Na, K ATPase activity during early postnatal development of the rat submandibular gland

The activity of the ouabain-sensitive Na, K ATPase was measured in membrane fractions of the submandibular gland of 1-, 7-, 14- and 21-day-old rats. This activity increased with age and reached adult levels by 21 days.     

Overexpression of copper-zinc superoxide dismutase in trisomy 21

Down's syndrome (DS), the most frequent of congenital birth defects, results from the trisomy of chromosome 21 in all cells of affected patients. This disease is characterized by developmental anomalies, mental retardation and features of rapid aging, particularly in the brain, where the occurrence of Alzheimer's disease is observed in trisomy 21 patients over the age of 35. Copper-zinc superoxide dismutase (CuZnSOD) is one of the proteins encoded by chromosome 21 (21q22.1). As a consequence of gene dosage excess, CuZnSOD activity is increased by 50% in all DS tissues. This work reports the SOD activity of a population of DS patients with complete trisomy 21, partial trisomy 21, translocations and mosaicism, in order to confirm the gene dosage effect of SOD on the clinical features of DS, and to help to establish which is the critical region of chromosome 21 in DS. CuZnSOD was measured in red blood cells using the Minami and Yoshikawa method. In the population with complete trisomy 21, SOD activity was increased by 42%; in the population with partial trisomy 21, translocations and mosaicism, SOD activity was normal. In the population diagnosed as DS, but not karyotyped, SOD activity was increased by 28%. No differences between sexes or among ages were found. We conclude that the 21q22.1 segment is not the critical region responsible for DS, as we have found normal SOD activity in patients with the clinical features of DS.     

Novel substrates for the kinetic assay of esterases associated with insecticide resistance

Naphthalene thioesters were synthesized as substrates for a continuous, non- disruptive kinetic assay of general carboxylesterase activity. The continuous nature of the assay is based on the production of a soluble dianion chromophore from the reaction of naphthalene thio with 5,5-dithiobis (2-nitrobenzoic acid). Applications with 1- and 2-naphthalene thioacetates demonstrated their use in a fast accurate kinetic microassay of esterase activity, using porcine carboxylesterase as a model. These novel esters proved to be useful as substrates for the spectrophotometric assay of insecticide-resistance in two aphid species and may be applicable to other esterasebased diagnostic procedures.     

Esculentin(1-21), an amphibian skin membrane-active peptide with potent activity on both planktonic and biofilm cells of the bacterial pathogen Pseudomonas aeruginosa

Pseudomonas aeruginosa is an opportunistic bacterial pathogen that forms sessile communities, named biofilms. The non-motile forms are very difficult to eradicate and are often associated with the establishment of persistent infections, especially in patients with cystic fibrosis. The resistance of P. aeruginosa to conventional antibiotics has become a growing health concern worldwide and has prompted the search for new anti-infective agents with new modes of action. Naturally occurring antimicrobial peptides (AMPs) represent promising future template candidates. Here we report on the potent activity and membrane-perturbing effects of the amphibian AMP esculentin(1-21), on both the free-living and sessile forms of P. aeruginosa, as a possible mechanism for biofilm disruption. Furthermore, the findings that esculentin(1-21) is able to prolong survival of animals in models of sepsis and pulmonary infection indicate that this peptide can be a promising template for the generation of new antibiotic formulations to advance care of infections caused by P. aeruginosa.     

Der Einfluss einiger C21-Steroide auf die Wassereinlagerung in wachsenden embryonalen Knorpeln in vitro

Summary Embryonic chick cartilages were cultivated in a synthetic nutrient medium. In this experimental procedure cortisol is known to reduce the water uptake of the cartilages. It was found that some C₂₁-steroids exert a similar activity. Those of the steroids investigated, which inhibit water uptake, have in common a ⁴-3-keto group, an 11-hydroxy group, and a keto group in position 20; oxygen functions in positions 21 and 17 seem to be of minor importance for this activity. The possibility of these steroids being inactivated by cellular enzymes is discussed.     

