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Mitochondrial ATPase inhibitor protein (IF1) reacts reversibly with complex V and inhibits up to 90% of its ATPase activity. Both the rate and extent of inhibition are pH and temperature dependent and increase as the pH is lowered from pH 8 tp 6.7 (the lowest pH examined) or as the temperature is increased from 4 to 36 degrees C. Nucleotide triphosphates plus Mg2+ ions are required for inhibition of complex V ATPase activity by IF1. In the presence of Mg2+ ions, the effectiveness order of nucleotides is ATP greater than ITP greater than GTP greater than UTP. Highly purified complex V, which requires added phospholipids for expressing ATPase and ATP-Pi exchange activities, cannot be inhibited by IF1 plust ATP-Mg2+ unless phospholipids are also added. This indicates that the active state of the enzyme is necessary for the IF1 effect to be manifested, because F1-ATPase, which does not contain nor require phospholipids for catalyzing ATP hydrolysis, can be inhibited by IF1 plus ATP-Mg2+ in the absence of added phospholipids. The IF1-inhibited complex V, but not IF1-inhibited F1-ATPase, can be reactivated by incubation at pH greater than 7.0 in the absence of ATP-Mg2+. The reactivation rate is pH dependent and is influenced by temperature and enzyme concentration. Complex V preparations contain small and variable amounts of IF1. This endogenous IF1 behaves the same as added IF1 with respect to conditions described above for inhibition and reactivation and can result in 25-50% inhibition in different complex V preparations. However, complex V lacking endogenous IF1 can be reconstituted from F0, F1, oligomycin sensitivity conferring protein, and phospholipids. Inhibition of this reconstituted preparation in the presence of ATP-Mg2+ depends entirely on addition of IF1. In general, the ATP-Pi exchange activity of complex V is more sensitive to the chemical inhibitors of F1-AtPase tha its ATPase activity. This is not so, however, for IF1. Under conditions that IF1 caused approximately 75% inhibition of ATPase activity of complex V, no more than 10% of the ATP-Pi exchange activity was inhibited.

We prospectively studied 12 patients to compare fibreoptic enteroscopy and methylene blue injection through superselectively prepositioned angiographic catheters in intra-operative localization of bleeding small intestinal lesions. Four patients were excluded because the lesions were easily detected by gross examination. Eight patients were subjected to study by these two methods. As methylene blue injection required a positive angiography, it was possible only in five patients. In two of these five patients, non-bleeding lesions were present outside the intestinal segments localized by this method. Enteroscopy was simple, accurate and reliable in seven patients. Complete enteroscopy was impossible in one patient with advanced lymphoma because the intestine and its mesentery were matted together. Fortunately, the bleeding ulcers were still within the reach of the endoscope. Enteroscopy localized bleeding small intestinal lesions more exactly than methylene blue injection so that the length of gut resection could be shorter. We, therefore, conclude that enteroscopy is the better intra-operative localization procedure.

A young man developed acute renal failure and hypercalcaemia following severe burns. The hypercalcaemia was initially controlled by haemodialysis, but it persisted after return of renal function. Plasma PTH was inappropriately elevated, but the nephrogenous cyclic adenosine monophosphate level was low; thus the PTH was probably not biologically active, and may have been artefactually elevated by the moderate renal impairment. Bone histology, showed a normal resorbing surface, but a zero forming surface, implying that the bone dissolution leading to hypercalcaemia resulted from a failure of bone formation. Because of widespread infection and impaired renal function, the hypercalcaemia could not be treated by corticosteroid drugs, mithramycin or phosphate, and there was no response to salmon calcitonin. He was therefore treated with intravenous sodium sulphate, which increased urinary calcium excretion and reduced the plasma calcium. Sodium sulphate still has a role in the treatment of patients with hypercalcaemia.

The combined use of fentanyl-epinephrine-lidocaine in epidural analgesia was investigated in 60 mothers who underwent elective Cesarean section. They were randomly classified into 4 groups of 15 each. Group A received 20 mL of 2% lidocaine 3 mL normal saline. Group B received 20 mL of 2% lidocaine with freshly added 1:200000 epinephrine and 5 mL normal saline. Group C received 20 mL of 2% lidocaine and 2 mL of fentanyl in 3 mL normal saline. Group D received 20 mL of 2% lidocaine and 5 mL normal saline. The duration of action was nearly doubled in the groups with epinephrine added. The quality of the analgesia was assessed by the degree of intraoperative analgesia; 93% of the patients in group A were classified as "Excellent"; while only 67% in group B were classified as "Excellent". The difference in patient's acceptance was statistically significant. 93% vs 27% in Group A and Group C; 67% vs 6.7% in Group B and Group D rated "Excellent". The results suggest that both fentanyl and epinephrine improve intraoperative analgesia. Epinephrine offers a greater analgesic effect than fentanyl.

