An immunoblotting procedure for screening glycophorins and band 3 protein in the same blots: Identification of glycophorin and band 3 variant forms

An immunoblotting procedure is described which makes it possible to screen multiple blood samples for the presence of glycophorin and band 3 variant forms with altered electrophoretic mobility. The procedure can be simplified by using whole red blood cell hemolysates instead of membranes for SDS-polyacrylamide gel electrophoresis. The use of hemolysates also has the advantage that antigens sensitive to proteolysis are not degraded in vitro. The same nitrocellulose blots were used for immunoenzymatic detection of glycophorins with a set of anti-glycophorin monoclonal antibodies, and for autoradiographic detection of band 3-derived bands with ¹²⁵I-labeled anti-band 3 monoclonal antibody. The screening of 157 Caucasian blood samples revealed the presence of a slower-migrating form of band 3 in seven cases and variant glycophorin in one case. The variant glycophorin exhibited the features of hybrid glycophorin of B-A type.     

Down-regulation of V1a vasopressin receptors in the cerebellum after myocardial infarction

Vasopressin V1a receptors (V1aR) were found in the cerebellum but their functional role has not been determined. As V1aR are engaged in the central regulation of the cardiovascular system and anxiogenic behavior and their role increases in the heart failure and stress, we decided to find out whether expression of V1aR is altered after myocardial infarction and chronic stressing. RT-PCR and Western blot analysis were performed to determine V1aR mRNA and protein expression in the cerebellum of four groups of rats (control sham-operated, infarcted, chronically stressed and infarcted chronically stressed). The myocardial infarct was produced by left coronary artery ligation, and chronic stressing by exposing the rat for four weeks to different types of mild stressors. The rats were sacrificed four weeks after the myocardial surgery or sham operation. Expressions of V1aR mRNA and protein were significantly lower in the infarcted and infarcted chronically stressed rats than in the sham-operated controls and chronically stressed not infarcted rats. No significant differences were found between the sham-operated controls and chronically stressed rats and between the infarcted rats and infarcted rats exposed to chronic stressing. It is concluded that V1aR mRNA and protein expressions are significantly down-regulated in the rats with the post-infarct heart failure but they are not affected by mild chronic stressing.     

Hypopituitarism and hematological abnormalities mimicking myelodysplastic syndrome. Report of four cases

Insufficiency of the pituitary gland and hematological abnormalities may coexist in the context of two syndromes. In the course of some hematopoietic neoplasms, particularly acute leukemias, the pituitary insufficiency may be caused by destruction of the gland either by direct neoplastic infiltration or occlusion of vessels. Alternatively, thy pituitary dysfunction may be associated with but not caused by hematological abnormalities, usually mild peripheral cytopenias. We present four cases of the latter type (1. M/33, pituitary tumor, hypogonadism, hyperprolactinemia, anemia, mild leukopenia with leukocytosis, 2. F/54, pituitary tumor, hyperprolactinemia, thyreotropic and corticotropic insufficiency, anemia, thrombocytopenia, mild neutropenia, 3. F/27, pituitary tumor, diabetes insipidus, hypogonadism, sideropenic anemia, leukopenia, thrombocytopenia, 4. M/24, primary multihormonal insufficiency of the anterior portion of the pituitary gland, neutropenia, microcytosis). Trephine and aspiration bone marrow biopsies revealed topographic and cytological abnormalities partially resembling these found in myelodysplastic syndromes (MDS). Bone marrow cellularity varied markedly between and within the cases, and in three patients abnormal aplastic areas were found. The percentage of hematopoietic stem cells (CD34+) was low in three cases and normal in one case. Pituitary dysfunction may be associated with hematological abnormalities simulating MDS, but showing different, less aggressive clinical course. The proliferative potential of hematopoietic cells is low, the peripheral blood abnormalities are stable, and no patient developed acute leukemia. Detailed bone marrow examination, including the trephine bone marrow biopsy, is useful in differentiation with true MDS, which was also reported in the literature in the patients with pituitary insufficiency.     

