SOME ULTRAVIOLET PHOTOMICROGRAPHS OF B. SUBTILIS

1. Type-specific antipneumococcus immunity has been induced in rabbits by immunization with antigen prepared by combining a specific derivative of the capsular polysaccharide of Type III Pneumococcus with globulin from horse serum. 2. Rabbits immunized with this antigen acquire active immunity against infection with virulent Type III pneumococci. 3. The sera of the immune rabbits contain type-specific antibodies which precipitate the Type III capsular polysaccharide, agglutinate Type III pneumococci, and specifically protect mice against Type III infection. 4. The experimental data are discussed with reference to: (1) the concurrence in the immune sera of type-specific antibodies for Pneumococcus and precipitins for horse globulin; (2) the determining influence of the capsular polysaccharide on the specificity of the antigen as a whole; (3) the unity of the type-specific precipitins, agglutinins, and protective antibodies induced by a single compo of the pneu mococcus in chemical union with an unrelated, animal protein.     

Thoracic causes of acute abdominal pain

The origin of abdominal pain may be extra-abdominal, caused by a thoracic illness. This article illustrates the various thoracic disorders that may present with acute abdominal pain. An erroneous focus on the abdomen alone can easily lead to misdiagnosis and incorrect treatment. In cases of unexplained acute abdominal pain, radiologists should be aware of also viewing beyond the borders of the abdomen. The key to most of these thoracic diagnoses is detection of pulmonary consolidation, pleural fluid or pericardial fluid.     

Exhaled carbon monoxide in mechanically ventilated critically ill patients: influence of inspired oxygen fraction

OBJECTIVE: To assess the feasibility of exhaled carbon monoxide (CO) measurements in mechanically ventilated critically ill adult patients and to determine the influence of inspired oxygen fraction on this measurement. DESIGN: Prospective physiologic study. SETTING: Medical ICU in a community hospital. PATIENTS: The study was performed on nine mechanically ventilated patients with varying diagnoses. MEASUREMENTS AND RESULTS: Carbon monoxide concentration was determined with an infrared CO analyzer on exhaled breath collected at the outlet of the ventilator. We assessed the stability of exhaled carbon monoxide concentration over a 4-hour period and determined its course during a 7-hour period after inspired oxygen fraction had been abruptly increased from baseline to 1. Carbon monoxide was detected in exhaled breath in each patient at a higher concentration than in inspired gas (0.64 +/- 0.1 ppm vs 0.25 ppm, approximately). Exhaled carbon monoxide did not vary during a 4-hour period in five hemodynamically stable patients. When inspired oxygen fraction was increased from baseline (0.52 +/- 0.04) to 1, exhaled carbon monoxide concentration increased abruptly from baseline (0.63 +/- 0.13 ppm) to a peak value of 1.54 +/- 0.16 ppm within 15 min and returned slowly to baseline values within 7 h. CONCLUSION: CO was easily detected in the exhaled breath of mechanically ventilated patients and CO lung excretion was markedly but transiently dependent on inspired oxygen fraction. Other studies are warranted in order to determine the different factors that might influence CO lung excretion in critically ill patients.     

Accurate assessment of in situ isometric contractile properties of hindlimb plantar and dorsal flexor muscle complex of intact mice

An isometric torque sensor for measuring in situ contractions of plantar or dorsal flexors of intact mouse hindlimb has been developed and evaluated. With this device, muscle torque can be accurately measured within the range of -14 mN.m to +14 mN.m. Special attention was paid to fixation of the mouse hindlimb to the measurement device. Halothane-anaesthetized Swiss wild-type mice were positioned on the thermostatic measurement platform, and fixated with a hip and foot fixation system. The novel fixation unit was evaluated by measuring knee and ankle displacements during a contraction. A mathematical muscle model was used to quantify the effects of these displacements on the contractile parameters. Measured ankle and knee displacement, due to non-absolute fixation. resulted in a calculated muscle fibre shortening of 2.5%. Simulations of a contraction with this degree of fibre shortening, using the mathematical muscle model, showed only minor effects on maximal torque generation and the temporal parameters (half-relaxation time and 10-50% rise time). Furthermore, we showed that muscle torque in our set-up is hardly affected by eccentricity between ankle and measurement axis. Measured tetanic muscle torques of intact dorsal and plantar flexors were 3.2+/-0.4 mN.m and 11.8+/-1.6 mN.m, respectively. The half-relaxation time of plantar flexors was significantly higher than that of dorsal flexors (12.9+/-2.7 ms versus 8.8+/-1.2 ms), whereas the 10-50% rise time was longer in plantar (14.9+/-0.6 ms) than in dorsal (11.8+/-2.0 ms) flexors.     

