Selection of susceptible and refractory lines of Glossina morsitans centralis for Trypanosoma congolense infection and their susceptibility to different pathogenic Trypanosoma species.

In a single generation of selection, two lines of Glossina morsitans centralis were established that differed significantly in susceptibility to Trypanosoma congolense clone IL 1180. Reciprocal crosses demonstrated that susceptibility was a maternally inherited trait. Differences between the lines, to all phases of the trypanosome infection, were maintained for eight generations, whereas differences in susceptibility to midgut infections were maintained for twenty-eight generations. Thereafter, the lines did not differ in susceptibility to Trypanosoma congolense IL 1180. Susceptibility to infections with Trypanosoma congolense IL 1180 was only a weak predictor of susceptibility to T. congolense clones IL 13-E3 and K60/1, as well as clone T. brucei brucei STIB 247-L. However, the susceptible and refractory lines displayed these phenotypes when tested with Trypanosoma vivax, indicating that the factors that affect susceptibility to trypanosomes are expressed both within and outside the midgut.

LIS-lnterlink-connecting laboratory information systems to remote primary health-care centres via the Internet.

A pilot study was performed to evaluate the feasibility of using the Internet to securely deliver patient laboratory results, and the system has subsequently gone into routine use in Poland. The system went from design to pilot and then to live implementation within a four-month period, resulting in the LIS-Interlink software product. Test results are retrieved at regular intervals from the BioLink(TM) LIS (Laboratory Information System), encrypted and transferred to a secure area on the Web server. The primary health-care centres dial into the Internet using a local-cell service provided by Polish Telecom (TP), obtain a TCP/IP address using the TP DHCP server, and perform HTTP 'get' and 'post' operations to obtain the files by secure handshaking. The data are then automatically inserted into a local SQL database (with optional printing of incoming reports)for cumulative reporting and searching functions. The local database is fully multi-user and can be accessed from different clinics within the centres by a variety of networking protocols.

Sensitive and specific detection of Trypanosoma vivax using the polymerase chain reaction.

The nucleic acid probes that are currently in use detect and distinguish Trypanosoma vivax parasites according to their geographic origin. To eliminate the need for using multiple DNA probes, a study was conducted to evaluate the suitability of a tandemly reiterated sequence which encodes a T. vivax diagnostic antigen as a single probe for detection of this parasite. The antigen is recognized by monoclonal antibody Tv27 currently employed in antigen detection ELISA (Ag-ELISA). A genomic clone which contained a tetramer of the 832-bp cDNA sequence was isolated and shown to be more sensitive than the monomer. Oligonucleotide primers were designed based on the nucleotide sequence of the 832-bp cDNA insert and used in amplifying DNA sequences from the blood of cattle infected with T. vivax isolates from West Africa, Kenya, and South America. The polymerase chain reaction (PCR) product of approximately 400 bp was obtained by amplification of DNA from all the isolates studied. The oligonucleotide primers also amplified DNA sequences in T. vivax-infected tsetse flies. Subsequently, PCR was evaluated for its capacity to detect T. vivax DNA in the blood of three animals experimentally infected with the parasite. T. vivax DNA was detectable in the blood of infected animals as early as 5 days post-infection. Blood and serum samples from the three cattle and from six other infected animals were also examined for the presence of trypanosomes and T. vivax-specific diagnostic antigen. Trypanosomes appeared in the blood 7-12 days post-challenge, while the antigenemia was evident on Days 5-20 of infection. Analysis of the data obtained in the three animals during the course of infection revealed that the buffy coat technique, Ag-ELISA, and PCR revealed infection in 42, 55, and 75% of the blood samples, respectively. PCR amplification of genomic DNA of T. vivax is thus superior to the Ag-ELISA in the detection of T. vivax. More importantly, both the T. vivax diagnostic antigen and the gene encoding it are detectable in all the T. vivax isolates examined from diverse areas of Africa and South America.

Recombinant human interleukin-3 in refractory severe aplastic anaemia: a phase I/II trial.