Progesterone inhibits human endothelial cell proliferation through a p53-dependent pathway

Previous studies have shown that progesterone inhibits endothelial cell proliferation through a nuclear receptor-mediated mechanism. Here, we further demonstrate that progesterone at physiologic levels (5 – 500 nM) dose- and time-dependently inhibited DNA synthesis of cultured human umbilical vein endothelial cells (HUVEC). The mRNA and protein levels of p21, p27, and p53 in HUVEC were increased by progesterone. The formation of CDK2-p21 and CDK2-p27 were increased and the CDK2 activity was decreased in the progesterone-treated HUVEC. The progesterone-inhibited [3H]thymidine incorporation was completely blocked when the expressions of p21 and p27 were knocked-down together. Transfection of HUVEC with dominant negative p53 cDNA prevented the progesterone-induced increases in p21 and p27 promoter activity and protein level, decreases in thymidine incorporation, and capillary-like tube formation. Matrigel angiogenesis assay in mice demonstrated the antiangiogenic effect of progesterone in vivo. These findings demonstrate for the first time that progesterone inhibited endothelial cell proliferation through a p53-dependent pathway. Received 28 July 2008; received after revision 25 September 2008; accepted 26 September 2008     

A thermosensitive inhibitor of the replication of polyomavirus, an extract from BHK2I cells transformed by the Tsa mutant of this virus

BHK21 cells transformed by wild type or Ts3 mutant polyoma virus contain an inhibitor of polyoma virus replication when grown at permissive (36 degrees C) as well as non-permissive temperature (39 +/- 0.5 degrees C). Cells transformed by the Tsa mutant contain the inhibitor at the permissive but not at the non-permissive temperature. The inhibitor reappears in the latter cells however, upon shift from the non-permissive to the permissive temperature. If a reversible protein inhibitor (methionyl-adenylate, reversible inhibitor of the aminoacyl-t-RNA synthetase) is applied during the temperature shift experiments, the inhibitor does not reappear indicating that new protein synthesis is required for the recovery of its activity.     

Metabolic implications in the elevation of serum activity of intestinal alkaline phosphatase in chronic renal failure

The activity of intestinal isoenzyme of serum alkaline phosphatase was evaluated in 21 non-dialyzed patients with advanced renal failure and in 52 patients on regular hemodialysis. In patients without hepatopathy, a significant inverse correlation was found between the enzyme activity and serum calcium levels. Hepatopathy was the most significant variable influencing the enzyme activity in patients on dialysis. Secondary hyperparathyroidism and a decreased rate in enzyme elimination should be assessed for the above-normal activities of intestinal ALP in serum in chronic renal failure.     

Metabolic implications in the elevation of serum activity of intestinal alkaline phosphatase in chronic renal failure

Summary The activity of intestinal isoenzyme of serum alkaline phosphatase was evaluated in 21 non-dialyzed patients with advanced renal failure and in 52 patients on regular hemodialysis. In patients without hepatopathy, a significant inverse correlation was found between the enzyme activity and serum calcium levels. Hepatopathy was the most significant variable influencing the enzyme activity in patients on dialysis. Secondary hyperparathyroidism and a decreased rate in enzyme elimination should be assessed for the above-normal activities of intestinal ALP in serum in chronic renal failure.     

Metabolic implications in the elevation of serum activity of intestinal alkaline phosphatase in chronic renal failure

The activity of intestinal isoenzyme of serum alkaline phosphatase was evaluated in 21 non-dialyzed patients with advanced renal failure and in 52 patients on regular hemodialysis. In patients without hepatopathy, a significant inverse correlation was found between the enzyme activity and serum calcium levels. Hepatopathy was the most significant variable influencing the enzyme activity in patients on dialysis. Secondary hyperparathyroidism and a decreased rate in enzyme elimination should be assessed for the above-normal activities of intestinal ALP in serum in chronic renal failure.     

Metabolic implications in the elevation of serum activity of intestinal alkaline phosphatase in chronic renal failure

Summary The activity of intestinal isoenzyme of serum alkaline phosphatase was evaluated in 21 non-dialyzed patients with advanced renal failure and in 52 patients on regular hemodialysis. In patients without hepatopathy, a significant inverse correlation was found between the enzyme activity and serum calcium levels. Hepatopathy was the most significant variable influencing the enzyme activity in patients on dialysis. Secondary hyperparathyroidism and a decreased rate in enzyme elimination should be assessed for the above-normal activities of intestinal ALP in serum in chronic renal failure.     

Effect of abrupt withdrawal of chronically administered β-blocking drugs on cardiac sensitivity in the rat

Summary Increased tachycardia to isoprenaline was observed in pithed rats 2 days after withdrawal of propanolol but not after withdrawal of atenolol (a cardioselective drug) or LL 21-945 (a long acting beta-blocking drug).Acknowledgments. We are grateful to Sandoz Ltd for a gift of LL 21-943 and to I.C.I. for gifts of propranolol and atenolol.     

