Biomedical subjects

J Szkudlarek

Publications and source records attributed to J Szkudlarek.

At least 19 recordsLinked to original sources

PubMed2001

Effect of high-dose growth hormone and glutamine on body composition, urine creatinine excretion, fatty acid absorption, and essential fatty acids status in short bowel patients: a randomized, double-blind, crossover, placebo-controlled study.

BACKGROUND: Positive effects of high dose growth hormone and glutamine (GH + GLN) on body composition in short bowel patients have been described. Lack of effects on intestinal absorption found in some studies has been ascribed to concomitant essential fatty acid (EFA) deficiency. This study describes changes in body weight (BW) and composition, 24-h urine creatinine excretion, intestinal fatty acid absorption (total, saturated, unsaturated and EFA), and EFA status in relation to treatment with GH + GLN in 8 short bowel patients. METHODS: A double-blind, crossover study between placebo and growth hormone (mean, 0.12 mg/kg/day) plus oral (mean, 28 g/day) and parenteral glutamine (mean, 5.2 g/day) for 28 days. Body composition was measured by dual-energy absorptiometry (DEXA) scans. Intestinal fatty acid absorption was evaluated in balance studies, and EFAs were measured in plasma phospholipids by gas liquid chromatography. RESULTS: Active treatment did not increase BW, lean body mass (LBM), fat mass (FM) and bone mass significantly compared with placebo treatment, but BW increased 1.03 kg (1.7%, P < 0.05), LBM 2.93 kg (8.7%, P < 0.001) and FM decreased 2.41 kg (10.6%, P < 0.001) in comparison with baseline. Twenty-four-hour urine creatinine excretion did not differ between study periods. No changes in intestinal absorption of fatty acids were seen, and no changes in EFAs measured in plasma phospholipids were observed. Only 1 of 8 patients, who did not receive parenteral lipids, had a Holman index above 0.2, indicative of EFA deficiency. All developed peripheral oedema. CONCLUSIONS: Combined high dose growth hormone and glutamine administered for 4 weeks, did not improve absorption of fatty acids or EFA status in short bowel patients. No changes in BW or composition were seen when comparing treatment to placebo periods. The increase in LBM measured by DEXA scan, comparing treatment and baseline periods, was not accompanied by an increase in the 24-h urinary creatinine excretion and is suspected to be associated with an accumulation in extracellular fluids.

Absorptiometry, Photon↗

PubMed2001

Juvenile hormone binding protein and transferrin from Galleria mellonella share a similar structural motif.

It has been previously suggested that juvenile hormone binding protein(s) (JHBP) belongs to a new class of proteins. In the search for other protein(s) that may contain structural motifs similar to those found in JHBP, hemolymph from Galleria mellonella (Lepidoptera) was chromatographed over a Sephadex G-200 column and resulting fractions were subjected to SDS-PAGE, transferred onto nitrocellulose membrane and scanned with a monoclonal antibody, mAb 104, against hemolymph JHBP. Two proteins yielded a positive reaction with mAb 104, one corresponding to JHBP and the second corresponding to a transferrin, as judged from N-terminal amino acid sequencing staining. Transferrin was purified to about 80% homogeneity using a two-step procedure including Sephadex G-200 gel filtration and HPLC MonoQ column chromatography. Panning of a random peptide display library and analysis with immobilized synthetic peptides were applied for finding a common epitope present in JHBP and the transferrin molecule. The postulated epitope motif recognized by mAb 104 in the JHBP sequence is RDTKAVN, and is localized at position 82-88.

Amino Acid Motifs↗

PubMed2000

Effect of high dose growth hormone with glutamine and no change in diet on intestinal absorption in short bowel patients: a randomised, double blind, crossover, placebo controlled study.

