Haematopoiesis in snakes (Ophidia) in early postnatal development.

The occurrence of haematopoiesis has been studied in various parts of the spine and in the ribs in four species of snakes (Boa constrictor L., Elaphe guttata L., Lamprophis fulaginosus Boie., Bothrops jararaca Wied.) from hatching until 150 days of postnatal development. Marrow spaces are formed by chondrolysis with various time frames depending on the studied species. Marrow cells egress to the general circulation in two ways: via migration through the endothelial cells lining the venous sinuses or by the rupture of protrusions. Erythroblasts are present in the lumen of marrow sinuses suggesting their final maturation there. Various relationships of the spleen to the pancreas have been found. No myelopoietic foci occur in the spleen, liver or kidney of any of the studied species. However, erythropoiesis (sparse islets) has been observed in Bothrops jararaca spleen.

Haematopoiesis and a new mechanism for the release of mature blood cells from the bone marrow into the circulation in snakes (Ophidia).

This is the first description of haematopoiesis in snakes. Studies were carried out on the following species belonging to Ophidia: Bothrops jararaca, Bothrops jararacusu, Waglerophis merremii, Elaphe taeniura taeniura, Boa constrictor,and Python reticulatus. Smears of the peripheral blood and histological preparations from the vertebrae, ribs, liver, and spleen were studied under a light and electron microscope. Myeloid cells were present in the following locations in the vertebrae: the neural spine, zygoapophysial processes, floor of the neural canal, lacunae in the bodies of vertebrae and also inside the ribs. Although the vascular system was well developed, especially around the ribs, vessels inside the marrow cavities were scarce, both in the ribs and elsewhere where haematopoiesis was found. Venous sinuses were well developed in the vertebrae and in the rib regions from their costal head towards the middle area. They consisted of one layer of fine endothelial cells. Mature cells in the process of migration into the general circulation were only sporadically encountered when venous sinuses were studied on perfusion-fixed specimens. In contrast, almost every sinus venosus contained protrusions directed towards the lumen, filled mostly with mature and immature blood cells. Various stages of their formation were seen in the cross sections of venous sinuses ranging from small, newly formed to large, elongated ones, filled with many fully developed and some maturing blood cells. In many cases the apices of the protrusions were ruptured, and mature blood cells, as well as a few immature ones, were seen in their vicinity. This observation led us to a new hypothesis that blood cells are released from the extravascular space into the lumen of venous sinuses. In snakes, these cells are released into the systemic circulation mainly via the rupture of protrusions filled with mature blood cells and, to a lesser degree, by transcytosis as known in mammals. In the spleens from young specimens, 1-2 foci of haematopoiesis were encountered where lymphopoiesis predominated. Haematopoiesis was not detected in the liver.

Hematopoiesis in snakes (Ophidia).

Locations of the hematopoietic tissue have been described in the following ophidian species: Bothrops jararaca, Bothrops jararacusu, Waglerophis merremii, Elaphe teniura teniura, Boa constrictor, and Python reticulatus. Studies were carried out on perfusion fixed vertebrae, ribs, spleen, liver, thymus, and kidney. Routine histological technique was applied using both light and electron microscopy. Hematopoietic tissue was found in the following locations of the vertebrae: neural spine, neural arch, postzygophysis processes, hypapophysis, vertebral centre. Moreover, intense hematopoiesis was found inside the ribs. In the spleen and thymus, only lymphopoiesis was found. Hematopoietic islets in the spleen were sporadically found only in young specimens. No hematopoiesis was observed in the liver and kidney. In the studied species, there were no differences in the location of hematopoietic tissue. A new model of mature and immature blood cell release to the lumen of marrow sinuses different from that known to operate in higher vertebrates is proposed.

[The efficacy of non-pharmacological intervention in obese patients with newly diagnosed diabetes mellitus type II]. / Skutecznosc interwencji niefarmakologicznej u chorych z nowo rozpoznana cukrzyca typu 2 i towarzyszaca otyloscia.

