Crystallization and preliminary X-ray diffraction analysis of the light-harvesting protein phycocyanin from the thermophilic cyanobacterium Synechococcus elongatus.

The crystallization and preliminary crystallographic study of phycocyanin from the thermophilic cyanobacterium Synechococcus elongatus is reported. Phycocyanin is composed of alpha- and beta-subunits consisting of 162 and 172 amino-acid residues, respectively. These associate to form an alphabeta heterodimer, which further associates to give a ring-shaped trimer (alphabeta)(3). Two trimers bind head-to-head to form a hexamer (alphabeta)(6). Phycocyanin crystals have been obtained by the sitting-drop vapour-diffusion method with a precipitant solution containing 30%(w/v) PEG 4000 and 100 mM MES pH 7.5-8.0. Using synchrotron radiation, the crystals diffract to 2.0 A resolution. They belong to the trigonal space group R32, with unit-cell parameters a = b = 186.75 (3), c = 59.75 (4) A, alpha = beta = 90, gamma = 120 degrees. Assuming that the crystallographic triad is identical to the threefold axis of the hexamer and with three (alphabeta)(6) molecules in a unit cell, the calculated molar volume (V(M)) is 2.64 A(3) Da(-1). This value corresponds to a solvent content of approximately 53%, with one alphabeta heterodimer occupying the asymmetric unit.

Effect of Mao-bushi-saishin-to on infection of ofloxacin resistant bacteria and of unknown origin.

When seven elderly patients with fever due to bacterial infection failed to respond to ofloxacin 300 mg/day for 3 days, we prescribed Mao-bushi-saishin-to for 7 days. Bushi, one of the components of Mao-bushi-saishin-to, is a complex of various alkaloids, which is known to stimulate the conducting system of the heart, so half of the standard dose was prescribed for these patients. We assessed changes in the patients' body temperatures and serum C-reactive protein concentrations. As a result, the patients' body temperatures all fell to under 37 degrees C and C-reactive protein levels decreased from an average of 3.06 mg/dl to an average of 0.36 mg/dl (P< 0.02). In cases where a fever does not decrease through dosage of an antibiotic such as ofloxacin after about 3 days, half of the standard dose of Mao-bushi-saishin-to for 7 days should be considered as a complementary therapy.

Decrease in serum cortisol during yoga exercise is correlated with alpha wave activation.

We examined changes in brain waves and blood levels of serum cortisol during yoga exercise in 7 yoga instructors and found that alpha waves increased and serum cortisol decreased. These two measures were negatively correlated (r = -.83). Comparison with a control group of nonpractitioners is desirable.

Improvement of C-reactive protein levels and body temperature of an elderly patient infected with Pseudomonas aeruginosa on treatment with Mao-bushi-saishin-to.

OBJECTIVE: To examine the effectiveness of Mao-bushi-saishin-to (Ma-Huang-Fu-Zi-Xi-Xin-Tang in Chinese medicine) (Tochimototenkaido Co. Ltd., Osaka, Japan), one of the traditional herbal medicines, against resistant bacterial infection. SETTING: The Nursing Center Himawari, Izumo, Japan DESIGN, PATIENT, AND PREPARATION: Half of the standard dose of Mao-bushi-saishin-to was prescribed for 7 days to one elderly patient with fever and positive C-reactive protein (CRP) levels suffering from drug resistant Pseudomonas aeruginosa. The daily standard dose of Mao-bushi-saishin-to is prepared from 1200 mg of dried extract obtained from three crude drugs, Ephedrae Herba (4 g), Asiasari Radix (3 g), and Aconiti Tuber (1 g). It is certified by the Japanese Ministry of Health and Welfare. RESULTS: The patient's fever and CRP level returned to normal levels. CONCLUSIONS: In cases in which the fever does not fall in response to antibiotics for at least 3 days, half of the standard dose of Mao-bushi-saishin-to for 7 days might be worth trying to induce remission, especially for elder patients.

Use of photic feedback as an adjunct treatment in a case of Miller Fisher syndrome.

Photic Feedback treatment of a patient diagnosed with Miller Fisher syndrome has resulted in the rapid and permanent remission of symptoms. During Photic Feedback treatment, the CD20 appeared to be slightly increased. This may have been associated with changes in humoral immunity. The present clinical observation of a single patient suggests that Photic Feedback treatment should be investigated as a possible adjunct therapy for patients who suffer from polyneuropathies, such as Miller Fisher syndrome, within a carefully controlled clinical trial.

