Field evaluation of the MM3-SERO ELISA for detection of anti-Fasciola IgG antibodies in milk samples from individual cows and bulk milk tanks

We carried out a field evaluation of the MM3-SERO ELISA for the diagnosis of Fasciola hepatica infection, by analysing serum and milk samples from individual cows and samples from bulk milk tanks. The diagnostic performance of the assay was assessed with serum samples from all 257 cows in eight fluke-free herds, and 240 cows with natural fasciolosis (diagnosed in vivo and/or post-mortem). Assay performance for individual milk samples was determined by analysis of paired serum and milk samples from 947 lactating cows from 33 F. hepatica-infected farms. The diagnostic usefulness of the assay for bulk tank milk was evaluated by analysis of bulk milk from infected (33) and non-infected (35) farms. For serum samples, the sensitivity, specificity and diagnostic accuracy of the assay were respectively 99.2% (95% CI: 97.0%–99.9%), 100% (95% CI: 98.6%–100%) and 0.997 (95% CI: 0.987–1.000). The only two infected animals in which serum antibodies were not detected had very low parasitic burdens (with only 2 and 3 flukes observed). The performance of the MM3 SERO ELISA for individual milk samples was similar to that for serum samples, and the stepwise linear regression revealed a strong correlation between the results for the milk samples and the serum samples (R² = 0.84; p < 0.001). The agreement between results obtained with pairs of serum and milk samples was very high: there was matching classification in 96% (910/947) of paired samples (kappa = 0.92; p < 0.001). Individual milk samples may therefore be used, instead of serum samples, in the MM3-SERO ELISA, for reliable detection of seropositive cows. Testing bulk tank milk samples enabled detection of infected herds, even when the within-herd prevalence of infection was as low as 12%. We conclude that the MM3-SERO ELISA is a sensitive and highly specific test for serodiagnosis of bovine fasciolosis, and can be used with individual samples of either serum or milk. Use of the assay with bulk milk samples enables estimation of the within-herd prevalence of infection.     

Comparison of clinical findings,microbiological results,and farming parameters in goat herds affected by recurrent infectious mastitis

A total of 1388 goats from 31 farms distributed throughout Sardinia, Italy, were investigated in this study. The goat population of Sardinia accounts for a quarter of the entire goat population of Italy. In this work, data and information concerning each farming site were collected, a detailed clinical examination of the udder of lactating animals was carried out, and microbiological examination of milk was performed. Results were then subjected to statistical analysis in order to highlight possible correlations among all findings. Significant results were obtained by comparing clinical findings with bacteriological positivity of milk.Intra-mammary infections were detected in 22.7% of all goats examined. Staphylococcus spp., Streptococcus spp., and Mycoplasma spp. were detected in 73.5%, 9.7%, and 4.7% of positive milk cultures, respectively. Other bacterial genera showed a prevalence lower than 3%. Milk positivity to Staphylococcus aureus was significantly associated to presence in the udder of pustules, ulcers, nodules, and rubor (p < 0.05). Staphylococcus caprae was associated to manual vs mechanical milking, age, udder edema, and absence of mammary secretions (p < 0.05). Staphylococcus epidermidis was positively associated to age (p < 0.05). Streptococcus uberis was associated to mechanical milking, atrophic udder texture, and reactive mammary lymph nodes (p < 0.05). Several other factors showed statistically significant correlations. Taken together, the results presented in this work highlight the specificity, the usefulness, and the practical value of performing a clinical examination of the udder, and suggest its relevance as the most important diagnostic tool for complementing microbiological examination of milk.     

Seroprevalence and risk factors associated with zoonotic parasitic infections in small ruminants in the Greek temperate environment

A cross-sectional serological study was carried out to screen the sheep and goat population of Thessaly, Greece for evidence of infection with Toxoplasma, Toxocara, Leishmania, and Echinococcus and to determine the risk factors related to herd characteristics, herd management practices, farmer status, and the bioclimatic variables associated with these zoonotic parasitic infections. A total of 540 sheep and goat serum samples were examined. The seroprevalence of infection in all examined animals was 24.5% for Toxoplasma, 32% for Toxocara, 0% for Leishmania and 85.9% for Echinococcus. The final logistic regression model showed that the species of small ruminant, herd size, anthelmintic treatment, class of anthelmintic treatment, grazing with other herds, educational level of farmer, elevation of farm location, and generalized land cover were associated with Toxoplasma gondii infections, while the species of small ruminant, farm type, anthelmintic treatment, class of anthelmintic treatment, rotation of grazing, age of farmer, elevation of farm location, and generalized land cover were associated with Toxocara canis infections. Antibodies to T. gondii were detected in 102 (28.3%) of 360 sheep and in 30 (16.8%) of 179 goats. Animals in small flocks (150–300 animals) had an approximately 0.42-fold lower risk of having positive cases of T. gondii among animals compared with large flocks (> 300 animals). Antibodies to T. canis were found in 155 (42.9%) of 361 sheep and 18 (10.1%) of 179 goats. The later finding constitutes the first report of seropositive goats to Toxocara. The risk of positivity for T. canis was 7.71-fold higher in sheep than in goats. Geographically, animals from plain areas had 2.9 and 2.01-fold higher risk of having positive cases of T. gondii and T. canis respectively. The significant bioclimatic variables (p < 0.05) associated with the occurrence locations of T. gondii infection were related to higher temperature, lower precipitation, and lower elevation compared to the absence locations of T. gondii. The significant bioclimatic variables (p < 0.05) associated with occurrence locations of T. canis infection were related to lower temperature and higher precipitation compared to absence locations of T. canis. These findings are useful to formulate appropriate control strategies for zoonotic parasites of sheep and goats in Greece and other areas with similar climatic conditions.     

