Effect of heat or chemical treatment on leptospiral antigens.

The use of bacterins is one of the primary methods in the control of leptospirosis in domestic animals, especially cattle, swine, and dogs. Bacterins have been made using chemically treated or heat-inactivated organisms. Many of the initial studies indicated that the bacterins gave complete protection, whereas later reports found that some inactivation methods resulted in bacterins that protect against death but not against kidney infection and renal shedding. This raised the possibility that some of the antigens had been altered. To investigate this possibility, we heat or chemically treated Leptospira interrogans serovar icterohaemorrhagiae CF-1, L. interrogans serovar canicola Moulton, L. interrogans serovar grippotyphosa SC4397, L. interrogans serovar hardjo Hardjoprajitno, and L. interrogans serovar pomona MLS. Temperatures used to determine alterations in antigens were 50, 56, 80, 100, and 121 degrees C. The chemicals used were formaldehyde, phenol, and thimerosal. Analysis was done using Laemmli gels and Western blots. We found that heating at 50 or 56 degrees C had the smallest effect on antigens, whereas heating at 80 or 100 degrees C caused the appearance of high-molecular-weight bands on Western blots. Heating at 121 degrees C caused almost all the bands to disappear, in both the stained gel and the Western blot. Both phenol and Formalin treatment altered proteins, whereas thimerosal treatment appeared to have little effect compared with heating at 50 degrees C.     

The morphological and functional changes in rat bladder following photodynamic therapy with phthalocyanine photosensitization

We have studied photodynamic therapy (PDT) in the rat bladder with a new photosensitizer, aluminium sulfonated phthalocyanine (AlSPc) given intravenously and intravesically. The microscopic distribution of photosensitizer fluorescence in the bladder wall was studied by laser fluorescence microscopy. Prior to PDT the bladder capacity and compliance were assessed by filling cystometry. Intravesical red light (675 nm.) from a copper vapour pumped dye laser was used to activate the photosensitizer using light doses of 20 to 200 J/cm2. Urodynamic and histologic changes were studied at intervals for up to three months. The fluorescence studies showed that AlSPc was eliminated from the deeper muscle layers more quickly than from the superficial layers of the bladder wall so that by 24 hours there was four times as much fluorescence from the mucosa and lamina propria compared to the deeper muscle. Control bladders illuminated with laser light alone showed no effects at these light doses. Animals treated 24 hours after sensitization showed a reduction in bladder capacity of up to 78% (20 J/cm2. light and 1.5 mg./kg.AlSPc). An initial reduction in compliance recovered in two weeks after low doses (0.5 mg./kg.) of AlSPc but was still abnormal at three months after higher doses (1.5 mg./kg.); though there was no long term histologic abnormality seen. Aluminium sulfonated phthalocyanine is a promising photosensitizer for bladder photodynamic therapy and using low doses of the drug it is possible to produce a superficial necrosis without muscle damage across a range of light doses. This heals by epithelial regeneration with no long term functional impairment. Direct absorption of this photosensitizer following intravesical administration seems unreliable.     

Scalp electrical recording during paralysis: quantitative evidence that EEG frequencies above 20 Hz are contaminated by EMG.

OBJECTIVE: To identify the possible contribution of electromyogram (EMG) to scalp electroencephalogram (EEG) rhythms at rest and induced or evoked by cognitive tasks. METHODS: Scalp EEG recordings were made on two subjects in presence and absence of complete neuromuscular blockade, sparing the dominant arm. The subjects undertook cognitive tasks in both states to allow direct comparison of electrical recordings. RESULTS: EEG rhythms in the paralysed state differed significantly compared with the unparalysed state, with 10- to 200-fold differences in the power of frequencies above 20 Hz during paralysis. CONCLUSIONS: Most of the scalp EEG recording above 20 Hz is of EMG origin. Previous studies measuring gamma EEG need to be re-evaluated. SIGNIFICANCE: This has a significant impact on measurements of gamma rhythms from the scalp EEG in unparalysed humans. It is to be hoped that signal separation methods will be able to rectify this situation.     

Retinal pigment epithelial tear associated with subretinal hemorrhage: a case report

Tears of the retinal pigment epithelium (RPE) are a recognized complication of pigment epithelium detachments associated with age-related macular degeneration (ARMD). The RPE tears are often associated with a poor visual prognosis and typically do not occur with subretinal hemorrhage. We report the case of a 76-year-old man with ARMD who developed an RPE tear associated with subretinal hemorrhage, without a demonstrable subretinal neovascular membrane. Temporally, there was a marked decrease in his vision that returned to his earlier baseline visual acuity over the course of six months of observation.     

Mechanisms responsible for the thermal sensitivity of adrenal microsomal monooxygenases.

