Legionella pneumonia due to exposure to 24-hour bath water contaminated by Legionella pneumophila serogroup-5

A 79-year-old man was admitted to hospital from his nursing home for treatment of pneumonia, but died 7 days after admission. Legionella pneumonia was diagnosed after isolation of Legionella pneumophila serogroup-5 from sputum culture. The environment of the nursing home was investigated; only water specimens from the 24-hour bath were positive by culture for Legionella pneumophila serogroup-5. Subsequent analysis by pulsed-field gel electrophoresis revealed an identical pattern in isolates from both sputum culture and 24 hour bath water culture. Among 123 inpatients and staff of the nursing home, 17 were found to be seropositive for Legionella pneumophila serogroup-5.     

Mass spectrometric identification of citrullination sites and immunohistochemical detection of citrullinated glial fibrillary acidic protein in Alzheimer's disease brains

Peptidylarginine deiminases (PADs) are posttranslational modification enzymes that convert protein arginine to citrulline residues in a calcium ion‐dependent manner. Previously, we reported the abnormal accumulation of citrullinated proteins and the increase in the amount of PAD2 in hippocampi from Alzheimer's disease (AD) patients. Moreover, glial fibrillary acidic protein (GFAP), an astrocyte‐specific marker protein, and vimentin were identified as citrullinated proteins by using two‐dimensional gel electrophoresis and MALDI‐TOF mass spectrometry. To clarify the substrate specificity of PADs against GFAP, we prepared recombinant human (rh)PAD1, rhPAD2, rhPAD3, rhPAD4, and rhGFAP. After incubation of rhGFAP with rhPAD1, rhPAD2, rhPAD3, and rhPAD4, citrullinated (cit‐)rhGFAP was detected by Western blotting. The citrullination of rhGFAP by rhPAD2 was unique, specific, and time dependent; additionally, rhPAD1 slightly citrullinated rhGFAP. We then generated eight anti‐cit‐rhGFAP monoclonal antibodies, CTGF‐125, ?128, ?129, ?1212, ?1213, ?1221, ?122R, and ?1224R, which reacted specifically with cit‐rhGFAP. Two of those eight monoclonal antibodies, CTGF‐122R and ?1224R, reacted with both cit‐rhGFAP and rhGFAP in Western blots. By using the CTGF‐1221 antibody and a tandem mass spectrometer, we identified the two independent citrullination sites (R270Cit and R416Cit) of cit‐rhGFAP. Immunohistochemical analysis with CTGF‐1221 antibody revealed cit‐GFAP staining in the hippocampus of AD brain, and the cit‐GFAP‐positive cells appeared to be astrocyte‐like cells. These collective results strongly suggest that PAD2 is responsible for the citrullination of GFAP in the progression of AD and that the monoclonal antibody CTGF‐1221, reacting with cit‐GFAP at R270Cit and R416Cit, is useful for immunohistochemical investigation of AD brains. © 2015 Wiley Periodicals, Inc.     

Bioavailability of vitamin C from mashed potatoes and potato chips after oral administration in healthy Japanese men

Potato (Solanum tuberosum) tubers contain vitamin C (VC) and commercial potato chips have more VC content per wet weight by dehydration during frying. However, intestinal absorption of VC from orally ingested potatoes and its transfer to the blood remains questionable. The present study was designed to determine whether the dietary consumption of potatoes affects VC concentration in plasma and urinary excretion of VC in human subjects. After overnight fasting, five healthy Japanese men between 22 and 27 years of age consumed 87 g mashed potatoes and 282 g potato chips. Each portion contained 50 mg of VC, 50 mg VC in mineral water and mineral water. Before and after a single episode of ingestion, blood and urine samples were collected every 30 min or 1 h for 8 h. When measured by subtraction of the initial baseline value before administration of potatoes from the values measured throughout the 8 h test period, plasma VC concentrations increased almost linearly up to 3 h. Subsequently, the values of potato-fed subjects were higher than those of water, but did not differ significantly from those of VC in water (P = 0·14 and P = 0·5). Less VC tended to be excreted in urine during the 8 h test than VC in water alone (17·0 (sem 7·5) and 25·9 (sem 8·8) v. 47·9 (sem 17·9) μmol/mmol creatinine). Upon human consumption, mashed potatoes and potato chips provide VC content that is effectively absorbed in the intestine and transferred to the blood. Clearly, potatoes are a readily available source of dietary VC.     