BACKGROUND: High dose growth hormone, glutamine, and a high carbohydrate diet may improve intestinal function in short bowel patients. AIMS: To investigate if growth hormone with glutamine and no change in diet improved intestinal function. PATIENTS AND METHODS: Eight short bowel patients were randomised in a double blind crossover study between placebo and growth hormone (mean 0.12 mg/kg/day) with oral (mean 28 g/day) and parenteral glutamine (mean 5.2 g/day) for 28 days. Balance studies were performed at baseline and five days after placebo and treatment were terminated. Dietary energy, carbohydrate, and fat were maintained as usual. RESULTS: Growth hormone with glutamine did not improve intestinal absorption of energy (baseline, placebo, treatment, mean: 46%, 48%, 46% of oral intake, respectively), carbohydrate (71%, 70%, 71%), fat (20%, 15%, 18%), nitrogen (27%, 18%, 19%), wet weight (37%, 39%, 31%), sodium (-16%, -16%, -36%), potassium (43%, 47%, 33%), calcium (-16%, -16%, -15%) or magnesium (-3%, 4%, 2%) compared with placebo or baseline (p>0.05) five days after treatment was terminated. All patients experienced adverse effects. CONCLUSIONS: Combined high dose growth hormone and glutamine administered for four weeks did not improve intestinal absorption five days after treatment was terminated in short bowel patients on their usual diet.

Adult↗

PubMed2000

[Developmental osteopenia: decrease of bone mineral density or systemic impairment?].

In the quantitative evaluation of bone osteopenia is defined as a decrease of mineral density by more than 1 SD from the established normal values (age, sex, peak bone mass...). The border of osteopenia and osteoporosis is demarcated by -2.5 SD (T-score) in adults, while in children the most proper is considered to be -2.0 SD (Z-Score). The aim of the study was to determine whether developmental osteopenia is accompanied by biochemical abnormalities and what are clinical symptoms concomitant with this condition. The studies include 28 children aged 5-17 years, in whom no chronic disease, especially of locomotor system, was found. The basis for diagnosis was densitometric examination of bone, with DEXA method (densitometer by Lunar), vertebral column (Spine) in the pediatric program or for adults. The most frequent causes for referring to the examination were pain in the spine, limbs or history of multiple bone fractures. In the performed biochemical examinations hypomagnesemia, decreased concentration of 25OHD and PTH in blood serum, increased activity of bone isoenzyme of alkaline phosphatase as well as increased excretion of hydroxyproline in urine, were found in several children. In about 1/3 of the children low body mass, and in some cases also retardation of the bone age was revealed. The results of our studies allow a conclusion, that in children with certain clinical abnormalities from locomotor system osteopenia may take place. This disturbance is concomitant with various deviations in calcium-phosphate metabolism and requires adequate therapy. It may be supposed, that in the majority of children, osteopenia was caused by low dietary calcium intake, together with reduced physical activity and vitamin D deficiency. The observations and conclusions from the study are of important practical significance, because children with osteopenia are the risk group for the appearance of osteoporosis in their future life.

Adolescent↗

PubMed1998

Differential effects of somatostatin and its analog on protein tyrosine kinases activity in the rat pituitary and the murine colonic tumors.

The effects of the native somatostatin-14 (SST-14) and of its analog octreotide (OCT) on the activity of protein tyrosine kinases (PTK) in the normal rat anterior pituitary gland, diethylstilbestrol (DES)-induced rat pituitary tumor and murine colonic cancer Colon 38 were studied in vitro. PTK activity was estimated in tissue homogenates using gamma-[32P]ATP and poly (Glu80, Tyr20) as a substrate. It was found that both SST-14 and OCT suppressed the PTK activity in all examined tissues. The suppressive effect was more pronounced in DES-induced pituitary tumor than in normal anterior pituitary gland, and in the former, OCT was more effective than SST-14. In contrast, SST-14 stronger suppressed PTK activity in colonic cancer than OCT. We hypothesize that SST-14 acts on PTK activity in colonic cancer mainly via SSTR-1 subtype of somatostatin receptors.

Animals↗

PubMed1996

Immunoaffinity purification of juvenile hormone-binding protein from Galleria mellonella hemolymph.

Previously described methods of purification of hemolymph juvenile hormone-binding protein (hJHBP) from Lepidoptera were tedious and required multiple steps. These methods resulted in low protein yield (Kramer et al., 1976; Goodman et al., 1978; Peterson et al., 1982; Park et al., 1993; Ozyhar & Kochman, 1987). In this report a simple method of purification of hJHBP from Galleria mellonella (L.) larvae is described. Monoclonal antibodies against hJHBP were obtained and crosslinked to CNBr-activated Sepharose 4B. The hemolymph of G. mellonella was centrifuged and then chromatographed on Sephadex G-200 gel filtration column. Juvenile-hormone-binding activity containing material from Sephadex G-200 column was subjected to purification on an immunoaffinity column. Bound protein was eluted from anti-hJHBP Sepharose 4B gel by lowering pH to 3.0 with 200 mM citric acid 200 mM Na2HPO4 buffer. This method resulted in 320-fold purification of G. mellonella hJHBP with 56% yield.

