Search

MilkProtChip is an oligonucleotide microarray based on the arrayed primer extension (APEX) technique, allowing genotyping of single nucleotide polymorphisms (SNPs) in genes of interest for bovine milk protein biosynthesis. APEX consists of a sequencing reaction primed by an oligonucleotide anchored with its 5'end to a glass slide and terminating one nucleotide before the polymorphic site. The extension with one fluorescently labeled dideoxy nucleotide complementary to the template reveals the polymorphism. A total of 75 SNPs were selected among those associated directly or potentially with milk protein content. Among the 75 SNPs, 4 did not produce a positive signal. Most of the remaining SNPs produced a signal for both strands, except for 4 (one strand). In the validation step, 12 Polish Holstein bulls, 1 Polish Red bull, 1 bison (Bison bonasus), 11 Jersey cows and 25 Polish Holstein cows were screened to validate SNPs. Among the 71 selected SNPs--26 were found monoallelic, the rest showing at least two genotypes for the entire population under study. All the animals were earlier genotyped for 2-5 SNPs by PCR-RFLP and PCR sequencing and all showed complete concordance with APEX genotyping. APEX reactions showed relatively high signal frequencies: more than 0.9, 0.9-0.8 and below 0.8, for 65, 4 and 2 DNA samples, respectively. The primary application of the MilkProtChip is the simultaneous genotyping of dozens of SNPs to reveal and clarify the genetic background of milk protein biosynthesis. The chip may possibly be used for dairy cattle identification and paternity analysis, evolutionary studies, the evaluation of genetic distances between wild and domestic cattle breeds and the domestication history of bovine species.

Milk protein genes expression in cows' mammary epithelial cells is regulated mostly by the action of prolactin mediated through the STAT5A transcription factor. The STAT5A gene is a potential quantitative trait locus (QTL) and genetic marker of production traits in dairy cattle. The sequence of the bovine STAT5A gene was analysed in this study to investigate if mutations in this sequence might be responsible for quantitative variations in milk yield and composition. Ten PCR fragments representing most important functional domains of STAT5A were screened for polymorphism. Using the SSCP method a new SNP (A/G) was found, located in intron 9 at position 9501 (GenBank AJ237937). The frequencies of alleles were estimated in 186 Black-and-White cows (0.52 and 0.48 for A and G, respectively) and in 138 Jersey cows (0.58 and 0.42 for A and G, respectively). For Black-and-White cows with different STAT5A genotypes no significant associations between STAT5A genotypes and milk performance traits were found. Statistically significant differences in the first and second lactations for milk yield, fat and protein content were found in Jersey cows. Cows with the GG genotype showed the highest milk yield, while cows with genotypes AA and AG showed higher protein contents when compared to cows with the GG genotype. Interestingly, cows with genotype AG showed significantly higher protein yields in comparison to cows with the AA genotype. For fat content, cows with genotype AA showed the highest level of this trait in the 1st and 2nd lactation. Further studies are necessary to evaluate an allele substitution effect in the population of sib-families of STAT5A heterozygous bulls.

This study demonstrates the usefulness of PLA and nested-PCR methods for the detection of cattle infection with enzootic bovine leukemia virus. Serological control with PLA was by 1.7% more sensitive than reference ELISA examination. Both the DNA of env-BLV gene in NCR, V-NCR, FLK and in recombinated AG clone cells, as well as in the whole blood and in blood isolated leukocytes of dairy cattle, were detected with the nested-PCR.

It is currently debated whether identification of ESR (estrogen receptor) genotypes should be introduced into breeding programs of Large White pigs. The aim of this study was to evaluate the possible relations between ESR/Ava I polymorphism and carcass performance traits in Polish Large White boars. We examined 103 boars originating from one herd in NE Poland. ESR/Ava I genotypes were determined by the PCR-RFLP method. By the use of the Duncan test, we found highly significant differences (P < 0.01) between WW and MW genotypes, as well as significant differences (P < 0.05) between WW and MM genotypes for meatiness. No significant differences were found for daily gain and selection index.

A method allowing simultaneous genotyping of two loci: ryanodine receptor 1 (RYR1) and estrogen receptor (ESR) is presented. In multiplex PCR amplification, two amplicons were simultaneously produced: a 272 bp fragment of RYR1 gene and a 185 bp fragment of ESR gene and were then subjected to "one-tube" restriction enzyme digestion with Hin6 I and Ava I, respectively. A total of 122 Polish Large White and Polish Landrace pigs were genotyped by this method, demonstrating its reliability, convenience and lower costs. This method may be useful in the wide-scale genotyping of both loci in pig breeding programmes.

Case records of 200 patients operated on in 1998/99 for herniated lumbar disc in Neurosurgery Dept. showed that 95 patients (47.5%) had been treated previously by 148 alternative medical or non-medical procedures. The authors discuss the problem of non-conventional treatment methods applied for herniated lumbar disc by professionals or non professionals. The procedures are often dangerous.

Haplotypes of three bovine casein loci were analyzed as a way of determining genetic linkage. A total of 330 individual sperm cells from a triply heterozygous bull were selected by means of a new method involving fixing the sperm cells into low-melting-point agarose gels. The method is simple and very accurate with an efficiency close to 100% for picking sperm cells and producing amplifiable DNA templates, circumventing the complicated statistical analysis mandated by automated cell sorting. The DNA was amplified in a two-step polymerase chain reaction (PCR) to achieve the necessary high specificity of amplification. In the first reaction, primers flanking the polymorphic site at each locus were used. The PCR product from the first reaction was then reamplified in a second PCR using primers that create allele-specific restriction sites (ACRS) in the PCR products for two of the three loci, allowing the alleles to be determined by gel electrophoresis. No recombinants were found among the 330 single sperm cells analyzed, giving a lod score higher than 30 and proving a very strong linkage between bovine casein genes.

