The feasibility of sentinel lymph node biopsy for gastric cancer: the experience from Serbia.

PURPOSE: The prediction of outcome for patients with gastric cancer is determined largely by the presence of lymph node metastases, which could be detected by sentinel lymph node (SLN) biopsy (SLNB). The purpose of this work was to determine the feasibility of SLNB in patients with gastric cancer for the assessment of regional lymph node status, including performing immunohistochemical (IHC) staining of SLN tissue. METHODS: We reviewed 137 consecutive patients with operable gastric cancer over a 10-year period using a retrospective (to examine skip metastases) and prospective (to evaluate successful mapping) study design. SLNs were mapped, biopsied and subsequently explored by routine hematoxylin & eosin (H&E) staining and by IHC staining using a cytokeratin 8/18 antibody. RESULTS: The retrospective study showed a low incidence of skip metastases (3.7%). Mapping of SLNs in the prospective study was highly successful (98.2%). During the prospective study, IHC examination of SLNs from 56 patients showed statistically significant change in disease stage. CONCLUSION: This study demonstrated highly successful mapping and biopsy of SLNs (98.2%), as well as highest specificity (100%), sensitivity (100%) and accuracy (100%) to predict metastasis in the surrounding lymph nodes of gastric carcinoma. In addition, we believe that IHC study might enable "ultra staging" and additional selection of patients for further cancer treatment.

Baseline and therapy-induced chromosome damages in peripheral blood lymphocytes of breast cancer patients assessed by the micronucleus assay.

PURPOSE: Radiotherapy (RT) alone or in combination with chemotherapy (CT) leads nearly always to increase of DNA damage in cancer patients. The purpose of this study was to determine the variability rate and individual sensitivity of breast cancer (BC) patients to the applied RT and RT in combination with CT. METHODS: The analysed sample included 30 women with histologically confirmed BC. The frequency of micronuclei (MN) was estimated in peripheral blood lymphocytes (PBL) by using the cytokinesis-block micronucleus (CBMN) assay before the administered therapy and one month later. RESULTS: The mean therapy-induced MN value was significantly higher (p < 0.001) compared with mean baseline MN. Both therapies (RT and combined RT+CT) significantly increased the MN frequency in patients' lymphocytes (p<0.001), but without significant differences in the therapy-induced MN frequency between these two groups (p > 0.05). The administered therapy induced significant difference in cell kinetics (p < 0.05). The results showed a wide range of inter-individual variability in both baseline and the therapy-induced MN frequency. CONCLUSION: The applied therapies increased the MN frequency in PBL in BC patients, and the presented data indicate absence of synergistic effect of these two therapies. None of the variation factors (age, smoking and therapy type) had influence on the noticed variability.

Effect of captopril on serum lipid levels and cardiac mitochondrial oxygen consumption in experimentally-induced hypercholesterolemia in rabbits.

Angiotensin converting enzyme inhibitors are widely used in therapy of cardiovascular diseases. However, the consensus on effects of these inhibitors in control of myocardial oxygen consumption during the process of experimental hypercholesterolemia and under the condition of endothelial dysfunction has not been reached. Here we examined effects of captopril, an angiotensin converting enzyme inhibitor, on serum lipid levels and oxygen consumption rate in mitochondria isolated from heart of rabbits treated by hypercholesterolemic diet. During the twelve-week period, the Chinchilla male rabbits were daily treated by saline (controls); 1 % cholesterol diet; 5 mg/kg/day captopril or 1 % cholesterol + 5 mg/kg/day captopril. Total- and high-density lipoprotein cholesterol and triglyceride in serum were measured spectrophotometrically. The left ventricle mitochondrial fraction was isolated and myocardial oxygen consumption was measured by Biological Oxygen Monitor. Mitochondria isolated from hearts of rabbits exposed to hypercholesterolemic diet showed significantly reduced respiration rates (state 3 and state 4) with altering adenosine diphosphate/oxygen ratio, whereas the respiratory control ratio was not affected when compared to controls. Mitochondria from cholesterol/captopril-treated animals showed significantly reduced respiration rates without altering adenosine diphosphate/oxygen ratio index or respiratory control ratio. Although captopril did not exert the favorable effect on serum lipid levels in cholesterol-treated animals, it restored the mitochondrial oxygen consumption. Further studies should be performed to define the underlying physiological and/or pathophysiological mechanisms and clinical implications.

In vitro evaluation of the genotoxicity of ritodrine and verapamil in human lymphocytes.