Bile and pancreatic juice replacement ameliorates early ligation-induced acute pancreatitis in rats.

BACKGROUND: In healthy rats, combined bile and pancreatic juice diversion from gut has a synergistic rather than additive effect on stimulation of exocrine pancreatic protein secretion. We hypothesized that exclusion of combined bile and pancreatic juice from gut exacerbates bile and pancreatic-duct ligation-induced acute pancreatitis in rats to a greater extent than exclusion of either bile or pancreatic juice alone. METHODS: Bile and pancreatic juice (obtained fresh from donor rats) were replaced, separately or together, via a duodenal fistula beginning immediately before 6 hours of duct ligation. Pancreatic morphologic changes were evaluated with an acute pancreatitis histology score and morphometric quantitation of acinar-cell necrosis. Plasma amylase and cholecystokinin concentrations and pancreatic subcellular distribution of cathepsin B activity were determined. Characteristics of bile and pancreatic juice obtained from donor rats were also studied. RESULTS: Combined bile and pancreatic juice replacement limited the increase in acute pancreatitis histology score by 77%, acinar cell necrosis by 95%, hyperamylasemia by 77%, and hypercholecystokininemia by 99%, while preventing subcellular redistribution of cathepsin B. Amelioration of pancreatic morphologic changes was significantly greater with combined bile and pancreatic juice replacement than with replacement of either bile or pancreatic juice alone. CONCLUSION: In this experimental corollary of early gallstone-induced acute pancreatitis, combined bile and pancreatic juice exclusion from gut contributes to disease pathogenesis to a greater extent than exclusion of either bile or pancreatic juice alone.

Ligation-induced acute pancreatitis in rats and opossums: a comparative morphologic study of the early phase.

Bile and pancreatic duct ligation (BPDL) in rats does not induce severe acute pancreatitis but only mild inflammation, which is self-limiting and eventually leads to pancreatic atrophy. However, BPDL in opossums induces severe acute necrotizing pancreatitis which uniformly leads to death within 14 days. We compared pancreatic morphologic changes after 24 hr of BPDL in rats and opossums. Pancreatitis histology score and acinar cell ultrastructural changes were evaluated. In both species, BPDL was associated with significant increases in histology score compared to sham controls (5.0 +/- 0.3 vs 1.5 +/- 0.3 in rats, 5.3 +/- 0.4 vs 1.1 +/- 0.1 in opossums; mean +/- SEM, ANOVA, P < 0.05). However, there was no significant difference in histology score between rats and opossums following BPDL; histologic changes, such as white blood cell infiltration, acinar cell vacuolation, and focal acinar cell necrosis, were similar. Acinar cell ultrastructural changes after BPDL in both species included dilated endoplasmic reticulum and autophagic vacuole formation. These findings indicate that the early morphologic changes after BPDL in rats are quite similar to those seen early in the course of BPDL-induced acute necrotizing pancreatitis in opossums. As the rat is a more economical and convenient model to study than the opossum, this study supports the use of the rat model to conduct pilot studies of early events in the development of BPDL-induced acute pancreatitis. This study also suggests the potential for investigating mechanisms that may be present in the rat which protect against progressive and fatal acute necrotizing pancreatitis as observed in opossums after longer periods of BPDL.

A new model for study of pancreatic exocrine secretion: the tethered pancreatic fistula rat.

Diversion and recirculation of bile and pancreatic juice in rats are done in studies of pancreatic exocrine secretion. Previously, the modified Bollman cage was used to restrain rats with bile and pancreatic fistulas. To mimic physiologic conditions as closely as possible and to develop a more humane model, we designed a partial-restraint tethering system to study pancreatic exocrine secretion. Eight rats were prepared with biliary pancreatic, and duodenal fistulas, of which five were given enteral supplements via a gastric fistula and three were given saline supplements via a jugular venous line. Catheters exited at the nape of the neck and passed through the hollow of a cable coil that tethered the rat. On the fourth postoperative day pancreatic juice flow and protein output were studied. The tethering system allowed grooming, feeding, and ample mobility. This model of the tethered pancreatic fistula rat is a more human model for studies of pancreatic exocrine secretion in conscious rats, compared with the Bollman cage system of near total restraint.

Octreotide and cholecystokinin antagonist reduce edema in obstruction-induced acute pancreatitis.

