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Isolation and structure elucidation of radical scavengers from Thymus vulgaris leaves.

2,2-Diphenyl-1-picrylhydrazyl radical (DPPH*) scavenging activity-guided fractionation of a leaf extract of Thymus vulgaris led to the isolation of the radical scavengers rosmarinic acid 1, eriodictyol, taxifolin, luteolin 7-glucuronide, p-cymene 2,3-diol, p-cymene 2,3-diol 6-6'-dimer, carvacrol, thymol, and a new compound, 2. The fractionation was considerably facilitated by using an on-line HPLC detector for radical scavenging activity. In this detector activity is monitored as the disappearance of the color of a postcolumn added stable radical after reacting with radical scavengers in a reaction coil. Compound 2, which consists of rosmarinic and caffeic acid moieties linked via a C-3'-C-8' ' ether bridge, was mainly elucidated by various NMR techniques and CD. Phenylpropanoid trimer 2 was a weaker and stronger radical scavenger than rosmarinic acid 1 in off-line TEAC and DPPH* assays, respectively.

Biphenyl Compounds↗

Ganglioside composition of rabbit thymus.

Gangliosides of rabbit thymus were extracted and analyzed by the ganglioside-mapping procedure developed previously. 1 g of thymus contained 205.1 nmol of lipid-bound sialic acid and the relative amounts of monosialoganglioside and di- and trisialoganglioside fractions were 76.6 and 23.3%, respectively, as based on lipid-bound sialic acid. No ganglioside containing N-acetylgalactosamine was detected in rabbit thymus. The predominant component was N-glycoloylneuraminosyl lacto-N-norhexaosyl ceramide, NeuGc(alpha, 2-3)Gal(beta, 1-4)GlcNAc(beta, 1-3)Gal(beta, 1-4)GlcNAc(beta, 1-3)Gal(beta, 1-4)Glc(beta, 1-1) ceramide, which constituted 38.4% of the total gangliosides. The other major gangliosides were N-glycoloylneuraminosyl lacto-N-neotetraosyl ceramide (31.3%), GD3 containing N-glycoloylneuraminic acid (11.8%), GM3 containing N-glycoloylneuraminic acid (10.6%) and GM3 containing N-acetylneuraminic acid (6.4%). C-18 sphingosine was the only long-chain base in all the gangliosides. Palmitic acid was the major fatty acid of thymus gangliosides and chromatographically more polar gangliosides contained higher proportions of palmitic acid: 46.3% in GM3, 47.5% in GD3, 60.2% in N-glycoloylneuraminosyl lacto-N-neotetraosyl ceramide and 89.6% in N-glycoloylneuraminosyl lacto-N-norhexaosyl ceramide.

Animals↗

Local transrectal enzymatic treatment for chronic nonbacterial prostatitis and prostatodynia: initial clinical experience.

We report our initial clinical experience with local transrectal application of enzymatic treatment for chronic nonbacterial prostatitis and prostatodynia in 20 patients. Using a specially designed symptom score for evaluation of subjective treatment parameters, a statistically significant improvement of symptoms was found in the areas of pain, micturition, and recreational activities. No statistically significant differences were noted in laboratory values before and after treatment. Minimal local side effects were seen in only one patient. A favorable clinical response was noted in 75% of patients, whereas the remaining 25% showed only moderate improvement of symptoms. No patient experienced complete treatment failure.

Adjuvants, Immunologic↗

Immunological detection of a conserved structure for terminal deoxynucleotidyltransferase.

The polypeptide structure of terminal transferase in crude extracts of thymus or cultured cells was examined by electrophoresis on polyacrylamide gels in the presence of sodium dodecyl sulfate, electrophoretic transfer of separated peptides to nitrocellulose, specific labeling with rabbit anti-calf thymus terminal transferase, and visualization with an immunoperoxidase reaction. The major form of terminal transferase detected in crude extracts or enzyme fractions after phosphocellulose chromatography is a single 58,000- to 60,000-dalton peptide for calf thymus, rat thymus, mouse thymus, chicken thymus, cat thymus, human lymphoblastoid cells, and mouse lymphoblastoid cells. Since the anti-calf thymus terminal transferase antibody was prepared against a homogeneous calf thymus enzyme consisting of two polypeptide chains in a hydrodynamic structure of Mr = 32,000, these results suggest that the homogeneous calf thymus enzyme preparation is a proteolytically degraded form of the 58,000-dalton peptide. Terminal transferase peptides from human, bovine, rat, chicken, cat, and mouse are immunologically related and have similar conserved polypeptide structure.

