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1.
Benef Microbes ; 10(4): 385-393, 2019 Apr 19.
Article in English | MEDLINE | ID: mdl-30882242

ABSTRACT

Vaginal discharge is one of the common reasons for gynaecologist consultation, as bacterial vaginosis and candidiasis are the main causes of discharge. These patients frequently experience numerous problems due to recurrent infections, side effects and drug resistance therefore alternative drugs are needed. Our primary aim was to evaluate safety and tolerability of the potentially probiotic Lactobacillus crispatus strains in volunteer women considering themselves healthy. We also monitored the effects of these strains on vaginal health parameters and lactobacilli counts in vagina and intestine. Forty women were recruited into trial. Absence of chronic diseases was confirmed by questionnaire and blood analysis at screening visit. In randomised double-blind placebo-controlled crossover study the eligible participants were randomly allocated to one of four groups and had to consume one of the two study products (Pro I or Pro II) - a capsule containing 3 strains, 109 cfu per strain, or placebo for 1 week. Treatment period was followed by 2-week washout period and continued with second treatment and washout period. Individuals receiving firstly probiotic, received later placebo and vice versa. Blood, vaginal and faecal samples were collected, and self-reported questionnaires were applied. Thirty subjects completed the trial. The probiotic capsules were well-tolerated. The Pro II intake resulted in a significant decrease in Nugent score (from median 3.0 to 2.0, mean 3.9 to 2.6, P=0.002) and reduction in Gardnerella vaginalis counts (log10 3.57 to 2.38; P=0.027). Reduction of total vaginal bacterial counts was revealed in Pro I group (log10 7.99 to 7.72; P=0.048). In conclusion, the selected vaginal L. crispatus strains are well tolerable and Pro II mixture is prospectively effective in reducing Nugent score and vaginal counts of G. vaginalis. Therefore, these strains seem to be promising candidates for development of novel evidence-based well-focused probiotics to target female urogenital tract disorders.


Subject(s)
Lactobacillus crispatus/physiology , Probiotics/administration & dosage , Vaginosis, Bacterial/drug therapy , Administration, Oral , Adult , Cross-Over Studies , Double-Blind Method , Female , Gardnerella vaginalis/drug effects , Gardnerella vaginalis/growth & development , Gastrointestinal Microbiome , Humans , Lactobacillus/growth & development , Safety , Treatment Outcome , Vagina/microbiology , Young Adult
2.
NPJ Vaccines ; 3: 30, 2018.
Article in English | MEDLINE | ID: mdl-30083396

ABSTRACT

Koala retrovirus (KoRV) infects the majority of Australia's koalas (Phascolarctos cinereus) and has been linked to several life-threatening diseases such as lymphoma and leukemia, as well as Chlamydia and thus poses a threat to the continued survival of this species. While quarantine and antiretroviral drug treatment are possible control measures, they are impractical, leaving vaccination as the only realistic option. In this study, we examined the effect of a recombinant envelope protein-based anti-KoRV vaccine in two groups of South Australian koalas: KoRV infected or KoRV free. We report a successful vaccination response in the koalas with no vaccine-associated side effects. The vaccine induced a significant humoral immune response as well as the production of neutralizing antibodies in both groups of koalas. We also identified B-cell epitopes that were differentially recognized in KoRV-infected versus KoRV-free koalas following vaccination. Importantly, we also showed that vaccination had a therapeutic effect on koalas infected exogenously with KoRV by reducing their circulating viral load. Together, this study highlights the possibility of successfully developing a vaccine against KoRV infection in koalas.

