Effects of Confluent, Roscovitine Treatment and Serum Starvation on the Cell-cycle Synchronization of Bovine Foetal Fibroblasts

The present study was designed to examine the effects of cell-cycle synchronization protocols, such as confluent, roscovitine treatment and serum starvation, in bovine foetal fibroblasts on synchronization accuracy at G0/G1, viability, apoptosis, necrosis and ploidy for use as a nuclei donor. The cells in 5-10 passages were randomly allocated into three treated groups. Cells were cultured either in Dulbecco's modified Eagle's medium (DMEM) + 10% foetal bovine serum (FBS) until 90% confluent (group 1, confluent), in DMEM + 10% FBS + 30 microM roscovitine for 12 h (group 2, roscovitine), or in DMEM + 0.5% FBS for 5 days (group 3, serum starvation). Most of the cells (>80%) in all groups were arrested at the G0/G1 stage. Although the rates did not differ, cells in group 1 showed an increased cell population arrested at the G0/G1 phase. Significantly (p < 0.05) higher rates of apoptosis occurred in group 3 than in group 1 and 2 (10% vs 6% and 6%, respectively). No differences in chromosomal abnormality were observed among groups. However, by increasing the number of cell culture passages up to 15, significantly (p < 0.05) higher chromosomal abnormality was observed than in 5 and 10 passages (39% vs 28% and 23%, respectively) in group 1. The results clearly indicated that bovine foetal fibroblasts could be effectively synchronized at G0/G1 stages by all the three different treatments, confluent, roscovitine and serum starvation. However, cells in confluent showed reduced apoptosis and necrosis when they underwent 5-10 passages, exhibiting increased percentage of cells with stable chromosome diversity. Hence, cells in confluent merit further studies before they could be used as nuclear donors.     

Concepts and Direction of Induced Systemic Resistance in Plants and its Application

Resistance to plant disease is often specific and metabolites and receptors contributing to this specificity may have specific structures. However, simple, structurally-unrelated compounds induce systemic resistance in unrelated plants to diverse pathogens including fungi, bacteria and viruses. Both resistance and induced systemic resistance (ISR) are associated with the rapid accumulation of the same structurally unrelated putative defense compounds that have diverse functions. It has been suggested that cultivar (race)-specific resistance is initiated by the specific interaction of a pathogen product (or pathogen induced product) and a plant receptor. However, restricted infection by pathogens can result in ISR and many different compounds can cause ISR. It is thus evident that there are both specific and non-specific routes to the master switch for ISR and there may be more than one master switch. Are reactive oxygen species and free radicals regulating the master switch(es) via both routes? It is also evident there are many switches, other than the master switch. Adding to the complexity of resistance and ISR are the observations that different compounds and pathways may mediate different biochemical resistances. Activation of one of the pathways may antagonize or enhance the activation or effectiveness of another. The review will address these complexities and questions and propose directions of research which require high priority. Factors which encourage and suppress the application of ISR in agriculture will also be addressed.     

The Characterization of Heavy Metals in the Grapevine (Vitis vinifera) Cultivar Rkatsiteli and Wild Blackberry (Rubus fruticosus) from East Serbia by ICP-OES and BAFs

Inductively coupled plasma optical emission spectroscopy, in combination with calculation of various biological factors, was used in this study not only for a simple illustration of the present status of some essential and non-essential metals such as copper (Cu), zinc (Zn), nickel (Ni), lead (Pb), arsenic (As), and cadmium (Cd) in the grapevine cultivar Rkatsiteli and wild blackberry from East Serbia but also for the estimation of plant bioaccumulation potentials. Great majority of detected metal concentrations were in normal ranges except in few cases: Zn concentrations were at the level of deficiency in almost all plant parts, and some Cu and As concentrations were at the levels that could be considered phytotoxic. Biological accumulation factors provided very informative data on metal accumulation and translocation in both plants and pointed to low accumulation rates of metals except in the case of Cu, Zn, and As in some wild blackberry tissues.     

Maize defence mechanisms against the European corn borer,Ostrinia nubilalis Hübner (Lepidoptera: Crambidae)

Maize is arguably the most widely grown crop in the world, but it is often associated with one of its major insect pests, the European corn borer (ECB). The damage caused by this species to maize production is generally variable, but in many cases it is economically significant. This review paper provides an overview of the research findings on the natural plant defence mechanisms against ECB larvae published till now. What is resistance and how it is achieved, what is the chemical response of maize plants to insect feeding and how tolerance can be increased. A short introduction in breeding for resistance and a discussion if the mentioned traits can be used in conventional breeding in order to create maize hybrids less affected by ECB are given.     

