Jaagsiekte Sheep Retrovirus Can Reach Peyer's Patches and Mesenteric Lymph Nodes of Lambs Nursed by Infected Mothers.

Ovine pulmonary adenocarcinoma (OPA) is a contagious lung cancer of sheep caused by jaagsiekte sheep retrovirus (JSRV). It is generally accepted that transmission by the respiratory route occurs under natural conditions. However recent studies strongly indicate that JSRV can also be transmitted to lambs perinatally via colostrum and milk (C/M). The aim of this work was to confirm that C/M can transmit JSRV infection to lambs under natural conditions and investigate the initial events associated with this transmission route. We have analyzed the presence of JSRV in C/M samples from 22 naturally infected, asymptomatic ewes throughout a lactation period, and in various tissues collected from a group of 36 of their lambs that were fed naturally. The lambs were euthanized at 12, 24, 48, and 72 hours and at 5 and 10 days after birth. We detected JSRV-provirus by PCR in the somatic C/M cells from 10/22 ewes (45.45%). The virus was also detected in 9/36 lambs (25%). JSRV-infected cells, with lymphoreticular-like morphology, were observed by immunohistochemistry (IHC) and in situ hybridization (ISH) in Peyer's patches (PP) from the small intestine of the youngest lambs and in mesenteric lymph nodes (MLN) from lambs older than 72 hours. The virus was also detected by PCR in white blood cells (WBC) in 2/36 lambs (5.5%). These results confirm colostral transmission of JSRV to lambs under natural conditions. Infected lymphoreticular cells contained in C/M appear to be involved. These cells can cross the intestinal barrier of newborn lambs, reach the MLN and enter into circulation.

Ultrasound and microwave techniques for assisting ageing on lees of red wines.

Ageing on lees is a slow process that carries microbiological and economic risks in the wineries. This study evaluates the possibility of enhancing the extraction of different compounds from the lees, using combined strategies, such as ultrasound (US) or microwaves (MW) and the addition of inactive dry yeasts (IDY), to reduce the lees ageing time. The complete chemical analysis of the wine was done, amino acids, polysaccharides, colour and volatile compounds, together with the sensory analysis. The combined treatments increased the release of total polysaccharides, mannoproteins and total monosaccharides in the wines, and some amino acids like proline. However, wines treated with US and MW, with and without lees, showed a decrease in tannins and colour intensity, and in some volatile compounds like fatty acid esters, acetates and terpenes. The wines treated with IDY and MW were the best valued for their floral and red berry flavours and less astringency.

Effect of addition of soybean trypsin inhibitor to colostrum on immunological status in goat kids.

The aim of the study was to evaluate the influence of soybean trypsin inhibitor (TI) on immunoglobulin G (IgG) serum levels and growth in neonatal goat kids. Twenty-four newborn kids were fed with natural colostrum (group A), and 24 kids received the same colostrum with 1 g of TI per litre (group B). Blood samples were obtained at birth and on days 1, 2 and 4 of life to analyze serum proteins, IgG and haematological parameters. There were no clinical signs of disease and no significant differences in body weight between the groups. Haematological parameters were not affected by treatment. The peak of serum IgG was reached at 24 h of life, but no effects of soybean TI was observed on serum IgG levels. The apparent efficiency of absorption of IgG was similar in both groups (group A 24.5% vs. group B 25.2%, p > 0.05). The addition of TI to colostrum did not change the concentration of serum proteins and their fractions in goat kids. The correlation between serum IgG and gamma-globulin was positive and significant (p < 0.01, r = 0.64) in group A, but not in group B (p > 0.05, r = 0.08), suggesting a negative influence of soybean TI on gamma-globulin absorption. These results show that addition of soybean TI to colostrum did not improve the performance or immunological status in goat kids.

Fining with purified grape pomace. Effect of dose, contact time and varietal origin on the final wine phenolic composition.

