Whole-Body Barometric Plethysmography Characterizes Upper Airway Obstruction in 3 Brachycephalic Breeds of Dogs.

BACKGROUND: A novel test using whole-body barometric plethysmography (WBBP) was developed recently to diagnose brachycephalic obstructive airway syndrome (BOAS) in unsedated French bulldogs. HYPOTHESIS/OBJECTIVES: The hypotheses of this study were: (1) respiratory characteristics are different between healthy nonbrachycephalic dogs and brachycephalic dogs; and among pugs, French bulldogs, and bulldogs; and (2) obesity and stenotic nares are risk factors for BOAS. The main objective was to establish a diagnostic test for BOAS in these 3 breeds. ANIMALS: A total of 266 brachycephalic dogs (100 pugs, 100 French bulldogs, and 66 bulldogs) and 28 nonbrachycephalic dogs. METHODS: Prospective study. Exercise tolerance tests with respiratory functional grading, and WBBP were performed on all dogs. Data from WBBP were associated with functional grades to train quadratic discriminant analysis tools to assign dogs to BOAS+ and BOAS- groups. A BOAS index (0-100%) was calculated for each dog. Receiver operating characteristic (ROC) curves were used to evaluate classification ability. RESULTS: Minute volume was decreased significantly in asymptomatic pugs (P = .009), French bulldogs (P = .026), and bulldogs (P < .0001) when compared to nonbrachycephalic controls. Respiratory characteristics were different among breeds and affected dogs had a significant increase in trace variation. The BOAS index predicted BOAS status for each breed with 94-97% (95% confidence interval [CI], 88.9-100%) accuracy (area under the ROC curve). Both obesity (P = .04) and stenotic nares (P = .004) were significantly associated with BOAS. CONCLUSIONS AND CLINICAL IMPORTANCE: The WBBP can be used as a clinical tool to diagnose BOAS noninvasively and objectively.

The highly conserved, N-terminal (RXXX)8 motif of mouse Shadoo mediates nuclear accumulation.

The prion protein (PrP)-known for its central role in transmissible spongiform encephalopathies-has been reported to possess two nuclear localization signals and localize in the nuclei of certain cells in various forms. Although these data are superficially contradictory, it is apparent that nuclear forms of the prion protein can be found in cells in either the healthy or the diseased state. Here we report that Shadoo (Sho)-a member of the prion protein superfamily-is also found in the nucleus of several neural and non-neural cell lines as visualized by using an YFP-Sho construct. This nuclear localization is mediated by the (25-61) fragment of mouse Sho encompassing an (RXXX)8 motif. Bioinformatic analysis shows that the (RXXX)n motif (n=7-8) is a highly conserved and characteristic part of mammalian Shadoo proteins. Experiments to assess if Sho enters the nucleus by facilitated transport gave no decisive results: the inhibition of active processes that require energy in the cell, abolishes nuclear but not nucleolar accumulation. However, the (RXXX)8 motif is not able to mediate the nuclear transport of large fusion constructs exceeding the size limit of the nuclear pore for passive entry. Tracing the journey of various forms of Sho from translation to the nucleus and discerning the potential nuclear function of PrP and Sho requires further studies.

Distinct hydration properties of wild-type and familial point mutant A53T of α-synuclein associated with Parkinson's disease.

The propensity of α-synuclein to form amyloid plays an important role in Parkinson's disease. Three familial mutations, A30P, E46K, and A53T, correlate with Parkinson's disease. Therefore, unraveling the structural effects of these mutations has basic implications in understanding the molecular basis of the disease. Here, we address this issue through comparing details of the hydration of wild-type α-synuclein and its A53T mutant by a combination of wide-line NMR, differential scanning calorimetry, and molecular dynamics simulations. All three approaches suggest a hydrate shell compatible with a largely disordered state of both proteins. Its fine details, however, are different, with the mutant displaying a somewhat higher level of hydration, suggesting a bias to more open structures, favorable for protein-protein interactions leading to amyloid formation. These differences disappear in the amyloid state, suggesting basically the same surface topology, irrespective of the initial monomeric state.

Epidemiological modelling of risk factors of human papilloma virus in women with positive cytology in the county of Csongrád.

This study was carried out to determine the prevalence and risk factors of genital HPV infection in women diagnosed with non-negative cytology in Southeastern Hungary. Cervical samples were collected for cytology and HPV testing from women seen at gynaecological outpatient clinics and diagnosed with non-negative cytology. The observed overall average HPV infection rate was found to be 61%. A smoking habit was the only risk factor in the logistic regression analysis that related significantly to exposure to HPV infection. Thus, prevention strategies should focus on the regular clinical cytological screening of HPV-infected patients and on the reduction of smoking.