History and results of investigations in 34 patients with cerebello-pontine angle tumours are presented. Tumour was diagnosed in the first, audiological stage of the disease only in 4 patients. In 7 more patients additional symptoms of central nervous system focal disorders occurred. The remaining 23 patients were in the third stage of the disease with symptoms of increased intracranial pressure. The main reason for such a late diagnosis of the tumour was neglecting of the early audiological symptoms of the disease by patients and by general practitioners as well.

The authors describe in detail the technique of fronto-temporo-sphenoidal craniotomy of Yasargil and stress its advantages in surgical treatment of aneurysms of the circle of Willis. This method was used in surgical treatment of 82 patients with intracranial aneurysms, expanding lesions situated in the vicinity of the sella and ala parva of the sphenoid bone, and with other intracranial processes, and very good conditions for operation were obtained. The authors believe that fronto-temporo-sphenoidal craniotomy is the most favourable approach for the management of aneurysms of the circulus arteriosus.

The authors present the results of surgical treatment of 48 aneurysms situated on the anterior communicating artery operated upon by the classic and microsurgical methods. The latter method has made it possible to raise the proportion of very good and good results from 23% in the classic method to 70%, and to reduce the postoperative mortality in groups I and II of condition severity from 13.6% to 4%. Independently of the method used Korsakoff's syndrome was observed in 17% of cases. The high value of Yasargil's surgical approach is stressed.

The authors present a group of 38 patients with middle cerebral artery aneurysms in whom 48 aneurysms were excluded from the cerebral circulation by microsurgical techniques. In 11 cases multiple aneurysms were found. In 2 cases the wall of the aneurysm was strengthened. In the light of this material and a survey of the pertinent literature the authors analyse the relationship between the results of surgical treatment and various factors. A significant increase in postoperative mortality was found with more severe condition of the patients by the Botterell scale. The factors worsening the prognosis were also: presence of intracerebral haematoma and aneurysm location in the bifurcation of an artery. Arteriospasm increased the incidence of neurological deficits.

The author analyses the clinical data and pathological manifestations in 29 cases of intraspinal tumours in which the results of preoperative lumbar tap were equivocal or excluded presence of vertebral canal obliteration. On the other hand, myelography showed presence of complete or incomplete contrast stop at the level of the tumour, despite misleading information of the diagnostic lumbar tap.

A prospective, randomized trial evaluates the effects of two postoperative treatment regimens on survival in 198 adult patients with supratentorial gliomas. All patients were irradiated with 6,000 rads after possibly radical removal of tumors. CCNU administration in the doses of 100 mg/sq m of body surface every 6-8 weeks following surgery proved to have no significant effect on the survival of patients. The median survival time in patients receiving radiation therapy alone was 61 +/- 7 weeks, while in those receiving additional chemotherapy was 56 +/- 4 weeks. Tumor histological malignancy and patients age were found to be the only important prognostic factors, irrespective of the treatment modality.

Therapeutic results are reported in 115 patients operated upon for intracranial aneurysms using the microsurgical technique from Yasargil's approach. Elimination of the aneurysm from the circulation by clipping its pedicle was done in 94 cases (82%), trapping of the afferent artery (12%), in 7 cases (6%) only the wall of the aneurysmatic sac was strengthened by wrapping. A good therapeutic result was obtained in 68 patients (59.1%), satisfactory in 6 (5.2%), poor in 13 (11.3%). The overall mortality was 24.4% (28 cases) but after taking into account 4 patients dying due to extracerebral causes, the mortality caused by complications related to exclusion of the aneurysm from cerebral circulation was 20%. The factors influencing the results of surgical treatment of cerebral aneurysms are discussed in the light of the own material.

The authors analyzed the signs and clinical data from the preoperative period in 39 patients with tumours of the conus terminalis and cauda equina. Most patients had been treated for a long time for lumbar discopathy with sciatic pains. Most symptoms and neurological signs were in many cases suggestive rather of lumbar disc prolapse than spinal tumour. Radiological examinations of the spine failed in a overwhelming majority of cases to provide clues for correct diagnosis, only the type of tendon jerks loss, results of Queckenstedt's test and protein level in the cerebrospinal fluid suggested spinal tumor as a cause of the disease. Contract radiological examination of the vertebral canal gave definite answer in all cases.

Thirteen patients treated surgically for multiple intracranial aneurysms are described. They accounted for 12% of all patients with intracranial aneurysms diagnosed by arteriography. The incidence of multiple intracranial aneurysms, their location, surgical tactics and results of surgical intervention are discussed in the light of an analysis of own results and a survey of the pertinent literature.

The authors describe a case of a giant intracranial aneurysm of the left internal carotid artery in which signs of raised intracranial pressure predominated. In view of presence of internal hydrocephalus Holter's valve was implanted. The aneurysm was excluded from the circulation by trapping the afferent vessel. A good therapeutic result was achieved.

On the basis of control angiographies after surgical treatment of 59 intracranial aneurysms full effectiveness of aneurysm elimination from the circulation was assessed and found to be fully effective in 93.2% of cases. In 3 cases the aneurysmal sac was partially visualized, and in 2 cases the metal clip was found to have slipped off from the aneurysm neck. One of these patients was reoperated successfully. The investigations demonstrated in 17 cases (29%) presence of postoperative vasospasm, in 8 cases internal hydrocephalus following past bleeding and in 2 cases accumulation of cerebrospinal fluid in the subdural space. On the basis of literature reports and own experiences the authors stress the necessity of possibly early control angiography which should be accepted as a routine method. In case of internal hydrocephalus demonstration insertion of a ventriculoatrial shunt is suggested.