In Poland like in other countries we observe an increasing number of diabetes mellitus cases with about half of the patients in whom the disease remains undiagnosed. Therefore it seems necessary to improve early diagnoses and prevention of the disease. The aim of the study was to assess the efficiency of a 3-month non-pharmacological intervention based on diet and increased physical activity in patients with newly diagnosed diabetes type 2 (diagnosis based on oral glucose tolerance test (OGTT), WHO 1999). We investigated 37 newly diagnosed diabetic type 2 patients, 16 men (aged 52.4 +/- 5.4) and 21 women (aged 51.0 +/- 5.7). Anthropometric and biochemical measurements were performed before and after intervention. Two-kilometres Walking Test with an intermediate estimation of VO2max and fitness index (FI) was performed before and after intervention. Total abdominal fat volume (measured from diaphragm to pubis): visceral fat volume (VFV) and subcutaneous fat volume (SFV) (mm3) were assessed according to the standard protocol of NMR abdominal examination. Patients completed 12 weeks of supervised intervention focused on weight reduction, increase of physical activity, changes of nutritional habits. Students t-test, Mann-Whitney test and Spearman's correlation were used for statistical analysis. In women the average weight reduction was 4.7 kg (5.8% of initial body weight), whereas in men 5.9 kg (5.9% of initial body weight). In women VO2 max increased from 23.7 +/- 6.4 to 24.9 +/- 4.8 (ns), and fitness index increased from 78.7 +/- 11.7 to 83 +/- 14.7 (ns). In men VO2 max increased from 22.5 +/- 6.7 to 26.6 +/- 8.6 (ns) and fitness index increased from 55.1 +/- 12.5 to 64.8 +/- 13.7 (p < 0.05). In women the level of fasting glycaemia decreased from 6.47 +/- 1.2 to 4.84 mmol/l +/- 0.6 (p < 0.01) and the level of glycaemia at 120 minutes of OGTT decreased from 13.2 +/- 2.5 to 6.76 +/- 2.7 mmol/l (p < 0.01). The decrease of plasma glucose was accompanied by the decrease of fasting insulin from 19.2 +/- 15.5 to 8.53 +/- 93.2 uj/ml (p < 0.01) and in 120 minutes of OGTT from 148.8 +/- 86.2 to 58.4 +/- 41.0 uj/ml (p < 0.01). In men the level of fasting glycaemia decreased from 8.63 +/- 2.0 to 7.07 mmol/l +/- 2.4 (p < 0.05) and the level of glycaemia at 120 minutes of OGTT decreased from 15.76 +/- 3.2 do 9.3 +/- 5.7 mmol/l (p < 0.01). The decrease of plasma glucose was accompanied by the decrease of fasting insulin from 21.99 +/- 12.6 to 10.1 (3.8 uj/ml (p < 0.05) and at 120 minutes of DGTT from 81.5 +/- 52.7 do 41.6 +/- 21.0 uj/ml (p < 0.05). After the intervention 45% of the patients (57% of women and 31% of men) were non-diabetic (correct OGTT). In men visceral fat volume (VFV) was greater than in women (7642.6 +/- 1774.6 and 4789.9 +/- 1242.0 mm3 respectively (p < 0.01). Subcutaneous fat volume (SFV) was smaller in men than in women (7116.5 +/- 2048.5), in men and 10533.9 +/- 3478.3 respectively (p < 0.01). In women and men a strong (p < 0.01) correlation between waist circumference and visceral fat volume (VFV) (r = 0.573 (p < 0.01) and r = 0.833 (p < 0.01) respectively) and subcutaneous fat volume (SFV) (r = 0.900 (p < 0.01) and r = 0.790 (p < 0.01) respectively) was found. The results of the study confirm that in newly diagnosed diabetic type 2 patients body weight reduction and increased physical activity result in the improvement of biochemical indices. In about one half of patients the early phase of the disease might be reversible due to weight reduction and increased physical activity. The non-pharmacological intervention should be the first intervention undertaken in newly diagnosed diabetic type 2 patients.