A survey for Echinococcus spp. of carnivores in six wildlife conservation areas in Kenya

To investigate the presence of Echinococcus spp. in wild mammals of Kenya, 832 faecal samples from wild carnivores (lions, leopards, spotted hyenas, wild dogs and silver-backed jackals) were collected in six different conservation areas of Kenya (Meru, Nairobi, Tsavo West and Tsavo East National Parks, Samburu and Maasai Mara National Reserves). Taeniid eggs were found in 120 samples (14.4%). In total, 1160 eggs were isolated and further analysed using RFLP-PCR of the nad1 gene and sequencing. 38 of these samples contained eggs of Echinococcus spp., which were identified as either Echinococcus felidis (n = 27) or Echinococcus granulosus sensu stricto (n = 12); one sample contained eggs from both taxa. E. felidis was found in faeces from lions (n = 20) and hyenas (n = 5) while E. granulosus in faeces from lions (n = 8), leopards (n = 1) and hyenas (n = 3). The host species for two samples containing E. felidis could not be identified with certainty. As the majority of isolated eggs could not be analysed with the methods used (no amplification), we do not attempt to give estimates of faecal prevalences. Both taxa of Echinococcus were found in all conservation areas except Meru (only E. felidis) and Tsavo West (only E. granulosus). Host species identification for environmental faecal samples, based on field signs, was found to be unreliable. All samples with taeniid eggs were subjected to a confirmatory host species RLFP-PCR of the cytochrome B gene. 60% had been correctly identified in the field. Frequently, hyena faeces were mistaken for lion and vice versa, and none of the samples from jackals and wild dogs could be confirmed in the tested sub-sample. This is the first molecular study on the distribution of Echinococcus spp. in Kenyan wildlife. The presence of E. felidis is confirmed for lions and newly reported for spotted hyenas. Lions and hyenas are newly recognized hosts for E. granulosus s.s., while the role of leopards remains uncertain. These data provide the basis for further studies on the lifecycles and the possible link between wild and domestic cycles of cystic echinococcosis in eastern Africa.     

Breeding management in goat farms of Lombardy,northern Italy: Risk factors connected to gastrointestinal parasites

This study is the first parasitological survey dealing with goats at a regional scale in northern Italy. In particular, we characterized goat farms in Lombardy and investigated the risk factors of parasite infections there. Within our study area, both the breeding system and the size of the flocks varied. In fact, large flocks are mainly located down the valleys of Alpine areas and in the Prealps at low altitudes, such animals are reared under intensive husbandry. Medium-sized flocks are mostly found in mountainous areas where goats are reared under extensive or semi-extensive systems. Autochthonous races, then, are important in some marginal areas where a farm economy is supported by the promotion of traditional goat products, e.g. typical cheese and meat derivatives. Significant factors associated with the presence of parasites are (i) breeding system, extensive for Moniezia benedeni, semi-extensive for Strongyloides; (ii) size of the farm, over 100 goats for Strongyloides; (iii) altitude, above 500 m a.s.l. and up to 1000 m a.s.l. for M. benedeni and Strongylida; (iv) pasture, Strongylida; (v) season, autumn for M. benedeni; (vi) separation of goats into groups, (Skrjabinema). Goats in first lactation were significantly more infected with Strongylida and Trichuris than multiparous ones (both p < 0.001). Eggs excretion of Strongyloides was higher in pregnant females (p < 0.001). Differences emerged also for Nematodirus and Strongylida when eggs excretion was related to pregnancy (p < 0.05 and p < 0.001, respectively), but epg values of these nematodes were significantly higher in not pregnant individuals. Heavier infection with S. was found in lactating goats rather than in dry goats (p < 0.001), while the latter ones were more infected with Trichuris and Strongyloides (p < 0.05 and p < 0.001, respectively). In conclusion, according to the results of our survey, as to extensive husbandry we highly recommend correct practices in pasture management associated with a targeted programme of anthelminthic treatments for Stronylida and Cestoda. As to the rearing of large flocks almost entirely indoor, we noted a higher presence of other parasites, i.e. Strongyloides, probably due to a high density of heads associated with a scarce sanitary management. Concerning, then, young goats in first lactation and lactating females our data confirmed that they both are more vulnerable to the infection with Strongylida, while pregnant females are more prone to the infection with Strongyloides spp. This can be partially due to the synchronization of the reproductive cycle, as performed in most farms, and to the occurrence of the period of pregnancy in winter, when the animals are stabled indoor.     

