Quantification of metabolites in the indole alkaloid pathways of catharanthus roseus: implications for metabolic engineering

In this article, we present a review of the current state of metabolic engineering in Catharanthus roseus. A significant amount of research has contributed to characterization of several individual steps in the biosynthetic pathway of medicinally valuable alkaloids. However, knowledge of the regulation of these pathways is still sparse. Using hairy root cultures, we studied the responses of alkaloid metabolism to environmental stimulation such as light and elicitation. Through precursor feeding studies, the putative rate-limiting steps of the terpenoid pathway in hairy root cultures also have been examined. Relating this knowledge to specific events at the molecular level, and the cloning of corresponding genes are the next key steps in metabolic engineering of the C. roseus alkaloids. Copyright 1998 John Wiley & Sons, Inc.     

In vitro evaluation of leptin fragments activity on the ob receptor.

In an attempt to identify regions in the leptin molecule responsible for its bioactivity, we tested six related-leptin peptide fragments denoted: Ac-hLEP(23-47)-NH(2) (I), Ac-hLEP(48-71)-NH(2) (II), Ac-hLEP(72-88)-NH(2) (III), Ac-hLEP(92-115)-NH(2) (IV), Ac-[Ser(117)]-hLEP(116-140)-NH(2) (V), Ac-hLEP(141-164)-NH(2) (VI) and their correspondent disulfide bridged dimer forms. The activity of the fragments was evaluated in comparision to leptin, by their ability to interact with leptin receptor using a cytosensor microphysiometer. Our results indicated that the fragments IV and V and [D-Leu(4)]-OB(3) and its human sequence analog were recognized by leptin receptor present in HP-75 cells, in agreement with the results obtained by other workers, validating that this region of the molecule contain the functional epitope of the leptin molecule.     

A novel quality control slide for quantitative immunohistochemistry testing.

We introduce a novel quality control technology that may improve intra- and interlaboratory immunohistochemistry (IHC) standardization. The technology involves the creation of standardized antibody targets that are attached to the same slides as the patient sample. After IHC staining, the targets turn the same color as the stained cells or tissue elements. Unlike current clinical practice, our proposed targets are neither cells nor tissue sections. To create reproducible standards that are available in unlimited supply, we use short constrained peptides as antibody targets. These peptides are attached directly to the glass slide. We show that these peptides simulate the portion of the native antigen to which the antibody binds. They are useful in detecting subtle changes in IHC staining efficacy. Moreover, the peptides do not degrade after deparaffinization or antigen retrieval treatments. This technology may be valuable in creating nationally standardized controls to quantify IHC analytical variability.     

The height-for-age of Indian children

The height for age of children is used as an indicator of the prevalence of undernutrition, i.e., 'stunting'. This study uses unit-record data on over 20,000 rural children, from 16 states of India, to explain variations in their height-for-age. Previous studies of children's heights have focused exclusively on the mean of the distribution of heights-for-age using ordinary least squares (OLS) regression. Instead, this paper uses quantile regression--which analyses different parts of the height for age distribution--and permits a richer set of conclusions to be drawn. The analysis shows the importance of locating malnourished children in terms of their place in the distribution of nutritional outcomes and, then, studying the differential impact of the determining variables on outcomes for children in different locations of the distribution.     

On the incidence of diarrhoea among young Indian children

Diarrhoea, claiming over three million young lives in the world every year, is the second biggest killer of children in developing countries. Using data for over 13,000 children in rural India, under the age of 3 years, this paper examines the relative effects of the different factors--inter alia the quality of the water supply, mother's literacy, housing conditions, and the level of development of the villages in which the children lived--contributing to diarrhoea. The paper highlights the importance of two factors: that children born to undernourished mothers may be more susceptible to infection than children whose mothers are well nourished, and that good hygienic practices within the home, such as washing hands with soap before feeding a child, can reduce the incidence of diarrhoea. The paper also quantifies the relative strength of the factors that determine whether mothers do so. The results emphasize the importance of mothers being literate, of household affluence and of institutional support (through the availability of trained midwives and mother and child centres in villages) in promoting domestic hygiene.     

A photoswitchable GPCR-based opsin for presynaptic inhibition

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Effect of short-term heat treatment of rice seedlings on sensitivity of thylakoid membranes to photoinhibition

The after-effects of 24 h high temperature (35 or 45 °C) treatment on the photochemical activities and photooxidative lipid peroxidation, subsequent to their irradiation were studied in 7-d-old etiolated rice (Oryza sativa) seedlings. Photosystem (PS) 1 and PS 2 mediated photoreactions of thylakoids isolated from the seedlings exposed to high temperature did not differ significantly from the thylakoids isolated from control seedlings (25 °C). Hence, all kinds of tested thylakoids were equally efficient in capturing and utilizing radiant energy. The high irradiance induced loss in PS 2 activity and lipid peroxidation measured in terms of malondialdehyde production was more rapid in thylakoids isolated from stressed seedlings as compared to that of control seedlings. Thus the thylakoids isolated from the stressed seedlings were more prone to photodamage than those from the control seedlings.     

Electroporation-mediated gene transfer into intact nodal meristemsin planta

Transient expression and stable integration and expression of transgenes were observed in the tissues and offspring of certain leguminous plants after electroporation of DNA into intact nodal meristemsin planta. The method described in this article thus allows the study of transgene expression in tissues differentiating from meristematic cells present in the treated buds. In addition, transgenic plants can be recovered in the offspring of electroporated individuals. Therefore, this technique allows the production of transgenic legumious plants without the need for in vitro tissue culture, often a major hurdle with this family.     

Transgenic grain legumes obtained byin planta electroporation-mediated gene transfer

Electroporation-mediated gene transfer into intact plant tissues was demonstrated in pea, cowpea, lentil, and soybean plants. Transient expression of a chimericgus reporter gene was used to monitor the uptake and expression of the introduced DNA in electroporated nodal axillary buds in vivo. The branches that grew out of the nodal meristems were chimeric and expressed the introduced gene up to 20 d after electroporation. Transgenic R₁ pea, lentil, and cowpea plants were recovered from seeds originating on these chimeric branches as shown by Southern blot hybridization and GUS expression. Transgenic R₂ soybean and lentil plants were also obtained. Segregation ratios in these populations showed a strong bias against transgene presence or expression.