hydroxymethyl)cyclohexyl]-2-propene-1,2-diol could pass PhysChem, FAFDrugs and AdmetSAR filter. The designed ligands were non-substrate and non-inhibitor of CYP450 and nontoxic in AMES test. LD50 of RD-6-2 was 793mg/kg with the toxicity class of four and The LD50 of RD-7-1 was calculated as 5000mg/kg within the toxicity class of five. The designed molecules are introduced as the new theoretical small molecules, which can theoretically inhibit 4- maleylacetoacetate isomerase in a competitive manner.


A Boltzmann Sampler for 1-Pairs with Double Filtration.
Recently, a framework considering RNA sequences and their RNA secondary structures as pairs led to some information-theoretic perspectives on how the semantics encoded in RNA sequences can be inferred. This pairing arises naturally from the energy model of RNA secondary structures. Fixing the sequence in the pairing produces the RNA energy landscape, whose partition function was discovered by McCaskill. Dually, fixing the structure induces the energy landscape of sequences. The latter has been considered originally for designing more efficient inverse folding algorithms and subsequently enhanced by facilitating the sampling of sequences. We present here a partition function of sequence/structure pairs, with endowed Hamming distance and base pair distance filtration. This partition function is an augmentation of the previous mentioned (dual) partition function. We develop an efficient dynamic programming routine to recursively compute the partition function with this double filtration. Our framework is capable of dealing with RNA secondary structures as well as 1-structures, where a 1-structure is an RNA pseudoknot structure consisting of "building blocks" of genus 0 or 1. In particular, 0-structures, consisting of only "building blocks" of genus 0, are exactly RNA secondary structures. The time complexity for calculating the partition function of 1-pairs, that is, sequence/structure pairs where the structures are 1-structures, is O(h3b3n6), where h, b, n denote the Hamming distance, base pair distance, and sequence length, respectively. The time complexity for the partition function of 0-pairs is O(h2b2n3).


Adapting algebraic diagrammatic construction schemes for the polarization propagator to problems with multi-reference electronic ground states exploiting the spin-flip ansatz.
For the investigation of molecular systems with electronic ground states exhibiting multi-reference character, a spin-flip (SF) version of the algebraic diagrammatic construction (ADC) scheme for the polarization propagator up to third order perturbation theory (SF-ADC(3)) is derived via the intermediate state representation and implemented into our existing ADC computer program adcman. The accuracy of these new SF-ADC(n) approaches is tested on typical situations, in which the ground state acquires multi-reference character, like bond breaking of H2 and HF, the torsional motion of ethylene, and the excited states of rectangular and square-planar cyclobutadiene. Overall, the results of SF-ADC(n) reveal an accurate description of these systems in comparison with standard multi-reference methods. Thus, the spin-flip versions of ADC are easy-to-use methods for the calculation of "few-reference" systems, which possess a stable single-reference triplet ground state.


Cryopreservation of human spermatozoa decreases the number of motile normal spermatozoa, induces nuclear vacuolization and chromatin decondensation.
Even though cryopreservation of human spermatozoa is known to alter sperm motility and viability, it may also induce nuclear damages. The present study set out to determine whether or not cryopreservation alters motile sperm morphology under high magnification and/or is associated with chromatin decondensation. For 25 infertile men, we used high-magnification microscopy to determine the proportions of various types of motile spermatozoa before and after freezing-thawing: morphometrically normal spermatozoa with no vacuole (grade I), ≤ 2 small vacuoles (grade II), at least 1 large vacuole or >2 small vacuoles (grade III), and morphometrically abnormal spermatozoa (grade IV). The spermatozoa's chromatin condensation and viability were also assessed before and after freezing-thawing. Cryopreservation induced sperm nuclear vacuolization. It decreased the proportion of grade I + II spermatozoa (P < .001). It induced a decrease in the sperm viability rate (P < .001) and increased the proportion of sperm with noncondensed chromatin (P < .001). The latter parameter was strongly correlated with sperm viability (r = 0.71; P < .001). However, even motile sperm presented a failure of chromatin condensation after freezing-thawing, because the proportion of sperm with noncondensed chromatin was correlated with high-magnification morphology (r = -0.49 and 0.49 for the proportions of grade I + II and grades III + IV, respectively; P < .001). Cryopreservation alters the organelle morphology of motile human spermatozoa and induces sperm chromatin decondensation. High-magnification microscopy may be useful for evaluating frozen-thawed spermatozoa before use in assisted reproductive technology procedures (such as intrauterine insemination, in vitro fertilization, and intracytoplasmic sperm injection) and for performing research on cryopreservation methods. If frozen-thawed sperm is to be used for intracytoplasmic sperm injection, morphological selection under high magnification may be of particular value.


Expression of bovine beta-lactoglobulin/human erythropoietin fusion protein in the milk of transgenic mice and rabbits.
We have generated several transgenic mouse lines and rabbits expressing efficiently (up to 0.3 mg/ml in mice and up to 0.5 mg/ml in rabbits) human erythropoietin in their milk as bovine beta-lactoglobulin fusion protein. Human erythropoietin cDNA was inserted in frame into exon 5 of the bovine beta-lactoglobulin gene with a linker oligonucleotide encoding the cleavage site for bacterial IgA protease. RNA analysis performed on one lactating transgenic mouse and one transgenic rabbit revealed that the fusion gene was expressed almost exlusively in the mammary gland, although low amounts of transgene-derived RNA were detectable in salivary glands and uterus or in the kidney. The fusion protein was specifically cleaved with IgA protease. The erythropoietin part obtained upon digestion had a lower molecular mass than recombinant erythropoietin produced in Chinese hamster ovary cells. By deglycosylation analysis it was shown that the difference in size was due to a different type of glycosylation. Biological activity of the fusion protein, as determined by growth stimulation of TF-1 erythroleukemia cells, was less than 15% of that of human recombinant erythropoietin. Upon digestion of the fusion protein with IgA protease, biological activity comparable to that of the recombinant erythropoietin was recovered. Transgenic males and virgin females did not show signs of enhanced erythropoiesis, but lactating females expressing the transgene displayed transient increases in their hematocrit values.


[Perinatal mortality at the Gynecology-Obstetrical Clinic of the Clinical Center in Sarajevo in 2001].
Work results of the gynecology-obstetrics clinic Sarajevo maternity ward have been shown. Number of deliveries reached 3289, which iz significantly less then in the year 200. Prematurity reached 6.6%. Cesarian Sections were carried out in 19.3% cases and this represents a remarkable increase in comparison with the previous year 2000, when it reached 16.8%. Perinatal mortality iz 11.1@1000. Perinatal mortality shows a decrease in comparison with the period before the war. Late fetal mortality has not been significantly changed. Neonatal mortality shows an apparent decrease. Total perinatal mortality of mature newborns iz 6.9@1000 and has a constant tendency to decrease, reaching 10.5@1000 in 1991. Clinical presentation of deceased newborns predominantly presents with respiratory distress syndrome.