Animals↗

PubMed1993

Treatment of severe obesity with a highly selective serotonin re-uptake inhibitor as a supplement to a low calorie diet.

Inhibitors of serotonin (5HT) re-uptake have generally been successful in inducing modest but statistically significant weight reductions in clinical trials. Citalopram is a new, highly selective inhibitor of 5HT re-uptake. It is effective and safe in relieving major depression at doses up to 60 mg daily. In our study, 72 severely obese subjects (BMI > 44 kg/m2) were instructed in a 4500 kJ carbohydrate-rich (50%) diet. The initial two week run-in diet+placebo period was followed by a 12 week double-blind period of diet+citalopram or placebo. There were seven withdrawals during the initial two weeks. Of the remaining 65 patients, 45 were randomized to 60 mg citalopram daily and 20 received placebo. A trend towards a higher frequency of nausea at week 6 was noted in the citalopram group. Weight loss during the initial two weeks was 2.01 kg. In the following 12 weeks, mean weight loss was 3.78 kg in the citalopram group vs. 2.64 kg in the placebo group (P = 0.29), reductions occurring almost entirely during the initial four weeks of the treatment period. We conclude that citalopram is of no clinical value in the treatment of obesity when added to a 4500 kJ diet.

Adolescent↗

PubMed1991

Substance P and its fragments affect Ca2+/calmodulin-dependent synaptosomal membrane protein phosphorylation from rat cerebral cortex.

1. We have used synaptosomal membranes to study the influence of substance P and its fragments and analogues of its C-terminal fragment on Ca2+/calmodulin-dependent synapsin I endogenous phosphorylation. 2. SP1-11, SP1-4, [Tyr8]SP6-11 and [pGlu6, Tyr8]SP6-11 at 10(-3) M greatly inhibited synapsin I phosphorylation. 3. SP6-11 at all investigated concentrations and SP1-11, SP1-4, [Tyr8]SP6-11, [pGlu6, Tyr8]SP6-11 at 10(-4) and 10(-5) M were ineffective. 4. The results indicate that SP1-11 and its N-terminal fragment and analogues of its C-terminal fragment act on the phosphorylation of specific synaptic protein (synapsin I) and therefore may influence the release of neurotransmitters, membrane conductance and potentiation or inhibition of other signalling systems.

Animals↗

PubMed1990

The influence of substance P and its fragments on endogenous phosphorylation of synaptosomal membrane protein (synapsin) from cerebral cortex of rat brain.

1. The effects of substance P and its fragments and analogue of a C-terminal fragment on cyclic AMP-dependent phosphorylation of synapsin I in synaptosomal membranes (SM) from cerebral cortex were investigated. 2. SP(I-II) and SP(1-4) at 10(-3) M caused a marked stimulation of synapsin I phosphorylation. 3. A C-terminal fragment of SP (SP6-11) had no effect on phosphorylation of synapsin 1. 4. Analogue of C-terminal fragment [(Tyr8)SP6-11] at 10(-3) M distinctly inhibits phosphorylation of synapsin I. 5. These data suggest that SPI-II and its C- and N-terminal fragments have a modulator function against the phosphorylation of some rat brain proteins.

Animals↗

PubMed1988

Immunoreactivity of proteolytic degradation products of human prostatic acid phosphatase.

Prostatic acid phosphatase (EC 3.1.3.2) was fragmented by trypsin and papain in the presence of sodium dodecyl sulphate. Trypsin-catalysed cleavage gave a peptide of 33 kDa which was subsequently trimmed to 18 kDa, 15 kDa and 13 kDa peptides. Even the small tryptic fragments reacted with antiphosphatase antibodies from rabbit serum and with monoclonal antibody mAb-14. Papain treatment under these conditions resulted in the release of a 40 kDa peptide which was gradually reduced to a 18 kDa peptide. The monoclonal antibody mAb-14 to the prostatic phosphatase was bound exclusively to the 50 kDa subunit of the phosphatase and to the 40 kDa peptide. The results suggest that the monoclonal antibody mAb-14 binding site represents a "local" sequence rather than a "conformational" one and does not require an extensive tertiary folding of the antigen molecule.

