Modulation of lymphocyte subsets and humoral immune response by florfenicol administered to sheep red blood cell-immunized broiler chickens.

Florfenicol is a broad-spectrum bacteriostatic antibiotic commonly used for the treatment of systemic infections in farm animals. The aim of this study was to determine the effect of florfenicol on the percentage of T lymphocytes (CD3+, CD4+, CD8+, TCRgd+ cells) and B lymphocytes (Bu-1+ cells) and on total serum anti - sheep red blood cell (SRBC) haemagglutinin titer in the peripheral blood of SRBC-immunized broiler chickens. The study included three groups of broiler chickens differentiated by weight (0.5, 1.2, 2.4 kg). Florfenicol was administered orally at a dose of 30 mg/kg. The drug was administered eight times at 24 h intervals. The chickens were immunized with SRBC 24 h after administration of the third dose of florfenicol. Florfenicol increased the percentage of CD3+ blood lymphocytes with a corresponding decrease in the percentage of B lymphocytes in birds weighing 0.5 and 2.4 kg. Florfenicol reduced the production of total anti SRBC-haemagglutinins on day 5 after antigen injection in all three body weight groups of the broiler chickens. In conclusion, florfenicol exerted a modulating effect on the immune response of the birds and this should be taken into consideration when using this antibiotic for certain indications.

Immunotherapies based on PD-1/PD-L1 pathway inhibitors in ovarian cancer treatment.

Immunotherapies based on anti-programmed death 1/programmed death ligand 1 (PD-1/PD-L1) pathway inhibitors may turn out effective in ovarian cancer (OC) treatment. They can be used in combination with standard therapy and are especially promising in recurrent and platinum-resistant OC. There is growing evidence that the mechanism of the PD-1/PD-L1 pathway can be specific for a particular histological cancer type. Interestingly, the data have shown that the PD-1/PD-L1 pathway blockade may be effective, especially in the endometrioid type of OC. It is important to identify the cause of anti-tumor immune response suppression and exclude its other mechanisms in OC patients. It is also necessary to conduct subsequent studies to confirm in which OC cases the treatment is effective and how to select patients and combine drugs to improve patient survival.

High-dose therapy with hematopoietic stem cell rescue in patients with poor prognosis Ewing family tumors.

The purpose of the study was to evaluate the feasibility and safety of two cycles of high-dose chemotherapy (HDT) followed by autologous hematopoietic SCT (HSCT) in patients with poor prognosis Ewing family of tumors (EFT). Twenty patients with primary metastatic bulky disease or recurrent EFT were enrolled to a treatment protocol with two cycles of HDT and HSCT. Patients tolerated well the first (n=20) and second (n=13) cycles, with limited and predictable toxicities. Only one (5%) TRM occurred during the second cycle. Myeloid engraftment occurred at the median of 11 days after both cycles. At 3 years, the overall and EFS were 45% (confidence interval; CI 0.22, 0.69) and 47% (CI 0.25, 0.70), respectively, for the entire group and 58% (CI 0.30, 0.86) for patients who completed two cycles. Dose intensification with two cycles of HDT and HSCT is feasible and safe, with low and acceptable treatment-related morbidity and mortality. Adding a second course of therapy does not impair engraftment. However, only 65% of the patients were able to proceed to the second cycle. Further studies are required to define the optimal mode of delivery of HDT and HSCT in treatment of advanced EFT.

Results of IDA-FLAG programme in the treatment of recurrent acute myeloblastic leukaemia--preliminary report.

BACKGROUND: Unsatisfactory treatment results of acute myeloblastic leukaemia inspire the search for new drugs, characterised by higher efficiency and lower toxicity. The aim of the study was the assessment of the efficiency and side effects associated with the implementation of IDA-FLAG protocol. MATERIAL AND METHODS: The assessment of treatment results and undesirable effects was based on the material of 4 children with the relapse of acute myeloblastic leukaemia, after a total of 6 IDA-FLAG protocols. RESULTS: Complete remission was obtained in 2 (50%) children, with mean remission time of 13.5 months. One of the children underwent the transplantation of haematopoietic cells from unrelated donor. The main side effect observed was bone marrow aplasia. Leucopenia, granulocytopenia, thrombocytopenia and anaemia persisted for approx. 20 days. Apart from haematological symptoms, the following were also observed: the symptoms of intolerance after ARA-C, aspergillosis of paranasal sinuses and lungs and severe coagulation disorders. CONCLUSION: IDA-FLAG protocol may be recommended for use in children with AML relapse and the undesirable effects observed are acceptable.