Two mifepristone doses and two intervals of misoprostol administration for termination of early pregnancy: a randomised factorial controlled equivalence trial.

OBJECTIVE: To compare the efficacy of 100 mg and 200 mg of mifepristone and 24- and 48-hour intervals to administration of 800 microg vaginal misoprostol for termination of early pregnancy. DESIGN: Placebo-controlled, randomized, equivalence trial, stratified by centre. SETTING: 13 departments of obstetrics and gynecology in nine countries. POPULATION: 2,181 women with 63 days or less gestation requesting medical abortion. METHODS: Two-sided 95% CI for the risk differences of failure to complete abortion were calculated and compared with 5% equivalence margin between two doses of mifepristone and two intervals to misoprostol administration. Proportions of women with adverse effects were compared between the regimens using standard testes for proportions. OUTCOME MEASURES: Rates of complete abortion without surgical intervention and adverse effects associated with the regimens. RESULTS: Efficacy outcome was analysed for 2,126 women (97.5%) excluding 55 lost to follow up. Both mifepristone doses were found to be similar in efficacy. The rate of complete abortion was 92.0% for women assigned 100 mg of mifepristone and 93.2% for women assigned 200 mg of mifepristone (difference 1.2%, 95% CI: -1.0 to 3.5). Equivalence was also evident for the two intervals of administration: the rate of complete abortion was 93.5% for 24-hour interval and 91.7% for the 48-hour interval (difference -1.8%, 95% CI: -4.0 to 0.5). Interaction between doses and interval to misoprostol administration was not significant (P = 0.92). Adverse effects related to treatments did not differ between the groups. CONCLUSIONS: Both the 100 and 200 mg doses of mifepristone and the 24- and 48-hour intervals have a similar efficacy to achieve complete abortion in early pregnancy when mifepristone is followed by 800 micrograms of vaginally administered misoprostol.

Genotype screening for hereditary hemochromatosis among voluntary blood donors in Hungary.

Hereditary hemochromatosis (HH) is a common genetic disorder. Although it is inherited in an autosomal recessive manner, heterozygous individuals are believed to be protected against iron deficiency. Screening to estimate the prevalence of HH was frequently performed among blood donors, not considering that carriers of the HH gene mutations may be present in higher proportion in this population. To examine the allele frequencies of the HH gene (HFE) point mutations, C282Y and H63D genotyping was carried out in 996 consecutive, first-time, and regular Hungarian blood donors by PCR-RFLP techniques. Iron parameters of the first-time donors and the identified C282Y heterozygotes and age, gender, and number of previous blood donation-matched wild-type donors were also determined. We were not able to demonstrate a significant increase in the frequency of C282Y and H63D alleles among regular blood donors, compared to first-time blood donors. However, there was a trend of higher C282Y allele frequency among women with higher number of previous blood donations (2.2 +/- 1.5% in female blood donors with 0-8 previous blood donations compared to 4.8 +/- 2.3% in women with more than 8 previous blood donations, P = 0.06). No detectable phenotypic differences were observed in serum iron, ferritin, and transferrin saturation values between C282Y wild-type and heterozygous groups. However, the single identified C282Y homozygous male (age 21) showed definite signs of iron overload. Our observations suggest that the protective effect of C282Y heterozygosity against iron deficiency may be less significant than other environmental (e.g., iron-rich diet) or genetic factors.

[A new method of molecular testing in the differential diagnosis of hereditary hemochromatosis]. / Uj, molekuláris genetikai módszer az öröklödö haemochromatosis differenciáldiagnosztikájában.

Hereditary hemochromatosis is an autosomal, recessive disorder of the iron metabolism. The hemochromatosis gene (HFE) was previously located on chromosome 6 and recently identified by positional cloning. A point mutation, C282Y, was found to be present in the HFE gene in homozygous form in 64 to 100% of patients with established hemochromatosis. The relationship of a second polymorphic variant of the HFE gene, H63D to the formation of iron overload is debated. Although hemochromatosis is one of the most common inherited disorders among Caucasians, in the absence of specific signs it is rarely diagnosed. In order to obtain comparable epidemiological data for Hungary, we tested 1271 and 277 randomly selected, unrelated, healthy subjects for C282Y and H63D respectively. In addition C282Y testing was carried out in 58 patients suffering from liver cirrhosis, and in 191 individuals with suspected hemochromatosis. For C282Y and H63D mutation analyses polymerase chain reaction technique followed by Rsa I and Bcl I restriction enzyme digestion was used. We developed an alternative method for the detection of C282Y based on an amplification-generated Kpn I restriction site. The allele frequencies were 3.8% and 12.3% for C282Y and H63D respectively in the normal Hungarian population. There was no significant difference in C282Y allele frequencies between liver disease patients (1.7%) and the normal population. We identified 15 homozygous and 25 heterozygous individuals among 191 individuals with suspected hemochromatosis. The C282Y and the H63D allele frequencies in the normal Hungarian population were found to be similar to the allele frequencies observed in other European populations, indicating that there is a large number of individuals susceptible for iron overload in Hungary (1:700). Mutation analysis is a novel, non-invasive method in the diagnostics of hereditary hemochromatosis, which increasingly becomes part of the routine clinical work.