The effects of hoarding habitat selection of Eurasian red squirrels (Sciurus vulgaris) on natural regeneration of the Korean pines

Cone-cores discarded by Eurasian red squirrels were used to study the habitat selection of Korean pine-seeds hoarding, in forest patch Nos. 16 and 19 in Liangshui Nature Reserve, China. Ten transects with a total length of 15 km were uniformly set, and data from 343 valid samples were collected in a 369 hm² area. One hundred and eighty four were hoarding samples which were determined according to the cluster analysis based on the number of the cone-cores, while the other 159 were control samples. The principal component analysis, using 11 habitat factors, suggested that the distance from Korean pine forest, forest type, number of Korean pine seedlings, density and type of bush significantly influenced the habitat selection of hoarding by Eurasian red squirrels. The results of Bailey’s method indicated that the squirrels showed (1) preference for natural coniferous forest, natural fir and spruce forest and planted spruce forest; (2) avoidance of planted Korean pine forest and planted larch forest; and (3) random use of natural Korean pine forest. Moreover the distance from the Korean pines in the range of 150–600 m showed no effect on the habitat selection of hoarding by the Eurasian red squirrels. More than 50% of the cone-cores were discarded in either fringe or gap of the Korean pine forest with more cone-cores found at <300 m than at 300 m away (One-Way ANOVA; df = 3, 183, F = 5.76, p = 0.0009). This demonstrated that the Eurasian red squirrels could take the cone-cores out of the Korean pine forest. The density of bushes in samples of hoarding area was significantly lower than that in control samples (Kruskal–Wallis test; df = 1, χ² = 83.99, p < 0.0001). The number of the Korean pine seedlings in samples of hoarding area was significantly higher than that in the control samples (Kruskal–Wallis test; df = 1, χ² = 104.13, p < 0.0001). This illustrated that the hoarding habitat favoured the germination of the Korean pine seedlings. In conclusion the behavior of hoarding Korean pine seeds by the Eurasian red squirrels can promote the regeneration and dispersal of the Korean pines.     

Effects of grassland conversion to cropland and forest on soil organic carbon and dissolved organic carbon in the farming-pastoral ecotone of Inner Mongolia

Effects of grassland conversion to cropland and forest on soil organic carbon (SOC), dissolved organic carbon (DOC) in the farming-pastoral ecotone of Inner Mongolia were investigated by direct field sampling. SOC content and DOC content in soil decreased after grassland were shifted to forest or cropland, in the sequence of grassland soil > forest soil > cropland soil. SOC stock declined by 18% after grassland shifted from to forest. Reclamation of cropland for 10 years, 15 years and 20 years lost SOC in 0–30 cm soil layer, by 34%, 14% and 18%, respectively, compared with that of grassland. DOC in 3 soil layers was within 21.1–26.5 mg/L in grassland, 12.1–14.6 mg/L in forest soil, and 8.0–14.0 mg/L in cropland soil. Correlation analysis indicated that SOC content and DOC content were positively dependent on total nitrogen content (p < 0.05), but negatively on bulk density or land use type (p < 0.05). DOC was positively correlated SOC (p < 0.01). Moreover, SOC content could be quantitatively described by a linear combination of land use types (p = 0.000, r² = 0.712), and DOC content by a linear combination of two soil-related variables, land use types and SOC (p = 0.000, r² = 0.861).     

Evaluation of Montandoniola confusa Streito and Matocq sp. nov. and Orius insidiosus Say (Heteroptera: Anthocoridae), for control of Gynaikothrips uzeli Zimmerman (Thysanoptera: Phlaeothripidae) on Ficus benjamina