In vitro tumor-pulsed or in vivo Flt3 ligand-generated dendritic cells provide protection against acute myelogenous leukemia in nontransplanted or syngeneic bone marrow-transplanted mice.

To determine whether immune stimulation could reduce acute myelogenous leukemia (AML) lethality, dendritic cells (DCs) were pulsed with AML antigens and used as vaccines or generated in vivo by Flt3 ligand (Flt3L), a potent stimulator of DC and natural killer (NK) cell generation. Mice were then challenged with AML cells. The total number of splenic anti-AML cytotoxic T-lymphocyte precursors (CTLPs) present at the time of challenge was increased 1.9-fold and 16.4-fold by Flt3L or DC tumor vaccines, respectively. As compared with the 0% survival of controls, 63% or more of recipients of pulsed DCs or Flt3L survived long term. Mice given AML cells prior to DC vaccines or Flt3L had only a slight survival advantage versus non-treated controls. NK cells or NK cells and T cells were found to be involved in the antitumor responses of Flt3L or DCs, respectively. DC vaccines lead to long-term memory responses but Flt3L does not. Syngeneic bone marrow transplantation (BMT) recipients were analyzed beginning 2 months post-BMT. In contrast to the uniform lethality in BMT controls given AML cells, recipients of either Flt3L or DC vaccines had a significant increase in survival. The total number of splenic anti-AML CTLPs at the time of AML challenge in BMT controls was 40% of concurrently analyzed non-BMT controls. Flt3L or DC vaccines increased the total anti-AML CTLPs 1.4-fold and 6.8-fold, respectively. Neither approach was successful when initiated after AML challenge. It was concluded that DC vaccines and Flt3L administration can enhance an AML response in non-transplanted or syngeneic BMT mice but only when initiated prior to AML progression.

TNFRs and IL-6R transmembrane receptors expression and release of their soluble forms by neutrophils and mononuclear cells from cancer patients.

TNF-alpha and IL-6 are multipotential mediators involved in the control of many host's reactions to tumour. Their biological effects are mediated through the membrane-bound receptors (TNFRp55, TNFRp75 and IL-16R respectively) which can exist in soluble forms. In the present study we compared release of soluble sTNFRp55, sTNFRp75 and sIL-6R with expression of their membrane-bound on PMN and PBMC. Cells were isolated from patients with cancer diseases with a different location and histological classification. We have found that alterations of membrane-bound TNFRp75 expression and in the secretion of soluble TNFRp75 form, as opposed to other receptors examined, are characteristic features of neutrophils and mononuclear cells isolated from cancer patients. The similar changes observed in the expression of TNFRp75 by PMN and PBMC appear to confirm a significant role of PMN in the tumour response mediated by TNF-alpha. Furthermore, the altered membrane-bound TNFRp75 expression and sTNFRp75 secretion appear to depend on the tumour type.

[Flow cytometry: diagnostic, therapeutic and prognostic usefulness in solid tumors]. / Cytometria przeplywowa--przydatnosc diagnostyczna, lecznicza i rokownicza w litych guzach.

Flow cytometry is a new technique measuring several optical parameters of particular cells in cell suspension. These parameters are: optical density, light scatter and fluorescence emission. Optical assay conjugated with digital data processing permit the precise characterisation of chosen cellular subpopulations. The optical signs transformed by photomultipliers as analogue signals are presented as descriptive cytograms. Digitally converted signals analysed by multichannel analyser are stored as histigrams. Flow cytometry is widely used in the diagnostics of blood neoplastic diseases and, in a less routine manner, in the diagnostics of solid tumours. The latter requires accurate disaggregation of tumour tissue to obtain homogenous cell suspension. Different, more or less efficient, mechanical or enzymatic disaggregation techniques are used to prepare appropriate cell suspensions. Flow cytometry DNA analysis with simultaneous determination of several protein proliferation factors are heavily used in research and clinical laboratories to follow tumour cell proliferation, efficacy of cancer therapy and finally as prognostic factors. Although these assays yield promising results in some specific tumours, their value and accuracy seems still equivocal.

[The evaluation of lymphocyte subpopulation by flow cytometry in healthy children]. / Ocena subpopulacji limfocytów oznaczanych metoda cytometrii przeplywowej u zdrowych dzieci.