Animals↗

[Clinical trial of the treatment of rheumatoid arthritis with TFX (thymus factor X)].

The results of rheumatoid arthritis treatment with the preparation TFX (extract of calf thymus) are reported. The drug was given to 11 patients with intense inflammatory process treated previously in typical fashion without evident improvement and in whom immunological studies demonstrated disturbances of cell-mediated and humoral response. The duration of treatment and observation was 6 months. In 9 patients clinical improvement was achieved. We observed no correlation between clinical improvement and immunological indices besides statistically significant changes of skin tests. The results (assessment of health state, laboratory investigations) encourage to TFX use in the treatment of rheumatoid arthritis.

Adult↗

Responses to immunization in the thymus of the adult mouse.

The rate of synthesis of RNA in the thymus glands of adult mice increased after immunization with sheep red blood cells (SRBC). The specific activity of some fractions of RNA, separated first by density gradient centrifugation and then by polyacrylamide gel electrophoresis, was 16-fold higher on day 3 after immunization than control mice not injected. RNA synthesis in the thymus was inhibited by rabbit anti-mouse thymus serum, injected along with antigen. A material was found in RNA extracts from the thymus glands of mice immunized with SRBC which converted a small proportion of either spleen cells or peritoneal cells from nonimmunized mice to form sheep cell hemolysins. Neither extracts from the glands of nonimmunized mice nor the livers of immunized mice were active. Extracts from the thymus glands of mice immunized with rabbit red blood cells (RRBC) were inactive and activity was destroyed by ribonudease. The residual antigen content was not determined. Biologically active extracts from the thymus had a different electrophoretic mobility from active extracts from the spleen.

Animals↗

Relaxant effect of Thymus vulgaris on guinea-pig tracheal chains and its possible mechanism(s).

Thymus vulgaris for the treatment of respiratory diseases is indicated widely, and relaxant effects on smooth muscle have been shown previously. In the present study, the relaxant effects of macerated and aqueous extracts of Thymus vulgaris on tracheal chains of guinea-pigs were examined using cumulative concentrations of macerated and aqueous extracts in comparison with saline (as the negative control) and theophylline (as the positive control). The relaxant effects of four cumulative concentrations of macerated and aqueous extracts (0.25, 0.5, 0.75 and 1.0 g %) in comparison with saline (as the negative control) and four cumulative concentrations of theophylline (0.25, 0.5, 0.75 and 1.0 mm; as the positive control) were examined for their relaxant effects on precontracted tracheal chains of guinea-pig by 60 mm KCl and 10 microm methacholine in two different conditions: non-incubated tissues and incubated tissues with 1 microm propranolol and 1 microm chlorphenamine. There were significant correlations between the relaxant effects and the concentrations for both extracts and theophylline in all experimental groups (p < 0.01 to p < 0.001). These results demonstrated a potent relaxant effect of Thymus vulgaris on guinea-pig tracheal chains that was comparable to theophylline at the concentrations used.

Animals↗

Detection and differentiation of chicken anemia virus isolates by using the polymerase chain reaction.

Complementary oligonucleotide primers which flank a 675-bp DNA fragment encompassing part of the putative gene for the capsid protein of chicken anemia virus (CAV) were used for the enzymatic amplification of CAV DNA by the polymerase chain reaction (PCR). Application of a dot blot hybridization assay by using a 32P-labeled cloned CAV DNA probe allowed PCR product amplified from as little as 0.1 fg of the target DNA sequence to be detected. When it was used for PCR amplification, DNA extracted from thymus tissue by a guanidine isothiocyanate-based method proved to be more efficient than that extracted by methods involving phenol or boiling. DNAs specified by 14 CAV isolates originating in the United Kingdom, Ireland, Germany, Sweden, the United States, Japan, and Australia were amplified. Restriction endonuclease analysis of the PCR-amplified DNAs with the enzymes HaeIII, HinfI, and HpaII indicated that the 14 CAV isolates can be assigned to seven groups, with isolates from different countries usually exhibiting the greatest number of restriction site differences.