Since its discovery in Florida in 2003, the weeping fig thrips, Gynaikothrips uzeli Zimmerman has spread rapidly throughout the southeastern United States in shipments of ornamental Ficus benjamina L. Concurrently, there have been reports of an invasive anthocorid, Montandoniola confusa (=moraguesi) Streito and Matocq sp. nov., widely associated with G. uzeli populations in landscape plantings of ornamental Ficus spp. We evaluated M. confusa and a commercially available anthocorid, Orius insidiosus Say, as biological control agents of G. uzeli. Prey preference studies revealed that eggs were the numerically preferred host stage for both predator species (representing 92% and 94% of all prey taken in ‘no choice’ and ‘choice’ tests, respectively). Females of both predator species consumed significantly more eggs than males (83–91 versus 25–35 per 48 h period, respectively), and (in the absence of eggs) also more larvae (4.1–5.5 versus 2.1–2.5). Fecundity of M. confusa was significantly higher than for O. insidious, 10.6 ± 1.5 eggs per 48 h versus 5.0 ± 1.4, respectively. Greenhouse tests on heavily infested F. benjamina revealed that M. confusa was a highly effective predator of G. uzeli. Evaluations with three F. benjamina cultivars showed that M. confusa reproduced throughout the year and reduced thrips populations ⩾95% and leaf galls by up to 77% within 5 weeks. By contrast O. insidiosus did not establish or significantly reduce populations of G. uzeli inside leaf galls. Methods to monitor and protect M. confusa in urban landscapes are discussed.     

A study on ectoparasites of sheep and goats in eastern part of Amhara region,northeast Ethiopia

A study on ectoparasites of small ruminants was carried out in three districts (woredas) of the eastern part of Amhara Regional State, Ethiopia, from November 2003 to March 2004, with the objectives of determining the prevalence of ectoparasites and identifying the potential risk factors associated with the problem. Out of 752 sheep and 752 goats examined, 50.5% of sheep and 56.4% of goats were infested with one or more ectoparasites. The ectoparasites identified in sheep were Damalina ovis (38.5%), Melophagus ovinus (12.5%), tick infestations (3.4%) and Linognathus spp. (2.4%). In goats, parasites such as Linognathus spp. (28.3%), ticks (22.2%), sarcoptic mites (6.1%) and Ctenocephalides spp. (8.1%) were identified. The prevalence of M. ovinus and D. ovis infestations in sheep were significantly (p < 0.05) higher in the highlands than both lowlands and midlands, which were almost similar. In goats, the risk of sarcoptic mange infestation in lowland and midland was 4.6 and 5.0 times higher than the highlands, respectively. The prevalence of Linognathus spp. in goats was significantly (p < 0.05) higher in the highlands than the lowlands. Both in sheep and goats, no significant difference between young and adult age groups was observed for any ectoparasites except for D. ovis in sheep in which adults (42.2%) were more affected than young animals (29.9%). The goats with poor body condition were 4.3 times at risk for sarcoptic mange (OR = 4.3, p < 0.05), Linognathus spp. (OR = 2.1, p < 0.05) and tick (OR = 1.6, p < 0.05) infestation than goats of good body condition. On the contrary, no significant variation (p > 0.05) in ectoparasite infestation of sheep was observed in relation to body condition. Favorable climates, poor management, poor awareness of farmers and poor animal health extension services are believed to have contributed for the widespread occurrence of ectoparasites.     

Association between interleukin 15 receptor, alpha (IL15RA) polymorphism and Korean patients with ossification of the posterior longitudinal ligament

ObjectivesRecently, a number of evidences have been reported concerning the genetic factor involved in the development of ossification of the posterior longitudinal ligament (OPLL). The purpose of this study was to investigate single nucleotide polymorphisms (SNPs) of the interleukin 15 receptor, alpha (IL15RA) gene as a risk factor in Korean patients with OPLL.DesignTo investigate the genetic association, two coding SNPs (rs2296139, Thr73Thr; rs2228059, Asn182Thr) in IL15RA were genotyped in 166 OPLL patients and 230 control subjects. SNPStats, SNPAnalyzer, and Helixtree programs were used for association analysis.ResultsIn the present study, we found the association between a missense SNP (rs2228059) and the risk of OPLL in codominant (p = 0.0028, OR = 1.58, 95% CI = 1.17–2.14), dominant (p = 0.0071, OR = 1.82, 95% CI = 1.17–2.82), and recessive models (p = 0.036, OR = 1.79, 95% CI = 1.04–3.09). The frequency of rs2228059 allele was significantly associated with the susceptibility of OPLL (p = 0.0043, OR = 1.52, 95% CI = 1.14–2.02). After Bonferroni correction, the missense SNP (rs2228059, Asn182Thr) still had significant correlations (p = 0.0056 in codominant model; p = 0.0142 in dominant model; p = 0.0086 in allele analysis). Haplotype variation in IL15RA was associated with OPLL (global haplotype test, p = 0.025).ConclusionsThese results suggest that IL15RA polymorphism may be associated with the susceptibility of OPLL in Korean population.     