Seeing that amount of experiences concerning the cytometry of lymphocyte subpopulation in particular age ranges is very low we considered that undertaking this subject is useful. By the method of flow cytometry 49 children at the age from 2 months to 15 years were studied. According to the results of our studies we found, starting from the third year of life decrease of mean percentage values of cells CD19+, progressive in relation to the age, increase of mean percentage values of cells CD2+, increase of percentage values of cells CD8+ and cells CD56++ and the decrease of mean values of the indicator Th/Ts. Remaining CD3, CD4 did not show any statistically significant differences in investigated age ranges. Determined by the method of flow cytometry the percentage and complete values of lymphocyte subpopulations in peripheral blood in investigated age ranges can make the reference values in the assessment of immunological state.

Relationship of plasma pharmacokinetics of high-dose oral busulfan to the outcome of allogeneic bone marrow transplantation in children with thalassemia.

We analyzed plasma pharmacokinetics of busulfan in 64 children and young adults (age 2.8-26; median 11 years) with homozygous beta-thalassemia transplanted with bone marrow from HLA-identical sibling donors. A uniform conditioning regimen was employed, using busulfan 14 or 16 mg/kg in 12 divided doses, and cyclophosphamide 120 or 200 mg/kg. Three sets of parameters were examined in this homogenous patient population: (1) factors that affect the plasma kinetics of busulfan, such as age and pre-transplant liver status defined by liver function tests, ferritin levels and liver biopsy; (2) busulfan-related toxicity: occurrence of veno-occlusive disease, seizures and idiopathic interstitial pneumonitis; and (3) the relationship between busulfan exposure and transplant outcome: engraftment delay or rejection, aplasia, occurrence of mixed chimeras and mortality. Kinetic analysis of first and 10th dose (using area under the curve (AUC), maximum and minimum concentration) as comparable, showing no sign of accumulation or decline in busulfan plasma levels over time. Age and liver status did not influence busulfan metabolism. No relationship was found between busulfan exposure and toxicities or transplant outcome. We conclude that busulfan monitoring is not predictive in children and young adults with homozygous beta-thalassemia receiving busulfan and high-dose cyclophosphamide along with histocompatable sibling donor marrow.

Unrelated donor bone marrow transplantation for acute leukemia in patients with Down's syndrome.

The incidence of hematologic disorders in patients with Down's syndrome (DS) is significantly increased, and includes neonatal transient abnormal myelopoiesis and acute leukemias. Treatment of children with DS and leukemia has been controversial because of toxicity and associated congenital cardiac and other abnormalities. The role of BMT, particularly from an unrelated donor (URD), remains undefined in this population. We report two children with DS and acute leukemia successfully treated with intensive chemotherapy and matched URD bone marrow transplantation. One child was transplanted in third remission of ALL and has been disease free for 8 months. A second child with AML was transplanted in second remission and is disease free 15 months post-BMT.

[Anomalies of the cervical spine in children]. / Anomalie w zakresie kregoslupa szyjnego u dzieci.

Two children are presented with congenital anomalies of the cervical spine and associated malformations of the nervous and urinary systems. The attention is paid to the usefulness of the cervical spine assessment in children in cases of recurrent cerebellar manifestations of unclear aetiology or combined malformations of the urinary system.

TAR syndrome with orofacial clefting.

A case of TAR syndrome with bilateral cleft lip and palate is presented. Bilateral symmetric focomelia, normal thumbs among five fingers of hands, synostosis of IVth and Vth metacarpal bones and some defects of lower limbs with associated thrombocytopenia were noted. Dysmorphic facial features included hypertelorism, epicanthus, blue sclerae, broad nasal root, micrognathia, low-set ears, sparse blond hair. To our knowledge this patient represents an unusual association of TAR syndrome with orofacial clefting. A common background of TAR and Roberts/SC syndrome is suggested.

[The course of acute HCV infection in patients with chronic non-A, non-B hepatitis]. / Przebieg ostrego zakazenia HCV u chorych na przewlekle zapalenie watroby NANB.

