Detection of 1-phenyl-N-methyl-1,2,3,4-tetrahydroisoquinoline and 1-phenyl-1,2,3,4-tetrahydroisoquinoline in human brain by gas chromatography—tandem mass spectrometry

1-Phenyl-N-methyl-1,2,3,4-tetrahydroisoquinoline and 1-phenyl-1,2,3,4-tetrahydroisoquinoline were detected for the first time in parkinsonian human brain using gas chromatography-tandem mass spectrometry (GC-MS-MS). Since these compounds are structural analogues of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) that produces parkinsonism in humans, they might be candidates for endogenous MPTP-like neurotoxins.     

An early and massive wave of germinal cell apoptosis is required for the development of functional spermatogenesis.

Transgenic mice expressing high levels of the BclxL or Bcl2 proteins in the male germinal cells show a highly abnormal adult spermatogenesis accompanied by sterility. This appears to result from the prevention of an early and massive wave of apoptosis in the testis, which occurs among germinal cells during the first round of spermatogenesis. In contrast, sporadic apoptosis among spermatogonia, which occurs in normal adult testis, is not prevented in adult transgenic mice. The physiological early apoptotic wave in the testis is coincident, in timing and localization, with a temporary high expression of the apoptosis-promoting protein Bax, which disappears at sexual maturity. The critical role played by the intracellular balance, probably hormonally controlled, of the BclxL and Bax proteins (Bcl2 is apparently not expressed in normal mouse testis) in this early apoptotic wave is shown by the occurrence of a comparable testicular syndrome in mice defective in the bax gene. The apoptotic wave appears necessary for normal mature spermatogenesis to develop, probably because it maintains a critical cell number ratio between some germinal cell stages and Sertoli cells, whose normal functions and differentiation involve an elaborate network of communication.     

Simple and rapid method for determining nicotinamide adenine dinucleotide synthetase activity by high-performance liquid chromatography

A method is described for the determination of nicotinamide adenine dinucleotide synthetase (NADS) activity in human blood. Using high-performance liquid chromatography (HPLC), the formed NAD is separated from the substrates and the other blood components in less than 13 min. The activity of NADS determined by HPLC is closely correlated with that determined by the conventional spectrophotometric method, which requires two steps of enzyme reaction. The present method is simple and reliable and facilitates the routine analysis of NADS activity.     

Evaluation of Antiherpetic Compounds Using a Gastric Cancer Cell Line: Pronounced Activity of BVDU against Herpes Simplex Virus Replication

We developed a rapid and simple method for the screening of antiviral agents against herpes simplex virus (HSV) in a model of gastrointestinal herpetic infection in vitro. This method was based on inhibition of HSV-induced cytopathogenicity in gastric adenocarcinoma MKN-28 cells, as monitored by an MTT colorimetric assay. From the various compounds that were evaluated for their activity against HSV-1 and HSV-2, brivudine (BVDU) emerged as the most effective. When the 50% effective concentration (EC₅₀) values of the antiherpes agents in MKN-28 cells were compared with those in human embryo lung MRC-5 cells, all compounds, except for BVDU, showed higher EC₅₀ values in MKN-28 cells. For BVDU the EC₅₀ values in MKN-28 cells were 0.8 (HSV-1) and 0.036 (HSV-2) times the EC₅₀ values in MRC-5 cells. Thus BVDU was 27.5 times more active against HSV-2 in MKN-28 cells than in MRC-5 cells. The MKN-28 gastric cancer cells may be useful for the rapid screening of anti-HSV agents and, in particular, those that may be useful in therapy of gastrointestinal HSV infections in gastrointestinal herpetic infection.     

Effect of the presence of trophectoderm vesicles on blastocyst in relation to in vitro hatching,clinical pregnancy,and miscarriage rates

Trophectoderm vesicles (TVs) are observed in some blastocysts that penetrate cells from the zona pellucida to the outer margin. Therefore, we compared this incidence in relation to hatching, pregnancy, and miscarriage rates between conventional in vitro fertilization (c-IVF) and intracytoplasmic sperm injection (ICSI). Vitrified/warmed blastocysts (n = 112) were derived from surplus embryos. The blastocysts were then observed using time-lapse cinematography to resolve the relationship between hatching and implantation. Another study was conducted that comprised 681 embryo transfer cycles in 533 patients who received a single vitrified/warmed blastocyst from our clinic. The incidence of TV was significantly higher in embryos inseminated by ICSI compared with c-IVF [ICSI: 51/56 (91 %); c-IVF: 25/56 (45 %); P < 0.01]. The successful hatching rate was significantly lower in ICSI than in c-IVF [ICSI: 11/56 (20 %); c-IVF: 29/56 (52 %); P < 0.01]. In addition, the hatching rate was significantly lower when TVs were present (14/76; 18 %) than in non-TV embryos (26/36; 72 %) (P < 0.01). In regard to the clinical study results, no significant differences were found between the groups in the pregnancy rate (TV present group: 107/183, 58.5 %; TV absent group: 273/498, 54.8 %) and miscarriage rate (TV present group: 21/107, 19.6 %; TV absent group: 53/273, 19.4 %). In vivo, we hypothesized that hatching and hatched would occur naturally by assisting protease action in the uterus; therefore, these results suggest that the presence of TV has no effect on pregnancy rates in the clinical setting.     