Prostaglandins are involved in acetylcholine- and 5-hydroxytryptamine-induced,nitric oxide-mediated vasodilatation in human forearm

Both acetylcholine (ACh) and 5-hydroxytryptamine (5HT) are used to examine nitric oxide (NO)-mediated vasodilatation in humans. Animal data suggest that both substances can also induce the release of prostacyclin (PGI ). This study was designed to investigate the role of the prostaglandin pathway in Ach- and 5HT-induced vasodilation in humans. The experiments were done in three groups of healthy male volunteers. In group 1 (n = 6), ACh (100-1,000 ng/kg/min) and sodium-nitroprusside (10-100 ng/kg/min) were infused into the brachial artery alone, together with a continuous infusion of indomethacin (1.3 micro g/kg/min) and during a combined infusion of indomethacin and the competitive NO synthase inhibitor N -monomethyl-l-arginine (l-NMMA; 30 micro g/kg/min). In group 2 (n = 5), 5HT (0.3-1.0 ng/kg/min) was infused alone and together with a continuous infusion of indomethacin and l-NMMA. In group 3 (n = 6), the synthetic prostaglandin analog iloprost (0.5-4.5 ng/kg/min) was infused together with a continuous infusion of saline, l-NMMA, and l-NMMA with indomethacin, respectively. The infusions of indomethacin and l-NMMA started 10 min before the infusion of ACh, 5HT, iloprost, and sodium nitroprusside. Forearm blood flow was measured using computerized venous occlusion plethysmography. Both the Ach- and 5HT-induced vasodilator responses were significantly attenuated by indomethacin (p < 0.05 for both), but not further influenced by a concomitant infusion of l-NMMA. The vasodilatation induced by iloprost was significantly inhibited by l-NMMA (p < 0.05) and not affected by indomethacin. The sodium nitroprusside-induced vasodilation was influenced by neither l-NMMA nor indomethacin. It is concluded that in the human forearm, the prostaglandin pathway is involved in both the Ach- and 5HT-induced NO-mediated vasodilatation.     

Mutations inducing divergent shifts of constitutive activity reveal different modes of binding among catecholamine analogues to the beta(2)-adrenergic receptor

1. We compared the changes in binding energy generated by two mutations that shift in divergent directions the constitutive activity of the human beta(2) adrenergic receptor (beta(2)AR). 2. A constitutively activating mutant (CAM) and the double alanine replacement (AA mutant) of catechol-binding serines (S204A, S207A) in helix 5 were stably expressed in CHO cell lines, and used to measure the binding affinities of more than 40 adrenergic ligands. Moreover, the efficacy of the same group of compounds was determined as intrinsic activity for maximal adenylyl cyclase stimulation in wild-type beta(2)AR. 3. Although the two mutations had opposite effects on ligand affinity, the extents of change were in both cases largely correlated with the degree of ligand efficacy. This was particularly evident if the extra loss of binding energy due to hydrogen bond deletion in the AA mutant was taken into account. Thus the data demonstrate that there is an overall linkage between the configuration of the binding pocket and the intrinsic equilibrium between active and inactive receptor forms. 4. We also found that AA mutation-induced affinity changes for catecholamine congeners gradually lacking ethanolamine substituents were linearly correlated to the loss of affinity that such modifications of the ligand cause for wild-type receptor. This indicates that the strength of bonds between catechol ring and helix 5 is critically dependent on the rest of interactions of the beta-ethanolamine tail with other residues of the beta(2)-AR binding pocket.     

5'-Deoxy congeners of 9-(3-amido-3-deoxy-beta-D-xylofuranosyl)-N(6)-cyclopentyladenine: new adenosine A(1) receptor antagonists and inverse agonists

The synthesis and structure-activity relationship of N(6)-cyclopentyl-3'-substituted-xylofuranosyladenosine analogues with respect to various adenosine receptors were explored in order to identify selective and potent antagonists and inverse agonists for the adenosine A(1) receptor. In particular, the effects of removal of the 5'-OH group and introduction of selected substituents at the 3'-NH(2) position of 9-(3-amino-3-deoxy-beta-D-xylofuranosyl)-N(6)-cyclopentyladenine were probed. A solid phase-assisted synthetic approach was used to optimize the 3'-amide functionality. In view of the general concern of the presence of a 5'-OH moiety with regard to cellular toxicity, the present study describes 5'-deoxy compounds with reasonable affinity for the human adenosine A(1) receptor. Interestingly, this study shows that optimization of the 3'-"up" amide substituent can substantially compensate for the drop in affinity for the adenosine A(1) receptor, which is generally observed upon removal of the 5'-OH group. The fact that for several 3'-amido-substituted (5'-deoxy)-N(6)-cyclopentyladenosine derivatives, guanosine 5'-triphosphate-induced shifts in K(i) values were significantly lower than 1 implies that these analogues behave as inverse agonists. This is further supported by their 1,3-dipropyl-8-cyclopentylxanthine-like capacity to increase forskolin-induced adenosine cyclic 3',5'-phosphate production.