In a prospective open-labelled phase I/II trial we tested efficacy and tolerability of recombinant human interleukin-3 (rhIL-3) alone in patients with refractory severe aplastic anaemia (SAA). 15 patients with idiopathic (12 patients) or secondary (one post-hepatitic, one drug induced, one dyskeratosis congenita) SAA, refractory or relapsing after one to three courses of antilymphocyte globulin were included. 14 patients were transfusion dependent (RBC 14, platelet 12). RhIL-3 was planned for three patients each at five escalating dose levels of 1, 2, 4, 8 and 16 micrograms/kg, given daily as 24 h continuous infusion for 21 d. RhIL-3 was prematurely withdrawn at days 10 and 11 for adverse events in two patients. 9/15 patients showed an increase in WBC; 2/6 at the 1-2 micrograms/kg and 7/9 at the 4-16 micrograms/kg level, but no sustained effects were seen. No patient showed a response in platelet counts. Additionally, platelet and RBC transfusion requirements were unchanged pre and post study. All patients experienced one or more adverse event, mainly fever (15 patients), bleeding (nine patients), and headache (six patients). Occurrence of adverse events was dose related and the maximum tolerated dose was reached with 8 micrograms/kg. Five patients suffered serious adverse events. RhIL-3 as single growth factor and used alone is of minimal benefit in severe aplastic anaemia.

[Reference values for peripheral blood morphology in countryside population of northern Poland]. / Wartosci referencyjne dla parametrów morfologii krwi obwodowej w populacji wiejskiej Polski pólnocnej.

Reference values for hematological parameters were determined in countryside population living in the area surrounding Zarnowieckie Lake. Randomly selected population of 1065 individuals were divided into 11 groups: children from 0 to 3 years, 4 to 7 years and 8 to 11 years; and males and females from 12 to 20 years, 21, to 40 years, 41 to 55 years and older than 56 years. In the paper are presented reference values estimated on Technicon H1 analyser for 8 red cells parameters, 3 platelets parameters and absolute values for white blood cell and differential counts. Red cell parameters values were changing with age and sexes, and were similar to determined by the others. In examined population white blood cell counts were higher then in earlier reports. Moreover in males over 40 WBC were higher than in females. Neutrophil counts were parallel to white blood cells. Lymphocyte and eosinophil counts were decreasing with age, and were not depending on sex. Platelet counts were decreasing with age, with concommitant MPV increase.

A study on the maturation of procyclic Trypanosoma brucei brucei in Glossina morsitans centralis and G. brevipalpis.

Teneral Glossina morsitans centralis and G. brevipalpis were fed in vitro upon medium containing procyclic Trypanosoma brucei brucei derived from the midguts of G. m. centralis or G. brevipalpis which had immature trypanosome infections. The tsetse were then maintained on rabbits and, on day 31, were dissected to determine the infection rates. In G. m. centralis the midgut and salivary gland infection rates by T. b. brucei were 46.0% and 27.0% with procyclic trypanosomes from G. m. centralis, and 45.4% and 24.7% with procyclic trypanosomes from G. brevipalpis, respectively. In G. brevipalpis the rates were 20.2% and 0.0% with procyclic trypanosomes from G. m. centralis, and 28.0% and 0.0% with procyclic trypanosomes from G. brevipalpis, respectively. Teneral G. m. centralis and G. brevipalpis were also fed similarly upon procyclic T. b. brucei derived from G. m. centralis or G. brevipalpis on day 31 of infection, the former tsetse species had mature infections while the latter were without infections in the salivary glands. In G. m. centralis the infection rates in the midgut and salivary glands were 48.9% and 17.0%, and 38.0% and 17.0% when fed on procyclic trypanosomes from G. m. centralis and G. brevipalpis, respectively. In G. brevipalpis the rates were 21.5% and 0.0%, and 10.7% and 0.0% with procyclic trypanosomes of G. m. centralis and G. brevipalpis origin, respectively. Thus, procyclic T. b. brucei from susceptible G. m. centralis could not complete cyclical development in refractory G. brevipalpis, whereas those from G. brevipalpis developed to metatrypanosomes in the salivary glands of G. m. centralis.(ABSTRACT TRUNCATED AT 250 WORDS)

[Essential thrombocythemia--clinical course from personal material]. / Nadplytkowosc samoistna--analiza przebiegu klinicznego w materiale wlasnym.

The article presents clinical course analysis of essential thrombocythemia in 17 patients aged 29-82. The diagnostic criteria were the same as described by Polycythemia Vera Study Group. Mean platelet level of diagnosis was 1680 x 10(9)/l. Haemorrhagic complications were observed in 42% of the patients, while thrombotic ones or embolisms in 35%. In two cases both types of complications occurred. Asymptomatic course of the disease was observed in 5 patients. The statistical analysis proved that the patients with platelet count between 900-1900 x 10(9)/l are in danger of developing thrombotic episodes and thus antiaggregation treatment should be considered. If platelet level exceeds 1900 x 10(9)/l the risk of haemorrhage increases, so antiaggregation treatment is contraindicated and thrombocytapheresis is advised instead. The patients was started on treatment when platelet count was above 1000 x 10(9)/l in asymptomatic cases or with lower platelet level in symptomatic ones. The treatment consisted of busulphan, hydroxyurea or interferon alpha (in one of the patients) until lowering platelet level below 600 x 10(9)/l.