The influence of low-power laser energy on red blood cell metabolism and deformability.

The aim of this study was to test the hypothesis that exposure of red blood cells (RBC) to low-power laser energy can modulate their metabolism and deformability. The effects of exposure to a He-Ne (lambda=632.8 nm), GaAlAs (lambda=780 nm) and GaAs (lambda=904 nm) lasers have been examined. Red cells diluted to a hematocrit of 45% were incubated in a humidified atmosphere of 95% air and 5% CO(2) at 37 degrees C, and exposed to three different laser beams held 5 cm from the target area to yield a spot surface area of 2 cm(2). Three red cell suspensions belonging to the experimental groups were treated with each laser beam by 5, 15 and 30 minutes, respectively. The temperature was constant during the exposure's time. Three control suspensions of RBC were kept for the same time as sham-irradiated groups. The erythrocyte elongation index (EEI) was evaluated using a Rheodyn SSD (Myrene, Roetgen, Germany). The enzyme activities of glucose-6-phosphate dehydrogenase (G-6-PD), glutathione peroxidase (GSH-Px) and glutathione reductase (GR) were assayed in each sample spectrophotometrically. Adenosine triphosphate (ATP) and diphosphoglicerate (2,3-DPG) levels were also assessed. No statistical differences were observed in the erythrocyte elongation index at shear stresses of 0.30, 0.60, 1.20, 3.00, 6.00, 12.00, 30.00 and 60.00 Pa after being irradiated for 5 and 15 minutes as compared to not irradiated ones. At 30.00 and 60.00 Pa a decrease (p<0.03 and p<0.05, respectively) in EEI has been observed after 30 min exposure to all three wavelengths of laser light when compared to the control. The antioxidant enzyme activities showed no significant changes following 5, 15 and 30 min of irradiation by all three laser wavelengths laser tested. Similarly, erythrocyte organic phosphate levels (ATP and 2,3-DPG) showed no significant changes following treatment by laser radiation. This study revealed that the low-power laser at 632.8, 780 and 904 nm wavelengths have little biological effects on red blood cells in vitro.

Rheological properties of red blood cells (including reticulocytes) in patients with chronic renal disease.

Hemodialysis is a method of treatment of patients suffering from terminal renal disease and consists in removing uremic toxins from blood. The influence of hemodialysis on rheological properties of red blood cells was ascertained. We studied the deformability of RBC after density gradient separation of whole blood drawn from patients with renal disease before and after dialysis. The improvement in deformability of cells from the top layers was observed in postdialysis samples.

Disparity between dietary iron intake and iron status of children aged 10-12 years.

Iron status was assessed in a representative sample of 188 adolescents living in a medium-sized city in Poland. Dietary intakes were evaluated using records of diet over a period of seven consecutive days. Subjects were considered to be iron deficient when two or more of the following parameters were abnormal: serum ferritin, transferrin saturation or mean corpuscular haemoglobin concentration. Based on this definition, the prevalence of iron deficiency in the investigated sample of children aged from ten to twelve years was 12.7%. Iron deficiency anaemia was defined using the following criteria: haemoglobin values less than 12.0 g. dl (-1) in girls or less than 12.2 g. dl(-1) in boys, combined with an iron deficiency. With such a definition, the prevalence of iron deficiency anaemia in all subjects was 6.3%. Four boys (3.9%) and six girls (6.8%) were diagnosed as anaemic. The values for Hb in the anaemic boys ranged from 10.9 to 12.2 g. dl (-1) and in anaemic girls from 8.7 to 12.0 g. (-1). It was found that the majority of the individuals studied had a dietary haem-iron intake lower than that recommended. No relationship was found between the level of serum ferritin and total iron and vitamin C dietary intake, but there was positive correlation between serum ferritin and intake of haem iron. A seven-day dietary history questionnaire correctly identified children at risk of iron deficiency anaemia.