Acid Phosphatase↗

PubMed1988

Characterization of anti-prostatic acid phosphatase monoclonal antibody and its medical significance.

Monoclonal antibody to purified prostatic acid phosphatase from seminal plasma was produced by fusion of spleen cells from immunized mice with the Sp2/O-Ag 14 cell line. This hybridoma-derived antibody, designated MAb-14, was classified as IgG1 immunoglobulin. The apparent affinity constant of phosphatase.MAb-14 complex formation calculated by using the Langmuir isotherm is 2.4 x 10(-8) M. The molecular weight of the complex formed under the condition of antibody excess was found to be 350K, which suggests that 2 molecules of prostatic phosphatase bind to 1 molecule of the antibody. The MAb-14 antibody bound to phosphatase had a negligible effect on the catalytic activity of the enzyme. All isoenzymatic forms of catalytically active prostatic phosphatase, resolved by isoelectric focusing or by chromatofocusing in different pH gradients, reacted with the monoclonal antibody. Several peptides of Mr 25K to 76K and of 13K to 76K were adsorbed from the prostatic tissue extract and from seminal fluid, respectively, on an MAb-14-Sepharose column. The MAb-14 monoclonal antibody was applied to the immunohistochemical investigation of prostatic phosphatase distribution in normal human prostate gland, in nodular hyperplasia, and in adenocarcinoma of the prostate. Immunostaining was observed in prostatic secretory epithelium, within the luminal content of prostatic glands, and in the neighborhood of prostatic cancer cells. Metastatic prostatic carcinoma was also strongly immunoreactive with the antibody. There was no cross-reactivity with leukocytes, kidney, liver, pancreas, spleen, breast, stomach mucosal, and colon tissues.

Acid Phosphatase↗

PubMed1986

Effects in vitro of L-glutamate and kainic acid on the ATPase activities of synaptosomal membranes from different areas of rat brain.

Changes in the activities of enzymes responsible for the active transport of Na+, K+, Ca2+, Mg2+ in synaptosomal membrane (SM) preparations from the cerebral cortex, hippocampus and thalamus with hypothalamus after incubation with L-glutamate (Glu) or kainic acid (KA) were investigated. Glu stimulated Ca,Mg- and Na,K-ATPase activities in cortex but reduced the activities of all the investigated ATPases, except Na,K-ATPase in the hippocampus and thalamus with hypothalamus. KA reduced distinctly the activity of ATPases in the cortex and only slightly in the thalamus with hypothalamus, but stimulated the enzyme activities in the hippocampus. Both, Glu and KA in vitro altered the processes of active transport of cations in SM preparations.

Adenosine Triphosphatases↗

PubMed1986

Detection of prostatic tissue and seminal plasma peptides reacting with human seminal fluid acid phosphatase antibodies.

IgG1 monoclonal antibody to purified seminal fluid phosphatase was raised by fusion of spleen cells from immunized mice with cell line Sp2/O-Ag 14 using simple method of screening for antiphosphatase antibody secreting clones. All molecular forms of catalytically active seminal fluid phosphatase and prostatic tissue phosphatase, resolved by chromatofocusing in pH gradient, react with this monoclonal antibody and with rabbit antiserum to purified seminal fluid phosphatase. Peptides of Mr 25,000 to 76,000 and of Mr 13,000 to 76,000 were adsorbed from the prostatic tissue extract and from seminal plasma on the monoclonal antibody-Sepharose column.

Acid Phosphatase↗

PubMed1986

Quantitative analysis of tissue distribution of Ly10-like murine alloantigen(s) with antisera and monoclonal antibodies.

Tissue distribution of non-Lyt1.1 ("Ly10-like") antigen or antigens encoded by short chromosomal segment differentiating B6-Ly-1a congenic strain from B6 strain of mice was studied by quantitative absorption of (BALB/c X B6)F1 anti B6-Ly-1a antiserum and by direct cytotoxicity of Ly-10-132-12-26 monoclonal antibody on lymphoid cell populations. Identical strain but not tissue distribution pattern does not allow to conclude whether antiserum and monoclonal antibody detect the same or closely linked antigens. Absorption experiments revealed the highest antigen content in the brain tissue, lower in testis and kidney, still lower in lymphoid organs and the lowest in liver and lung. Among lymphoid cells, bone marrow cells had highest absorbing capacity, followed by thymus, spleen and lymph nodes. Monoclonal antibody lysed almost 100% of thymocytes, 30% bone marrow cells and 10-20% of spleen and lymph node cells (both T-cell and B-cell enriched populations contained the same proportion of positive cells). Cortisone resistant thymocytes showed the same sensitivity as cortical thymocytes to Ly-10-132-12-26 antibody which is distinguishable characteristics of medullary thymocytes from peripheral T cells. Mitogen activated lymphocytes exhibited significantly higher expression of Ly10-like antigen than resting peripheral lymphocytes.

