Inhibition by Brefeldin A of the envelopment of nucleocapsids in herpes simplex virus type 1-infected Vero cells

Summary Inhibition by Brefeldin A (BFA) of the multiplication of herpes simplex virus (HSV) type 1 in Vero cells was characterized quantitatively. The yield of infectious progeny virus decreased exponentially with increasing concentrations of BFA while the yield of enveloped virus particles decreased less steeply to the level of approximately one fifth of the yield in the untreated cells; the level then remained constant even at higher BFA concentrations. The yield of nucleocapsids was not markedly affected by the drug. These results suggest that there are two different (i.e., BFA-sensitive and -insensitive) pathways for the formation of enveloped particles in the HSV-1-infected cells and that the infectious progeny virus arises exclusively from the BFA-sensitive pathway. Addition of BFA at various times after infection showed that the agent inhibited the increase in the amount of enveloped particles and of infectious progeny virus immediately after the addition. Single-step growth experiments suggested that, even in the presence of mature viral envelope proteins and of nucleocapsids, the increase in the amount of enveloped particles was completely inhibited by the addition of BFA at a late stage of infection. These results are consistent with the concept that the Golgi complex, the most BFA-sensitive organelle, is the major envelopment site of HSV-1 nucleocapsids leading to the formation of the infectious progeny virus.This work was presented in The 16th International Herpesvirus Workshop, Pacific Grove, California on July 7–12, 1991.     

Dose and Load Studies for Subcutaneous and Oral Delivery of Poly(lactide-co-glycolide) Microspheres Containing Ovalbumin

Poly(lactide-co-glycolide) microspheres containing different loads of OVA (0.05, 0.1, 0.5 and 1.0% w/w) were manufactured by a w/o/w emulsion/solvent evaporation method. Low load efficiencies of less than 20% were observed. Normal size distributions with mean volume diameters ranging from 3.7 to 4.7 µm were obtained for different batches. The in vitro release of OVA from different loaded microspheres showed an expected burst release with all batches. The in vivo dose study (1, 10, 25, 50 µg of OVA) was performed by subcutaneous and oral inoculation in mice by single (0 week) or double (0 and 3 weeks) administration of PLGA 50/50 microspheres containing 0.1% OVA. Subcutaneous administration showed an immune response (serum Ig levels by ELISA) statistically (Fishers paired t-test; P < 0.05) above OVA saline negative controls at 3, 6 and 12 weeks after administration. Oral administration of microspheres produced statistically higher systemic immune responses at the higher doses. Single and double inoculation orally and subcutaneously produced similar serum antibody levels. The in vivo load study was performed by subcutaneous and oral administration to mice of 25 µg OVA contained in various loaded (0.05, 0.1, 0.5 and 1.0% w/w) microspheres. Serum immune responses at 3, 6, and 12 weeks after inoculation were statistically above OVA saline controls and were inversely proportional to the OVA load using either route. This observation suggested a relationship between the number of microspheres delivered and the in vivo serum response. Single subcutaneous administration of 0.05 or 0.1% OVA loaded PLGA 50/50 microspheres induced larger immune responses compared with complete Freunds adjuvant.     

Identifying costs of intravenous solution wastage

A monitoring programme to systematically evaluate and decrease wastage of intravenous (i.v.) solutions was developed. Documentation of type and quantity of wasted solutions can help identify potential causes of waste. Subsequent systems changes intended to decrease waste can then be evaluated. Pharmacy technicians daily recorded the number, identity and locations of discarded i.v. solutions. A computer spreadsheet application was developed and used to produce a monthly summary identifying the number, drug cost and solutions wasted by hospital unit, thus establishing specific areas to target for i.v. wastage control. Investigators conducted a 12-month retrospective evaluation of i.v. wastage, revealing that over 14,000 i.v. preparations were destroyed at an estimated $US90,000 in drug costs. This equates to 2.5% of total drug costs wasted in small volume parenterals. Target programmes were implemented for drugs that were frequently wasted and systems changes implemented that decreased overall wastage of i.v. solutions.     