Fining, which involves the addition of adsorptive material in order to reduce or eliminate certain unwanted components, is a common winemaking practice. Fining agents affect the wine phenolic compounds, some of which may be reduced. When this reduction is experimented by the tannins, a positive effect may result by decreasing astringency in the wine, although a decrease in the wine color may also take place when the anthocyanins are involved, affecting its quality. Recently, grape cell wall material has been tested as a potential fining agent in wines, since it shows a high affinity for tannins so that its use could reduce wine astringency. In this work, the use of purified grape pomace as fining agent is proposed and the effect of different doses and contact times on wine chromatic characteristics was investigated as well as how differences in the composition of the purified pomace could alter the phenolic composition of a red wine. The results showed that a Monastrell purified grape pomace dose of 6 mg/ml and a contact time of 5 days could be suitable for decreasing the wine tannin content without producing great changes in the wine chromatic characteristics. When comparing the effect of purified pomaces from four grape varieties, some differences in their capacity to interact with the wine tannins and anthocyanins were found, however, the results confirm that the purified grape pomace, a byproduct of the enology industry could be a new interesting fining material.

Influence of methyl jasmonate and benzothiadiazole on the composition of grape skin cell walls and wines.

Phenolic compounds are very important in crop plants, particularly in grapes. The different strategies to increase their levels include the use of elicitors such as methyl jasmonate (MeJ) and benzothiadiazole (BTH). In an attempt to improve the quality of wines, our aim was to evaluate the effect of preharvest application of these elicitors on the composition and structure of the skin cell walls of Monastrell, Merlot and Cabernet Sauvignon grapes, and to ascertain any relationship with the extractability of phenolic compounds during winemaking. The results indicated that the exogenous application of MeJ and BTH during veraison caused significant changes in several components of the skin cell walls, such as phenolic compounds, proteins and structural sugars. However these changes manifested themselves in different proportions in each variety and year, pointing to the varietal and meteorological dependence of the response to the application of these elicitors. The treatments delayed the maturation process in all varieties when rainfall was low. This observation, together with the observed increase in proteins and phenols in the skin cell wall of Monastrell and Cabernet Sauvignon, could contribute to the strength necessary to maintain the integrity of berries and to increasing resistance to fungal pathogens as the phenolic compounds evolve, thus improving the phenolic profile. However, the structural integrity of Merlot variety tended to decrease in the same conditions.

Application of enzyme preparations for extraction of berry skin phenolics in withered winegrapes.

Postharvest withering of grapes strongly affects the content and extractability of phenolic compounds in the production of sfursat, fortified and passito wines. This work evaluated the effectiveness of enzymes applied individually and/or in multi-enzyme blends, on the extraction of anthocyanins, oligomeric flavanols and polymeric flavanols from withered grape skins during simulated maceration. The study was performed on Vitis vinifera L. cv. Nebbiolo and Barbera because of their different skin phenolic profile and cell wall composition. Our findings highlight that the relationship between skin mechanical properties (berry skin break force and energy) and extraction yield of phenolic compounds is variety dependent. Significant correlations were found between the skin softening associated with cell wall degradation and the extraction of anthocyanins and flavanols in Nebbiolo, for which polygalacturonase, individually or in multi-enzyme blends, plays an important role. In Barbera, the extractability of phenolic compounds was not affected by the presence of exogenous enzymes.

In-situ demonstration of mitogen-activated protein kinase Erk 1/2 signalling pathway in contagious respiratory tumours of sheep and goats.

Ovine pulmonary adenocarcinoma (OPA) and enzootic nasal adenocarcinoma (ENA) are two contagious neoplastic diseases of secretory epithelial cells in the respiratory system of sheep and goats. Jaagsiekte sheep retrovirus (JSRV) is the aetiological agent of OPA, and enzootic nasal tumour virus (ENTV) is associated with ENA. The genomes of these retroviruses do not contain known oncogenes but products of the env gene are important in the generation of transforming stimuli. However, the cell signalling pathways activated in vivo are not completely understood. This study was based on the use of activation stage antibodies specifically detecting proteins of the extracellular signal regulated kinase Erk 1/2 cell signalling pathway and transcription factors. Tissue sections were collected from four natural cases of OPA, four experimentally induced OPA tumours, four ENA tumours in sheep, four ENA tumours in goats, two normal sheep lungs and two lungs with chronic inflammation. Routine immunohistochemical procedures with phosphorylation stage-specific antibodies were carried out. Representative proteins of the Erk1/2 pathway (Raf-1, Mek1/2 and p44/42MAPK) were activated in natural cases of OPA and ENA in sheep and goats and also in experimentally induced OPA. Transcription factors 90Rsk and Elk-1 were activated in OPA and ENA tumours. However, c-Myc was activated only in OPA tumours. In contagious respiratory neoplasms of sheep and goats the Erk1/2 pathway appears to be important for the in-vivo generation of the transforming stimuli.