The aim of this study was to investigate the genotoxic effects of ritodrine and verapamil on human peripheral lymphocytes in vitro using micronucleus (MN) test. Also, fluorescence in situ hybridization (FISH) with a centromeric probe was performed to determine the origin of the induced MN. Cells were treated with 8.4 × 10(-6) M - 25.2 × 10(-4) M concentrations for ritodrine and 0.56 - 11 × 10(-5) M concentrations for verapamil, separately and combined. The MN frequencies showed increase after all treatments, but the difference between treated cells and untreated controls were found to be statistically significant only in the concentration range from 8.4 × 10(-5) M - 4.5 × 10(-4) M for ritodrine, 1.1 - 3.3 × 10(-5) M for verapamil, and in combined treatment with concentrations 8.4 × 10(-5) M + 1.1 × 10(-5) M for ritodrine and verapamil. The highest tested concentrations of both medicaments showed cytotoxic effect. Both medicaments decreased the nuclear division index (NDI) in tested concentrations. The results of FISH analysis suggest that verapamil, separately or combined with ritodrine, shows to a larger extent aneugenic than clastogenic effect.

[New trends in neuromonitoring patients with with aneurysmal subarachnoid haemorrhage].

Neurointensive care of patients with subarachnoid haemorrhage is based on the theory that clinical outcome is the consequence of the primary haemorrhage and a number of secondary insults in the acute post haemorrhage period. Several neuromonitoring techniques have been introduced or accomplished into clinical practice in the last decade with the purpose of monitoring different but related aspects of brain physiology, such as cerebral blood flow (CBF), pressure within the cranial cavity, metabolism, and oxygenation. The aim of these techniques is to obtain information that can improve knowledge on brain pathophysiology, and especially to detect secondary insults which may cause permanent neurological damage if undetected and untreated in "real time", at the time when they can still be managed. These techniques include intracranial pressure (ICP) measurements, jugular venous oxygen saturation, near-infrared spectroscopy, brain tissue monitoring, and transcranial Doppler. The available devices are limited because they measure a part of complex process indirectly. Expense, technical difficulties, invasiveness, limited spatial or temporal resolution and the lack of sensitivity add to the limitation of any individual monitor. These problems have been partially addressed by the combination of several monitors known as multimodality monitoring. In this review, we describe the most common neuromonitoring methods in patients with subarachnoidal hemorrhage that can assess nervous system function, cerebral haemodynamics and cerebral oxygenation.

[Difference between values of gas analysis of arteries and internal jugular vein in patients with cerebral infarction].

INTRODUCTION: Cerebral infarction is more and more frequently present by massive participation and high percentage of mortality even in young population. It is notified as very hard, vitally endangered disease. OBJECTIVE: To prove whether there is a difference in the gas analyses of the arterial and cerebral venous blood between the patients with the cerebral infarction of the left and right hemisphere and why it is significant for the treatment of those patients. MATERIAL AND METHODS: Sixty-five (65) patients of both sexes who were diagnosed by CT as: cerebral infarction, cerebral multiinfarctions and cerebral haemorrhagic infarctions were investigated. Under the same conditions their radial artery and left and right internal jugular vein were tapped--on the fourth, eighth and tenth day of their disease. Gas analysis, calculation of oxygen content and statistical comparison of testing results were performed from the samples of arterial and venous blood. RESULTS: By continuous monitoring of gas analysis parameters of arterial and cerebral venous blood from internal jugular vein, we can follow up the phases of disease, the velocity of changes of gas analysis parameters and adequacy of brain circulation and to correct therapy adequately for the purpose of preventing basic disease complications and to estimate the results of treatment. CONCLUSION: There is a significant difference between the results of gas analysis of arteries and the internal jugular vein, in patients with infarctions of the left and right hemisphere of the brain.

[The role of myomectomy in female infertility].

The aim was to examine which is the role of myoma in women infertility. We analyzed 100 patients with infertility that underwent classic abdominal myomesctomy from 2000. to 2003. year. Frequency of conception was 46%. Most patients were over 30-years-old. Conception happened in 80% patients aged 30-39 years. In 69,6% patients with secundar infertility happened conception. Pregnancies occured more often in infertility shorter than 36 months, in front wall myoma and in intramural-subserose or subserose type. In 11 patients with intramural-submucose myomas, uteral cave was opened and only two of them got pregnant. Recidives of myoma happened in 18%, and postoperative adhesions in 29% of pacients. Age, duration of pre-operative infertility and characteristics of myoma did not have statistically significant influence on the conception. Miomectomy is the important method in treatment of infertility, especially if the other possible causes were excluded.