3.
J Appl Microbiol ; 123(2): 524-534, 2017 Aug.
Article in English | MEDLINE | ID: mdl-28574626

ABSTRACT

AIM: To assess antibiotic susceptibility of vaginal lactobacilli strains and provide the data required for assessing the potential of antibiotic resistance risk of new strains selected as probiotic. METHODS AND RESULTS: Potential probiotic vaginal lactobacilli used in the study included 31 vaginal strains of Lactobacillus crispatus (n = 27), Lactobacillus gasseri (n = 3) and Lactobacillus jensenii (n = 1) obtained from the collection of Competence Centre on Health Technologies. Two commercial probiotic strains were used as controls (Lactobacillus rhamnosus GR-1 and Lactobacillus fermentum RC-14). The phenotypic and genotypic antibiotic resistances of the strains were determined by E-test and PCR methods. The location (chromosomal DNA or plasmid) of antibiotic resistance genes was also detected. All lactobacilli strains expressed high level of resistance to kanamycin, metronidazole, norfloxacin and trimethoprim/sulphamethoxazole. Some of the strains also expressed resistance to other antibiotics (chloramphenicol, vancomycin) indicating acquired resistance. I class integrons were found in 20% (6/31) of the strains. The RPP (ribosomal protection protein) gene was found to be positive in 30% (9/31) of the strains. Only one L. jensenii strain was determined with tet(M) gene. The tet(K) gene was positive in 26·7% (8/31) and erm(B) gene in 43·3% (13/31) of strains. Three RPP and both four tet(K) and erm(B) genes were located in plasmids. CONCLUSIONS: High antibiotic resistance to clinically important antibiotics was demonstrated, including metronidazole, sulphonamides, aminoglycoside and quinolones. In addition, acquired tetracycline and erythromycin resistance genes were detected in either plasmid or chromosomal DNA of certain isolates, in some of the cases for the first time in the literature. It appears that antibiotic resistance genes erm(B) and tet(K) are widely spread in vaginal lactobacilli. SIGNIFICANCE AND IMPACT OF THE STUDY: This study provides new data about antimicrobial resistance and genotypic diversity of vaginal Lactobacillus isolates. In addition, it provides data assessing the potential of antibiotic resistance risk of new strains selected as probiotic.


Subject(s)
Anti-Bacterial Agents/pharmacology , Drug Resistance, Bacterial , Lactobacillus/drug effects , Lactobacillus/isolation & purification , Vagina/microbiology , Erythromycin/pharmacology , Female , Genotype , Humans , Lactobacillus/classification , Lactobacillus/genetics , Microbial Sensitivity Tests , Phenotype , Tetracycline/pharmacology , Vancomycin/pharmacology
4.
Benef Microbes ; 6(3): 233-43, 2015.
Article in English | MEDLINE | ID: mdl-25524863

ABSTRACT

The blood pressure-lowering effect of dairy products holds the potential to decrease the risk of cardiovascular disease (CVD). An open question is if the successful expression of functional properties of the probiotic strain depends on host biomarkers and/or food matrix properties. The probiotic Lactobacillus plantarum strain TENSIA® (DSM 21380) is a novel microorganism with antimicrobial and antihypertensive functional properties. The aim of this study was to characterise the functional properties of the probiotic L. plantarum TENSIA and compare its effects on host anthropometric, clinical, and blood biomarkers when consumed with cheese or yoghurt. This study involved two double-blinded randomised placebo-controlled exploratory trials (ISRCTN15061552 and ISRCTN79645828) of healthy adults over a three-week period. The three-week consumption of probiotic L. plantarum TENSIA in a daily dose of 1×1010 cfu in probiotic cheese or a daily dose of 6×109 cfu in yoghurt with different content of carbohydrates, proteins, and lipids did not significantly change the body mass index (BMI), plasma glucose and lipid levels, or inflammatory markers in the blood. Reduced lowered systolic and diastolic blood pressure values were detected, regardless of food matrix or baseline values for blood pressure and BMI. In conclusion, our study showed that three-week consumption of the probiotic L. plantarum TENSIA either in cheese or yoghurt lowered diastolic and systolic blood pressure regardless of food matrix and baseline values of blood pressure and BMI, confirming the impact of the functional properties of the probiotic strain in decreasing CVD risk.


