Stability of nitroblue tetrazolium-based alkaline phosphatase substrates.

This report demonstrates the stability of NBT substrate after multiple exposures to alkaline phosphatase. Perhaps more important than the ability to reuse substrates, the report provides some insight into the mechanisms by which tetrazoliums are reduced and evidence for the formation of an intermediary product, i.e., a half-formazan that is reduced more rapidly. (J Histochem Cytochem 49:1189-1190, 2001)     

Expression of the Chlamydia trachomatis major outer membrane protein-encoding gene in Escherichia coli: role of the 3' end in mRNA stability

The major outer membrane protein (MOMP)-encoding gene (omp1) of Chlamydia trachomatis has been cloned into Escherichia coli and partially sequenced. This recombinant gene expresses a full-length 40-kDa product, which is recognized by a monoclonal antibody directed against the species-specific epitope of MOMP. The recombinant omp1 is expressed in either insertion orientation, indicating that it utilizes its own promoter system. The endogenous omp1 promoter possesses a relatively low activity despite the high level of MOMP expression. Deletion of a 520-bp fragment at the 3' end encoding 39 amino acids (aa) at the C terminus and the remainder of the noncoding region leads to a significant decrease in mRNA stability and loss of protein synthesis. When the MOMP-encoding plasmid was introduced into E. coli minicells, it expressed 40- and 43-kDa proteins; however, inhibition of post-translational processing by ethanol revealed only a 43-kDa protein. These data indicate that the unprocessed omp1 gene product contains a 22-aa leader sequence which is cleaved during translocation to the outer membrane, to yield a processed 40-kDa protein. The recombinant MOMP was localized to the outer membrane E. coli fraction, comparable to the location of the native C. trachomatis protein.     

Activation of mitochondrial glycerol 3-phosphate dehydrogenase by cadmium ions

Mitochondrial glycerol 3-phosphate dehydrogenase (EC 1.1.2.1.) requires Ca2+ ions for its activity. Cadmium ions also have activatory effect on the enzyme. They activate the glycerol 3-phosphate dehydrogenase in a very narrow concentration range (1-2 mmol/l). As contrasted with calcium, strong inhibitory effect occurred at higher concentrations (3-4 mmol/l). The inhibition induced by cadmium ions was completely reversible by washing of the mitochondria.     

Extractive bioconversion in aqueous two-phase systems

Summary The transformation of hydrocortisone to prednisolone was studied in aqueous two-phase systems, as a model for the extractive bioconversion of fine chemicals. The bacterium, Arthrobacter simplex, was able to grow in the two-phase system and the cells could be revitalized after a period of use. Use of aqueous two-phase systems made it possible to operate the reaction at higher substrate concentrations than in pure buffer solution. An adsorptive method to remove the product from the top phase was tested and shown to be both efficient and compatible with the overall process. In order to reduce the costs of operation in aqueous two-phase systems, a cheaper starch-based polymer, Reppal-PES, was successfully used as a substitute for dextran.Dedicated to Professor Dr. Georg Manecke on the occasion of his 70th birthday     

The goldfish pituitary

Summary In the goldfish, Carassius auratus, morphological and functional aspects of the pituitary gland were studied at the ultrastructural level and six cell types could be distinguished in the pars distalis. Acidophilic cells of the rostral pars distalis were identified as prolactin cells, the chromophobic cells of the rostral pars distalis as ACTH cells, the non-globular basophilic cells of the rostral and the proximal pars distalis as TSH cells, the globular basophils of the proximal pars distalis as gonadotropic cells and the acidophils of the proximal pars distalis as somatotrophs.Besides some of the well established criteria of morphological and functional identification of different cell types, two new approaches have been used in the present study. One was to express the electron density of secretory granules objectively by means of a photometric method. It was found that both types of acidophilic cells which produce the proteohormones prolactin and somatotropin respectively, had granules with the highest electron densities. The basophilic cells producing the glycoproteins gonadotropin and TSH respectively, possessed granules of intermediate electron density whereas the chromophobic cells storing the peptide hormone ACTH had granules of lowest densities. The second new approach was the administration of the synthetic mammalian releasing hormones LH-RP and TRF, which helped in identifying gonadotropic and thyrotropic cells respectively. In the goldfish there is evidence for the presence of only one type of gonadotropic cell.Supported by a grant of the Science Research Council of Great Britain to Professor Sir Francis Knowles, F.R.S. The electron microscope used was provided by the Medical Research Council of Great Britain. The integrating photometer IPM2 was kindly on loan from Messrs. Carl Zeiss, Oberkochen, Germany. For technical advice we are greatly indebted to Mr. P. K. Kaul, B. E., M.I. Struct. E., C. Eng.     

Reproduction in the Indian Desert Gerbil Meriones hurrianae Jerdon

In the present study the whole sample of 772 desert gerbils was collected from the semi-arid areas in and around Jaipur (Rajasthan, India). The male exhibits active spermatogenesis and well maintained accessory reproductive organs at any time of the year. Reproductive organs of the male gerbil are briefly described. The vaginal and urinary openings are independent in the gerbil. There are four pairs of mammae. Follicular development takes place cyclically. Maximum atresia of follicles is noticed during metoestrus. A well developed Graafian follicle measures 590 μ across its widest diameter. The average diameter of the ovum is 65 μ. At the time of ovulation formation of stigma occurs which gives way to the ovum. The corpus luteum is formed from the remaining granulosa cells of the ovulated follicle. Fully formed corpus luteum of the cycle (during dioestrus) measures 740 μ. Maximum development of the corpus luteum is seen during late pregnancy (22–25 days) when it measures 1250 μ. During pregnancy large Graafian follicles are absent. After parturition the ovary shows corpora lutea of lactation. Histological changes in the uterus and vagina during the oestrous cycle of the gerbil are described. There is less proestrus distension of the uterus as compared with the rat. During oestrus the uterine endometrium hypertrophies and there are some degenerative changes in the epithelial lining. The changes in the vaginal epithelium are clear cut and are correlated with the ovarian activity. During proestrus proliferation of the epithelium occurs and at ‘heat period’ or oestrus the epithelium shows formation of the stratum corneum and the shedding of cornified cells. In the dioestrus the vaginal epithelium is heavily infiltrated by leucocytes. There are three peaks in the incidence of pregnancy rate, i.e., February to April, July, and September to November. Pregnancy rate is lowest during summer (May and June) and winter (December and January). Average monthly litter size shows almost similar changes. The average length of gestation period is 30 days. There is post-partum heat in this animal. Formation of a vaginal plug is noticed following copulation. Litter size varies from 2–7. The average body weight of the new born young is 3.5 and 3.4 gm respectively for the male and the female. Weaning of the young is observed after about 20 days. Descent of the testes into the scrotal sacs occurs around day 28. Vagina opens after about 75 days.