A comparison of the effects of carbon dioxide and medical air for abdominal insufflation on respiratory parameters in xylazine-sedated sheep undergoing laparoscopic artificial insemination

AIMS: To determine if abdominal insufflation with medical air will improve oxygenation and ventilation parameters when compared to insufflation with CO₂ in xylazine-sedated sheep undergoing laparoscopic artificial insemination (AI).

METHODS: Forty-seven sheep underwent oestrus synchronisation and were fasted for 24 hours prior to laparoscopic AI. Each animal was randomised to receive either CO₂ or medical air for abdominal insufflation. An auricular arterial catheter was placed and utilised for serial blood sampling. Respiratory rates (RR) and arterial blood samples were collected at baseline, after xylazine (0.1?mg/kg I/V) sedation, 2 minutes after Trendelenburg positioning, 5 minutes after abdominal insufflation, and 10 minutes after being returned to a standing position. Blood samples were collected in heparinised syringes, stored on ice, and analysed for arterial pH, partial pressure of arterial O₂ (PaO₂), and CO₂ (PaCO₂). The number of ewes conceiving to AI was also determined.

RESULTS: Repeated measures ANOVA demonstrated temporal effects on RR, PaO₂, PaCO₂ and arterial pH during the laparoscopic AI procedure (p<0.001), but no difference between insufflation groups (p>0.01). No sheep experienced hypercapnia (PaCO₂>50?mmHg) or acidaemia (pH<7.35). Hypoxaemia (PaO₂<70?mmHg) was diagnosed during the procedure in 14/22 (64%) ewes in the CO₂ group compared with 8/23 (35%) ewes in the medical air group (p=0.053). Overall, 15/20 (75%) ewes in the CO₂ group conceived to AI compared with 16/22 (72.7%) in the medical air group (p=0.867).

CONCLUSIONS AND CLINICAL RELEVANCE: There were no statistical or clinical differences in RR, PaO₂, PaCO₂, pH, or conception to AI when comparing the effects of CO₂ and medical air as abdominal insufflation gases. None of the sheep experienced hypercapnia or acidaemic, yet 42% (19/45) of sheep developed clinical hypoxaemia, with a higher percentage of ewes in the CO₂ group developing hypoxaemia than in the medical air group. Based on the overall analysis, medical air could be utilised as a comparable alternative for abdominal insufflation during laparoscopic AI procedures.     

Equine pituitary pars intermedia dysfunction: current understanding and recommendations from the Australian and New Zealand Equine Endocrine Group

The purpose of this article is to provide a review of the current knowledge and opinions about the epidemiology, clinical findings (including sequelae), diagnosis, treatment and monitoring of equine pituitary pars intermedia dysfunction, particularly in the Australian context. This information and the recommendations provided will assist practitioners in making informed decisions regarding the diagnosis and management of this disorder.     

Evidence for genetic diversity of Toxoplasma gondii in selected intermediate hosts in Serbia

To contribute to the insight into the worldwide population structure of Toxoplasma gondii, we genetically characterized a total of eight strains isolated from intermediate hosts including humans, sheep and pigeons in Serbia. Although parasite DNA was detected in 28.2% (60/213) of the human samples from 162 patients serologically suspected of active toxoplasmosis, as well as in 5/7 seropositive pigeons and in 2/12 seropositive sheep examined, multilocus PCR-RFLP genotyping, using SAG1, 5′SAG2, 3′SAG2, GRA6, 5′GRA7 and 3′GRA7 as markers, was successful in only four human isolates (of which one was isolated from both the bronchoalveolar lavage fluid and blood samples of a single patient), one sheep and three pigeons. Of the eight isolates, five were type II (62.5%), one was type III, one was atypical, and one had a type I allele at GRA6 as the single locus genotyped. Although type II, as elsewhere in Europe, predominated, these results may suggest a higher genetic diversity of T. gondii in Serbia, reflecting local environmental contamination and also the geographical position of the country in South-East Europe.     

Intratesticular benign peripheral nerve sheath tumour in a ferret (Mustela putorius furo)

A domestic ferret was submitted for sterilisation because of right testis enlargement. Oestradiol and cortisol concentrations were within normal physiological ranges, but testosterone was below and progesterone above normal. Microscopically, the right testis, with the exception of a small part of the epididymis, was replaced with neoplastic tissue. The tumour was composed of streams and bundles of closely packed spindle to ovoid cells forming whorls around collagen and capillaries, and separated by a collagenous matrix. In some areas, cells were loosely arranged and separated by a pale myxomatous matrix. The left testis showed atrophy. The majority of neoplastic cells expressed vimentin and S-100 protein, while expression of collagen IV was moderate and there was no expression of glial fibrillary acid protein. On the basis of macroscopical and histopathological findings, and supported by immunohistochemical reactivity, the diagnosis of benign peripheral nerve sheath tumour was made. This is the first report of benign peripheral nerve sheath tumour in ferret testis.