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Biomedical subjects

R Valyasevi

Publications and source records attributed to R Valyasevi.

6 recordsLinked to original sources

Genotypic and phenotypic characterization of garlic-fermenting lactic acid bacteria isolated from som-fak, a Thai low-salt fermented fish product.

AIMS: To evaluate the importance of garlic for fermentation of a Thai fish product, and to differentiate among garlic-/inulin-fermenting lactic acid bacteria (LAB) at strain level. METHODS AND RESULTS: Som-fak was prepared by fermentation of a mixture of fish, salt, rice, sucrose and garlic. pH decreased to 4.5 in 2 days, but omitting garlic resulted in a lack of acidification. LAB were predominant and approximately one third of 234 isolated strains fermented garlic and inulin (the carbohydrate reserve in garlic). These strains were identified as Lactobacillus pentosus and Lact. plantarum. Randomly Amplified Polymorphic DNA (RAPD) analysis revealed one major RAPD type (29 strains) isolated from all stages of fermentation. CONCLUSION: Garlic was essential for acidification of som-fak and garlic-fermenting strains constituted a significant, homogeneous part of the LAB flora. SIGNIFICANCE AND IMPACT OF THE STUDY: The present study indicates the role of fructans (garlic/inulin) as carbohydrate sources for LAB. Fructan fermenters may have several biotechnological applications, for example, as probiotics.

Animals↗

Gene organization and transcription of a late-expressed region of a Lactococcus lactis phage.

The lactococcal phage bIL41 belongs to the small isometric-headed phages of the 936 quasi-species and is resistant to the abortive infection determined by abiB. A 10.2-kb segment from this phage, in which late transcription is initiated, has been sequenced. Thirteen open reading frames (ORFs) organized in one transcriptional unit have been identified. The location of two of them and the structural features of the proteins they code for are evocative of terminase subunits. Five other ORFs specify proteins which are highly homologous to structural proteins from the closely related phage F4-1. By comparing the phage bIL41 sequence with partial sequences available for four related phages, we were able to deduce a chimerical phage map covering the middle- and a large part of the late-expressed regions. Phages from this quasi-species differ by the insertion or deletion of either 1 to about 400 bp in noncoding regions or an entire ORF. Transcription was initiated 9 min after infection at a promoter with a -10 but no -35 consensus sequence. Synthesis of a phage activator protein was needed for initiation of transcription. A large 16-kb transcript covering all of the late-expressed region of the genome was synthesized. This transcript gave rise to smaller units. One of these units most probably resulted from a RNase E processing.

Bacteriophages↗

Thyroid dysfunction in HIV-infected children: is L-thyroxine therapy beneficial?

PURPOSE: To assess thyroid function in symptomatic HIV-infected infants and children with failure to thrive and to evaluate clinical response in patients with hyperthyrotropinemia to levothyroxine. METHODS: We evaluated 11 children (mean age: 1.5 years; males = 5, females = 6) with HIV infection and failure to thrive (weight and height > 2 SD below the mean) for presence of hypothyroidism. Thyroid-stimulating hormone (TSH) and thyroxine (T4) were measured. Four children with high basal TSH and one child with a normal basal TSH underwent the Thyrotropin-releasing hormone (TRH) stimulation test as well. Four children received treatment with L-thyroxine. RESULTS: Eight of the 11 children had high basal TSH levels. All patients had normal or increased T4 values. The TRH stimulation test showed an exaggerated response in all four patients with high basal TSH values and a normal response in one patient with normal basal THS values. All four patients who received treatment with L-thyroxine showed normalization of the TSH. Increased growth velocity was noted in 3 of 4 patients. The fourth patient died within two weeks following initiation of therapy for overwhelming candida sepsis. The autopsy of this patient showed atrophy of the thyroid gland. Two of the four patients with high TSH levels, who did not receive L-thyroxine, died; the other two did not show any improvement in their height z scores. Two patients with a high TSH also had high thyroid-binding hormone (TBG) levels. Both patients with the high TBG died within 2 to 3 months of the study. CONCLUSIONS: Hypothyroidism as indicated by elevated basal TSH and abnormal TRH response was common in the HIV-infected children in this study and may have contributed to failure of growth in these children. Replacement therapy resulted in correction of abnormal TSH and improvement of height z scores during the period of observation. The significance of these findings needs to be confirmed in a larger prospective study.

Child, Preschool↗

A membrane protein is required for bacteriophage c2 infection of Lactococcus lactis subsp. lactis C2.

Phage-resistant mutants, isolated from cultures of Lactococcus lactis subsp. lactis C2 infected with phage c2, did not form plaques but bound phage normally. The mutants were sensitive to another phage, sk1, although the number of plaques was reduced approximately 56% and the plaques were four times smaller. Binding to phage sk1 was reduced about 10%. Another group of phage-resistant mutants, isolated from cultures infected with phage sk1, bound normally to both phages c2 and sk1 but did not form plaques with either phage. Carbohydrate analyses by gas chromatography of the cell walls showed no significant differences in saccharide compositions between the wild-type and phage-resistant cells. However, a difference was observed in the interactions of the phage with the cytoplasmic membranes. Membranes from the wild-type cells, but not mutant cells, inactivated phage c2. Phage sk1 was not inactivated by membrane from either strain. Treatment of wild-type membranes with proteinase K eliminated the ability of the membrane to inactivate the phage, whereas treatment with mutanolysin had no effect. On the basis of this ability to inactivate the phage, a membrane protein was partially purified by gel filtration and ion-exchange chromatography. Under nondenaturing conditions, the phage-inactivating protein has an apparent Mr of approximately 350,000. The protein has an apparent subunit size of 32 kDa, which suggests that it normally exists as a multimer with 10 to 12 subunits or in association with other membrane components. It is proposed that this protein is required for phage c2 infection.

Bacterial Proteins↗

The bacteriophage kh receptor of Lactococcus lactis subsp. cremoris KH is the rhamnose of the extracellular wall polysaccharide.

A receptor for bacteriophages of lactic acid bacteria, including Lactococcus lactis subsp. cremoris KH, was found on the cell wall and not on the cell membrane, as determined by a phage-binding assay of sodium dodecyl sulfate- and mutanolysin-treated cell walls. The cell wall carbohydrates of L. lactis subsp. cremoris KH were analyzed by gas chromatography and mass spectrometry and found to contain rhamnose, galactose, glucose and N-acetylglucosamine. Similar analysis of mutants that were reduced in the ability to bind phages kh, 643, c2, ml3, and 1 indicated that galactose was essential for binding all phages. In addition, rhamnose was required for binding phages kh and ml3. Inhibition studies of phage binding by using two different lectins with a specificity for galactose indicated that phage kh may not bind directly to galactose. Rather, galactose may be an essential structural component located in the vicinity of the receptor. Incubation of any of the five phages with rhamnose or of phage kh with purified cell walls inactivated the phages. Inactivation required divalent cations and was irreversible. Inactivation of phages was stereospecific for rhamnose, as neither L-(+)- nor D-(-)-fucose (the stereoisomers of rhamnose) inhibited the phage. Furthermore, phage infection of a culture was completely inhibited by the addition of rhamnose to the medium. Therefore, the receptor for phage kh appears to be a rhamnose component of the extracellular wall polysaccharide.

Bacteriophages↗