Enzootic nasal adenocarcinoma of sheep and goats.

Enzootic nasal adenocarcinoma is a contagious tumour of the mucosal nasal glands affecting young adult sheep or goats. The disease occurs naturally in all continents except Australia and New Zealand. Clinical signs include continuous nasal discharge, respiratory distress, exophthalmos and skull deformations. The tumour is classified histologically as a low-grade adenocarcinoma. Nasal glands of both respiratory and olfactory muosal glands seem to be the origin of the neoplasia. It has been experimentally transmitted in sheep and goats using either tumour extracts or concentrated nasal fluids. Two distinct retroviruses are implicated in the aetiology of the neoplasia one in sheep (ONAV) and one in goats (CNAV). We suggest that jaagsiekte sheep retrovirus (JSRV), ONAV, CNAV, and their endogenous counterparts represent a unique family of retroviruses. The similarities between these viruses suggests that any control strategies, including vaccination, may be appropriate to both diseases. The differences, however, represent a unique resource for delineating the function of individual regions of the virus. It is intriguing that whilst ONAV and CNAV appear to be as different to each other as they are to JSRV, that they have very similar disease pathologies, distinct from that of OPA. Additionally, all three exogenous viruses manage to avoid instigating any apparent immune response. Whether this is indeed a result of tolerance induced by the endogenous counterparts or whether the viruses themselves have unique immunosuppressive properties will be an important finding.

Vaccination schedules in small ruminant farms.

Development and implementation of health management plans is the cornerstone of profitable farms; prevention of microbial diseases by means of vaccination is an integral part of such a plan. In every production type and management system in small ruminants, microbial diseases have a major significance, hence their proper control must be based in good health management practices, including use of effective and safe vaccines. Development of various types of vaccines is evolving very quickly in recent years and the improvement of new type of vaccines offers prospects. The article reviews and discusses vaccination programs and latest advances in development of vaccines against diseases that cause major economic losses in small ruminants. Specifically, vaccination schedules for the following diseases are reviewed: bacterial abortion (abortion associated with Brucella melitensis, Campylobacter spp., Chlamydophila abortus, Coxiella burnetii, Salmonella abortus ovis or Salmonella brandenburg), caseous lymphadenitis, clostridial diseases, colibacillosis, contagious echtyma, epididymitis caused by Brucella ovis, footrot, mammary diseases (contagious agalactia, mastitis), paratuberculosis and respiratory diseases (respiratory disease caused by Mannheimia haemolytica or other Pasteurellaceae).

Lack of a specific immune response against a recombinant capsid protein of Jaagsiekte sheep retrovirus in sheep and goats naturally affected by enzootic nasal tumour or sheep pulmonary adenomatosis.

Enzootic nasal tumour (ENT) and sheep pulmonary adenomatosis (SPA) are two contagious adenocarcinomas of the respiratory tract of sheep and goats. Both diseases are associated with related, but distinct, type-D-retroviruses (ENTV and JSRV respectively). No evidence of circulating antibodies has been described in animals affected by either ENT or SPA using antigens from natural sources. We evaluated the usefulness of a recombinant JSRV capsid protein (JSRV-CA) as antigen to study the antibody responses of animals naturally affected by ENT or SPA, using immunoblotting. Positive reactions were detected in the sera of both affected and unaffected sheep and goats. The reactivity was abolished completely by absorption with the GST fusion partner but not by JSRV-CA, suggesting that it was not specific. The results support prior observations indicating that sheep and goats infected by JSRV and ENTV do not develop specific humoral responses to these retroviruses.

Sheep pulmonary adenomatosis: characterization of two pathological forms associated with jaagsiekte retrovirus.

Pathological and immunohistochemical studies were performed on the lungs of 10 sheep with lesions of "classical" sheep pulmonary adenomatosis (SPA) and six sheep with "atypical" lung tumours. Lung tumour samples and other tissues from the same 16 animals were tested for the presence of jaagsiekte retrovirus (JSRV) by a polymerase chain reaction (PCR) that amplified a portion of the U3 long terminal repeat. The differences in the gross appearance of the classical and atypical forms paralleled the histopathological differences. The latter mainly concerned the stroma of the tumours which in the atypical cases was more heavily infiltrated by inflammatory cells and connective tissue fibres. JSRV major capsid protein was detected immunohistochemically in the epithelial transformed cells of both classical and atypical tumours, but the immune reactivity was slightly milder in atypical SPA. Proviral U3 sequences of JSRV were detected by specific PCR in all the tumour samples. Furthermore, the sequences of amplimers obtained from the two different pathological forms of the tumour were very similar. However, the dissemination of JSRV to other organs was greater in sheep with classical SPA than in those with atypical SPA. The pathological and virological features of these two forms of tumour are compared in an attempt to clarify whether classical and atypical SPA are two separate diseases or different expressions of a single disease spectrum.