Population-genetic study of Balkan endemic nephropathy in Serbia.

The study of Balkan Endemic Nephropathy (BEN) in the affected localities of southern Serbia shows population-genetic difference between samples of BEN affected individuals and control group consisting of non-affected individuals from the same localities. Detailed population-genetic study in village Chepure, which includes 20 large families where BEN is present in 646 (from first to fourth degree) relatives of probants, shows familial character of disease as well as significant genetic influences in expression of the illness. Our study of genetic homozygosity degree includes an analysis of the presence, distribution and individual combination of 20 to 30 selected genetically controlled morpho-physiological traits in the sample of BEN patients and in the control-healthy group. Assuming that BEN is genetically controlled disease, we made a hypothesis that an increased homozygosity level, as well as the changed variability among the patients, could be population-genetic parameter for the prediction of the illness. Taking into consideration our experience, as well as the experience of numerous scientists who studied the nature of the inheritance of mono- and oligo-genically controlled qualitative traits, we applied a methodology to estimate the proportion of such homozygously recessive characters (HRC-test). This population-genetic study did not only show statistically significant difference of the mean values of genetic homozygosity (BEN - 8.7 +/- 0.3; control - 7.6 +/- 0.3), but of the differences in the type of distribution too, as well as the differences in the presence of certain individual combinations of such traits.

The effect of combined therapy with ritodrine, erythromycin and verapamil on the frequency of micronuclei in peripheral blood lymphocytes of pregnant women.

The main aim of this study was to investigate the genotoxic effect of combined pharmacotherapy applied in post-operative treatment of cervical cerclage in pregnant women over six days. This study included 19 phenotypically healthy pregnant women in mid-trimester with a diagnosis of cervical insufficiency, mean age 28+/-5.33. The frequency of micronuclei (MN) was estimated in peripheral blood lymphocytes of patients before surgical intervention and after the end of applied pharmacotherapy by application of cytokinesis block micronucleus (CBMN) test. Mean value of baseline MN frequency was 6.84+/-2.91 MN/1000 binucleated cells, and after the end of the applied therapy, 10.32+/-4.27 MN/1000 binucleated cells (P<0.001) were found. The data of cell proliferation index showed that the combined therapy did not induce significant difference in cell kinetics (P>0.05). Our results showed that combined pharmacotherapy treatment over six days significantly increased the frequency of MN in peripheral blood lymphocytes of pregnant women.

Population-genetic study of Balkan endemic nephropathy in Serbia.

The study of Balkan endemic nephropathy (BEN) in the affected localities of southern Serbia shows population-genetic difference between samples of BEN affected individuals and control group consisting of non-affected individuals from the same localities. Detailed population-genetic study in village Chepure, which includes 20 large families where BEN is present in 646 (from first to fourth degree) relatives of probands, shows a familial character of disease as well as significant genetic influences in expression of the illness. Our study of genetic homozygosity degree includes an analysis of the presence, distribution and individual combination of 20 to 30 selected genetically controlled morphophysiological traits in the sample of BEN patients and in the control-healthy group. Assuming that BEN is genetically controlled disease, we made a hypothesis that an increased homozygosity level, as well as the changed variability among the patients, could be populationgenetic parameter for the prediction of the illness. Taking into consideration our experience, as well as the experience of numerous scientists who studied the nature of the inheritance of mono-and oligo-genically controlled qualitative traits, we applied a methodology to estimate the proportion of such homozygously recessive characters (HRC-test). This population-genetic study did not only show statistically significant difference of the mean values of genetic homozygosity (BEN: 8.7 ± 0.3; control: 7.6 ± 0.3), but of the differences in the type of distribution too, as well as the differences in the presence of certain individual combinations of such traits.

Genetic variability in the group of patients with congenital hip dislocation.

Our study of genetic homozygosity degree includes an analysis of the presence, distribution and individual combination of 20 selected genetically controlled morpho-physiological traits in the group of patients (N = 93) with congenital hip dislocation (CDH) and in control sample consisting of school children from Belgrade (N = 200). Assuming that CDH is genetically controlled disease, we made a hypothesis that an increased homozygosity level, as well as the changed variability among the patients, could be population-genetic parameter for the prediction of the illness. Taking into consideration our experience, as well as the experience of numerous scientists who studied the nature of the inheritance of mono- and oligo-genically controlled qualitative traits, we applied a methodology to estimate the proportion of such homozygously recessive characters (HRC-TEST). This population-genetic study did not only show statistically significant difference of the middle values of genetic homozygosity (CDH-7.1+/-0.2; control - 5.2+/-0.1), but of the differences in the type of distribution too, as well as the differences in the presence of certain individual combinations of such traits. The described methodology can be used in further analyses, with hope that it can be applied as an early prognosis for decreased resistance to different diseases. The frequencies of ABO blood types in the sample of CDH patients were similar to the average value of Serbian population, while the percentage of blood group A is slightly increased. Comparing frequencies of Rh blood groups, there is no difference between tested samples.