Subject(s)
Antihypertensive Agents/metabolism , Cardiovascular Diseases/microbiology , Cardiovascular Diseases/physiopathology , Cheese/microbiology , Lactobacillus plantarum/metabolism , Probiotics/metabolism , Yogurt/microbiology , Adult , Blood Glucose/metabolism , Blood Pressure , Cardiovascular Diseases/diet therapy , Cardiovascular Diseases/metabolism , Humans , Lipids/blood , Male , Middle Aged , Young Adult
5.
Clin Pharmacol Ther ; 94(3): 309-11, 2013 Sep.
Article in English | MEDLINE | ID: mdl-23963219

ABSTRACT

In April 2012, MIT's Center for Biomedical Innovation and the European Medicines Agency (EMA) cosponsored a workshop on legal foundations of adaptive pharmaceuticals licensing. Past and present attorneys from the US Food and Drug Administration (FDA), the EMA, and Health Sciences Agency Singapore (HSA) found that existing statutes provided authority for adaptive licensing (AL). By contrast, an attorney from Health Canada identified gaps in authority. Reimbursement during initial phases of adaptive approaches to licensing was deemed consistent with existing statutes in all jurisdictions.


Subject(s)
Drug Approval/legislation & jurisprudence , Licensure/legislation & jurisprudence , Canada , European Union , United States
6.
J Dairy Sci ; 95(10): 5495-509, 2012 Oct.
Article in English | MEDLINE | ID: mdl-22863096

ABSTRACT

Safety of the probiotic Lactobacillus plantarum strain Tensia (DSM 21380) was tested in vitro, in semihard Edam-type cheese, in an animal model and after consumption of the probiotic cheese in double-blind randomized placebo-controlled human intervention studies with different age groups. The susceptibility of L. plantarum Tensia to 8 antibiotics, and the presence of tetracycline (tet M, S, O, K, L) genes and class 1 integron was assessed by applying epsilometer-test and PCR-based methods. Production of biogenic amines by the probiotic strain in decarboxylation medium containing 1% of l-histidine, l-glutamine, l-ornithine, l-arginine, or l-lysine and in cheese was tested by gas chromatography. The biosafety of L. plantarum Tensia was evaluated on National Institutes of Health-line mice fed cheese containing Tensia at a concentration of 9.6 log cfu/g for 30 consecutive days. In human intervention trials in adults and the elderly, the effects of different doses of Edam-type cheese and the probiotic bacterium on BW, gut functionality indices, and host metabolism were evaluated. The strain L. plantarum Tensia was susceptible to all tested antibiotics and did not possess the tetracycline resistance-determining genes tet(L), tet(S) and tet(O), nor did it contain the integron (Int1) gene. However, the strain was tet(K) and tet(M) positive. Lactobacillus plantarum Tensia did not produce potentially harmful biogenic amines, such as histamine or cadaverine. The amount of tyramine produced in the cheese environment during ripening and after 15 wk of storage was below the clinically significant content. In the animal model, no translocation of the administered strain or other microbes into the blood or organs of mice was detected. No harmful effect was observed on body mass index, inflammatory markers, or serum lipidograms during human intervention trials with different age groups at a daily dose of 10.3 or 8.17 log cfu/serving for 3 wk. No negative effect on gastrointestinal welfare was observed, but the consumption of 100g/d for 3 wk caused hard stools from the second week of the trial. The content of total lactobacilli increased in feces, and the presence of the ingested probiotic strain was confirmed after the consumption of cheese. Thus, L. plantarum strain Tensia is suitable for generally recognized as safe (GRAS) and qualified presumption of safety (QPS) criteria because it did not have any undesirable characteristics. The regular semihard Edam-type cheese (fat content of 26%) with the probiotic additive at a daily dose of 50 g or in excess (100g) and with a probiotic daily dose of 10 log cfu for 3 wk was safe.