Adenyl cyclase activity of the cell membrane throughout the cell cycle: ultrastructural study of the cell line BHK 21/C 13]

Adenylate cyclase activity in the surface membrane of baby Hamster kidney (BHK 21/C13) cells is ultrastructurally studied as a function of the cell cycle. Cells are synchronized by a physical method suited to structural and functional investigations. The activity is highest through G1, decreases in S, markedly drops as cells traverse G2 and is very low or often seems not present at all in M.     

Protective effect of N-(2-propynyl)-2-(5-benzyloxy-indolyl) methylamine (PF9601N) on mitochondrial permeability transition

PF9601N, N-(2-propynyl)-2-(5-benzyloxy-indolyl) methylamine, an monoamine oxidase (MAO) B inhibitor, has shown neuroprotective properties against dopaminergic toxins. To elucidate the mechanisms involved in this protection, the effect of PF9601N on mitochondria was assessed. PF9601N prevents mitochondrial swelling, drop in the electrical potential and oxidation of sulfhydryl groups, glutathione and pyridine nucleotides induced by Ca²⁺. These observations demonstrate the protective effect of PF9601N on the induction of mitochondrial permeability transition. This protection is due to the interaction of the secondary protonated amino group in the molecule with pore-forming structures and to its antioxidant property, rather than to inhibition of MAO B activity. PF9601N also prevents the release of cytochrome c from mitochondria, suggesting its potential inhibitory effect on mitochondria-mediated apoptosis. The low IC⁵⁰ value for this inhibition, in comparison with deprenyl, make it a more efficient compound than propargylamines and other amines in protecting the bioenergetic functions of mitochondria. Received 9 March 2006; received after revision 10 April 2006; accepted 21 April 2006     

On the distribution of plasma L-asparaginase

The guinea-pig, Cavia porcellus, is unusual in possessing plasma L-asparaginase, an enzyme with anti-tumor activity, 21 additional species have been examined as to the presence of this enzyme: the results confirm and extend its remarkably limited species distribution.     

可溶性重组全长人PPARγ的表达、纯化及功能表征

目的 建立可溶性表达、纯化有活性的重组全长人PPARγ蛋白的方法.方法 构建pReceiver-B13-SUMO-PPARγ质粒转化至Escherichia coli BL21(DE3)细胞,进行诱导表达,优化细胞生长环境;利用亲和色谱法纯化可溶性重组全长人PPARγ蛋白;以罗格列酮为阳性配体,以GW9662为拮抗剂,采用分子排阻色谱-高效液相色谱法(SEC-HPLC)法测定重组全长人PPARγ蛋白与配体药物的结合活性.结果 在优化条件(16℃、IPTG浓度0.4 mM、诱导时间18～20 h)下能获得高水平、可溶性重组全长人PPARγ蛋白;经亲和色谱一次纯化,得到纯度约为90％的重组全长人PPARγ蛋白;该蛋白与罗格列酮特异性结合率约为70％,Kd为500nM.结论 本文所建立的方法能可溶性表达、纯化得到有活性、纯度较高的重组全长人PPARγ蛋白,为该蛋白功能研究和配体药物筛选奠定基础.     

Fetal hypophysis as the main source of serum TSH in fetal rat

Decapitation performed at days 17-18 leads to a drastic drop (82%) in blood TSH of 19 and 21-day-old rat fetuses below the mother's level. 125I-TSH injected at 21 days into the mother's bloodstream is not found in fetal blood. The fetal hypophysis is the main source of fetal plasmatic TSH.     

Serum activity inhibiting specific simian virus 40-induced transplantation resistance and its correlation with primary SV40 tumors appearance in hamsters.

Using the modified technique of transplantation test, ITR serum activity was found in most (14 out of 21) individual hamster sera obtained during the latent period of primary SV40 carcinogenesis (60 days after virus infection when newborn). On the other hand, as a rule, no ITR activity was observed in the sera of the same hamsters after tumor appearance and during their growth. ITR activity rapidly disappeared from sera of hamsters neonatally infected with SV40 after their successful immunization with the same virus during the latent period. There appears to be a correlation between the presence of ITR serum factor during the latent period and the subsequent primary SV40 tumor appearance in hamsters.