Quantitative bone histology was studied in 23 patients with malignant hypercalcemia (MH) due to carcinoma (16) or immunoproliferative disease (7). Plasma calcium was 3.37 +/- 0.47 (mean +/- SD) mmol/liter. Bone resorbing surface (RS) was measured using a sensitive histochemical stain to identify osteoclasts. In the MH patients with carcinoma, the RS was 3.1 +/- 2.6% compared to 1.0 +/- 0.3% in controls (P less than 0.02). In the myeloma patients it was 2.3 +/- 1.7%, and in normocalcemic patients with malignant disease 0.8 +/- 1.1%. RS did not correlate with serum PTH, and several high RS values were associated with undetectable PTH. RS correlated with forming surface (FS) in MH patients (r = 0.44, P less than 0.05) and controls (r = 0.68, P less than 0.005), but there was a greater RS relative to FS in MH patients than in controls (P less than 0.005). "Excess" RS in the MH patients was calculated by subtracting the RS accounted for by the measured FS, using the relationship defined by the controls. Bone loss, as reflected in urinary calcium excretion, correlated weakly with excess RS (r = 0.44, P less than 0.05) but was high even when excess RS was zero. Thus, the histological findings do not account for the bone loss, and additional resorption around bone metastases is likely; the results of this study are consistent with a humoral substance produced by the malignant tissue causing generalized bone resorption in addition to bone dissolution around metastases.

We studied the effect of age on bone composition and osteocyte viability in femoral heads from fifty-one subjects. The assessment included determination of: bone volume, ash weight, calcium, and phosphorus content; osteocyte viability in fresh sections stained for lactate dehydrogenase activity; microfractures in fresh sections after removal of marrow elements; bone area, the presence of metabolic bone disease, and the histology of microfractures in embedded calcified sections; and the extent of trabecular microfractures. Bone area and numbers of microfractures were also assessed in eight elderly hip-fracture patients. Bone volume decreased with age, but there was considerable variation in each age group, and no significant difference between men and women. Ash weight and the bone content of calcium and phosphorus also decreased with age, but were constant if corrected for bone volume. Almost all osteocytes were viable in subjects who were younger than twenty-five years, and thereafter viability progressively decreased to a mean of 74 per cent in the eighth decade of life. There was a significant negative correlation between osteocyte viability and age. There was no evidence of metabolic bone disease in any patient. The numbers of microfractures increased with age and correlated negatively with bone viability (r = -0.31, p less than 0.05); in simple linear correlation a relationship between bone area and microfractures could not be demonstrated but in multiple linear correlation, after the inclusion of bone viability, there was an additional negative correlation between numbers of microfractures and bone area (p less than 0.005). Bone area and numbers of microfractures in hip-fracture patients were similar to those in age-matched controls.

Congenital nystagmus may occur in a manifest or latent form. Since most patients with latent nystagmus also have nystagmus with both eyes open (manifest latent nystagmus), differentiation between these two entities frequently requires electronystagmographic wave form analysis. Such analysis shows distinctive characteristics for manifest and latent or manifest latent congenital nystagmus, suggesting different origins. Several compensation mechanisms exist by which congenital nystagmus is decreased and visual acuity improved. For manifest nystagmus, these mechanisms include version or convergence innervation or a rest point between primary position and maximal lateroversion. In latent or manifest latent nystagmus, the nystagmus decreases when the fixating eye is adducted. Patients with manifest congenital nystagmus may use several of these compensation strategies. Because of the difficulties involved in distinguishing between manifest and manifest latent nystagmus without electronystagmography, confusion has arisen regarding the etiologic relationship between congenital nystagmus and infantile esotropia. Such relationship clearly exists in some patients with manifest nystagmus and convergence dampening. However, in most patients with infantile esotropia and congenital nystagmus, the nystagmus is latent or manifest latent and there is no evidence of a causal relationship between these two entities.

75 patients with Paget's disease of bone were treated with a drug combination intended to increase the production of endogenous calcitonin and decrease that of parathyroid hormone. The first regimen of oral calcium, thiazide diuretic, aluminum hydroxide and low-phosphorus diet was given to 41 patients for a mean of 800 days. A simpler regimen of oral calcium and thiazide diuretic was given to 34 patients for a mean of 750 days. There was a similar fall in mean plasma alkaline phosphatase to 71 +/- 24 (SD)% of initial with the first regimen and 72 +/- 17% with the second at 150 days, with a gradual rise after 500 days. Urinary hydroxyproline fell from 165 +/- 111 to 112 +/- 93 mg/day. Plasma calcium rose slightly with both regimens and plasma inorganic phosphorus fell with the first. Serum parathyroid hormone and calcitonin levels were unchanged. Urinary calcium was not changed by the first regimen and rose by 40 +/- 54 mg/24 h with the second. Clinical improvement approximately paralleled biochemical improvement. It is suggested that, in view of its low cost and convenience, this treatment has a place in the management of Paget's disease of bone.