Coexistence of enzootic nasal adenocarcinoma and jaagsiekte retrovirus infection in sheep.

Ten sheep naturally affected with enzootic nasal adenocarcinoma (ENA), a disease associated with ovine enzootic nasal tumour virus (ENTV-1), were found also to be infected with jaagsiekte sheep retrovirus (JSRV), the causal agent of ovine pulmonary adenocarcinoma (OPA). Only one of the sheep showed OPA lung lesions. The animals belonged to 10 flocks located in a geographical area in which OPA is frequently seen. ENTV-1 was found in all the ENA tumours but only occasionally in extra-tumoral sites, confirming the results of a previous study. In contrast, JSRV had a disseminated tissue distribution, similar to that previously reported for animals infected with JSRV. However, the occurrence of JSRV in lymphoid tissues was clearly greater than in sheep infected with JSRV but with no lesions of ENA. The data suggested a synergistic relationship between ENTV-1 and JSRV, resulting in increased proliferation of JSRV.

[Transient neurological deficit secondary to a leptomeningeal cyst in an adult patient]. / Déficit neurológico transitorio secundario a quiste leptomeníngeo en un paciente adulto.

INTRODUCTION: An uncommon complication of fractures of the skull in infancy is a leptomeningeal cyst, which is also known as a growing skull fracture. A post traumatic leptomeningeal cyst may occur in adults, but is much rarer and may be attributed to a fracture of the skull which occurred in childhood. This complication is caused by a tear in the dura mater, through which pulsation of the cerebro spinal fluid forces the arachnoid layer to herniate. The commonest clinical finding is a soft tissue swelling or tumour appearing on the head. CLINICAL CASE: We report the case of a 47 year old man, with a past history of a head injury in childhood. He presented complaining of loss of sensation in the right arm and deviation of the mouth, from which he recovered within an hour. Neuro imaging studies showed irregular destruction of the right temporal bone and hypodensity of the underlying brain tissue. Surgical operation and histological study of the bone removed showed that it was a leptomeningeal cyst, associated with a cerebral infarct at the site of an old skull fracture. CONCLUSIONS: A post traumatic leptomeningeal cyst in an adult patient is caused by a tear in the dura mater caused by a skull fracture during childhood. It may be associated with a cerebral infarct. It may present with only transient focal neurological symptoms.

Solitary tumours associated with Jaagsiekte retrovirus in sheep are heterogeneous and contain cells expressing markers identifying progenitor cells in lung repair.

Ovine pulmonary adenocarcinoma (OPA) is a naturally occurring lung cancer of sheep caused by jaagsiekte sheep retrovirus (JSRV). This study examines immunohistochemically solitary lung nodules considered as early OPA lesions from 11 sheep infected naturally by JSRV. All 11 neoplastic nodules exhibited features of adenocarcinoma and in four of them mesenchymal growth was also observed. Both types of lesion were labelled with antibody specific for JSRV-Env. In two cases infiltrating lymphoreticular cells also contained JSRV-Env. All tumours had a high Ki67 labelling index and variably contained cells expressing CC10 (a marker of Clara cells (CCs)), SPC (a marker of type II pneumocytes), p63 and keratin 14 (markers for stem/progenitor cells of the lung airway epithelia). Tumours with mesenchymal growth had intense expression of vimentin and desmin, weak expression of smooth muscle actin and did not express pancytokeratin and p63. Both epithelial and mesenchymal proliferations did not express the stem cell markers CD90 and CD117, but some tumour infiltrating cells expressed CD133. Solitary OPA tumours can therefore be adenocarcinomas or mixed tumours and have a heterogeneous cellular composition, containing groups of cells expressing markers that characterize local progenitor cells involved in lung repair.

Multiple cranial neuropathy and intracranial hypertension associated with all-trans retinoic acid treatment in a young adult patient with acute promyelocytic leukemia.