Subject(s)
Cheese/microbiology , Lactobacillus plantarum/metabolism , Probiotics/adverse effects , Adult , Age Factors , Aged , Animals , Biogenic Amines/adverse effects , Biogenic Amines/analysis , Cheese/adverse effects , Cheese/analysis , Cross-Over Studies , Double-Blind Method , Feces/microbiology , Female , Food Safety , Genes, Bacterial , Hemolysis , Humans , Male , Mice , Microbial Sensitivity Tests , Polymerase Chain Reaction , Probiotics/analysis
7.
Vopr Pitan ; 81(3): 74-81, 2012.
Article in Russian | MEDLINE | ID: mdl-22888676

ABSTRACT

The probiotic L. plantarum strain TENSIA (DSM 21380) is a novel microorganism having antimicrobial and antihypertensive properties. The aim of the study was to test the efficacy of the consumption of the cheese, comprising the novel strain TENSIA on multiple health markers of humans. Human intervention trial showed that the blood pressure lowering effect of cheese, comprising L. plantarum TENSIA was evident in healthy volunteers with high normal blood pressure up to normal values. The 3-week consumption of the prohiotic cheese did not increase the CVD risk factors like BMI, the level of plasma lipids and glucose as well as inflammatory and immunological markers of human body.


Subject(s)
Blood Pressure/drug effects , Cheese/microbiology , Lactobacillus plantarum/growth & development , Lipid Metabolism/drug effects , Lipids/blood , Probiotics/pharmacology , Adolescent , Adult , Aged , Body Mass Index , Cross-Over Studies , Double-Blind Method , Female , Humans , Lipid Peroxidation/drug effects , Male , Middle Aged , Treatment Outcome , Young Adult
8.
Benef Microbes ; 2(1): 79-90, 2011 Mar.
Article in English | MEDLINE | ID: mdl-21831792

ABSTRACT

The aim of the study was to evaluate the safety and persistence of selected Lactobacillus strains in the gastrointestinal tract (GIT) of healthy adult volunteers after oral consumption of high doses of lactobacilli to identify potential candidates for probiotic and biotechnological applications. In the first phase of the study, nine individuals consumed capsules containing Lactobacillus gasseri 177 and E16B7, Lactobacillus acidophilus 821-3, Lactobacillus paracasei 317 and Lactobacillus fermentum 338-1-1 (each daily dose 1×1010 cfu) for 5 consecutive days. Data on gut health, blood parameters, and liver and kidney function were collected. The persistence of Lactobacillus strains was assessed by culturing combined with arbitrarily primed polymerase chain reaction (AP-PCR) and PCR-denaturing gradient gel electrophoresis (PCR-DGGE) on days 0, 5, 8, 10 and 20 from faecal samples. All strains survived gastrointestinal passage and were detected on the 5th day. L. acidophilus 821-3 was detected in four volunteers on the 8th day (4.3 to 7.0 log10 cfu/g) and in two on the 10th day (8.3 and 3.9 log10 cfu/g, respectively). In the second phase of the study, five additional volunteers consumed L. acidophilus 821-3 (daily 1×1010 cfu) for 5 consecutive days. The strain was subsequently detected in faeces of all individuals using real-time PCR on the 10th day (range 4.6-6.7; median 6.0 log10 cell/g) in both phases of the study for at least 5 days after discontinuation of consumption. The administration of high doses of different Lactobacillus strains did not result in any severe adverse effects in GIT and/or abnormal values of blood indices. Thus, the strain L. acidophilus 821-3 is a promising candidate for probiotic and biotechnological applications. Further studies will be performed to confirm the strain persistence and safety in a larger number of individuals.


Subject(s)
Gastrointestinal Tract/microbiology , Lactobacillus , Probiotics/administration & dosage , Administration, Oral , Adult , Colony Count, Microbial , Female , Humans , Male , Middle Aged , Probiotics/adverse effects , Probiotics/pharmacokinetics , Young Adult
9.
Lett Appl Microbiol ; 48(6): 797-800, 2009 Jun.
Article in English | MEDLINE | ID: mdl-19344357