Biochemical data and bone histology from 44 haemodialysis patients was compared using an histologic technique capable of evaluating separately the individual components of osteodystrophy. Hyperparathyroid bone disease was diagnosed by an elevated osteoclast count, and in advanced disease there was also fibrosis and woven bone. Osteomalacia, defined as an impairment in the rate of bone mineralisation, was present in two distinct forms: osteomalacia type I, characterised by wide osteoid seams, and osteomalacia type II, characterised by extensive thin, inactive osteoid. The histologic diagnoses were hyperparathyroid bone disease (15), osteomalacia type I (3), osteomalacia type II (6), hyperparathyroid bone disease and osteomalacia type I (12), hyperparathyroid bone disease and osteomalacia type II (6), normal (2). Aluminium was evident histochemically in 17 biopsies. Vitamin D metabolite levels were low in most patients and did not correlate with any biochemical or histological parameter. Parathyroid hormone levels were highly correlated with histological features of hyperparathyroid bone disease, and also correlated with plasma calcium, suggesting a degree of autonomy of parathyroid hormone secretion. Urea and creatinine were higher in the hyperparathyroid bone disease than the osteomalacia groups suggesting that poor dialysis contributes to the former. Statistical analysis showed that osteomalacia type I was associated with relatively low plasma calcium and phosphorus levels; osteomalacia type II was associated with increased bone aluminium and with the uraemic process itself, as reflected in the plasma creatinine level. This study shows relationships between renal osteodystrophy and plasma calcium and phosphorus levels, but no relationship with vitamin D metabolites. Aluminium appears to impair mineralisation even at relatively low levels of accumulation. However there are other unidentified factors associated with the uraemic process, contributing to all three components of renal osteodystrophy.

Study Objective: To determine whether the failure to attain normal bone mass in young adulthood contributes to the later development of osteoporotic fractures., Design: Case-control study., Setting: Referral-based bone clinic at a large teaching hospital., Patients: Sequential sample of 35 asymptomatic relatives, aged 19 to 59 years, of patients with osteoporotic fractures, and 24 patients with osteoporotic fractures., Measurements and Main Results: Bone mineral density in the spine was measured by quantitative computed tomographic scanning. Bone mineral content in the os calcis was measured in 19 of the relatives of osteoporotic patients by single-photon absorptiometry. The values for bone mineral density in the spine were corrected to age 50 years with the regression equation derived from the normal values in the controls. The values were lower in relatives of osteoporotic patients than in controls. In men, the mean values (+/- standard deviation [SD]) for relatives were 91 +/- 16 mg/cm3, and for controls, 129 +/- 21 mg/cm3 (P less than 0.001). In women, the mean values for relatives were 96 +/- 17 mg/cm3 and for controls, 126 +/- 19 mg/cm3 (P less than 0.001). In the osteoporotic patients, the corrected mean value for men was 53 +/- 12 mg/cm3, and for women, 77 +/- 20 mg/cm3. The os calcis values did not correlate with the spine values and were mostly well within the normal range., Conclusions: Mean bone mass is lower in apparently healthy young and middle-aged adult relatives of osteoporotic patients than in normal persons with no family history of osteoporosis. Our findings suggest that the failure to attain an adequate peak bone mass may play an important role in the later development of osteoporotic fractures. Relatives of osteoporotic patients should be advised to have measurements of bone mass taken. This measurement should be taken at the spine, because peripheral sites do not appear to provide adequate information about early osteoporosis.

The use of monoclonal antibody especially in immunophenotyping of lymphomas is firmly established in clinical practice. This has helped in refining the classification of non-Hodgkin's lymphomas and the updated Kiel Classification is now much favoured by the Europeans. Nonetheless, clinicians by and large preferred the National Cancer Institute Working Formulation Classification. This article explores the potential usage of monoclonal antibodies reactive to the so-called Clusters of Differentiation antigens on lymphoid cells and the role of a clinical pathologist in achieving an accurate clinical diagnosis. Such immunophenotyping is only complementary to the morphological diagnosis of lymphomas.

Sixty-four patients with nystagmus blockage syndrome (NBS) underwent surgery consisting of unilateral recession-resection, bimedial recession with and without posterior fixation suture, and bimedial recession with resection of one or both lateral recti. The results of the unilateral recession-resection and bimedial recession with or without posterior fixation were comparable. Over- and undercorrections occurred more frequently and the number of reoperations was higher than in a control group of essential infantile esotropia without nystagmus. These unpredictable surgical results in NBS present yet another feature that distinguish this condition from essential infantile esotropia without nystagmus. While restoration of normal binocular vision was not achieved in a single case, a functionally desirable result of subnormal binocular vision or microtropia occurred in 26%. An unusual complication consisting of postoperative asymmetric gaze nystagmus with an anomalous head posture was observed in five patients and required a Kestenbaum-Anderson operation in four to eliminate the head turn.