Antihypertensive effect of cardiovascular Ca2+-antagonist in hypertensive patients in the absence and presence of beta-adrenergic blockade

The effect of the administration of a cardiovascular Ca²⁺-antagonist (nifedipine) on arterial blood pressure, heart rate, and plasma renin activity was investigated.Blood pressure of normotensive healthy volunteers (n = 4) does not change significantly by administration of 10 mg. (from $\text{119}\text{78}\text{to}\text{120}\text{75}\text{mm. Hg}$) and 30 mg. (from $\text{114}\text{69}\text{to}\text{105}\text{66}\text{mm. Hg}$) of nifedipine. When nifedipine (30 mg.) is administered with propranolol (beta-blockade), a slight decrease of systolic and diastolic blood pressure was observed (from $\text{119}\text{73}\text{to}\text{104}\text{67}\text{mm. Hg}$). Blood pressure of hypertensive patients is significantly lowered by 10 mg. of nifedipine from 172.9 to 130.3 mm. Hg (25 per cent reduction) systolic, and from 112.9 to 86.6 mm. Hg (23 per cent) diastolic (n = 7). Thirty mg. nifedipine had a slightly stronger hypotensive effect, namely 27 per cent reduction in systolic and 28 per cent in diastolic values (n = 9). Combined administration of nifedipine and propranolol, resulted in lowering initial blood pressure by 32 per cent and 30 per cent reduction in systolic and diastolic (n = 8), respectively.The heart rate of normotensive patients hardly changes with administration of 10 and 30 mg. of nifedipine and combined medication. But in hypertensive subjects it is significantly increased by nifedipine; from 66.3 to 80.3 (p < 0.001) by 10 mg., and from 70.0 to 82.2 beats/minute (p < 0.01) by 30 mg. On the contrary, combined administration of nifedipine and propranolol causes no increase or only a slight decrease in heart rate (from 68 to 66 beats/minute).Plasma renin activity of normotensive and hypertensive subjects is increased by 30 mg. of nifedipine. Combined administration of nifedipine and propranolol decreases plasma renin activity in both normotensive and hypertensive patients.The antihypertensive effect of nifedipine is enhanced and prolonged by propranolol. The observed increase in heart rate and plasma renin activity with nifedipine is inhibited by propranolol, probably by inhibiting the cardiovascular effects of the activity of the sympathetic nervous system.The results of this study indicate that oral administration of nifedipine is very effective in lowering arterial blood pressure in hypertensive patients, especially when combined with propranolol.Administration of nifedipine with beta-blockade resulted in satisfactory management of hypertension with minimal adverse drug reactions which could be possibly attributed to the preparation, especially in the management of hypertensive emergencies including hypertension associated with acute myocardial infarction and coronary insufficiency.     

Direct dissolution of gallstones with methyl tert-butyl ether (MTBE) via endoscopic transpapillary catheterization in the gallbladder (ETCG)

In a pilot study of direct dissolution therapy of gallstones with methyl tert-butyl ether (MTBE), endoscopic transpapillary catheterization in the gallbladder (ETCG) was performed. Complete dissolution was seen in 8 out of 12 (66%) patients and partial dissolution was seen in 2 (16%) patients. In one of the 8 complete dissolution patients, combined extracorporeal shock wave lithotripsy (ESWL) and dissolution therapy was carried out successfully. These 8 patients were followed up for 12–20 months with regular ultrasonography. During this period, 1 patient underwent laparoscopic cholecystectomy due to stone recurrence. Thickening of the gallbladder wall was seen in 2 patients, but there were no other complications. Using Tsuchiya's classification based on ultrasound, complete dissolution was seen in type Ia stones. This pilot study suggests that the direct dissolution of gallstones with MTBE via ETCG might be a useful and safe non-invasive treatment in patients with cholesterol stones in preserved gallbladders.     