Animals↗

PubMed1984

Monoclonal anti-TL antibody recognizing a TL.2-like antigen.

We obtained a monoclonal anti-TL antibody-producing hybridoma by fusion of Sp2/0 hybrid cells with spleen cells from (B6 X A-Tlab) F1 mice immunized with ASL1 leukemia cells. The antibody, TL-22-27-17, reacts in complement-dependent cytotoxicity assay with ASL1 cells as well as with normal thymocytes of TL+ mouse strains (B6-Tlaa, A, BALB/c, DBA/2 and 129) but not with thymocytes from TL- mice (B6--Ly-1a). In addition, it did not react with lymph node cells or Concanavalin A-induced spleen blasts from B6-Tlaa mice; this result shows that TL-22-27-17 recognizes TL and not Qa-1 antigen. The positive reactions with BALB/c (TL.1-2+3-5-6-7+) and 129 (TL.1-2+3-5-6-7-) thymocytes suggest that antibody TL-22-27-17 reacts with TL.2 or TL.2-like antigenic determinant.

Animals↗

PubMed1984

Cell surface antigens of L-1210 leukemia recognized by monoclonal antibodies.

Conventional antisera to L-1210 leukemia were being prepared in our laboratory for nearly a decade and consistently the only specificity detectable was anti Mammary Leukemia antigen (ML). Serological analysis of five monoclonal antibodies obtained following the same immunization schedule showed more diverse pattern of reactivity. Two antigens detected belong to oncofetal category. The third one is differentiation antigen Ly-6 and the nature of two others, expressed on leukemic cells only, remains at present unclear. Thus none of the clones analysed produces antibodies to ML antigen. Our previous analysis of cell surface antigens of L-1210 leukemia with the use of conventional antisera has already been described. This paper presents the results of applying monoclonal antibodies in a comparable studies.

Animals↗

PubMed1981

Cell surface antigens of human renal cancer defined by mouse monoclonal antibodies: identification of tissue-specific kidney glycoproteins.

Seventeen monoclonal antibodies derived from fusions with spleen cells of mice immunized with established culture lines of renal cancers identified nine cell-surface antigenic systems. Six of the systems (gp160, S25, gp120r, gp120nr, gp115, and V1) represent antigens not previously described. The other three systems are related to HLA-A, -B, and -C heavy chain and A and B blood group antigens. The most restricted of the newly described antigens are gp160, S25, and gp120r. These determinants are found only on cells of renal origin, both normal and malignant, and represent differentiation antigens of human kidney. In addition to the difference in the molecular weight of two of these antigens, gp160, S25, and gp120r can be distinguished on the basis of differential expression on a panel of cultured renal cancers and normal kidney epithelium and fetal kidney cells. Glycoproteins bearing gp120r share a determinant with renal gp120nr (as indicated by sequential precipitations with monoclonal antibodies that detect gp120r and gp120nr), but gp120nr is found on a broader range of cell types, including fibroblasts and cell lines derived from lung, bladder, and colon cancers. The two other new systems, gp115 and V1, have characteristics of broadly occurring differentiation antigens but can be distinguished from each other and from gp120nr by differences in molecular weight, heat stability (V1 is a heat-stable determinant), and differential expression on cell types of diverse origin.

ABO Blood-Group System↗

PubMed1980

Lyt-1.1 antisera contain antibody against new cell surface alloantigen having wide tissue distribution.

Lyt-1.1 antisera produced in Lyt-1 congenic strain combination are contaminated with additional antibody detecting new cell surface alloantigen showing wide tissue distribution. The antibody was detected by complement independent inhibition of colony forming hemopoietic stem cells. Selective absorption experiments and differences in strain distribution prove that the discovered antigen is different from Lyt-1.

Animals↗