[Detection of human papillomavirus infection by the nucleic acid hybridization method (a multicenter study)]. / Human papillomavírus-fertözés kimutatás nukleinsav hibridizációs módszerrel (multicentrikus tanulmány).

The human papillomaviruses (HPV) are regarded as one of the important agents of cervical carcinoma. A multicentre study was organized to determine the prevalence of HPV in the fertile female population in Hungary. Parallel with the clinical sample collection, a questionnaire interview was performed to acquire data on the life style, socioeconomic status, sexual practice, etc. 1200 women were examined colposcopically and cervix samples were collected for cytology and the detection of HPV DNA. 17.4% of the samples were HPV-infected. 3.9% of the patients had acquired low-risk, and 10.1% 10.2% high-risk HPV types; 3.4% of the women were at the same time infected with both low-risk and high-risk HPV types. Simultaneously performance of cytology and the HPV hybrid capture assay contribute to recognise and treat the precancerous status and risk factors.

Synthetic peptides comprising defined sequences of CH-2 and CH-3 domains of human IgG1 induce prostaglandin E2 production from human peripheral blood mononuclear cells.

Synthetic peptides Y48 and Y75 comprising sequences at exposed sites within the CH-2 and CH-3 domains of human IgG1 at a concentration of 10(-5) M, increase PGE2 production by human peripheral blood mononuclear cell (PBMC) cultures. An increase of leukocyte migration inhibitory factor (LMIF) production in PBMC cultures--as a result of synthetic peptide treatment--was also observed. This LMIF activity, to some extent, is attributed to the PGE2 production by the cells; the inhibition of leukocyte migration being abolished by the presence of indomethacine or antibody to PGE2.

Silibinin (Legalon-70) enhances the motility of human neutrophils immobilized by formyl-tripeptide, calcium ionophore, lymphokine and by normal human serum.

Experiments reported here were designed to investigate the effect of silibinin (extracted from Silybum marianum) on human polymorphonuclear leukocyte (PMN) motility and on leukocyte immobilizing activity of lymphokine (leukocyte inhibitory factor, LIF), formyl-Met-Leu-Phe (fMLP), calcium ionophore A-23187 and human sera inactivated by heat (HI-S). In the in vitro experiments, silibinin (1-10 micrograms/ml) failed to influence the random motility of unstimulated PMNS in agarose droplet assay, but enhanced the motility of the PMNs immobilized by fMLP, calcium ionophore, LIF or by autologous human sera. In the in vivo study, silibinin (Legalon-70) two hours after the administration was effective in enhancing spontaneous motility of leukocytes obtained from health volunteers which action could be regarded as a consequence of the decrease of leukocyte immobilizing activity being present in normal human plasma.

Demonstration of leukocyte-immobilizing activity in normal human plasma.

The effect of normal human plasma on random migration of polymorphonuclear leukocytes (PMNs) in agarose micro-droplet assay was studied. Plasma, even at a concentration as low as 1%, inhibits leukocyte motility and this immobilizing activity seems to be related to a specific factor which present in human sera and is stable at 56 degrees C for 30 min. The immobilizing effect of heat-inactivated sera was abolished in vitro by indomethacin at a concentration of 0.1 micrograms/ml and it was significantly decreased by anti PGE2 antibody. Spontaneous migration of leukocytes failed to be enhanced by indomethacin (0.1 microgram/ml) alone. Anti-PGE2 alone moderately enhanced PMN motility. Immobilizing activity in human sera decreased 2 hours after indomethacin administration (Indomethacinum CHINOIN, 50 mg orally). No direct enhancing effect of indomethacin on leukocyte motility was shown in vivo. The data indicate a leukocyte-immobilizing activity of human sera. This effect is mediated by prostaglandin production induced by serum factor(s). The factor(s) responsible for the above activity may have a role in regulating leukocyte distribution and motility in vivo.

Short-term stimulation of lymphocyte proliferation by indomethacin in vitro and in vivo.

The effect of short-term (up to 24 h) in vitro and in vivo treatment with indomethacin was studied on the blastogenesis of mouse spleen cells. Indomethacin in itself induced a strong proliferation of the lymphocytes starting after 6 h treatment both in vitro and in vivo. Besides, significantly enhanced the blastogenesis of splenocytes in response to various doses of PHA and Con A. The stimulation of lectin-induced lymphocyte proliferation occurred after indomethacin treatment both in vitro and in vivo. Indomethacin had no major effect on the distribution of Lyt-1+ and Lyt-2+ subsets within the spleen cell population. An important role of the prostaglandins in the early phase of lymphocyte activation is suggested.