[Micronuclei and ageing in a sample of Yugoslavian population]. / Microiadra i starenie. Issledovanie na vyborke zhitelei Iugoslavii.

OBJECTIVE: Instability in the organization and expression of the genetic material has been hypothesized as the basic mechanism of ageing. Aim of this study was to quantify the effect of ageing on spontaneous micronuclei (MN) frequency in peripheral blood lymphocytes. METHOD: Analysis of Yugoslavian population sample (164 tested individuals, age 0-62 years) has performed by application of cytokinesis-block technique (CB). RESULTS: There was an increase of MN frequency with age, from newborns to 40-year-old persons. The total average of MN frequency per 1000 analyzed binuclear cells amounts to 8.03 +/- 0.42, with variation from 0 to 26 MNs. In a sample of 29 newborns the recorded average MN frequency was 6.91 +/- 0.81, while among 69 persons 25-39 years old, the MN frequency was 9.16 +/- 1.00. The lowest average MN frequency, however, was noted in the sample of 34 tested individuals 40 to 62 years of age. CONCLUSION: An increase with age in MN frequency has been observed in samples of studied individuals from newborns to 40-year-old persons. A decrease of MN frequency in older groups could be explained by a gradual decrease of proliferative cell capacities.

Variability of haptoglobin and alpha 2-macroglobulin in Albino Oxford and Dark Augustin inbred strains of rats before and after trauma.

Increased synthesis of acute-phase proteins (APP) in the liver represents the most prominent aspect of acute phase reaction (APR). Haptoglobin (Hp) and alpha 2-macroglobulin (alpha 2-M) are among the most important APP in rats. In this study AO (Albino Oxford) and DA (Dark Augustin) inbred strains of rats, with different resistance to trauma, were used to compare concentrations of Hp, alpha 2-M, IL-1, IL-6 and TNF, before and after tourniquet trauma. Cytokines IL-1, IL-6 and TNF have an important role in APP gene expression. Our results showed that before trauma, more resistant AO rats had higher concentrations of Hp and alpha 2-M compared to DA rats. The influence of genetic elements of these differences is significant. Tourniquet trauma caused an early increase of IL-1, IL-6 and TNF activities in the serum which preceded the increase in the Hp and alpha 2-M concentrations but differences between strains were not found.

Genetic load and coadaptation of chromosomal inversions. II. O-chromosomes in Drosophila subobscura populations.

We have analysed the inversion polymorphism and genetic load of O-chromosomes in three populations of D. subobscura from southeastern Europe. As expected for a central populations the inversion polymorphism was extensive. In a like fashion, the genetic load, in particular the frequency of lethals, was heavy in all three populations. There were significant differences in the frequency of moderately deleterious genes. These differences in viability can be attributed to balancing selection. A comparison of these two kinds of genetic polymorphism indicates that there are differences in mean viability among different gene arrangements of O-chromosomes in the three populations. The differences observed are due to an unequal distribution of various viability classes among O-chromosome gene arrangements. We here show for the first time a specific distribution of lethal genes among these arrangements within the Palearctic distribution area of D. subobscura. The lethal allelism test showed lethals are non-randomly associated with the Ost gene arrangement. The amount of genetic load is heavy in gene arrangements with a high frequency, in comparison with the ones with a low frequency. Lethal genes may be protected in combinations of low and moderate frequency gene arrangements that harbor more lethal genes, as the Ost in the one population. Some arrangements that are less protected against recombination have a higher load than ones that are more protected against recombination. This can be taken as evidence for coadaptation.

Analysis of background-dependent genetic interactions without inbred strains.