Studies were done to determine the mechanism(s) responsible for the thermal lability of adrenal microsomal monooxygenases. Preincubation of guinea pig adrenal microsomal suspensions at 37 degrees C caused large time-dependent declines in benzo(a)pyrene (BP) hydroxylase and benzphetamine (BZ) demethylase activities. Similar preincubations with hepatic microsomes had little effect on enzyme activities. The decreases in adrenal enzyme activities were completely prevented by co-incubation of microsomes with cytosol, but were not diminished by reduced glutathione, ascorbic acid, or bovine serum albumin. Partial protection was afforded by EDTA, suggesting that lipid peroxidation might be involved, but malonaldehyde production was not demonstrable and MnCl2, a potent inhibitor of lipid peroxidation, did not affect the decline in enzyme activities. The decreases in the rates of BP and BZ metabolism were prevented by including NADPH or NADP+ in the preincubation medium. The preincubation conditions causing losses of adrenal enzyme activities did not affect cytochrome P-450 concentrations or substrate binding to cytochromes P-450, as indicated by type I difference spectra. NADH-cytochrome c reductase activity also was not affected, but there were decreases in NADPH-cytochrome c reductase activity that were proportionately similar to the declines in drug-metabolizing activities. Direct assessment of NADPH-cytochrome P-450 reductase revealed similarly large decreases in enzyme activity resulting from preincubation of adrenal microsomes. The results demonstrate a need for extra caution when doing preincubation experiments with adrenal microsomal preparations, and suggest that the thermal lability of adrenal monooxygenases is attributable to effects at the active site of NADPH-cytochrome P-450 reductase.     

Effect of cirrhosis on accuracy of 99Tcm-tin colloid scintigraphy and ultrasound scanning in diagnosis of hepatocellular carcinoma

The effect of cirrhosis on the accuracy of ultrasonography and scintigraphy in the diagnosis of hepatocellular carcinoma (HCC) has not been established. We have therefore studied the findings on ultrasound and scintigraphy in four groups of patients. Fourteen had HCC and cirrhosis, 13 had HCC but no cirrhosis, 53 had cirrhosis but no HCC and 31 had evidence neither of cirrhotic nor malignant liver disease. The accuracies of a positive interpretation of HCC on ultrasound and scintigraphy in the absence of cirrhosis were 85% and 66% respectively (NS), but fell (significantly in the case of scintigraphy: p less than 0.05) to 80% and 31% respectively in the presence of cirrhosis (p less than 0.01). The poorer performance of scintigraphy in the presence of cirrhosis was a result both of its lower sensitivity (50% compared with 85%) and the greater frequency of false positives (28% vs 5%: p less than 0.001). The accuracy of a negative interpretation of HCC on scintigraphy in the absence of cirrhosis was 96% but fell to 84% in the presence of cirrhosis, whereas on ultrasound it remained the same (96%). In conclusion, we recommend ultrasound in preference to scintigraphy as a more accurate technique for the diagnosis of HCC in the presence of cirrhosis.     

Changes in bladder function following a surgical alteration in outflow resistance

The changes in bladder function occurring after a surgical alteration in bladder outflow resistance were studied in 20 males undergoing transurethral prostatectomy (TURP) and in 20 females undergoing an endoscopic bladder neck suspension (EBNS). Serial cystometrograms (CMG) were performed before operation, and on alternate days after spontaneous micturition was re-established, for 5 days in the males and for 21 days in the females. CMGs were repeated at 3 months and 1 year after operation. After TURP voiding pressures (P_det) fell rapidly from a mean of 118 cm H₂O before operation to 57 cm H₂O at 5 days, with an increase in flow rate during this time from 10.5 ml/s to 24 ml/s. Detrusor instability that had been present in 14 patients resolved within 2 days in 12. There was no further significant urodynamic change over the 1-year study period. After EBNS, there was an early rise in voiding pressure (P_det rose from 26 cm H₂O before operation to 42 cm H₂O at 3 days). This continued to increase up to 21 days particularly in those patients with initial large residuals. Three patients developed detrusor instability. Flow rates were greatly reduced at first (27 ml/s before operation and 13 ml/s at 3 days), and gradually increased in line with voiding pressures, yet were still diminished 1 year after operation. The urodynamic changes following a reduction in bladder outflow resistance by TURP are immediate and sustained and unlikely to be the result of structural changes within the bladder wall. EBNS produces an increase in outflow resistance and it can be several weeks before balanced voiding is achieved, with significantly increased detrusor pressures needed to achieve complete bladder emptying at a reduced flow rate.     

Case report 733

Calcific tendinitis at the site of origin of the rectus femoris muscle is a rare lesion. Presumed to be formed by deposition of hydroxyapatite crystals, this entity may be confused with other lesions such as os acetabuli or posttraumatic abnormalities. The characteristic location and appearance of the calcifications in a symptomatic patient with no history of trauma should allow diagnosis and subsequent symptomatic therapy.