[Platelet defects in chronic myeloproliferative disorders]. / Zaburzenia plytkowe w przewleklych zespolach mieloproliferacyjnych.

Platelet abnormalities are common in patients with chronic myeloproliferative disorders. In this study we report abnormalities in platelets morphology and function in 45 patients with chronic myeloproliferative disorders: 15 with chronic myelogenous leukaemia (CML), 8 with polycythemia rubra vera (PRV), 20 with essential thrombocythemia, and 2 with myelofibrosis (ME). We investigated flow cytometric features of platelets as measured with Technicon H1 technology, VIZ, mean platelet volume (MPV), plateletocrit), platelet distribution width (PDW), and modal platelet volume (PLT Mode) Platelet aggregation in response to ADP, epinephrine and collagen was used as functional test. In patients with ET, PRV and MF we found a significant decrease in platelet volume (both MPV and PLT MODE). Decrease in platelet aggregation and secretion in response to ADP, epinephrine and collagen was the most frequent abnormality in platelets function and was observed in most of patients with thrombocythemia in chronic myeloproliferative disorders.

Flowcytometric pattern of leucocyte recovery after therapy induced aplasia.

33 aplastic episodes in 24 patients undergoing bone marrow ablative therapy for the treatment of haematological malignancies were monitored with automated flowcytometric counts with particular emphasis to BLAST, MONO (monocytes) and LUC (large unstained cells) determined by Technicon H1, and MFR and HFR reticulocytes determined by Sysmex R1000. We found regular pattern of appearance of flowcytometric parameters, which were predictive for leucocyte recovery. A transient increase of BLAST to > 10% and MONO+LUC to > 0.08 x 10(9)/l were predictive for leukocyte recovery after aplastic episodes following ABMT, BMT and high dose AraC therapy, but not after conventional chemotherapy. The prognostic value of MFR+HFR reticulocytes increase over 5% was limited to ABMT and BMT. This report demonstrates the capabilities of automated flowcytometric analysis in early prediction of haematological recovery.

A comparison of African buffalo, N'Dama and Boran cattle as reservoirs of Trypanosoma vivax for different Glossina species.

Teneral Glossina morsitans centralis were fed on the flanks of African buffalo, N'Dama or Boran cattle infected with Trypanosoma vivax IL 2337. The infected tsetse were maintained on goats and on day 25 after the infected feed, the surviving tsetse were dissected to determine the infection rates. The mean mature infection rates (% +/- S.E.) in the tsetse fed on buffalo, N'Dama and Boran cattle were 34.3 +/- 9.9, 33.7 +/- 13.4 and 58.9 +/- 7.1, respectively. Logistic regression analysis indicated that infection rates in the labrum and hypopharynx of the tsetse were significantly lower when fed on the infected buffalo or N'Dama than Boran cattle. Similarly, the risk of infection was significantly lower in male than female tsetse. When teneral G. m. centralis, G. pallidipes, G. p. gambiensis, G. brevipalpis and G. longipennis were fed simultaneously on either the buffalo cow, the N'Dama bull or the Boran steer infected with T. vivax IL 2337, the mature infection rates were higher in the two morsitans group than the two fusca group tsetse, whilst G. p. gambiensis was relatively refractory to the infection, irrespective of the host species on which they fed. Logistic regression analysis indicated that the infection rates in the labrum and hypopharynx were significantly different amongst the five tsetse species for each of the three infected host animals. Nevertheless, the trypanotolerant African buffalo and N'Dama cattle may serve as reservoirs of T. vivax infection as can trypanosusceptible Boran cattle.

Vector competence of Glossina pallidipes and G. morsitans centralis for Trypanosoma vivax, T. congolense and T. b. brucei.

