Differentiation of Bacteroides from Sphaerophorus and Fusobacterium

This paper is concerned with new findings in the comparative study of the genera, Bacteroides, Sphaerophorus and Fusobacterium. Victoria blue 4R and barbiturates were found to be effective in differentiating the genera by their discriminative inhibition of the growth. A new character, glutamic acid decarboxylation, was found in Bacteroides but neither in Sphaerophorus nor in Fusobacterium. The procedure for this test was presented. In the classification of newly isolated fecal strains this test and other differential features were compared. An improved method for the threonine deamination test was reported. The use of these characters and tests would be a step in the improvement of the present incomplete classification of gram-negative anaerobic non-sporulating rods.     

Increase of Catecholamines in Mouse Brain by Systemic Administration of γ-Glutamyl L-3,4-Dihydroxyphenylalanine

We investigated the effect of systemic administration of gamma-glutamyl L-3,4-dihydroxyphenylalanine (gamma-Glu-DOPA) on catecholamine contents in the brain. gamma-Glu-DOPA was transformed to dopamine (DA) in vitro with brain homogenate by the sequential action of gamma-glutamyl transpeptidase and aromatic L-amino acid decarboxylase. Intraperitoneal injection of gamma-Glu-DOPA to mice increased DA markedly and noradrenaline (NA) moderately in the brain. The increase of endogenous DA was followed by elevation of the main DA metabolites (3,4-dihydroxyphenyl-acetic acid and homovanillic acid). These increases were in a dose-dependent manner. The maximal elevation of DA was observed within 30 min after administration of gamma-Glu-DOPA, but a substantial increase of NA was observed 2 h after the administration. These results suggest that gamma-Glu-DOPA may be applicable to the treatment of Parkinson's disease.     

Simple purification of aromatic L-amino acid decarboxylase from human pheochromocytoma using high-performance liquid chromatography

We purified aromatic L-amino acid decarboxylase (AADC) homogeneously and rapidly from human pheochromocytoma using high-performance liquid chromatography. HPLC with gel permeation and hydrophobic columns was highly effective, and the entire purification could be finished within 3 days. Purified AADC showed a single band with an Mr of 50,000 on sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and decarboxylated L-3,4-dihydroxyphenylalanine, L-5-hydroxytryptophan, and L-threo-3,4-dihydroxyphenylserine (a synthetic precursor of natural norepinephrine). Amino acid analysis of purified AADC was performed.     

Variance component analysis of bristle characters in local populations of Drosophila melanogaster.

The genetic variabilities of sternopleural and abdominal bristle numbers existing in local natural populations were assessed. Using second chromosome lines of Drosophila melanogaster extracted from three natural populations in Japan (the Ishigakijima, Ogasawara and Aomori populations), experiments were conducted to estimate the components of genetic variances, additive and dominance variances. The following results were obtained: For both sternopleural and abdominal bristle numbers, the additive genetic variances (sigma 2A) were much larger than the dominance variances (sigma 2D) especially in the southern populations. For example, in the Ishigakijima population, for females sternopleural bristle numbers of the inversion-free chromosome group, the additive and dominance variances were estimated to be 1.255 +/- 0.2034 and 0.0552 +/- 0.0180, respectively. The magnitudes of the estimates of additive genetic variances were nearly equal from north to south. By comparing the additive genetic variances of the inversion-free chromosome group with those of the In(2L)t-carrying chromosome group, it was inferred that sufficient number of generations to achieve the equilibrium state has not passed since the introduction of a single or a small number of the In(2L)t-carrying chromosomes to the Ishigakijima population.     

NOS3 deficiency augments hypoxic pulmonary vasoconstriction and enhances systemic oxygenation during one-lung ventilation in mice.

Nitric oxide (NO), synthesized by NO synthases (NOS), plays a pivotal role in regulation of pulmonary vascular tone. To examine the role of endothelial NOS (NOS3) in hypoxic pulmonary vasoconstriction (HPV), we measured left lung pulmonary vascular resistance (LPVR), intrapulmonary shunting, and arterial PO2 (PaO2) before and during left mainstem bronchus occlusion (LMBO) in mice with and without a deletion of the gene encoding NOS3. The increase of LPVR induced by LMBO was greater in NOS3-deficient mice than in wild-type mice (151 +/- 39% vs. 109 +/- 36%, mean +/- SD; P < 0.05). NOS3-deficient mice had a lower intrapulmonary shunt fraction than wild-type mice (17.1 +/- 3.6% vs. 21.7 +/- 2.4%, P < 0.05) during LMBO. Both real-time PaO2 monitoring with an intra-arterial probe and arterial blood-gas analysis during LMBO showed higher PaO2 in NOS3-deficient mice than in wild-type mice (P < 0.05). Inhibition of all three NOS isoforms with Nomega-nitro-L-arginine methyl ester (L-NAME) augmented the increase of LPVR induced by LMBO in wild-type mice (183 +/- 67% in L-NAME treated vs. 109 +/- 36% in saline treated, P < 0.01) but not in NOS3-deficient mice. Similarly, systemic oxygenation during one-lung ventilation was augmented by L-NAME in wild-type mice but not in NOS3-deficient mice. These findings indicate that NO derived from NOS3 modulates HPV in vivo and that inhibition of NOS3 improves systemic oxygenation during acute unilateral lung hypoxia.     