Obstruction-induced acute pancreatitis in rats is associated with increased plasma cholecystokinin (CCK) levels. Duodenal replacement of bile reduces severity of pancreatitis and limits CCK increase. We investigated the role of CCK in the pathogenesis of obstruction-induced acute pancreatitis by pretreating rats with the somatostatin analog octreotide and the CCK antagonist L-364,718. Octreotide inhibits duodenal CCK release, and L-364,718 competitively blocks CCK receptors. We studied 31 rats after (1) sham operation (n = 7), (2) bile and pancreatic duct obstruction (BPDO) (n = 12), (3) BPDO plus octreotide (20 micrograms/kg IP and then 5 micrograms/kg/hr IV) (n = 6), and (4) BPDO plus L-364,718 (1 mg/kg IP and then 0.25 mg/kg/hr IV) (n = 6). Rats were killed after 18 hours. Pancreas weight, acute pancreatitis histology score, and plasma amylase and CCK levels were determined. Octreotide and L-364,718 limited the increase in pancreas weight. Octreotide also limited the rise in plasma CCK levels. These findings suggest that CCK may play a role in the pathogenesis of obstruction-induced acute pancreatitis.

Increased circulating cholecystokinin in obstruction-induced acute pancreatitis. II. Pancreatic duct obstruction with and without bile duct obstruction.

Pancreatic exocrine stimulation by cholecystokinin (CCK) has been implicated in the pathogenesis of experimental acute pancreatitis. Bile exclusion from the gut stimulates duodenal CCK release and exacerbates obstruction-induced acute pancreatitis. Pancreatic and bile duct obstruction increases circulating CCK concentration. We hypothesized that acute pancreatitis induced by pancreatic and bile duct obstruction would be ameliorated when bile was returned to the duodenum. As many small pancreatic ducts drain into the bile duct in rats, preservation of bile flow required the use of a bile shunt. We studied acute pancreatitis and the time course of circulating CCK increase in three groups of rats after: (1) sham operation (dissection, no obstruction), (2) bile and pancreatic duct obstruction, and (3) bile and pancreatic duct obstruction with bile shunt. The rats were killed at 3-, 6-, and 18-hr intervals after operation. Their blood was collected for measurement of CCK, amylase, and bilirubin concentrations. The pancreata were excised, weighed, and processed for histological examination. The shunting of bile back to the duodenum ameliorated the acute pancreatitis along with a simultaneous limitation of the rise in CCK concentration. This suggests that bile duct obstruction, another form of bile exclusion, exacerbates pancreatic duct obstruction-induced acute pancreatitis. The elevation in CCK concentration showed an early peak indicating that the potential role of CCK in the pathogenesis of obstruction-induced acute pancreatitis is predominantly in the early phase of its development.

Increased circulating cholecystokinin in obstruction-induced acute pancreatitis. I. Bile duct obstruction with and without pancreatic duct obstruction.

Bile exclusion from the gut exacerbates pancreatic duct obstruction-induced acute pancreatitis. We hypothesized that obstruction-induced acute pancreatitis involves an increase in circulating cholecystokinin (CCK), as bile and pancreatic juice exclusion from the gut stimulates duodenal CCK release. We studied 54 rats after the following operations: (1) sham operation (n = 18), (2) hepatic bile duct obstruction alone (n = 18), (3) hepatic bile duct and common bile-pancreatic duct obstruction (n = 18). Rats recovered and were killed in subgroups of six rats each at 3, 6, and 18 hr after operation; blood was collected for measurement of plasma CCK and amylase concentrations. Each pancreas was excised, weighed, and processed for histological examination; an acute pancreatitis score was determined. Combined bile and pancreatic duct obstruction induced acute pancreatitis and was associated with a marked increase of circulating CCK concentration. Bile duct obstruction alone did not induce acute pancreatitis but was associated with an increase of circulating CCK of lower magnitude. The time course of circulating CCK increase showed an early peak. These findings support our hypothesis and suggest that CCK plays a role in the pathogenesis of obstruction-induced acute pancreatitis.

Investigation on the drying and decomposition of sodium oxalate.

The drying of sodium oxalate at various temperatures was investigated by the micro-determination of the residual water in the heated sample and of the carbon monoxide produced by thermal decomposition. Sodium oxalate heated for 2hr above 200 degrees and cooled contains less than 20 ppm of water, and may be used as a standard for titrimetry. The decomposition of sodium oxalate into sodium carbonate and carbon monoxide was investigated by non-aqueous titrimetric micro-determination of carbon monoxide. The decomposition begins at 290 degrees and heating between 200 degrees and 250 degrees is recommended for the dehydration of sodium oxalate. The decomposition is complete between 750 degrees and 800 degrees within 20 min and the sodium carbonate obtained begins to decompose at above 810 degrees .