The 2,3-DPG levels of human red blood cells during an incremental exercise test: relationship to the blood acid-base balance.

The aim of this study was to evaluate the influence of exercise with the intensity progressively increasing from rest until maximal oxygen uptake (VO2max) on 2,3-DPG levels in red blood cells (RBC) in relation to the changes in the acid-base balance and plasma lactate concentration. Six healthy young men (age 22.5+/-1.5 years, VO2max 3.48+/-0.20 l/min) participated in this study. The subjects performed an incremental exercise test on a cycloergometer until exhaustion. Blood samples were tested for acid-base balance indices (pH, HCO3-, BE), plasma lactate and RBC 2,3-DPG concentration. Gas exchange variables were measured continuously breath-by-breath. In this paper we present data concerning 2,3-DPG, plasma lactate, pH, HCO3- and BE measured at rest, at the power output corresponding to the lactate threshold (PO LT), at the power output at maximal oxygen uptake (PO VO2max), as well as 5, 15 and 30 min after finishing the incremental test. Increase of power output above the lactate threshold to the PO VO2max was accompanied by a significant (p<0.01) increase of plasma lactate from 2.58+/-0.78 mmol/l to 10.22+/-3.04 mmol/l. This was also accompanied by a significant drop (p<0.01) in blood pH value from 7.352+/-0.025 at the PO LT to 7.294+/-0.041 at the PO VO2max. No significant changes of the RBC 2,3-DPG level were observed at any of the analysed stages of the exercise. The RBC 2,3-DPG level expressed in relation to the changes of haematocrit showed only minor changes during the exercise period and after 15 min of recovery vs. resting value (3.21+/-1.19). However, after 30 min of recovery, RBC 2,3-DPG decreased to the value of 2.32+/-1.19 micromol/ml. We conclude that, during an incremental test, no increase in RBC 2,3-DPG concentration is required to reach the maximal oxygen uptake level. Moreover, a rapid decrease in blood pH, developing during a single bout of exercise, is not a stimulus powerful enough to cause significant changes in the RBC 2,3-DPG level during short-term exercise.

Relationship among reduced level of stored iron and dietary iron in trained women.

The aim of the study was to assess the relationship between dietary iron intake (both haeme- and non-haeme-iron) and its status in trained female subjects. Forty female athletes and forty non-trained women of the same age participated in this study. Blood samples were taken to assess haematological (red blood cell count-Er, haemoglobin concentration-Hb, packed cell volume-PCV) and iron related parameters (serum ferritin concentration-SF, serum iron concentration-SI, total iron binding capacity-TIBC). A self questionnaire was used to record food intake for seven days and diets were analyzed on the basis of mean daily nutrient intake, energy values, iron intake and sources of dietary iron. According to established clinical criteria for iron deficiency some athletes and control subjects shown iron depletion (20% and 10%, respectively), iron deficiency erythropoiesis (10% and 7.5%, respectively) and iron-deficient anaemia (10% and 7.5%, respectively). There was no difference in the mean total dietary iron intake between the two female groups, while the mean intake of haeme-iron was significantly lower in the control group. The findings in the present study are: (1) the significant relation exists between serum ferritin level and quantity of haeme-iron intake (but not with total iron intake), and (2) 10% of female athletes have iron deficiency.

The relationship between iron status and perceived exertion in trained and untrained women.

The subjects for this study were 24 female, well-trained distance runners and 41 sedentary, female university students. On the basis of haematological measurements, the subjects were divided into four groups: a group with iron deficiency, non-anemic (normal haemoglobin and red cell count but serum ferritin less than 20 micrograms degrees 1-1) runners (IDR, n = 11), a group of runners with normal iron status (NR, n = 13), a group of sedentary women with normal iron status (NC, n = 26) and a group of iron deficient, non-anemic sedentary subjects (IDC, n = 15). Maximal oxygen uptake (VO2max) and lactate thresholds were determined during incremental bicycle ergometer exercise testing and on separate occasions the ratings of perceived exertion (RPE) were assessed at 25, 50, 75 and 90% VO2max using Borg's scale. There were no differences in the relationship between RPE and work load groups of runners or sedentary subjects with normal and deficient iron status. Iron deficiency did not affect VO2max and lactate thresholds. However, low body iron stores (serum ferritin level below 20 micrograms degrees 1-1) without overt anemia was associated with increased venous blood lactate concentration after maximal exercise load. This study demonstrated that physical work capacity and RPE were not affected by iron deficiency without anemia either in trained or untrained women.