Senile plaques in an aged two-humped (Bactrian) camel (Camelus bactrianus)

Senile plaques with -protein as a major constituent are a conspicuous feature in the brains of aged humans, monkeys, dogs, and bears. We found cerebral senile plaques of the diffuse and primitive type, but not the classical type, in an aged female camel of more than 20 years old. The senile plaques and a few cortical capillaries were immunoreactive with anti--protein serum. Congophilic amyloid deposition was detected in a small number of the capillaries, but not in the senile plaques. We believe this to be the first detailed report of senile plaques in a herbivore, and these findings suggest the ossibility of senile plaque formation in a wide variety of mammlian species.     

Cohort study of the total leukocyte count and periodontal disease among company employees

There have been few epidemiological studies of the relationship between the leukocyte count and dental disease. In the present study, therefore, we investigated the relationship between oral health indicated by the Community Periodontal Index of Treatment Needs (CPITN) and the total leukocyte count in the cohort study. The 1,035 subjects were male factory workers employed at a chemical factory in Osaka, Japan. Their oral conditions were recorded as the CPITN score. The relationship between the total leukocyte count and the oral condition of the subjects classified according to their smoking habits was investigated over a 5-year period. Among the current smokers, the total leukocyte count was highest each year for the group with CPITN level 4, followed by those with CPITN levels 3 and 2 in descending order, showing that the total leukocyte count was reduced as the periodontal diseases ameliorated. Among the nonsmokers, the total leukocyte count was high every year in the group with CPITN levels 4 or 3 compared to that of the group with CPITN level 2. The fluctuations of the total leukocyte count in current smokers and nonsmokers whose CPITN score increase or decreased in five years was investigated. There were no significant differences among the all groups.     

Histological factors of 1-h biopsy influencing the delayed renal function and outcome in cadaveric renal allografts

The morphological characteristics of a kidney biopsy specimen taken 1 h after reperfusion of blood into the graft (1-h biopsy) during a cadaveric transplant operation were studied. The aim of the 1-h biopsy is to evaluate the pre-transplant risk factors for the delayed graft function, assess the renal function of the graft, and predict long-term graft survival. The total number of 1-h biopsies was 113, consisting of 86 male and 27 female donors. The mean age of the donors is 39.5 +/- 17.3 yr. Arteriosclerosis (AS) and tubulo-interstitial injury (TI) were both estimated using a semi-quantitative scale. AS score was graded into four categories, according to the severity of the thickening of interlobular artery: 0: none, 1: mild, 2: moderate and 3: severe. No biopsy revealed severe AS of grade 3. The TI score was graded from 0 to 5, according to the morphological injury: 0: none, 1-2 non-specific tubulo-interstitial injury (NSTI), and 3-5 compatible with acute tubular necrosis (ATN) in terms of pathological diagnosis. The mean ages of donors showing as AS of score 0, 1, and 2 were 30.6 +/- 14.6, 49.7 +/- 13.5, and 56.9 +/- 6.30, respectively. The mean donor age of the AS 1 group and AS 2 group was significantly lower than for the AS 0 group. The lowest serum creatinine values after operation (best Cr) of recipients with AS scores of 0, 1, and 2 were 1.31 +/- 0.45, 1.60 +/- 0.70, and 1.84 +/- 0.71 mg/dL, respectively; the best Cr of AS scores of 1 and 2 was significantly higher than in the AS score 0 group. The mean creatinine level at the final point of the AS 0 group was significantly lower than in the combined AS 1 and AS 2 group (serum creatinine 1.44 +/- 1.03 vs 1.87 +/- 1.53 mg/dL: p < 0.01). The duration of severe hypotension less than 50 mmHg or 80 mmHg was significantly shorter in the NSTI group than in the ATN group (less than 50 mmHg was 29.7 +/- 124 vs 72.5 +/- 174, less than 80 mmHg 105 +/- 234 vs 193 +/- 261 min: p < 0.01). The post-operative (po) day expressing diuresis in excess of 1000 mL of urine per d was 8.28 +/- 17.5 and 13.7 +/- 23.3 (p < 0.01) in the NSTI and ATN group, respectively. The po-d of the last hemodialysis and the po-d showing serum creatinine less than 2.0 mg/dL in NSTI and ATN group was 7.74 +/- 17.4 and 13.3 +/- 23.2 (p < 0.01), and 25.0 +/- 30.5 and 38.0 +/- 35.2 (p < 0.01), respectively. We concluded that 1-h renal biopsy is useful for assessing the outcome of renal allograft. AS of a donor kidney is one of the most important risk factors for both short and long-term outcome of the graft. The TI score was useful to predict the outcome of delayed graft function.     