Elucidating the DDK‐dependent step in replication initiation

By phosphorylating specific replication factors, cell cycle kinases ensure that eukaryotic DNA replication is initiated once and only once per mitotic cell division. New work in The EMBO Journal now reveals how DDK‐mediated phosphorylation of Mcm2‐7 helicase subunits is read out by Sld3, which provides further integration with CDK phosphorylation.     

The position of the disulfide bonds in human plasma alpha 2 HS-glycoprotein and the repeating double disulfide bonds in the domain structure

The positions of the inter- and intra-chain disulfide bonds of human plasma alpha 2 HS-glycoprotein were determined. alpha 2 HS-glycoprotein was digested with acid proteinase and then with thermolysin. The disulfide bonds containing peptides were separated by reversed-phase HPLC and detected by SBD-F (7-fluorobenzo-2-oxa-1,3-diasole-4-sulfonic acid ammonium salt) method. One inter-disulfide bond containing peptide and five intra-disulfide bond containing peptides (A-chain) were purified and identified as Cys-18 (B-chain)--Cys-14 (A-chain), Cys-71--Cys-82, Cys-96--Cys-114, Cys-128--Cys-131, Cys-190--Cys-201 and Cys-212--Cys-229, respectively. The location of the intra-disulfide bonds revealed that the A-chain of alpha 2 HS-glycoprotein is composed of three domains. Two domains were shown to possess intramolecular homology judging from the total chain length of the domains, size of the loops formed by the S--S bonds, the location of two disulfide loops near the C-terminal end of domains A and B, the distance between two S--S bonds of each domain, the amino acid sequence homology between these two domains (22.6%), number of amino acid residues between the second S--S loops and the end of domains A and B, and the positions of the ordered structures.     

The development of porcine zona pellucida using monoclonal antibodies: II. Electron microscopy

Following our previous study on the immunohistochemistry of porcine zonae pellucidae (ZP), we undertook the present study to localize the components of the ZP with immunoelectron microscopy, using three types of anti-porcine-ZP monoclonal antibodies (Mabs), named STA-1, STA-2, and STA-3. Some organelles of the oocyte were seen to react with STA-2 and STA-3 prior to ZP formation. As soon as a follicle began to mature, STA-2 and STA-3 reacted with the perinuclear space and the endoplasmic reticular membrane of the oocyte. The follicle first reacted with STA-1 at the secondary follicle stage. At this stage, the positive reaction involved the follicular cell layer as well as the oocyte and ZP. Positive reaction was scattered within and limited to the interfollicular cell space and was never found in the cytoplasm of follicular cells. At the antral follicle stage, the oocyte was surrounded by a thick, electron-dense ZP. A strong reaction was observed in the outer layer, but no significant reaction occurred in the inner layer. The convex and ragged outer margin of the ZP was characterized by the strongest reaction.     

Social life factors affecting the mortality, longevity, and birth rate of total Japanese population: effects of rapid industrialization and urbanization

To expand upon the findings that lower mortality was found in Japanese urban areas in contrast to the Western model where in the US and Britain the risk of death was higher in metropolitan areas and conurbations, 22 social life indicators are examined among 46 prefectures in Japan in terms of their effect on age specific mortality, life expectancy, and age adjusted marriage, divorce, and birth rates. The effects of these factors on age adjusted mortality for 8 major working and nonworking male populations, where also analyzed. The 22 social life factors were selected from among 227 indicators in the system of Statistical Indicators on Life. Factor analysis was used to classify the indicators into 8 groups of factors for 1970 and 7 for 1975. Factors 1-3 for both years were rural or urban residence, low income and unemployment, and prefectural age distribution. The 4th for 1970 was home help for the elderly and for 1975, social mobility. The social life indicators were classified form 1 to 8 as rural residence in 1970 and 1975, urban residence, low income, high employment, old age, young age, social mobility, and home help for the elderly which moved from 8th place in 1970 to 1st in 1975. Between 1960-75, rapid urbanization took place with the proportion of farmers, fishermen, and workers declining from 43% in 1960 to 19% in 1975. The results of stepwise regression analysis indicate a positive relationship of urban residence with mortality of men and women except school-aged and middle-aged women, and the working populations, as well as life expectancy at birth for males and females and ages 20 and 40 years for males. Rural residence was positively associated with the male marriage rate, whereas the marriage rate for females was affected by industrialization and urbanization. High employment and social mobility were positively related to the female marriage rate. Low income was positively related to the divorce rate for males and females. Rural residence and high employment were positively related to the birth rate. The birth rate is higher in rural areas. Mortality of professional, engineering, and administrative workers was slightly lower than the total working population, while sales workers, those in farming, fishing, and forestry, and in personal and domestic service had significantly higher mortality. The mortality of the nonworking population was 6-8 times higher than sales, transportation, and communication, and personal and domestic service as well as the total population.