Effects on immune reactions of non-steroidal anti-inflammatory drugs and of two new pyrido-pyrimidine derivatives (CHINOIN 127 and CHINOIN 105).

The effect of three non-steroidal anti-inflammatory agents (NSAIs) and of two new pyrido-pyrimidine derivatives, CHINOIN 127 (1-6-dimethyl-4-oxo-1,6,7,8,9)a-hexahydro-4H-pyrido (1,2-2)pyrimidine-3-carboxamide) and CHINOIN 105 (1-6-dimethyl-4-oxo-1,6,7,8-tetrahydro-4H pyrido (1,2-a) pyrimidine-3-carboxamide) was compared in immune tests using human cells in vitro including T and B lymphocyte proliferation induced with mitogens, spontaneous motility of polymorphonuclear leukocytes, lymphokine (LIF) production, antibody-dependent cell-mediated cytotoxicity (ADCC) and natural killer cell (NK) activity. Indomethacin (INDO) at therapeutic concentrations enhanced the proliferation of lymphocytes stimulated by both Con A and PWM. Acetylsalicylic acid (ASA) increased proliferation of lymphocytes stimulated by Con A and at high concentration inhibited DNA synthesis induced by PWM. Phenylbutazone (PhB) increased DNA synthesis induced by PWM but not by Con A. CHINOIN 127 and CHINOIN 105 inhibited the proliferation of T lymphocytes but failed to influence DNA synthesis of B cells. Indomethacin, ASA and PhB inhibited the spontaneous motility (migration) of polymorphonuclear leukocytes. LIF production of lymphocytes was inhibited by INDO, ASA, PhB, CHINOIN 127 and CHINOIN 105. Phenylbutazone and ASA inhibit ADCC and NK. INDO, Chinoin 127 and Chinoin 105 failed to influence the effector cells of ADC and NK cytotoxicity.

Ketoconazole in vitro inhibits mitogen-induced blastogenesis, antibody-dependent cellular cytotoxicity, natural killer activity and random migration of human leukocytes.

Ketoconazole at concentrations of 1-10 micrograms/ml dose dependently inhibits mitogen-induced blastogenesis, antibody-dependent and spontaneous cytotoxic (natural killer) activity of human lymphocytes and random migration of human leukocytes. Lectin-dependent cytotoxic activity is not affected by the drug.

Inhibitory effect of tumour cells on leukocyte motility.

Hep-2 cells inhibit the random migration of human polymorphonuclear leukocytes in a tumour cell-leukocyte microenvironmental model, which is a special form of co-migration. The inhibition is indomethacin-sensitive. It is very likely that prostaglandins, produced as a consequence of interaction between tumour cells and leukocytes, are the mediators of this phenomenon. The tumour-induced inhibition of leukocyte motility may represent a tumour-protective mechanism.

Effect of prostaglandins on polymorphonuclear leukocyte motility.

The effect of prostaglandins (PGE1, E2, F2 alpha on LIF production, LIF activity, and on the random migration of polymorphonuclear leukocytes was investigated. Concanavalin A-induced LIF production was tested in an indirect LIF assay using agarose microdroplet technique; PGE1 and PGE2 inhibited the LIF production. PGE1, PGE2, and F2 alpha in physiological concentration, decreased the random migration of leukocytes, they also interfered with the LIF activity of supernatants, decreasing their inhibitory effect on cell migration; but the opposite explanation that LIF-treated PMNs escaped the inhibitory effect of PGs, cannot be ruled out. Macrophages produce supernatant factor(s) in the course of phagocytosis which inhibit the motility of polymorphonuclear leukocytes. The active component of supernatant seems to be PG, since its production can be blocked by indomethacin. These data indicate that prostaglandins may have different sites of attack in the local regulation of leukocyte motility.

Sensitivity of preincubated lymphocytes to suppressor regulation.

DNA synthesis of human peripheral blood lymphocytes increases if Con A is added to the culture after 24 h preincubation at 37 degrees C. During preincubation the mitogen reactive lymphocytes lose their sensitivity to the suppressive effect of autologous mitomycin-treated mononuclear leukocytes, and of supernatants of autologous preincubated cells. The reactive lymphocytes preincubated at 4 degrees C retain their sensitivity to the suppressive effect of regulatory cells and their supernatants. It is assumed that the enhancement of mitogen response after preincubation at 37 degrees C is caused by a decrease of suppressor regulation of human lymphocytes. Prostaglandins may be regarded as one of the mediators of the suppression.