ABSTRACT

AIM: To evaluate the impact of the consumption of a synbiotic product on the antioxidative activity markers of blood in asymptomatic H. pylori-colonized persons. METHODS AND RESULTS: Fifty-three healthy adult volunteers without gastric symptoms participated in a randomized, double-blind placebo-controlled study. The crossover consumption of the enterocoated capsules containing antioxidative Lactobacillusfermentum ME-3, Lact. paracasei 8700:2 and Bifidobacterium longum 46 with Raftilose P95 lasted for 3 weeks and did not change the H. pylori colonization. In H. pylori-positive subjects the sera values of total antioxidative status (TAS) were significantly lower compared to H. pylori-negative subjects (0.97 vs 1.05 mmol l(-1), P = 0.008). After the consumption of the synbiotic, TAS values (0.97 vs 1.03 mmol l(-1), P = 0.004) increased, while the ratio between oxidized and reduced glutathione (0.035 vs 0.030, P = 0.016) decreased in H. pylori-positive subjects. CONCLUSIONS: The consumption of a synbiotic containing an antioxidative probiotic strain improved the reduced systemic antioxidative activity in H. pylori-colonized asymptomatic subjects. SIGNIFICANCE AND IMPACT OF THE STUDY: A synbiotic product containing an antioxidative probiotic strain may be useful in the reduction of systemic oxidative stress in H. pylori infection.


Subject(s)
Antioxidants/metabolism , Blood/metabolism , Helicobacter Infections/metabolism , Helicobacter pylori/growth & development , Probiotics/administration & dosage , Adult , Bifidobacterium/physiology , Female , Glutathione/metabolism , Helicobacter Infections/microbiology , Humans , Limosilactobacillus fermentum/physiology , Male , Middle Aged , Young Adult
10.
J Appl Microbiol ; 100(6): 1324-32, 2006 Jun.
Article in English | MEDLINE | ID: mdl-16696680

ABSTRACT

AIM: To develop in vitro assays for comparing the antagonistic properties and anti-oxidative activity of probiotic Lactobacillus and Bifidobacterium strains against various entero- and urinary pathogens. METHODS AND RESULTS: The antagonistic activity of five probiotic lactobacilli (Lactobacillus rhamnosus GG, Lactobacillus fermentum ME-3, Lactobacillus acidophilus La5, Lactobacillus plantarum 299v and Lactobacillus paracasei 8700:2) and two bifidobacteria (Bifidobacterium lactis Bb12, Bifidobacterium longum 46) against six target pathogens was estimated using different assays (solid and liquid media, anaerobic and microaerobic cultivation) and ranked (low, intermediate and high). Bacterial fermentation products were determined by gas chromatography, and the total anti-oxidative activity of probiotics was measured using linolenic acid test. Pyelonephritic Escherichia coli was highly suppressed by GG and both bifidobacteria strains. Lactobacilli strains 8700:2, 299v and ME-3 were the most effective against Salmonella enterica ssp. enterica in microaerobic while ME-3 and both bifidobacteria expressed high activity against Shigella sonnei in anaerobic milieu. Lact. paracasei, Lact. rhamnosus and Lact. plantarum strains showed intermediate antagonistic activity against Helicobacter pylori under microaerobic conditions on solid media. The highest anti-oxidative activity was characteristic for Lact. fermentum ME-3 (P < 0.05). No efficient antagonist against Clostridium difficile was found. The positive correlations between the pH, lactic acid production and anti-microbial activity for all tested probiotics were assessed. CONCLUSIONS: Developed experimental assays enable to compare the anti-microbial and -oxidative activity of Lactobacillus and/or Bifidobacterium probiotics, which have been claimed to possess the ability of suppressing the growth of various enteric and urinary pathogens. SIGNIFICANCE AND IMPACT OF THE STUDY: Screening Lactobacillus and Bifidobacterium sp. strains according to their activity in various environmental conditions could precede the clinical efficacy studies for adjunct treatment with probiotics in cure of different gastrointestinal and urinary tract infections.