Anemia↗

[Thymic Kurloff inclusion. Action on the ovogenesis of the rat].

Grafts and injections of cellular extracts from thymus and spleen of guinea pigs, were carried out in puberal and impuberal female rats. These organs are especially rich in Foa-Kurloff cells after oestrogentherapy. Kidney served as a reference material for the same experimental conditions. Controls received cellular injections and grafts from guinea pig organs without treatment. In all cases, ovarian histological and biometric study of recipient animals shows no alteration of the ovarian cycle. On the other hand, grafts and injections of cellular extracts from organs with high concentration of Foa-Kurloff cells induced hyperactivity of internal theca folliculi and of ovarian interstitial cells, only in puberal female rats.

Animals↗

Acceleration by thymosin of the development of resistance to murine sarcoma virus-induced tumor in mice.

The development of resistance to progressive tumor growth and the effect of a thymic extract upon this developmental process were examined in CBA/Wh mice. Inoculating mice of various ages with murine sarcoma virus (Moloney) permitted the assessment of the period of time postnatally at which the animals developed a threshold level of resistance to progressive tumor growth. Resistance in CBA/Wh mice, at the virus dose used, was first detected at approximately two weeks of age and was completely developed at five weeks.Thymosin, a soluble calf thymic fraction, when administered to neonatal mice beginning on the second day of life, significantly accelerated the rate of development of resistance to progressive tumor growth. The data suggest a humoral role for the thymus in the development of tumor immunity, and that the normal development of cell-mediated immunological competence can be accelerated through the use of thymus-derived extracts.

Age Factors↗

Chemical studies on a basic peptide preparation derived from calf thymus.

A substance possessing antimycobacterial activity under certain conditions in vitro has been prepared from aqueous extracts of calf thymus. Chemical studies have demonstrated that the activity of this substance is due to a basic peptide or a mixture of basic peptides. Although this thymus fraction has been shown to be essentially free of compounds other than peptides, it has not been obtained in a homogeneous state. The thymus peptide preparation is soluble in water and in the lower alcohols. Its solubility is minimal between pH 10 and 11, suggesting that its isoelectric point may be in this vicinity. The microbiological activity of thymus peptide is destroyed by acid or alkaline hydrolysis and also by trypsin digestion, but is unaffected by pepsin digestion. Cellulose membranes are permeable to thymus peptide. The most noteworthy finding concerning the amino acid composition of thymus peptide is the preponderance of the basic amino acids lysine and arginine, which together account for about 40 per cent of the weight of this substance. No cystine, and only trace amounts of other amino acids containing sulfur, are present in the thymus peptide preparation.

Amino Acids↗

Nonspecific immunostimulation with low doses of cyclophosphamide (LDCY), thymostimulin, and Echinacea purpurea extracts (echinacin) in patients with far advanced colorectal cancers: preliminary results.

Outpatients (n = 15) with metastasizing far advanced colorectal cancers received immunotherapy consisting of low-dose cyclophosphamide (LDCY) 300 mg/m2 every 28 days i.v., thymostimulin 30 mg/m2, days 3-10 after low-dose cyclophosphamide i.m. once daily, then twice a week, and echinacin 60 mg/m2 together with thymostimulin i.m. All patients had had previous surgery and/or chemotherapy and had progressive disease upon entering the study. Two months after onset of therapy a partial tumor regression was documented in one and a stable disease in 6 other patients by abdominal ultrasonography, decrease of the tumor markers carcinoembryonic antigen (CEA), CA 19-9, CA 15-3, and/or chest roentgenography, which may also be attributed to the natural course of disease. Mean survival time was 4 months, 2 patients survived for more than 8 months. Immunotherapy was well tolerated by all patients without side effects.

Adjuvants, Immunologic↗