On the existence and separation of the follicle stimulating hormone releasing hormone from the luteinizing hormone releasing hormone.

Porcine hypothalamic fragments were extracted by 2M AcOH at 4°C, and the extractives were subsequently processed in the presence of one protease inhibitor and one anti-oxidant. Gel filtration was performed on Bio-Gel P-2, and supplementary [³H]-LHRH and [¹⁴C]- 3H]-LHRH, and was differentiated from [¹⁴C]- 相似文献   

Spatial and temporal patterns of temperature,alkalinity, dissolved oxygen and conductivity in an oligo-mesotrophic,deep-storage reservoir in central Texas

Impoundment behavior was determined for alkalinity, temperature, dissolved oxygen, and conductivity from stations located along the length of a bottom-draining, oligo-mesotrophic, hardwater, deep-storage reservoir located in central Texas. The epilimnion deepened the length of the reservoir throughout the summer as a result of drawdown. Bicarbonate alkalinity and conductivity exhibited both longitudinal and vertical stratification. Alkalinity and conductivity in the epilimnion decreased from the riverine reach downreservoir to the dam. This longitudinal progression was attributed to inflow and photosynthetically induced epilimnetic decalcification.Hypolimnetic anoxic conditions first occurred in the sedimentation zone in the upreservoir and riverine reaches and then developed in a downreservoir pattern as summer progressed as a result of drawdown. Alkalinity and conductivity in the hypolimnion increased during anoxic conditions and consequently increased in a downreservoir progression.     

Bacterial breakdown of benomyl. II. Mixed cultures

Experiments with mixed bacterial cultures grown in liquid media which contained the benzimidazole fungicide benomyl, with or without Na-lactate, as source of carbon provided circumstantial evidence for cleavage of the benzimidazole heterocyclic ring. Yet, neither 2-aminobenzimidazole (2-AB) nor benzimidazole, as sole source of carbon, supported any bacterial growth. Total ¹⁴C-balance analysis experiments conclusively showed production of ¹⁴CO₂ from [2-¹⁴C] methyl benzimidazol-2-yl carbamate (MBC), and thus cleavage of the benzimidazole nucleus; bioassays, however, showed that the actual rate of benomyl and MBC breakdown was only small, the parent compound benomyl being still recovered in substantial quantities after up to 80 days of incubation. Therefore, cleavage of the benzimidazole ring is probably a matter of cometabolism, n-butylamine which originates from the butylcarbamoyl side chain serving as the proper source of carbon.Besides radiolabelled 2-AB and CO₂, an unknown metabolite was isolated which showed characteristics of a 2-AB-nucleotide. Probably, 2-AB was incorporated into bacterial DNA, which upon lysis of the bacterial cells gave rise to the nucleotide in question. Therefore, 2-AB might exert its inhibitory action by interfering with the normal functioning of DNA.     

The active species of 'CO2' utilized by reduced ferredoxin:CO2 oxidoreductase from Clostridium pasteurianum.

Reduced ferredoxin:CO2 oxidoreductase (CO2 reductase) from Clostridium pasteurianum catalyzes the reduction of 'CO2' to formate with reduced ferredoxin, an isotopic exchange between 'CO2' and formate in the absence of ferredoxin, and the oxidation of formate to 'CO2' with oxidized ferredoxin. The active species of 'CO2', i.e. CO2 or HCO3 (H2CO3), utilized by the enzyme was determined. The method employed for the species identification was that of Copper et al. (1968). Both 'CO2' reduction to formate and the exchange reaction were studied. Data were obtained which are compatible with those expected if CO2 is the active species. The V and the dissociation constant Ks of the enzyme - CO2 complex in dependence of pH were determined from initial velocity studies of the exchange reaction. V was found to be only slightly affected by pH between 5.5 and 7.5. Ks was markedly dependent on pH; the constant increased with decreasing pH from 0.2 mM at pH 7.5 to 3 mM at pH 5.5.     

Postischaemic circulation disturbances.

Restoration of blood supply after ischaemic conditions in extremities and testes is inhibited by reversible intravasal aggregation of erythrocytes. This process is promoted by the increased permeability of the capillaries associated with the formation of oedema and the entailing increase of the haematocrit. For overcoming the stasis the increased structural viscosity caused by the aggregation of erythrocytes requires an increase in pressure as a starter effect which is not achieved by the flow pressure at once everywhere. Intravenously administered particles of Indian ink mark the formation and dissolution of aggregates. Even areas with originally normal blood supply may be obstructed by the later formation of aggregates. Thrombi on the walls of arterial and venous vessels and other lesions of the intima do not sufficiently explain the disturbance of perfusion. Oedema and extravasating leucocytes are found in the microcirculation. The parenchyma to be supplied shows formation of necrosis.     