Taurine conjugate of ursodeoxycholate plays a major role in the hepatoprotective effect against cholestasis induced by taurocheno-deoxycholate in rats

ABSTRACT— Rats which were taurine-deprived through ß-alanine administration and untreated rats were used to elucidate the mechanism of hepatoprotective effects of ursodeoxycholate (UDC). Animals were infused with taurochenodeoxycholate (TCDC, 0.4 μmol · min^-1 · 100 g^-1) alone or in combination with tauroursodeoxycholate (TUDC), or with UDC (both 0.6 μmol · min-1 · 100 g^-1) for 2 h. Ursodeoxycholate as well as TUDC prevented severe cholestasis and liver damage induced by TCDC infusion in both untreated and taurine-deprived rat groups. In untreated rats, however, UDC was less effective in hepatoprotection than TUDC as indicated by sequential changes in biliary LDH output during the period of 30 to 120 min (P<0.05). In rats receiving UDC and TCDC, total biliary output of LDH for 2 h was significantly higher in taurine-deprived rats than that in the control (73.40±10.10 vs 41.14±12.56: P < 0.05), suggesting that the difference became greater upon taurine deprivation. In contrast, in rats receiving TUDC and TCDC, the protective effect was comparable for the taurine-deprived and untreated rats. When the animals were infused with UDC and TCDC, taurine-deprived rats exhibited a biliary excretion rate for TUDC half that of control rats (P<0.05). Furthermore, a highly significant correlation was observed between the biliary excretion rate of TUDC and biliary output of LDH (r = –0.886, P<0.0001). These results suggest that UDC conjugates, especially TUDC, and not UDC may play a major role in the prevention of cholestasis and liver cell damage caused by TCDC infusion.     

Characterization of N-Linked Oligosaccharides Attached to Human Renin Expressed in COS Cells

ABSTRACT

To study the role of N-linked oligosaccharides attached to human renin, we generated three kinds of glycosylation-deficient renins in which one or both of two putative N-glycosylation sites was eliminated by amino acid replacement using site-directed mutagenesis. Examination of the three mutant renins (Asn-5 to Ala, Asn-75 to Ala, and both Asn-5 and -75 to Ala) expressed in COS cells demonstrated that both putative sites were certainly glycosylated with heterologous N-linked oligosaccharides. Moreover, the oligosaccharide chain attached at Asn-5 was different from that attached at Asn-75 in its molecular size. In addition, the secreted amount of the three mutant renins were different from one another, although the mutant and wild-type renins had practically the same specific activity. Our results suggest that the N-linked oligosaccharides have no effect on the enzymatic activity, but play an important role in stable secretion of human renin.     

Liquid formulation of a novel non-fluorinated topical quinolone, T-3912, utilizing the synergic solubilizing effect of the combined use of magnesium ions and hydroxypropyl-beta-cyclodextrin.

T3912 is a non-fluorinated topical quinolone antimicrobial agent currently under development. Its pK(a1) and pK(a2) values were determined as 5.6 and 7.5, respectively, and its intrinsic solubility at pH 7 was found to be approximately 2 microg/ml. This study evaluates the combined use of Mg(2+) ions, hydroxpyropyl-beta-cyclodextrin (HPbetaCD) and polyvinylpyrrolidone (PVP) in order to obtain a stable aqueous liquid formulation of T-3912. The use of Mg(2+) ions alone resulted in the improved solubility of T-3912 at physiological pH, however, it became unstable during storage. HPbetaCD increased T-3912 solubility with relatively high apparent association constant (K(1:1)=9.6 x 10(3) M(-1)) that was determined by a phase-solubility method at pH 7.5. Moreover, a binary system of Mg(2+) and HPbetaCD exerted a synergetic effect, such that the solubility of T-3912 increased a remarkable 500-fold. Furthermore, the addition of PVP prevented precipitation during storage, and as a result, the liquid formulation of T-3912 (1000 microg/ml) showed good stability under various conditions, i.e., in a refrigerator at 25 degrees C/60% RH and at 40 degrees C/75% RH for 6 months. The effect of light exposure of 1200000 lux.h was also tested. This combined system of Mg(2+) ions, HPbetaCD and PVP has potential as a liquid formulation of T-3912 for topical application, especially for opthalmic use.