Removal by chemicals of photoperiodic light requirements of Lemna gibba G3

Both the initial and the terminal 1 hr portions of the subjectiveday fraction, namely the L1- arid L2-phases, of a 24 hr daymust be illuminated in order for the day to be perceived asa long day in the min-LD determination by the long-day plant,Lemna gibba G3 (9). The light requirement of the L1-phase wassatisfied by a 10 min red light pulse given at the beginningof the phase. The red light effect was erased by a subsequent10 min far-red light, indicating phytochrome-mediated processesoccurring in the L1-phase. The light requirement of the L2-phasewas satisfied by blue or far-red light given during the terminal10 min period of the phase; there was no indication of phytochromeinvolvement. The light action on the L1-phase was replaced by10^–5 M of cyclic AMP or 10^–7 M of DL-isoproterenol.The isoproterenol action was antagonized by 10^–7 M ofDL-propranolol. Cyclic AMP (10^–5 M) combined with salicylicacid (10^–6 M), which can remove the light requirementof the L2-phase (10), rendered a completely dark day physiologicallyequivalent to a long day. Acetylcholine (10^–5 M) exertednyctomimetic action on the L1-phase of the second light day.The action of acetylcholine was antagonized by cyclic AMP (10^–5M). The L2-phase required no light in the presence of 10^–7M of DL-propranolol, and this propranolol action was not affectedby isoproterenol. These findings suggest changes in membranepermeability caused by the light given during the L1- and L2-phases. (Received July 7, 1976; )     

Characterization of three monoclonal antibodies which induce and modulate superoxide anion generation in guinea pig macrophages

Three monoclonal antibodies (Ab), termed KY 12, KY 22, and KY 25 and raised against guinea pig macrophages, induced superoxide anion (O2-) generation in the cells. Although each monoclonal Ab bound to macrophages, each had a different pattern of binding to other cell types. In response to each of the Ab, the amount of O2- generated by 5 X 10(5) macrophages was between 0.5 and 0.7 nmol/min and was augmented threefold to fivefold by the addition of F(ab')2 fragments of rabbit Abs to mouse Ig. When macrophages were pretreated with soluble immune complexes (I.C.) prior to stimulation by the monoclonal Ab, the O2- generation stimulated by KY 12 or KY 22 was reduced by more than 70%. In contrast, pretreatment of macrophages with I.C. did not reduce O2- generation in response to KY 25. KY 12 and KY 22 stimulated adenyl cyclase activity in macrophages, but KY 25 did not. Pretreatment of the cells with soluble I.C. did not interfere with the enhancing effect of the monoclonal Ab on adenyl cyclase activity. Pretreatment of macrophages with KY 12 reduced by over 60% of subsequent generation of O2- in response to wheat germ agglutinin, I.C., formyl-methionyl-leucyl-phenylalanine, phorbol myristate acetate, KY 22, or KY 25. KY 22 or KY 25 did not suppress the generation of O2- in response to other stimuli. These results suggest that KY 22 and KY 25 activate O2- generation in a manner that differs from that of KY 12. These monoclonal Ab should prove useful in examining the regulation of O2- production.     

The cyclic GMP-induced inward current in neuron R14 of Aplysia californica: similarity to a FMRFamide-induced inward current

Injecting cGMP into Aplysia neuron R14 induced an inward current similar to one elicited by application of FMRFamide to the outside of that cell. In contrast, injection of cAMP into R14 caused a long-lasting outward current and conductance increase. Phosphodiesterase inhibitors increased the cGMP and FMRFamide-induced inward currents in R14. The cGMP-induced inward current is voltage dependent and is largely carried by Na+. It is also strongly and inversely dependent on both external [Ca2+] and [Cl-], although these ions are not significant current carriers. Changing external [K+] had no effect. Voltage and ion dependencies of the cGMP-induced inward current are similar to those of an inward current induced by FMRFamide. Thus cGMP may be a second messenger to FMRFamide in producing a slow inward current in R14. cGMP does not appear to be a second messenger to FMRFamide in most Aplysia neurons.