Haematological and iron-related parameters of male endurance and strength trained athletes.

To obtain more information on the effects of long-lasting endurance and strength training on the constituents of the blood, several haematological and iron-related parameters were measured at rest in 39 male athletes from the Polish team who participated in the Olympics in Seoul in 1988. The athletes were divided into two groups: endurance-trained subjects (group E, cyclists, canoeists and rowers; n = 22) and strength-trained subjects (group S, wrestlers and judo; n = 17). The control group was composed of untrained male subjects (n = 48). Blood samples were taken from an antecubital vein with the subject at rest for determinations of haemoglobin concentration ([Hb]), packed cell volume (PCV), erythrocyte (RBC) and reticulocyte count, plasma free haemoglobin concentration, haptoglobin concentration, serum iron, transferrin concentration and ferritin concentrations ([Ferr]); red blood cells were used for estimation of glutamato-oxalate transaminase (GOT) activity and free erythrocyte protoporphyrin concentration ([FEP]). The mean [Hb], PVC, RBC measured in the E athletes were significantly lower than in the control group but were comparable to those obtained in the S atheletes. There were no significantly differences in the haematological indices [mean corpuscular volume (MCV), mean copuscular haemoglobin and mean corpuscular haemoglobin concentration] between the groups of atheletes and the control group. A significant increase in reticulocytosis and GOT activity was observed in the endurance-trained athletes. No impairment of erythropoiesis was observed as indicated by several sensitive markers of haemoglobin formation (FEP, MCV and inspection of blood smears) in the athletes.(ABSTRACT TRUNCATED AT 250 WORDS)

Blood flow in different regions of bone marrow after short-term exercise.

Total and regional blood flow was studied in femoral bone marrow of rats exposed to a short-term exhaustive exercise. The 10 micron microspheres labeled with 58Co were injected into femoral artery and the subsequent radioactivity yield was measured in 3 regions of the bone marrow (proximal metaphysis, diaphysis and distal metaphysis). Following exercise a rise of blood flow in the metaphyseal regions was accompanied by a fall in the diaphyseal area. The changes did not alter, however, the total blood flow rate in bone marrow which remained constant irrespective of the heavy exercise load.

Influences of physical training on the functional changes of young and old red blood cells.

Eleven male elite endurance-trained athletes and 10 male elite strength-trained athletes were compared to a non-trained group of men, to determine the effect of training on some haematological parameters and some indicators of red cell membrane properties. Erythrocytes were age-fractionated by centrifugation in Percoll gradients. It has been found that in the reticulocytes and young erythrocytes of endurance trained athletes activity of acetylcholinesterase (AChE) and concentration of glutathione (GSH) were higher than in strength-trained athletes and control. The red cell osmotic fragility (RCOF) and glycerol lysis time (GLT) of young cells were similar in all investigated groups. The endurance training indicating chronic adaptation mechanisms in significant changes of red cell metabolism but non membrane properties.

The red blood cell glutathione reductase activity in anaemic rats.

The enzyme activity of glutathione reductase (GR, EC 1.6.4.2) was measured in rat red blood cells separated centrifugally in a density gradient of Percoll. The animals were divided into three groups: Groups IPHH, anaemia was induced by administration of phenylhydrazine hydrochloride; Group IITr, anaemia was induced by intense physical training; and Group C, control group. The activity of active and inactive forms of GR were higher in erythrocytes of animals from groups IPHH and IITr, both in young and old forms of cells, than in red cells of control rats, the degree of decrease in activity of both forms of GR in senescent red cells is similar in different kinds of anaemia.