Subject(s)
Antibiosis/physiology , Bifidobacterium/physiology , Digestive System Diseases/microbiology , Lactobacillus/physiology , Probiotics , Urination Disorders/microbiology , Antioxidants , Bacteriology , Culture Media , Digestive System Diseases/therapy , Fatty Acids, Volatile/metabolism , Humans , Hydrogen-Ion Concentration , Species Specificity , Statistics, Nonparametric , Urination Disorders/therapy
11.
J Infect Dis ; 193(11): 1528-35, 2006 Jun 01.
Article in English | MEDLINE | ID: mdl-16652281

ABSTRACT

BACKGROUND: Carriage of hepatitis B virus (HBV) is a major risk factor for liver cirrhosis and hepatocellular carcinoma. Infant vaccination has been effective in preventing horizontal transmission during early childhood. It is unknown whether protection is maintained into early adulthood. METHODS: In 1984, early childhood vaccination was introduced in 2 rural Gambian villages. In 2003, serological assessment of 81.5% of 1,350 eligible participants 1-24 years old was done, to determine vaccine efficacy against infection and carriage. RESULTS: Overall vaccine efficacy against infection and carriage was 83.4% (95% confidence interval [CI], 79.8%-86.6%) and 96.5% (85% CI, 93.9%-98.9%), respectively. Vaccine efficacy against infection was similar when restricted to primary responders (85.3%), but a significant effect of peak antibody concentration was found. Both vaccine efficacy and levels of hepatitis B surface antibody (anti-HBs) decreased with age, resulting in a vaccine efficacy against infection and carriage among 20-24-year-old participants of 70.9% (95% CI, 60.4%-80.5%) and 91.1% (95% CI, 75.8%-100%), respectively. Fifteen years after vaccination, fewer than half of the vaccinees had detectable anti-HBs. The prevalence of carriage in the unvaccinated population was similar to the prevalence 20 years earlier. CONCLUSIONS: HBV vaccination early during life can provide long-lasting protection against carriage, despite decreasing antibody levels. The role played by subclinical boosting and the necessity of a booster need to be evaluated.


Subject(s)
Carrier State/prevention & control , Hepatitis B Vaccines/immunology , Hepatitis B/prevention & control , Adolescent , Adult , Age Factors , Child , Child, Preschool , Gambia , Hepatitis B Antibodies/blood , Hepatitis B Surface Antigens/blood , Hepatitis B Vaccines/administration & dosage , Humans , Infant
12.
J Dairy Sci ; 87(7): 2017-23, 2004 Jul.
Article in English | MEDLINE | ID: mdl-15328213

ABSTRACT

The aim of our study was to develop an original probiotic cheese based on the Estonian open-texture, smear-ripened, semisoft cheese "Pikantne." Cheese was produced by two methods using cheese starter cultures (Probat 505) in combination with 0.04% of probiotic Lactobacillus fermentum strain ME-3 (10(9) cfu/mL) with high antimicrobial activity and antioxidative properties. The probiotic Lactobacillus was added into milk simultaneously with starter cultures (cheese A) and into drained curd (cheese B). After addition of probiotic L. fermentum ME-3, the cheese composition, flavor, and aroma were comparable to the control cheese (score values = 4.5, 4.2, and 3.7 for control cheese, cheese A, and cheese B, respectively). Cheese A, which had good sensory properties, was chosen for further testing of viability and probiotic properties. The probiotic strain was found to withstand the technological processing of cheese, surviving and sustaining moderate antimicrobial and high antioxidative activity throughout ripening and storage (the ripened cheese contained approximately 5 x 10(7) cfu/g viable ME-3 cells), although the viability of the ME-3 strain incorporated into the cheese showed a slight decrease between d 24 and 54 after cheese preparation. Semisoft cheese "Pikantne" serves as a suitable carrier of antimicrobial and antioxidative L. fermentum ME-3.