Among 52 patients with chronic non-A, non-B hepatitis, observed for many years in the Department of Infectious Diseases of Pomeranian Medical Academy, retrospectively diagnosed towards HCV infection, 45 proved to be anti-HCV positive. Sera stored in the bank of sera were examined using 2nd generation tests: ABBOTT HCV EIA (Abbott), ORTHO RIBA (Ortho Diagnostics) and UBI HCV EIA (Organon), showing 85% of positivity. Mostly HCV infection was connected with the blood transfusion. The course of acute phase of HCV infection was mild, short lasting, with no or sporadic extrahepatic symptoms; the activity of aminotransferases and the bilirubin level were of average value. The only characteristic feature of the acute HCV infection was fluctuating aminotransferase activity, which can be the good sign of progression.

[TAR syndrome (congenital thrombocytopenia and aplasia of the radial bones) in a 5-months-old boy]. / Zespól TAR (wrodzonej trombocytopenii i braku kosci promieniowych) u 5-miesiecznego chlopca.

A boy aged 5 months had the TAR syndrome (thrombocytopenia and aplasia of radial bones) of autosomal recessive inheritance. Bilateral reduction of the length of the upper extremities included both forearm bones and humerus, so that the shoulder girdle was connected directly to the hand, with all fingers and thumb normally shaped. These skeletal anomalies were associated with thrombocytopenia. Various other abnormalities were present, with the most controversial anomaly being bilateral cleft of the lip and secondary palate. It may be an accidental coexistence of abnormalities or may suggest a common mechanism of the development of the TAR syndrome, Roberts syndrome and SC-phocomelia belonging to hereditary reduction of the extremities with identical mode of inheritance.

[A case od rare location of Ewing's sarcoma in a 6-year-old boy]. / Przypadek rzadkiej lokalizacji miesaka Ewinga u 6-letniego chlopca.

A 6 boy is described treated for two months for pneumonia. In view of absent improvement the diagnosis was verified and malignant tumour was recognized. Radiological examination and clinical observation showed presence of a tumour in the 4th rib with pulmonary metastases. Histological examination of a biopsy specimen from the rib demonstrated primary microcellular Ewing's sarcoma. Of importance are: the age of the patient, rarely described location and diagnostic difficulties in the initial stage of the disease.

[A particular role of free fatty acids in the fetus and newborn and during pregnancy and labor]. / Szczególna rola wolnych kwasów tluszczowych u plodu i noworodka oraz w okresie ciazy i porodu.

Free fatty acids account for a small fraction of plasma lipids. The author describes their significant role in the energy-yielding processes in the organism, and their participation in many important metabolic functions in the tissues during intrauterine life, neonatal period, pregnancy and labour.

Microheterogeneity forms of alpha 1-acid glycoprotein as indicators of rheumatoid arthritis activity.

The microheterogeneity of alpha 1-acid glycoprotein (AGP) has been studied in the sera of 48 patients with rheumatoid arthritis and of 12 healthy individuals. For each rheumatoid patient the disease activity has been assessed and each patient has been assigned to one of four activity grades: I, inactive; II, mildly active; III, moderately active; and IV, severe. Concanavalin A-affinity electrophoresis revealed three microheterogeneity variants of AGP: non-reactive with Con A, weakly reactive with Con A and strongly reactive with Con A. The relative amounts of AGP-variants observed in the healthy donors were similar to those observed in the patients with activity grade I, but differed significantly from patients with grades II, III and IV. The differences between the AGP-patterns of patients with activity grades II, III and IV were also statistically significant.

Concentration of free fatty acids (FFA) in amniotic fluid and maternal and cord serum in cases of intrauterine growth retardation.

There were no efficacious methods of diagnosing the syndrome of intrauterine growth retardation (IUGR) before birth. Only about 50% of cases with this syndrome can be recognized by clinical examinations. The aim of our investigations was an attempt to determine free fatty acid level (FFA) in amniotic fluid as a biochemical test in prenatal diagnosis. The investigations were carried out in 49 pregnant women and their newborns in whom IUGR was diagnosed clinically. The control group consisted of 30 pregnants who had an uneventful pregnancy and normal labor. Samples of amniotic fluid were taken in the first stage of parturition (dilatation of cervical os +/- 4-5 cm) by vaginal amniocentesis or amniotomy during cesarean section. The concentration of FFA in samples of amniotic fluid and serum were determined by Ducombe's method. It was found that during pregnancy complicated by IUGR, the free fatty acid concentration in amniotic fluid was almost three times higher than in normal pregnancy (0.355 mmol/l and 0.125 mmol/l) respectively. These differences are statistically significant (t = 12.58 p less than 0.001). Thus, determination of FFA can be used as a biochemical test in prenatal diagnosis of IUGR.