All-trans retinoic acid (ATRA) induces complete remission in 64-100 % of patients with acute promyelocytic leukemia (APL), and is considered to be a safe agent. Pseudotumor cerebri is a neurological side effect of ATRA reported in pediatric patients, and which is characterized by raised cerebrospinal fluid pressure in the absence of any intracranial pathology or secondary causes of intracranial hypertension. Involvement of cranial nerves other than II and VI is very uncommon in idiopathic intracranial hypertension (IIH); peripheral facial nerve palsy is exceptional and has rarely been described in the context of treatment with ATRA. We describe the case of a 15-year-old female patient with APL who developed an IIH and involvement of cranial nerves (bilateral papilledema, left facial and right sixth nerves) after receiving induction therapy including ATRA. Viral infections and other causes of secondary cranial nerve lesions were excluded. Symptoms completely subsided with the temporary withdrawal of ATRA and did not recur after reintroducing the drug. To date, the patient has managed to receive the treatment as per protocol. In conclusion, we report an atypical presentation of IIH that merits consideration, especially with respect to young patients with APL receiving ATRA; our most important observation is that the drug could be safely reintroduced once the symptoms had resolved.

The distribution of immune cells in the lungs of classical and atypical ovine pulmonary adenocarcinoma.

Ovine pulmonary adenocarcinoma (OPA) is a contagious tumour caused by infection of sheep with Jaagsiekte sheep retrovirus. Two forms of OPA have been identified, classical and atypical, which can be distinguished clinically and pathologically. Most notably classical OPA is progressive until death, while atypical OPA remains subclinical. In the present study the local immune responses in the lungs of cases of atypical OPA were compared with those from classical cases by immunohistochemistry using a panel of mouse anti-sheep mAbs. Distinct differences in the distribution of immune cell subsets in the two forms of OPA were observed. In particular there was an intratumoural influx of T cell subsets and MHC Class II expression on the tumour cells in atypical OPA, neither of which was seen in classical OPA. It is possible that these differences may contribute, at least in part, to determining the progressive course of classical OPA compared with the subclinical nature of atypical OPA.

Filter-based infrared detectors for high temperature size exclusion chromatography analysis of polyolefins: calibration with a small number of standards and error analysis.

Infrared detection has been shown to be very appropriate for high temperature analysis of polyolefins. After some early reports in which dispersive or single-band filter-based detectors were applied, Fourier transform detectors have been described for this application, in order to improve the method sensitivity. Modern simple filter-based detectors prove, however, comparable sensitivity while providing a number of practical advantages when coupled to chromatographic systems: reduced cell volume, simplified hardware, continuous generation of absorbance chromatograms, as well as simpler data collection and processing. A practical method for calibration, using multiple-band signals obtained with filter-based detectors and a small number of reference materials, is here discussed. Calibration data are used to compare the performance of detectors based on different opto-electronic technologies and filter designs. A procedure for estimation of errors in the slice-by-slice measured methyl frequency, based on signal-to-noise ratio considerations, is described. The good accuracy provided by the filter-based IR detectors was noticeable, considering that it was obtained using a small set of reference materials. A minimal concentration of 0.009 mg/mL was estimated to be required at the detector cell, in order to keep the errors below one unit of methyl per one thousand total carbons. This low minimal concentration requirement allows using standard SEC conditions, without compromising the molar mass distribution accuracy and resolution.

Finding the best separation in situations of extremely low chromatographic resolution.

Samples with a large number of compounds or similarities in their structure and polarity may yield insufficient chromatographic resolution. In such cases, however, finding conditions where the largest number of compounds appears sufficiently resolved can be still worthwhile. A strategy is here reported that optimises the resolution level of chromatograms in cases where conventional global criteria, such as the worst resolved peak pair or the product of elementary resolutions, are not able to detect any separation, even when most peaks are baseline resolved. The strategy applies a function based on the number of "well resolved" peaks, which are those that exceed a given threshold of peak purity. It is, therefore, oriented to quantify the success in the separation, and not the failure, as the conventional criteria do. The conditions that resolve the same amount of peaks are discriminated by either quantifying the partial resolution of those peaks that exceed the established threshold, or by improving the separation of peaks below it. The proposed approach is illustrated by the reversed-phase liquid chromatographic separation of a mixture of 30 ionisable and neutral compounds, using the acetonitrile content and pH as factors.