Downstream bioprocessing of human pluripotent stem cell‐derived therapeutics

With the advancement in lineage‐specific differentiation from human pluripotent stem cells (hPSCs), downstream cell separation has now become a critical step to produce hPSC‐derived products. Since differentiation procedures usually result in a heterogeneous cell population, cell separation needs to be performed either to enrich the desired cell population or remove the undesired cell population. This article summarizes recent advances in separation processes for hPSC‐derived cells, including the standard separation technologies, such as magnetic‐activated cell sorting, as well as the novel separation strategies, such as those based on adhesion strength and metabolic flux. Specifically, the downstream bioprocessing flow and the identification of surface markers for various cell lineages are discussed. While challenges remain for large‐scale downstream bioprocessing of hPSC‐derived cells, the rational quality‐by‐design approach should be implemented to enhance the understanding of the relationship between process and the product and to ensure the safety of the produced cells.     

Purification of alpha-toxin from Staphylococcus aureus and application to cell permeabilization

Crude alpha-toxin was produced by Staphylococcus aureus, strain Wood 46. The amount of exotoxin was monitored during growth and all subsequent purification steps by determination of its hemolytic activity against rabbit erythrocytes. The culture supernatant was treated with ammonium sulfate (75% saturation). The resulting precipitate was dialyzed and subjected to cation-exchange chromatography. The fractions containing the hemolytic activity were further purified by gel chromatography. The final product was enriched by a factor of 8.5 compared to the crude toxin. In sodium dodecyl sulfate-polyacrylamide gel electrophoresis the purified toxin exhibited one major band. It caused the release of 86Rb+ and ATP from rat insulinoma (RIN A2) as well as pheochromocytoma cells (PC12) in culture, indicating efficient permeabilization of their plasma membranes for small molecules.     

Response of rats to the presence of stressed conspecifics as a function of time of day

The physiological response of nonrestrained rats to the presence of immobilized conspecifics during the beginning of the active period and the inactive period was studied. In immobilized animals concentrations of serum corticosterone (SCS), serum glucose, and liver glycogen, and the activity of liver tyrosine aminotransferase (TAT) during both the active and the inactive periods, were consistent with earlier studies. In nonrestrained rats the presence of immobilized conspecifics induced a significant increase in SCS during the active period, whereas it had no effect during the inactive period. The level of TAT was significantly elevated in the nonrestrained rats only during the inactive period and remained unchanged during the active period. The results demonstrate a physiological influence of stressed rats on unstressed conspecifics and provide evidence for regulation of TAT activity that is dependent on the situation and the time of day.     

Preferential release of oxytocin in response to vasoactive intestinal polypeptide in rats

Vasoactive intestinal polypeptide (VIP) was infused into the aorta of pentobarbitone-anesthetized rats (n = 12) in stepwise increasing doses of 0.001 to 10 micrograms/rat at rates varying from 0.3 pmol/min/kg to 3000 pmol/min/kg over 3 min. Blood was withdrawn from the vena cava inferior for the measurement of oxytocin (OT) and vasopressin (AVP) by RIA. The loss of blood was compensated for by infusion of isotonic saline (0.9% NaCl with 0.5% human serum albumin). Control rats received this solution only (n = 11). VIP infusions resulted in a dose-dependent increase in plasma OT which was significantly greater than the slight rise observed in the controls. The difference from controls was significant at infusion rates of 3 pmol/min/kg and more. Plasma AVP, on the other hand, did not rise in response to VIP infusions until the infusion rate was increased to 300 and 3000 pmol/min/kg. At these infusion rates, the increments in AVP were much smaller than those of OT, the levels during the highest infusion rates rising to 8.6 +/- 2.8 and 27.2 +/- 4.8 microU/ml, respectively (log normal means). The preferential release of OT in response to exogenous VIP in rats differs from the response in cats where intracarotid administration of VIP resulted in the release of proportionately more AVP than OT. Immunoreactive VIP is found in the hypothalamo-neurohypophyseal system of rats in close proximity of some of the magnocellular neurons as well as within the nerve terminals. This, together with our data, suggests that endogenous VIP may participate in the release mechanism for OT in rats.