Organ culture of pig cornea: biochemical analyses of mid-term-stored corneal tissue

Freshly prepared pig corneas were cultured at 22 degrees C in TC 199 medium supplemented with HEPES buffer, antibiotics and 10% sterile pig blood serum. The cultures were maintained in a closed system up to 30 days. The hydration as measured by wet and dry weights increased from 80 to 94% after 2 weeks of storage. Glucose decreased slowly and continuously during the culture time by 0.32 mmol in 30 days. This was equivalent to 11 mumol/day, 0.46 mumol/h and finally 0.264 mumol/h/cm2. Lactate increased in the culture accordingly by 0.34 mumol/h/cm2. In the corneal epithelium of the cultured tissues the glucose levels--normally at about 5 mumol/g--equilibrated with those of the medium during the first 10 days; later the epithelial glucose levels increased up to 20-30 mumol/g. The glycogen content of the epithelium remained constant for the first 10 days of storage. The ATP/ADP ratios in the corneal epithelium initially decreased but later recovered to values of about 12. The pO2 in the culture system decreased from 156 to 120 mm Hg, which is equivalent to an oxygen consumption of 7.41 microliters O2/h/cm2. The CO2 production initiating from a level of 16.6 mm Hg was 4.88 microliters/h/cm2. The data showed that the cornea was viable in this culture system for 30 days.     

Differences in leisure-time physical activity levels between blacks and whites in population-based samples: The Minnesota heart survey

Energy expenditure in leisure-time physical activity (LTPA) was measured using the Minnesota LTPA Questionnaire in 35- to 74-year-old black and white residents of Minneapolis-St. Paul, Minnesota. Estimates of the geometric mean LTPA energy expenditure were 129 and 204 kcal per day for black and white men (p<.05) and 91 and 123 kcal per day for black and white women (p<.05). The percentage of individuals expending 2000 kcal or more per week in LTPA was significantly lower in black men than white men (25 vs. 35%; p=.01) but was not different in black versus white women (18 vs. 17%). Although black men and women reported greater occupational physical activity than their white counterparts, LTPA and job activity were unrelated in all race and sex groups. In both races, LTPA energy expenditure declined with age. LTPA increased with level of formal education, and the largest LTPA difference between blacks and whites was observed in those who had a high-school diploma or less. Blacks had lower participation rates than whites in most of the individually assessed physical activities. Additional research is needed on the determinants and promoters of LTPA in population subgroups.The research was funded by National Heart, Lung, and Blood Institute Research Grant R01 HL 23727 and National Research Award T32 HL 07328-10.     

Effect of oral lactulose on clinical and immunohistochemical parameters in patients with inflammatory bowel disease: a pilot study

Background  

The prebiotic potential of lactulose is well established and preclinical studies demonstrated a protective effect of lactulose in murine models of colitis. The aim of the present study was to investigate the clinical and histological efficacy of lactulose in patients with inflammatory bowel disease (IBD), for which probiotic therapy yielded promising results.     

Evaluation of myocardial metabolism with microdialysis during bypass surgery with cold blood- or Calafiore cardioplegia.

BACKGROUND: For the first time, microdialysis was used to investigate in vivo and online the myocardial metabolism during and after cardiac surgery in patients treated with two different methods of myocardial protection. METHODS: Thirty patients underwent standard CABG with one of two different methods of myocardial protection. The patients were randomised to receive either cold blood (COLD group) or warm modified Calafiore cardioplegia (WARM group). Microdialysis probes were implanted into the myocardium of left ventricular apical region of the heart. Cardioplegia was given antegrade only. Microdialysis measurements were performed at time intervals before, during and 24 h after cardiopulmonary bypass and analysed for glucose, lactate, pyruvate and glycerol. RESULTS: Myocardial lactate concentrations were significantly higher in the WARM group compared with that of the COLD group, while serum lactate was comparable. Glycerol was significantly higher at the end of the clamping time in the WARM group. At the same time the glucose-lactate ratio as a marker of nutritional disorder had significantly lower levels in the WARM group. The cumulative CK-MB release over 24 h was significantly higher in those hearts protected with warm blood. CONCLUSIONS: The oxidative stress measured was significantly higher in patients undergoing CABG using modified Calafiore cardioplegia, whereas the cold cardioplegia minimised the effects of aortic clamping. The results indicate that cold cardioplegia offers superior protection of the heart, in terms of more rapid normalisation of myocardial metabolism. In elective myocardial revascularisation, intermittent antegrade warm blood cardioplegia is a comparable safe method of myocardial protection. However, in patients referring to a long clamping time, advantages of cold cardioplegia for myocardial revascularisation may be magnified.     