A forearm exercise screening test for mitochondrial myopathy

BACKGROUND: The authors hypothesized that impaired oxygen extraction in mitochondrial myopathy (MM) results in a high oxygen saturation in venous effluent blood from working muscle and that this phenomenon can be used as a diagnostic tool for MM. METHODS: Twelve patients with MM, 10 patients with muscular dystrophy, and 12 healthy subjects were studied. All subjects performed intermittent static handgrip exercise (1/2 Hz) at 40% of maximal voluntary contraction (MVC) for 3 minutes. Cubital venous oxygen saturation and brachial artery flow were measured in the exercised arm. RESULTS: Exercise-induced venous oxygen desaturation was smaller in patients with MM (Delta - 7 +/- 5%) than in subjects with muscular dystrophy (Delta - 38 +/- 2%; p = 0.00001) and healthy subjects (Delta - 43 +/- 2%; p = 0.0000002). MVC and exercise blood flow were similar in patients with MM (18 +/- 3 kg; 436 +/- 65 mL/min) and patients with muscular dystrophy (15 +/- 3 kg; 460 +/- 85 mL/min), but were higher in healthy subjects (32 +/- 4 kg; 630 +/- 58 mL/min; p < 0.03). In seven patients with MM and seven patients with McArdle disease, studied with a slightly different protocol, exercise-induced oxygen desaturation was also impaired in MM (Delta - +/- 5%) compared with McArdle disease (Delta - 26 +/- 3%; p = 0.007). CONCLUSION: Oxygen desaturation in venous blood from exercising muscle is markedly lower in patients with mitochondrial myopathy than in subjects with other muscle diseases and healthy subjects, suggesting that a forearm exercise test can be a diagnostic screening tool for mitochondrial myopathy.     

Iodine prophylaxis—the protective factor against stomach cancer in iodine deficient areas

BACKGROUND: Poland has one of the highest death rates for stomach cancer in Europe. Moderate iodine deficiency and in consequence high goitre prevalence led to the implementation in 1996 of a very efficient mandatory model of iodine prophylaxis, based on household salt iodisation (30 +/- 10 mg KI/1 kg of salt). AIM OF THE STUDY: The aim of the study was evaluation of incidence rate of stomach cancer and its possible relation to increased iodine consumption in the years 1992-2004. METHODS: Iodine supply and effectiveness of iodine prophylaxis were evaluated on the basis of comparative analysis of goitre prevalence and ioduria in schoolchildren. To allow comparison between time periods with varying population age structures, the incidence rates of stomach cancer were standardized for age, using the "world standard population". The direct standardization method has been applied. For each sex, the time-trend of incidence rates was shown in graphs over the years 1991-2004. RESULTS: Evident increase in iodine consumption in this period of time was proved by rise in percentage of schoolchildren (6-8 years old) with ioduria above 100 microg/l from 11.4% in 1992-1993 to 52.9.1% in 2003. It was correlated with the decrease in goitre prevalence from 18.8% to 3.2% respectively. The 24-h thyroid uptake of (131)I in investigated population fell from 45.5% in 1986 to 26.8% in 1998. In Krakow the standardized incidence ratio of stomach cancer for men decreased from 19.1 per 100,000 to 15.7 per 100,000, and for women from 8.3 per 100,000 to 5.9 per 100,000 in the years 1992-2004. A significant decline of average rate of decrease was observed in men and women (2.3% and 4.0% per year respectively). CONCLUSION: Observed association between improved iodine supply and decrease of incidence of stomach cancer could indicate the protective role against stomach cancer of iodine prophylaxis in iodine deficient areas--further studies are necessary.     