Contribution of epigenetic alteration of BRCA1 and BRCA2 genes in breast carcinomas in Tunisian patients

Objective: The aim of this study was to evaluate the contribution of the BRCA1 and BRCA2 promoter methylation in the pathogenesis of sporadic breast cancer in Tunisian patients. Methods: Breast carcinoma tissues (n = 117) and available paired normal breast tissues (n = 65) from Tunisian women who had no family history were investigated for the methylation status of BRCA1 and BRCA2 promoters using methylation-specific PCR. Breast specimens from women without carcinoma (16 fibroadenomas and 5 mastopathies) were used as control. Results: Hypermethylation of BRCA1 and BRCA2 promoters was detected respectively in 60.7% and 69.2% of the carcinoma tissues, and in only 7.7% and 4.6% of the paired normal breast tissues. None of the fibroadenomas and mastopathies showed hypermethylation. Correlations were found between BRCA1 and BRCA2 hypermethylation and decrease in their mRNA expression (p = 0.02 and p = 0.009, respectively). Moreover, BRCA1 methylation correlates with patients age (p = 0.01) and triple negative (ER?, PR?, HER2?) tumors (p = 0.01). Patients with methylated BRCA1 and/or BRCA2 had a significant prolonged survivals compared to those with unmethylated tumors (p = 0.002). Conclusion: Our results suggest an important role of BRCA1 and BRCA2 promoter methylation in breast cancer development in the Tunisian population.     

From chronic kidney disease to transplantation: the roles of obestatin

This study was designed to investigate the possible effect of sitagliptin on renal damage induced by renal ischemia reperfusion (I/R) in diabetic rats. T2DM in rats was induced by the administration of nicotinamide (230 mg/kg, i.p.), 15 min prior to a single dose of streptozotocin (65 mg/kg, i.v.). In vivo renal I/R was performed in both T2DM and normal rats. Each protocol comprised ischemia for 30 min followed by reperfusion for 24 h and a treatment period of 14 days before induction of ischemia. Sitagliptin treated diabetic rats that underwent renal I/R demonstrated significant decrease in the serum concentrations of aspartate aminotransferase (p < 0.01), urea nitrogen (p < 0.01) and creatinine (p < 0.001) compared to renal I/R in diabetic rats. Lipid peroxidation, xanthine oxidase activity, myeloperoxidase activity and nitric oxide level in renal tissue were significantly (p < 0.05, p < 0.001, p < 0.01, p < 0.05 respectively) decreased after renal I/R in sitagliptin treated rats compared to diabetic rats. Antioxidant enzymes like glutathione (p < 0.05), glutathione peroxidase (p < 0.001), superoxide dismutase (p < 0.05) and catalase (p < 0.001) were significantly increased after renal I/R in sitagliptin treated diabetic rats compared to non treated diabetic rats. The typical DNA laddering was observed when renal I/R performed in diabetic rats, which indicates cell apoptosis. Sitagliptin treated rats demonstrated a decrease in DNA fragmentation and apoptosis. Furthermore, renal histopathology preserved in sitagliptin treated rats verified protection against renal I/R in diabetes. The results of present investigation established sitagliptin treatment attenuated renal damage induced by renal I/R in diabetic rats.     

The effect of prostaglandin F(2α) administration at the time of insemination on the pregnancy rate of dairy cows

This paper addresses the question whether the pregnancy rate of dairy cows and heifers may be affected by administering prostaglandin F_2α at the time of artificial insemination. A field trial involving 1031 dairy cows and heifers distributed to a large number of small dairy farms in an area of extensive farming in central Germany provided evidence that intramuscular administration of 25 mg Dinoprost (Dinolytic^®) at the time of insemination has no effect on pregnancy rate (61% of the cows and heifers were pregnant in both prostaglandin F_2α-treated and saline control groups). On the other hand, deposition of 0.5 mL of a 0.5 mg/mL Dinoprost solution in the uterine lumen immediately after artificial insemination gave rise to a pregnancy rate of 66% as compared with 59% in saline controls. The increase in pregnancy rate of 229 prostaglandin F_2α-treated animals (66% pregnant) over that of 226 saline controls (59% pregnant) amounted to 12%. This improvement was not statistically significant (P = 0.12). Factors exerting a significant effect on pregnancy rate were parity (74% pregnancies in heifers versus 57% in cows, P < 0.01 and 65% pregnancies in first parity-cows versus 55% in older cows, P < 0.01) and season (57% during the barn season versus 64% during the pasture season, P < 0.05), whereas length of service period, level of milk production and serum or milk progesterone level at the time of insemination did not. A follow-up trial involving more animals will have to be conducted aimed at confirming the promising results obtained by intrauterine PGF_2α administration.     