Vector competence of Glossina pallidipes for pathogenic Trypanosoma species was compared to that of G. morsitans centralis. Cattle or goats were the hosts used to infect teneral tsetse, rabbits were used to maintain tsetse which were dissected on day 30. Mean infection rates of G. pallidipes and G. m. centralis by T. vivax isolated from a cow in Kenya were respectively 39.5 +/- 8.9% and 32.1 +/- 10.3% whilst for T. vivax isolated from a cow in Nigeria, they were 30.0 +/- 7.5% and 19.8 +/- 4.3%. Differences were not significant. Differences in infection rates between the sexes of flies were also not significant. Transmission capability to goats by either tsetse species was good for the two T. vivax isolates. Mean infection rates by T. congolense isolated from a lion in Tanzania were significantly lower in G. pallidipes (8.5 +/- 1.8%) than in G. m. centralis (22.5 +/- 2.0%). Males of either tsetse were more susceptible than females. Transmission rates to goats and mice by both tsetse species was 100%. G. pallidipes (3.5%) was less susceptible than G. m. centralis (25.1%) to T. congolense isolated from a cow in Nigeria, but transmission rates to goats and mice by either tsetse was 100%. Also, G. pallidipes (2.7 +/- 0.4%) was significantly less susceptible than G. m. centralis (18.4 +/- 1.1%) to T. b. brucei isolated from a hartebeest in Tanzania. Males of either tsetse species were more susceptible than females. Transmission rates to goats and mice by either tsetse was 100%. G. pallidipes (0%) was not susceptible to T. b. brucei isolated from a pig in Nigeria whilst G. m. centralis showed infection rate of 9.3%. When male G. pallidipes and G. m. centralis were fed every day for 27 days on a goat infected with this T. b. brucei from Nigeria, the infection rates were 8.7% and 20.2%, respectively. Transmission rates to mice by either tsetse species was 100%. In conclusion, G. pallidipes has a vector competence equal to that of G. m. centralis for T. vivax, whilst G. pallidipes has lower vector competence than G. m. centralis for T. congolense and T. b. brucei.

A comparison of African buffalo, N'Dama and Boran cattle as reservoirs of Trypanosoma congolense for different Glossina species.

Teneral Glossina morsitans centralis Machado were fed on the flanks of the African buffalo (Syncerus caffer Sparrman), N'Dama (Bos taurus L.) or Boran (Bos indicus L.) cattle infected with Trypanosoma congolense Broden. The infected tsetse were maintained on rabbits and on day 30 after the infected feed, the surviving tsetse were dissected to determine the infection rates. The mean infection rates (% +/- SE) in the midgut of tsetse fed on buffalo, N'Damas and Borans were 23.5 +/- 3.3, 31.6 +/- 2.7 and 33.7 +/- 4.6, respectively. The differences were not significant. However, the mean mature infection rate in tsetse fed on the buffalo (13.2 +/- 2.1%) was significantly lower compared to the rates in tsetse fed on the N'Dama (20.4 +/- 1.4) or the Boran cattle (21.4 +/- 1.1). When groups of teneral G.m.centralis, G.pallidipes Austen, G.p.gambiensis Vanderplank, G.f.fuscipes Newstead, G.brevipalpis Newstead and G.longipennis Corti were fed simultaneously on either an infected buffalo, an N'Dama or a Boran steer, the mature infection rates ranged from 0 to 16.1%. Irrespective of the host species used, the T.congolense infection rate was highest in G.m.centralis, lowest in the palpalis and fusca group tsetse, with G.pallidipes being intermediate. Nevertheless, the trypanoresistant African buffalo and N'Dama may serve as reservoirs of T.congolense as can trypanosusceptible Boran cattle.

Hematologic values of New Zealand white rabbits determined by automated flow cytometry.

Hematologic values of peripheral blood from normal adult New Zealand White rabbits were determined by five different automated flow cytometers in use in a routine clinical hematology laboratory: Technicon H1, Coulter Counter CC540, Coulter Counter VCS, Sysmex NE8000 and Sysmex R1000. The software designed for human blood analysis was used in all instances without adaptation. The total numbers of white blood cells, red blood cells, reticulocytes and platelets were measured with high precision and accuracy. Except for hemoglobin content, concordance was excellent for all measured and calculated values among the different automated flow cytometers. Determining the white blood cell differential count was more complex. Eosinophils and lymphocytes were quantified reliably by all the automated flow cytometers used. However, the results were rejected by Technicon H1 and Sysmex NE8000 in 50% of the cases. Rabbit basophils were recognized with accuracy by Technicon H1 only. The proportion of polymorphonuclear versus mononuclear white cells was identical when measured with Technicon H1 and Coulter Counter VCS. These results show that the new generation of automated flow cytometers designed for human blood can be used with some limitations for animal studies. They allow the standardization of normal values and comparison of results among or between laboratories. They also introduce new parameters, the value of which is as yet undefined.

[Subcutaneous administration of recombinant human erythropoietin (R-EPO) in the treatment of anemia in predialysis patients with chronic renal failure]. / Stosowanie podskórne rekombinowanej ludzkiej erytropoetyny (r-Epo) w terapii niedokrwistosci u chorych z przewlekla niewydolnoscia nerek leczonych zachowawczo.