Subject(s)
Anti-Infective Agents , Antioxidants , Cheese , Probiotics , Cheese/analysis , Cheese/microbiology , Lactobacillus/physiology , Smell , Taste
14.
Hemoglobin ; 22(1): 1-10, 1998 Jan.
Article in English | MEDLINE | ID: mdl-9494043

ABSTRACT

Among the causes of congenital methemoglobinemia, Hb M-Milwaukee-2 was one of the earliest described, in a patient who also had Hb E trait. The structure of Hb M-Milwaukee-2 has been elusive. DNA sequence analysis, as here reported, proves that this hemoglobin variant is due to the mutation CAC-->TAC at codon 92 of the beta-globin gene, corresponding to the substitution of tyrosine for histidine. This mutation is identical with that presumed to be the cause of Hb M-Hyde Park and Hb M-Akita. In addition, the DNA mutation of Hb E, GAG-->AAG at codon 26, was confirmed in this case.


Subject(s)
Codon , Globins/genetics , Hemoglobin M/genetics , Point Mutation , Adult , Aged , Female , Globins/chemistry , Hemoglobin M/chemistry , Humans , Sequence Analysis, DNA
15.
J Pediatr Hematol Oncol ; 19(4): 319-26, 1997.
Article in English | MEDLINE | ID: mdl-9256831

ABSTRACT

PURPOSE: Rhabdomyosarcomas (RMS) are heterogeneous in their clinical presentation, histology, and cytogenetics. The growth of some RMS cells has been found to be regulated by the tyrosine kinase insulin-like growth factor (IGF) type I receptor. However, RMS cells exhibit variable sensitivity to inhibitors of tyrosine kinases and IGF receptors. Collectively, these heterogeneous features suggest that differences exist in the growth regulatory pathways of RMS. The objective of this study is to identify active tyrosine kinase signal transduction pathways in embryonal and alveolar RMS cells. METHODS: RMS tumor samples and cell lines representing both embryonal and alveolar histologic subtypes have been analyzed by immunoprecipitation and immunoblotting techniques to characterize phosphotyrosyl protein patterns and to identify tyrosine phosphorylated proteins. RESULTS: RMS cells can be characterized based on the patterns of phosphotyrosyl proteins, including the phosphorylation status of the catenin-like protein Cas1 and the signal adapter protein SHC, and the activation of IGF type I receptor signaling cascades including the formation of SHC-GRB2 signal protein complexes and MAP kinase activation. CONCLUSIONS: Rhabdomyosarcomas, especially the embryonal histologic subtype, are heterogeneous at the level of tyrosine kinase signal transduction. It will be important to characterize the growth regulatory pathways active in individual RMS tumors before targeting molecular therapies to this malignancy.


Subject(s)
Adaptor Proteins, Signal Transducing , Cadherins/metabolism , Neoplasm Proteins/metabolism , Receptor, IGF Type 1/metabolism , Rhabdomyosarcoma/metabolism , Signal Transduction/physiology , Adolescent , Calcium-Calmodulin-Dependent Protein Kinases/metabolism , Catenins , Cell Adhesion Molecules/metabolism , Child , Child, Preschool , Enzyme Activation , Female , GRB2 Adaptor Protein , Humans , Infant , Phosphoproteins/metabolism , Phosphorylation , Phosphotyrosine/metabolism , Protein-Tyrosine Kinases/metabolism , Proteins/metabolism , Tumor Cells, Cultured , Tyrosine/metabolism , Delta Catenin
16.
Hemoglobin ; 21(3): 205-18, 1997 May.
Article in English | MEDLINE | ID: mdl-9140717

ABSTRACT

Hemoglobin variants with two amino acid substitutions affecting one globin chain are relatively rare. Hb T-Cambodia, a doubly substituted beta-globin variant, was characterized previously by amino acid sequencing as having sequence alterations in beta 26 (beta 8)Glu-->Lys and beta 121(GH4) Glu-->Gln (1). It is a variant that migrates cathodic to Hb A2 on alkaline electrophoresis and with Hb A on acid citrate agar electrophoresis. We report here the mutations of Hb T-Cambodia at the nucleotide level using DNA sequencing, in beta-globin gene codon 121 (GAA-->CAA) and in codon 26 (GAG-->AAG). These are the mutations of Hb D-Punjab and Hb E, respectively.