Different roles of zinc plus arachidonic acid on insulin sensitivity between high fructose- and high fat-fed rats

AimsThis study was to determine the effects of zinc plus arachidonic acid (ZA) treatment on the insulin action in the specific ZA target organs using hyperinsulinemic euglycemic clamp method.Main methods18 Sprague–Dawley rats weighing ~ 130 g were divided into 3 groups of 6 rats and treated them with 1) normal rat chow, 2) high fructose (60.0%) diet only, or 3) the same fructose diet plus drinking water containing 10 mg zinc plus 50 mg arachidonic acid (AA)/L. In a separate study, male Wistar rats weighing ~ 250 g were fed normal rat chow (n = 4) or high fat (66.5%) diet with drinking water containing zero (n = 9) or 10 mg AA plus 20 mg zinc /L (n = 9). After 4 week treatment, insulin action was assessed using the hyperinsulinemic eguglycemic clamp technique.Key findingsHigh fructose feeding impaired suppression of hepatic glucose output by insulin compared to controls during the clamp procedure (4.39 vs. 2.35 mg/kg/min; p < 0.05). However, ZA treatment in high fructose-fed rats showed a significant improvement of hepatic insulin sensitivity compared to non-treatment controls (4.39 vs. 2.18 mg/kg/min; p < 0.05). Glucose infusion rates in Wistar rats maintained on a high fat diet (HFD) were significantly lower compared to control rats (22.8 ± 1.3 vs. 31.9 ± 1.4 mg/kg/min; p < 0.05). ZA treatment significantly improved (~ 43%) peripheral tissue insulin sensitivity in HFD fed animals (26.7 ± 1.3 [n = 9] vs. 22.8 ± 1.3 mg/kg/min; p < 0.05).SignificanceThese data demonstrate that ZA treatment is effective in improving glucose utilization in hyperglycemic rats receiving either a high-fructose or a high-fat diet.     

Role of proinflammatory markers and NT-proBNP in patients with an implantable cardioverter-defibrillator and an electrical storm

BackgroundSeveral studies have attempted to identify risk factors for the development of an electrical storm (ES), which is defined as ⩾3 separate ventricular tachyarrhythmic (VT/VF) events, but in the majority of studies no triggers have been found. However, little is known about the role of inflammation and NT-proBNP in patients with ES. The aim of this study was therefore to assess the relationship of Interleukin-6 (IL-6), high-sensitivity C-reactive protein (hs-CRP) and NT-proBNP serum concentrations in ICD-patients with or without single spontaneous ventricular tachyarrhythmic events (VT/VF) and in ES.MethodsMarkers were determined in 51 patients without ICD-intervention, in 15 ICD-patients with single VT/VF-episodes during 9-months follow-up and in 20 ICD-patients with ES (blood sampling performed within 60 min after fulfilling ES criteria). VT/VF-episodes were analysed by stored ICD-electrograms.ResultsAll patients had idiopathic dilated cardiomyopathy (n = 23) or coronary artery disease (n = 63). Patients with ES revealed significantly higher mean serum concentrations of all markers (IL-6 15.19 ± 10.34 pg/mL, hs-CRP 20.12 ± 14.4 mg/L, NT-proBNP 4799 ± 4596 pg/mL) compared to baseline values of patients with single VT/VF-events during follow-up (IL-6 8.37 ± 5.8 pg/mL (p = 0.03), hs-CRP 4.7 ± 5.3 mg/dL (p < 0.001), NT-proBNP 1913 ± 2665 pg/mL (p = 0.04)) and compared to baseline values of ICD-patients without device intervention (IL-6 4.62 ± 3.66 pg/mL (p < 0.001), hs-CRP 4.1 ± 3.4 mg/L (p < 0.001), NT-proBNP 1461 ± 2281 pg/mL (p < 0.001)). In 9/20 patients presenting with ES (45%) baseline values were available. All markers were significantly higher during ES compared to event-free determination (IL-6 14.54 ± 10.43 vs. 7.03 ± 2.83 pg/mL (p = 0.04), hs-CRP 19.07 ± 16.07 vs. 6.5 ± 3.9 mg/L (p = 0.02), NT-proBNP 4218 ± 2561 vs. 2099 ± 1279 pg/mL (p = 0.03)).ConclusionsElectrical storm is associated with significantly elevated IL-6, hs-CRP and NT-proBNP serum concentrations in ICD-patients with structural heart disease. Thus, ES may be triggered by proinflammatory activity. Combined intraindividual elevation of determined markers might help to identify patients at risk of impending electrical storm.     