Depressed antigen presentation function and membrane interleukin-1 activity of peritoneal macrophages after laparotomy

Although major tissue trauma produces profound depression of cell-mediated immunity, it is not known whether surgical trauma (i.e., midline laparotomy) has any adverse effect on the antigen presentation function and membrane interleukin-1 (IL-1) activity of peritoneal macrophages. To study this, C3H/HEJ (endotoxin-tolerant) mice were anesthetized. An approximately 1-inch midline abdominal incision was made, followed by abdominal closure. On days 1, 3, 5, and 7, peritoneal macrophages were harvested by means of peritoneal lavage, and antigen presentation capability was tested by incubating various numbers of peritoneal macrophages with 2 X 10(4) D10.G4.1 cells per well in the presence of conalbumin (400 micrograms/ml). The T helper cell clone (D.10.G4.1) proliferates on recognition of conalbumin in the context of Iak and also proliferates in the presence of membrane-bound IL-1 plus concanavalin A. To measure membrane IL-1 expression in peritoneal macrophages, Concanavalin A (10 micrograms/ml) was substituted for conalbumin. Cultures were incubated for 72 hours, pulsed with tritiated thymidine, and harvested. Peritoneal macrophages from laparotomized mice induced significantly less T helper cell proliferation on days 1 and 3 in the antigen presentation assay (37% and 30%, respectively; p less than 0.05) and in the membrane IL-1 assay (14% and 10%, respectively; p less than 0.05) as compared with the control. This difference was not detectable on day 5. More effective antigen presentation capability (167% of control; p less than 0.05) was seen on day 7. Thus laparotomy by itself produces marked depression of both antigen presentation function and membrane IL-1 activity of peritoneal macrophages, which may enhance susceptibility to intra-abdominal sepsis.     

Intracerebral hemorrhage: natural history and rationale of ultra-early hemostatic therapy

Stroke is a major health problem worldwide, causing high morbidity and mortality. Intracerebral hemorrhage (ICH) accounts for 15% of stroke cases in the US and Europe and up to 30% in Asian populations. It is less treatable than other forms of stroke and causes higher morbidity and disability. Data suggest that early hematomy growth is the principal cause of early neurological deterioration after ICH. Prospective and retrospective studies indicate that early hematoma growth occurs in 18–38% of patients scanned within 3 h of ICH onset, and that hematoma volume is an important predictor of 30-day mortality. As hematoma growth in acute ICH is a dynamic process, intervention with ultra-early hemostatic therapy could lead to minimization and even prevention of early hematomy growth. Recombinant activated factor VII (rFVIIa, ‘NovoSeven’), a powerful initiator of hemostasis, is approved for the treatment of bleeding in patients with hemophilia and inhibitors and may also promote hemostasis in patients with normal coagulation. rFVa acts locally at the bleeding site without activating systemic coagulation and may be a valuable therapy during the hyperacute stage of ICH. A randomized, double-blind, placebo-controlled, dose-ranging trial is currently in progress to investigate the potential of rFVIIa as an ultra-early hemostatic therapy to prevent or minimize hematoma growth in ICH patients without coagulopathy.     

Erythromycin-Induced Immune Hemolytic Anemia

A 3-year-old female receiving Pediazole (erythromycin ethylsuccinate and sulfisoxazole) for tonsillitis and otitis media developed severe hemolytic anemia. No serum drug-dependent antibodies could be demonstrated with an in vitro 'immune-complex' method using Pediazole, pure erythromycin ethylsuccinate or pure sulfisoxazole. However, a method using red cells coated with erythromycin base showed in vitro lysis of the erythromycin-coated red cells. This is only the second case of immune hemolytic anemia associated with erythromycin and the first where in vitro drug-dependent hemolysis was demonstrable.