Subject(s)
Hemoglobins, Abnormal/chemistry , Hemoglobins, Abnormal/genetics , Mutation/genetics , Adult , Blood Protein Electrophoresis , Chromatography, High Pressure Liquid , Exons/genetics , Hemoglobin E/chemistry , Hemoglobin E/genetics , Hemoglobins/analysis , Hemoglobins, Abnormal/physiology , Humans , Male , Pedigree , Peptides/chemistry , Sequence Analysis, DNA , Trypsin
18.
Hemoglobin ; 20(4): 371-6, 1996 Nov.
Article in English | MEDLINE | ID: mdl-8936463

ABSTRACT

Approximately 700 hemoglobin variants have been reported, causing a variety of clinical manifestations, with the majority being clinically silent. We report a new hemoglobin variant, Hb Cook, that was found in combination with Hb E in a child of Thai origin. DNA sequencing of the beta-globin gene showed that the mutation is AAA-->ACA in codon 132, corresponding to beta 132 (H10)Lys-->Thr.


Subject(s)
Globins/genetics , Hemoglobins, Abnormal/genetics , Asia, Southeastern , Chromatography, High Pressure Liquid , DNA, Complementary , Female , Hemoglobins, Abnormal/analysis , Humans , Infant , Male , Oligonucleotide Probes , Point Mutation
19.
Mayo Clin Proc ; 71(9): 854-6, 1996 Sep.
Article in English | MEDLINE | ID: mdl-8790260

ABSTRACT

OBJECTIVE: To ascertain the usefulness of bone marrow and cerebrospinal fluid (CSF) examinations in identifying or predicting relapse in children with acute lymphoblastic leukemia (ALL) before discontinuation of chemotherapy. MATERIAL AND METHODS: We retrospectively reviewed the medical records of 113 children with ALL in first continuous complete remission who had undergone routine end-of-therapy bone marrow aspiration and CSF examinations. RESULTS: One patient had frank bone marrow relapse at the completion of therapy, which was evident by the presence of blasts in the peripheral blood. None of the other 112 patients had morphologic evidence of bone marrow relapse or positive CSF cytologic findings. The seven subsequent relapses could not have been predicted by the results of end-of-therapy bone marrow or CSF studies. CONCLUSION: Routine morphologic examination of the bone marrow and CSF at the completion of therapy for ALL has no diagnostic or prognostic value.


Subject(s)
Bone Marrow/pathology , Precursor Cell Lymphoblastic Leukemia-Lymphoma/cerebrospinal fluid , Precursor Cell Lymphoblastic Leukemia-Lymphoma/pathology , Adolescent , Cerebrospinal Fluid/cytology , Child , Child, Preschool , Female , Humans , Infant , Male , Precursor Cell Lymphoblastic Leukemia-Lymphoma/drug therapy , Predictive Value of Tests , Recurrence , Retrospective Studies
20.
Am J Hematol ; 52(4): 305-9, 1996 Aug.
Article in English | MEDLINE | ID: mdl-8701949

ABSTRACT

A 13-year-old African-American female with erythrocytosis and three different beta globins on electrophoresis beta A, beta S, and beta Osler, raised the possibility that one chromosome 11 might contain a duplicated beta globin gene, since there are normally only 2 beta globin genes. DNA sequence analysis showed GTG at codon 6 in exon 1, corresponding to Hb S and AAT at codon 145 in exon 3, indicating a substitution of Asn for Tyr. Thus, Hb Osler undergoes spontaneous post-translational deamidation, beta 145 Asn-->beta 145 Asp. Unmodified Hb Osler (Asn) co-migrates with Hb A on electrophoresis and co-elutes with Hb A on HPLC; therefore it has not been identified previously. All previous studies have incorrectly identified the mutation as being beta 145 (HC 2) Tyr-->Asp.


Subject(s)
DNA/chemistry , Globins/genetics , Hemoglobin, Sickle/genetics , Hemoglobins, Abnormal/genetics , Mutation , Adolescent , Base Sequence , Chromosomes, Human, Pair 11 , Exons , Female , Humans , Molecular Sequence Data , Sequence Analysis, DNA , Twins, Dizygotic
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