In situ cryopreservation of human embryonic stem cells in gas-permeable membrane culture cassettes for high post-thaw yield and good manufacturing practice

Current strategies for marine pollution monitoring are based on the integration of chemical and biological techniques. The sea urchin embryo-larval bioassays are among the biological methods most widely used worldwide. Cryopreservation of early embryos of sea urchins could provide a useful tool to overcome one of the main limitations of such bioassays, the availability of high quality biological material all year round. The present study aimed to determine the suitability of several permeant (dimethyl sulfoxide, Me₂SO; propylene glycol, PG; and ethylene glycol, EG) and non-permeant (trehalose, TRE; polyvinylpyrrolidone, PVP) cryoprotectant agents (CPAs) and their combination, for the cryopreservation of eggs and embryos of the sea urchin Paracentrotus lividus. On the basis of the CPAs toxicity, PG and EG, in combination with PVP, seem to be most suitable for the cryopreservation of P. lividus eggs and embryos. Several freezing procedures were also assayed. The most successful freezing regime consisted on cooling from 4 to −12 °C at 1 °C/min, holding for 2 min for seeding, cooling to −20 °C at 0.5 °C/min, and then cooling to −35 °C at 1 °C/min. Maximum normal larvae percentages of 41.5% and 68.5%, and maximum larval growth values of 42.9% and 60.5%, were obtained for frozen fertilized eggs and frozen blastulae, respectively.     

An extract based on the medicinal mushroom Agaricus blazei Murill stimulates monocyte-derived dendritic cells to cytokine and chemokine production in vitro

Background: There is a growing body of evidence that physical training exerts its potential benefits on the individual health status by modulating the immune system and the whole body metabolism. A better knowledge of the physiologic immune response to exercise may help to understand the benefits of physical exercise in healthy individuals and elite athletes. Aims: This study aims to analyse cardiotrophin-1 (CT-1) and Tumor Necrosis Factor-α (TNF-α) plasma levels at rest and during exercise in elite athletes and healthy controls. Methods: We studied 20 triathletes (TA) and 20 matched controls (CG). Chambers dimensions, left ventricular mass and left ventricular mass index were analysed by echocardiography. VO2 peak and VE/VCO2 were calculated by metabolic stress test. Blood samples were collected before the exercise session, at the exercise peak, and after the end of exercise. ELISA assays were used to measure CT-1 and TNF-α plasma levels. Results:Among TA and CG, no significant differences were found for CT-1 (0.25 ± 0.14 vs 0.20 ± 0.14 fm/l; p = 0.29) and TNF-α (10.8 ± 2.7 vs 9.7 ± 4.0 pm/l; p = 0.29) basal levels. In the TA, plasma levels of CT-1 were significantly different at rest and during exercise (basal 0.25 ± 0.13 pm/l; peak 1.07 ± 1.5 pm/l; post-exercise 0.67 ± 0.77 pm/l; p = 0.04). Conversely, no significant differences were found between basal, peak and post-exercise plasma values of TNF-α (basal 10.8 ± 2.7 pm/l; peak 11.7 ± 2.1 pm/l; post-exercise 11.4 ± 2.5 pm/l; p = 0.78) in TA. Conclusions: This study gives novel insights on the behavior of inflammatory cytokines during physical exercise in athletes and healthy individuals.     

The receptor for advanced glycation endproducts and its ligands in patients with myasthenia gravis

ObjectiveMyasthenia gravis (MG) is a T- and B-cell mediated autoimmune disorder affecting the neuromuscular junction. The receptor for advanced glycation endproducts (RAGE) plays a role in the amplification of chronic inflammatory disorders and autoimmune diseases. We sought to investigate the role of RAGE and its ligands in the pathophysiology of MG.MethodsIn this cross-sectional study we enrolled 42 patients with MG and 36 volunteers. We employed enzyme-linked immunosorbent assays to determine the concentration of soluble RAGE (sRAGE) and high mobility group box 1 (HMGB1) in serum of patients and volunteers. In a subpopulation of patients we measured the serum levels of endogenous secretory (es) RAGE and various RAGE ligands, such as S100B, S100A8 and advanced glycation endproducts (AGE-CML). Reported are means and standard error mean.ResultsWe found significantly reduced levels of the soluble receptors sRAGE and esRAGE in patients with MG compared to volunteers without MG (sRAGE [pg/ml] 927.2 ± 80.8 vs. 1400.1 ± 92.4; p < 0.001; esRAGE [pg/ml] 273.5 ± 24.6 vs. 449.0 ± 22.4; p < 0.001). Further categorization of patients with MG according to the distribution of muscle involvement revealed the following sRAGE concentrations: generalized MG 999.4 ± 90.8 and ocular MG 696.1 ± 161.8 (vs. control; One-way ANOVA: p < 0.001; Post hoc analysis: generalized vs. ocular MG: p = 0.264, generalized MG vs. control: p = 0.008, ocular MG vs. control: p = 0.001). In patients with detectable antibodies specific for acetylcholine receptors (Anti-AChR positive) the sRAGE concentration was 970.0 ± 90.2 compared to those without (seronegative) 670.6 ± 133.1 (vs. control; One-way ANOVA: p < 0.001; Post hoc analysis: Pos vs. Neg.: p = 0.418, Pos vs. control: p = 0.003, Neg. vs. control: p = 0.008). We next investigated the role of RAGE ligands in MG. The concentrations of RAGE ligands in patients with MG and controls were as follows: (HMGB1 [ng/ml] 1.7 ± 0.1 vs. 2.1 ± 0.2; p = 0.058; S100B [pg/ml] 22.5 ± 22.5 vs. 14.4 ± 9.2; p = 0.698; S100A8 [pg/ml] 107.0 ± 59.3 vs. 242.5 ± 103.6; p = 0.347; and AGE-CML [ng/ml] 1100.8 ± 175.1 vs. 1399.8 ± 132.8; p = 0.179).ConclusionsOur data suggest a role for the RAGE pathway in the pathophysiology of MG. Further studies are warranted to elucidate more about this immunological axis in patients with MG.     