The relationship of plasma ammonia and lactate concentrations to perceived exertion in trained and untrained women.

The purpose of this study was to investigate the covariance between perceived exertion (recorded using Borg's category-ratio scale CR-10) and the relative oxygen uptake, and lactate and ammonia concentrations in blood from a peripheral vein. Ratings of perceived exertion (RPE) at 25%, 50%, 75% and 90% maximal oxygen uptake and lactate and ammonia concentrations were compared in well-trained women distance runners (n = 22) and untrained women (n = 10). Ammonia concentrations in peripheral venous blood were significantly correlated with RPE (P less than 0.05), both in the trained and untrained women. Differences between the trained and untrained subjects occurred when the ammonia concentration increased to 148 mumol.l-1 in both groups investigated; similarly, the mean RPE correlated significantly with the lactate concentration (P less than 0.05), both in the trained and untrained women and there was a difference in RPE between groups when lactate concentration in the blood had risen to 4.4 mmol.l-1. It would seem that the correlation of blood ammonia and lactate concentrations with RPE during exercise could be a useful indicator of the development of fatigue.

Metabolism of red blood cells after short-term exercise in rats.

The aim of this study was to determine the changes of the basic parameters of the erythrocyte system and the activity of some red blood cell (RBC) enzymes prior to and after a single physical effort leading to exhaustion. The study was carried out on male Wistar rats subjected to running on an electric rotating drum at a speed of 25 m/min. A single exercise caused a decrease in the RBC count, haemoglobin concentration (Hb) and haematocrit (Hct) by 21.9, 16.7 and 16.1%, respectively, and an increase in the reticulocyte count (Ret) by 661.5%. The exercise triggered also changes in the activities of some erythrocytic enzymes: pyruvate kinase (PK) activity increased by 12.4%, glucose-6-phosphate dehydrogenase (G6PD) by 37.8%, glutathione reductase (GR) by 30.8% and acetylcholinesterase (AChE) by 248.7%. These increases in the activities of RBC enzymes can be explained by an increase in the red cells turn-over.

The effect of intrarenal nickel subsulfide injections upon the activity of selected erythrocyte and bone marrow enzymes in rats.

The experiments were carried out on Wistar rats. The animals were divided into five groups: Group 1: control group; Group 2: the animals received a single intrarenal dose of nickel subsulfide (Ni3S2) in saline; Group 3: the animals receiving a single intrarenal dose of saline; Group 4: the animals receiving a single intrarenal dose of Ag2S in saline; Group 5: the animals receiving a single intrarenal dose of activated charcoal in saline. Prior to the procedure peripheral blood indices were determined. The experiments included animals manifesting polycythemia. Samples of blood and bone marrow were collected and the activities of AchE, GR and G6PD were determined. An increase in the activity of the three studied enzymes was detected in Groups no. 2 and 5.

Post-exercise anaemia as a result of exercise overloading of the organism.

The aim of the present study was to determine the relation between the exercise load used in training and the changes in the basic parameters of the erythrocytic system. The experiments were carried out on 33 Wistar rats (females and males). The animals were forced to run at a velocity of 1.47 km/h inside an electrically-driven rotating drum. The experiments were performed for 35 days. In the 1st - (females) and 2nd - (males) group the exercise period was prolonged by 1 min per day on successive days, and by 2 min per day in the 3rd - (females) and 4th - (males) group. Prior to and after the training period the haemoglobin concentration, haematocrit, erythrocyte and reticulocyte counts were determined in the venous blood. Reticulocytes were divided into four age groups and the percentage each particular age group was calculated. All the trained groups manifested a decrease in haemoglobin concentration, hematocrit values and erythrocyte count and a concomitant elevation of the reticulocyte count with a shift towards youngest forms was found. Rats of the 3rd and 4th groups developed post-exercise anaemia.