Case study of paired cadaver renal allografts from the same donor: influence of local DIC kidney and concomitant acute rejection on early graft outcome

We report the clinical course of 2 recipients whose renal allografts were obtained from the same cadaver donor after cardiac arrest. The recipients showed different outcomes after transplantation. Graft biopsy after reperfusion revealed disseminated intravascular coagulation (so-called DIC kidney) and severe acute tubular necrosis (ATN) in both recipients. While one graft showed primary nonfunction, the other graft became functional after a post-operative anuric period. Serial graft biopsies performed during the oligo-anuric period revealed recovery of ATN and no intra-glomerular fibrin thrombi, but development of acute rejection was detected in both recipients. The left kidney graft showed more severe local DIC kidney than the right kidney, as well as more severe acute rejection in the oligo-anuric period. Despite aggressive anti-rejection therapy, the left kidney graft showed primary nonfunction. Therefore, severe acute rejection leading to primary nonfunction might have been related to more severe ischemic injury and more extensive local DIC kidney in the left kidney.     

Effect of histamine H2-receptor antagonist on the phosphorus-binding abilities of calcium carbonate and calcium lactate in hemodialysis patients

The effect of histamine H2-receptor antagonist (famotidine) on the phosphorus-binding abilities of calcium carbonate and calcium lactate were examined in 13 chronic hemodialysis patients. In seven patients receiving calcium carbonate, famotidine (20 mg/d) was given because of gastroduodenal disorders, and calcium carbonate was replaced with calcium lactate as a phosphorus binder after 4 wk of treatment with famotidine. With the 4-wk administration of famotidine accompanied by calcium carbonate, the serum phosphorus level increased from 6.3+/-0.9 to 7.1+/-0.5 mg/dl (P<0.05). However, with the substitution of calcium lactate, the serum phosphorus level decreased significantly when compared to that before substitution (6.3+/-0.2 and 6.0+/-0.9 mg/dl after 4 and 8 wk of substitution, respectively), despite continued administration of famotidine. Serum calcium, creatinine, alkaline phosphatase, high sensitive parathyroid hormone, blood urea nitrogen, arterial blood pH, and bicarbonate were not significantly altered during the trial period. In six control patients treated with calcium carbonate alone, there were no statistical changes in serum calcium and phosphorus levels after substitution of calcium lactate for calcium carbonate. These results suggest that famotidine significantly affects the phosphorus-binding ability of calcium carbonate, but not that of calcium lactate. A careful observation of changes in the serum phosphorus level should be required in hemodialysis patients receiving calcium carbonate and histamine H2-receptor antagonists. Calcium lactate may be useful as a phosphorus binder in such hemodialysis patients.     

Protective effect of nitric oxide against iron-induced neuronal damage

We investigated the effect of nitric oxide (NO) on iron-induced neuronal damage. Incubation of PC12 cells after the addition of FeCl2 induced rapid increases (within 1 hr) in lipid peroxidation and a concentration (0.1-2 mM)-dependent decrease in cell viability at 48 hr, both of which were blocked by deferoxamine and 2-methyl-6-(p-methoxyphenyl)-3,7-dihydroimidazo[1,2-a]pyrazine-3-o ne hydrochloride (MCLA) (a superoxide scavenger) but not by mannitol (a hydroxyl radical scavenger). Iron-induced cytotoxicity was also antagonized by superoxide dismutase with catalase. On the other hand, the NO donors S-nitroso-N-acetylpenicillamine (SNAP), 3-?(+/-)-(E)-ethyl-2'-[(E)-hydroxylamino]-5-nitro-3-hexenecarbo moyl?-pyridine (NOR-4), and 2,2'-(hydroxynitrosohydrazono)bis-ethanamine (NOC-18) decreased cell viability 48 hr after addition without increasing lipid peroxidation. However, when added with 1 mM FeCl2, NO donors including NOC-18, SNAP and NOR-4 (0.1-1 mM) inhibited lipid peroxidation in a concentration-dependent manner and suppressed cell death at lower concentrations. Addition of MCLA and NOC-18 also suppressed decreases in iron-induced [3H]thymidine incorporation. In rat brain homogenate, NOC-18 and SNAP both suppressed iron-induced lipid peroxidation. These findings suggest that NO has a dual effect on neuronal viability and can act as an antioxidant which protects neurons from iron-induced damage.