Determination of whole body circadian phase in lung cancer patients: melatonin vs. cortisol

Background: A quantifiable and reliable technique for the determination of body circadian phase applicable to non-laboratory studies would allow the evaluation of circadian dysregulation. In this study we evaluated feasible methodologies to individualize whole body circadian phase in lung cancer patients. Methods: Cortisol and melatonin serum levels were measured in blood samples collected every 4 h for 24 h from eleven male controls and nine men suffering from non-small cell lung cancer. Circadian rhythmicity was evaluated and the 4-hourly fractional variations (FV) were calculated to evaluate the dynamics of the rise and fall in serum levels. Results: Overall cortisol serum levels were higher in cancer patients (p < 0.001), and lower for melatonin, but not significantly (p = 0.261). Original serum levels of cortisol and melatonin each showed a prominent 24 h oscillation in both study groups, with highest values at night for melatonin and near awakening for cortisol. Using all data after normalization to percent of individual mean, ANOVA detected a significant time-effect (p < 0.001) and cosinor analysis detected a significant 24 h rhythm (p < 0.001) in each group. Overall fractional variation (FV) levels were lower for cortisol in cancer patients and higher for melatonin, but these differences were not significant. FV levels of cortisol and melatonin each showed a prominent 24 h oscillation in both study groups, with highest values prior to darkness onset for melatonin and near mid-dark for cortisol. ANOVA also detected a significant time-effect (p < 0.002) and cosinor analysis detected a significant 24 h rhythm (p < 0.001) for FV in each group, with maximal FV preceding maximal serum levels by ～3 h for melatonin and ～5 h for cortisol. Conclusions: A chronobiological evaluation of serum levels and fractional variations for cortisol and especially melatonin is a valuable methodology to define body circadian phase in lung cancer patients. It is possible to describe the complex process of hormone secretion with a methodology that allows the definition of both temporal characteristics and dynamic components. Impact: This kind of analysis might be useful in the study of hormone secretion(s) in cancer patients and other diseases and to guide therapeutic interventions. While lung cancer patients may have a negative prognostic value based upon stage and/or other hormonal aspects of their hypothalamus–pituitary–thyroid–adrenal axis function, patients that nevertheless maintain circadian rhythmicity in key body rhythm markers may still be eligible candidates for a chronotherapeutic approach of treatment(s).     

Prevalence and genotype identification of human papillomavirus in women undergoing voluntary cervical cancer screening in Molise,Central Italy

We examined the prevalence of HR- and LR-HPV by Linear Array genotyping test in 299 women aged 18–63 years who consecutively visited Molise Region main hospitals for routine Pap smear between February and August 2008. Ninety women were positive for any HPV (30.1%), and 66 for any HR-HPV (22.1%). The most prevalent HR-HPV types were HPV 16 (22.2% of all women with HPV infection), HPV 53 (14.4%), and HPV 66 (14.4%). HPV infections increased from 15.8% in the 18–20 years group to 50.0% in the 21–23 years group and then decreased to 9.1% in those aged 50 years or more (p = 0.008). Multiple HPV infections were observed in 15.7% of the study sample (52.2% of all HPV positive). There is a significantly higher prevalence of multiple infections in 18–32 years group women (24.5%) compared with females aged 33 years or more (6.8%) (p < 0.005). Current smokers were at increased risk of HPV infection (44.2% of HPV infections compared with 23.5% in never smokers, and 25.3% of multiple HPV infections compared with 11.3%; p = 0.001). HR-HPV infections were higher in women never been pregnant (27.1% compared with 7.7%; p = 0.001). Oral contraceptive use was completely unrelated to infection. Among the 122 women who had both cytological examination and HPV results, multiple HR-HPV types were found in 36.8% of those with abnormal cervical findings, and in 13.6% of those with normal cervical findings (p = 0.05). The results of the present investigation provide further evidence for the notion that cervical HPV infection is more widespread than previously suggested.