Reactions of Erythrocyte Glycoproteins and their Degradation Products with various Anti-I Sera

Abstract. Three fractions of erythrocyte glycoproteins obtained from Sepharose 4-B chromatography were tested for I activity with ten serologically differentiated anti-I sera. The most active was fraction I, eluted at the void volume and containing the lowest amount of alkali-labile oligosaccharide chains. The desialization of glycoproteins increased their activity toward anti-I^S and anti-I^D sera, and did not change or decreased the activity toward anti-I^F sera. The most abundant fraction II (major sialoglycoprotein of erythrocyte membranes) showed no or only a very weak I activity, but I-active glycopeptides were isolated from products of digestion of fraction II with trypsin. The major product of digestion, sialoglycopeptide IIT-2 showed I activity only after alkaline elimination of alkali-labile oligosaccharide chains. The results indicate that I receptors are present in hindered form on apparently I-inactive components of erythrocyte membrane.     

Effect of Modification of Amino Groups of Human Erythrocytes on M, N and NVg Blood Group Specificities

Modification of amino groups on the surface of human erythrocytes by acetic anhydride and trinitrobenzenesulfonic acid caused loss of agglutinability by human and rabbit anti-M and anti-N sera, was without any effect on agglutinability of N red cells by Vicia graminea agglutinin, and made M cells agglutinable by the same agglutinin. Modification of amino groups had no effect on agglutinability of red cells by anti-H and anti-A lectins and only slightly decreased agglutinability by human anti-H, anti-A and anti-I sera. The results suggest that the difference between M and N receptors is not determined by the structure of oligosaccharide chains.     

Changes in chromosome structure, mitotic activity and nuclear DNA content from cells of Allium Test induced by bark water extract of Uncaria tomentosa (Willd.) DC

The influence of water extract of Uncaria tomentosa (Willd.) DC bark on the meristematic cells of the root tips of Allium cepa L., e.g. cells of Allium Test, was investigated. The experiment was carried out in two variants: (1) continuous incubation at different concentrations (2, 4, 8 and 16 mg/ml) of the extract for 3, 6, 12, 24, 48 and 72h; and (2) 24-h incubation in three concentrations of the extract (4, 8 or 16 mg/ml), followed by post-incubation in distilled water for 3, 6, 12, 24 and 48h. During the continuous incubation, the mitotic activity was reduced (2 and 4 mg/ml) or totally inhibited (8 and 16 mg/ml), depending on the concentration of the extract. All the concentrations resulted in gradual reduction of the mitotic activity. In the concentration of 2 mg/ml, the mitotic activity reached its lowest value after 12h (2 mg/ml) and after 24h in 4 mg/ml, followed by spontaneous intensification of divisions during further incubation. Instead, in higher concentrations of the extracts (8 and 16 mg/ml), the mitotic activity was totally inhibited within 24h and did not resume even after 72h. Incubation caused changes in the phase index, mainly as an increase in the number of prophases. After 24h of incubation, in all phases, condensation and contraction of chromosomes were observed. During post-incubation, divisions resumed in all concentrations, reaching even higher values than the control. Cytometric analysis showed that the extract caused inhibition of the cell cycle at the border between gap(2) and beginning of mitosis (G(2)/M).     

Is there a time and place for health education in chain pharmacies? Perspectives of Polish community pharmacists

Pharmaceutical practice has evolved significantly from the provision of drugs to a more patient-centred model. However, the ownership structure of pharmacies may impact the level of cognitive services provided by them. The discrepancy may be observed between pharmacists’ opinions on how involved they should get and their actual involvement in health promotion and disease prevention. Given the growing market share of pharmacy chains in Europe, this study aimed to investigate the attitudes of pharmacists employed in them towards their role as health educators. It applies Ajzen's theory of planned behaviour to examine pharmacists’ perspectives with the use of semi-structured in-depth interviews. A total of 10 semi-structured face-to-face interviews were conducted in May 2017 among Polish pharmacists employed in chains. Obtained results showed that respondents welcomed the possibility of providing health education, but simultaneously they seek physicians’ and patients’ acceptance for undertaking this task. Competency gaps and unfavourable working conditions may also decrease pharmacists’ intention to serve as health educators. Including the role of a health educator to the scope of the pharmacists’ practice requires a precise definition of their competencies and responsibilities together with additional training aimed to fill potential gaps in their qualifications. The pharmacists’ image as a health educator should also be widely communicated to patients and other healthcare professionals.