Experimental analysis of background dependent effects of genetic interactions can be designed using strains generated by introgression of small genetic regions containing identical genotypes at loci in question into different inbred strains. We use a novel multilocus paradigm, denoted conditional intergenic functional association (CIFA), to simulate this procedure, with the trade-off of power for convenience that is affordable when sufficiently strong effects are present. We analyze nine enzyme loci at three chromosomes in groups of D. melanogaster with different developmental rates that showed similar allelic frequencies at individual loci. Results obtained suggest the presence of adaptive interaction between particular alleles at two loci when genetic variation at seven background loci is eliminated. Biochemical considerations show that, in the resulting developmental context, strong interaction between these genes may emerge from shifted control of the pentose phosphate pathway, with cascading effects on the glycolysis, TCA cycle, and biosynthetic pathways: one gene may assume control of the irreversible rate-limiting step in the pentose phosphate pathway, whereas the other gene may assume control of the NADP+ level that regulates the same rate-limiting step as an electron acceptor. The newly developing functional genomics research and the absence of inbreeding make CIFA directly applicable to complex human traits in large samples.

The Vogt-Koyanagi-Harada syndrome: association with autoimmune polyglandular syndrome type 1.

We describe a young woman with primary adrenal insufficiency, hypoparathyroidism (autoimmune polyglandular syndrome type 1), Graves disease, vitiligo, and alopecia universalis. Five years after the diagnosis, she presented with recurrent ophthalmological and neurological disorders as features of Vogt-Koyanagi-Harada syndrome. A marked therapeutic response was noted on systemic high-dose corticosteroid treatment. To the best of our knowledge, such a spectre of autoimmune diseases has not been reported previously.

[Soto's syndrome (cerebral gigantism)]. / Sotosov sindrom (cerebralni gigantizam).

In 1964, Sotos and co-workers defined a syndrome of advanced height and bone maturation, dating from birth, mental deficiency and unusual craniofacial appearance with acromegaloid features. About 200 cases have subsequently been reported, more often in males. Occurrence is sporadic, but inheritance may be dominant autosomal. The aetiology is unknown. A two- and a half-year-old boy with Sotos's syndrome is described. He was the first-born child of non-sanguineous healthy parents. The family history revealed no congenital abnormalities and no mental retardation. The proband was born at 36 weeks of gestation with a weight of 4100 g (> P97), length of 53 cm (> P90) and occipital-frontal circumference of 36 cm. His postnatal sornatic linear growth was excessive taking a curve above the 97th percentile. Skeletal maturation and bone age were accelerated. Premature eruption of teeth was observed. Clinical features included macroencephaly, dolichocephaly with a prominent forehead, down-slanting palpebral fissures, hypertelorism, high-arched palate and large hands and feet. His mental deficiency was mild (IQ = 60) with delay of early psychomotor development and expressive language. His karyotype was normal. The CT brain scans revealed mild ventricular dilatation and some cortical atrophy. Electroencephalographic records showed localised theta activity. Endocrinological and metabolic studies failed to show andy definite abnormality.

Adaptive significance of amylase polymorphism in Drosophila. X. Analysis of alpha-amylase activity of two amylase variants in individual Drosophila subobscura flies.

Experiments were designed to estimate the variation of alpha-amylase activity in Drosophila subobscura individuals homozygous for AmyS and AmyF alleles at the Amy locus. The measurements of enzyme activities in six groups of male progenies determined in each individual have shown that degree of variation differs between S and F strains. Variability of amylase activity among male progenies of S strain was substantially greater, which also had significantly higher specific amylase activity than in F strain. The analysis of amylase activity variance showed that this variance among the males is 40 times, and among the females 2.7 times smaller in F than in S strain.

The side-effects of clozapine: a four year follow-up study.

Clozapine, as the model agent for the atypical antipsychotic drugs, is currently recommended as effective regarding negative symptoms of schizophrenia and treatment-resistant schizophrenic patients. This study focuses on the clozapine-induced side-effects in 100 hospitalized schizophrenic patients (negative and therapy-resistant forms), followed-up for a four year period. Clinically relevant side-effects occurred in 73% of all patients. Tachycardia (67%), the increase of liver enzymes (36%), hypotension (29%) and sedation (27%) were most frequent. Tachycardia, hypotension and sedation disappeared during the initial phase of treatment (i.e. 4-6 weeks), as tolerance developed with continuation of therapy. The increase of liver enzymes appeared to be dose related, since the reduction of daily clozapine dose led to the normalization of transaminases values. The other side-effects (constipation, nausea and vertigo) were rare and transient. Leucopenia was not registered in any patient during the follow-up period. Therefore, clozapine is efficient and, with some precautions concerning hepatotoxicity, is safe for in- and outpatient long-term treatment in appropriately selected patients.