10 anemic (HB less than 9.0 g/dl) predialysis patients with chronic renal failure were treated for three months with s.c. administration of r-Epo. Blood morphological parameters were estimated using hematological autoanalyser Technicon H1. An increase of the mean hemoglobin (Hb) level from 8.39 to 10.57 g/dl was observed. In 8 patients Hb concentration after 3 months therapy ranged from 9.4 to 12.7 g/dl, but in the remaining two of them Hb was lower than 9.0 g/dl. Appearance of a high percentage of hypochromic erythrocytes is probably the most characteristic response to r-Epo treatment. This phenomenon was caused by iron deficiency. A significant increase of serum creatinine and BUN levels were observed in treated patients, without the concomitant decrease of endogenous creatinine clearance. No clinical symptoms suggesting deterioration of the renal function were observed. Subcutaneous therapy with r-Epo appeared an effective and convenient method of treatment of anemia in predialysis patients.

[Evaluation of erythropoiesis under the influence of recombinant human erythropoietin (R-EPO) in dialyzed patients]. / Ocena odnowy ukladu czerwonokrwinkowego pod wplywem rekombinowanej erytropoetyny (r-Epo) u pacjentów dializowanych.

5 deeply anemic (Hb less than 8 g/dl, Ht less than 25%) dialyzed patients with chronic renal failure were treated during four months with r-Epo. Blood cells morphological parameters were estimated using hematological autoanalyser Technicon H1. Satisfactory increase of the Hb levels and RBC counts were observed in 4 patients, in one the improvement was insignificant. We observed three types of response to r-Epo treatment: 1) macrocytic type, 2) hypochromic type, and 3) non-hypochromic type, without lasting macrocytosis. Our results suggest that type of erythropoiesis depends on other active biological substances (iron, folic acid, vit. B12) necessary for correcting erythropoiesis. r-Epo administration appeared to be a safe and effective method of anaemia treatment in dialyzed patients. Its administration eliminated blood transfusion for six months.

ATP citrate lyase in cholinergic nerve endings.

The activity of ATP-citrate lyase in homogenates of five selected rat brain regions varied from 2.93 to 6.90 nmol/min/mg of protein in the following order: cerebellum less than hippocampus less than parietal cortex less than striatum less than medulla oblongata and that of the choline acetyltransferase from 0.15 to 2.08 nmol/min/mg of protein in cerebellum less than parietal cortex less than hippocampus = medulla oblongata less than striatum. No substantial differences were found in regional activities of lactate dehydrogenase, pyruvate dehydrogenase, citrate synthase or acetyl-CoA synthase. High values of relative specific activities for both choline acetyltransferase and ATP-citrate lyase were found in synaptosomal and synaptoplasmic fractions from regions with a high content of cholinergic nerve endings. There are significant correlations between these two enzyme activities in general cytocol (S3), synaptosomal (B) and synaptoplasmic (Bs) fractions from the different regions (r = 0.92-0.99). These data indicate that activity of ATP-citrate lyase in cholinergic neurons is several times higher than that present in glial and noncholinergic neuronal cells.

The contribution of citrate to the synthesis of acetyl units in synaptosomes of developing rat brain.

The activities of pyruvate dehydrogenase, citrate synthase, and choline acetyltransferase in rat brain synaptosomes increased during ontogenesis by 3 and 14 times, respectively. Activity of ATP-citrate lyase decreased by 26% during the same period. Pyruvate consumption by synaptosomes from 1-day-old animals was 40% lower than that found in older rats; however, citrate efflux from intrasynaptosomal mitochondria in immature synaptosomes was over twice as high as that in mature ones. The rates on production of synaptoplasmic acetyl-CoA by ATP-citrate lyase were 1.03, 1.40, and 0.49 nmol/min/mg protein in 1-, 10-day-old, and adult rats, respectively. 3-Bromopyruvate (0.5 mM) inhibited pyruvate consumption by 70% and caused a complete block of citrate utilization by citrate lyase in every age group. Parameters of citrate metabolism in cerebellar synaptosomes were the same as those in cerebral ones. These data indicate that production of acetyl-CoA from citrate in synaptoplasm may be regulated either by adaptative, age-dependent changes in permeability and carrier capacity of the mitochondrial membrane or by the inhibition of synthesis of intramitochondrial acetyl-CoA. ATP-citrate lyase activity is not a rate-limiting factor in this process. Metabolic fluxes of pyruvate to cytoplasmic citrate and acetyl-CoA are presumably the same in both cholinergic and noncholinergic nerve endings. The significance of citrate release from intrasynaptosomal mitochondria as a regulatory step in acetylcholine synthesis is discussed.