The effects of uncouplers and valinomycin plus nigericin (in the presence of K+) were studied on the apparent Km for substrates and apparent Vmax of the following energy-linked reactions catalyzed by submitochondrial particles: oxidative phosphorylation, NTP-33Pi exchange, ATP-driven electron transfer from succinate to NAD, and respiration-driven transhydrogenation from NADH to 3-acetylpyridine adenine dinucleotide phosphate. In all cases, partially uncoupling (up to 90%) concentrations of uncouplers of valinomycin plus nigericin were found to decrease apparent Vmax and to increase apparent Km. Results plotted as ln (Vmax/Km) versus the concentration of uncouplers or ionophores showed a linear decrease of the former as a function of increasing perturbant concentration (i.e., decreasing free energy). Because Vmax/Km may be considered as a measure of the apparent first-order rate constant for enzyme-substrate interaction and reflects the affinity between enzyme and substrate to form a complex, the results are consistent with the interpretation that membrane energization leads to a change in enzyme conformation with the resultant increase in enzyme-substrate affinity and facilitation of the reaction rate under consideration. The significance of these findings with respect to the mechanism of action of the energy-transducing systems studied is discussed.

We have developed a technique for assessing bone viability by the histochemical demonstration of lactate dehydrogenase (LDH) activity in osteocytes. Fresh sawn and ground (75 microns) sections were prepared from femoral heads removed at operation from patients with osteoarthritis of the hip. The sections were decalcified overnight in cold 10% EDTA, pH 7.0. LDH activity was shown by the tetrazolium-formazan reaction with nitroblue tetrazolium as indicator and lithium lactate as substrate. Osteocytes were regarded as viable if their cytoplasm stained dark blue, indicating LDH activity; lacunae containing non-viable osteocytes could be identified by interference contrast illumination. Nearly all osteocytes were viable in the samples studied. Small trabecular fragments, such as could be obtained by needle biopsy, were also suitable for staining after grinding to approximately 50 microns. The method should have application both in research and in diagnosis of ischemic bone disease.

The survival of cells cultured in medium containing the chemotherapeutic drug methotrexate (MTX) is related directly to drug concentration. Changes in DNA resulting from a severe imbalance in the cells' nucleotide pools are thought to account for this cytotoxicity. We have attempted to clarify the gross biochemical changes that might lead to cell death. DNA strand breaks occur in cells treated with high concentrations of MTX but it is not clear that these are sufficient to account for cytotoxicity at lower doses. We observed dramatic changes in cytoskeletal morphology. Gross reorganization of the cytoskeleton is shown by immunolabelling but is high-lighted dramatically when cells are lysed to leave 'nucleoids'. The nature of the changes seen in MTX-treated cells is characteristic of the cells' general stress response, seen originally following heat shock. This study shows that other factors, such as changes in cytoskeletal function, must be considered together with any contribution from DNA damage, in order to account for the lethal effects of MTX.

Osteocyte viability was investigated in femoral head bone removed from 38 patients with chronic hip disease, with the use of a histochemical stain to demonstrate lactate dehydrogenase (LDH) activity in osteocytes. Where the osteocyte cytoplasm did not show LDH activity, the cell was considered dead; when several adjacent osteocytes were dead, the bone in that area was regarded as nonviable. The preoperative diagnoses were idiopathic osteoarthritis in 25, chondrocalcinosis in six, rheumatoid arthritis in two, Paget's disease in two, avascular necrosis in two, and congenital dislocation of the hip in one patient. In 16 of the patients with idiopathic osteoarthritis and the two with avascular necrosis, nonviable osteocytes were present in the central regions of many trabeculae, these areas usually being separated by cement lines from viable bone. The pattern suggested previous necrosis of part of the femoral head, with later new bone formation. The pattern was not observed in either control subjects, or patients with known articular disease, such as chondrocalcinosis. Bone collapse of variable severity was apparent radiographically in nine patients with histologic bone death, but not in other patients. Bone death is commonly present in idiopathic osteoarthritis and could be a cause rather than a result of the arthritis.

Bone mineral content (BMC) was measured in the os calcis of 232 normal subjects aged 17-82 years. The mean reproducibility (coefficient of variation) of the measurement was 1.8%. Substantial bone loss occurred between the ages of 20 and 50 years, and in females the menopause was associated with additional bone loss. There was no significant difference in the rate of bone loss in females and males, but the mean BMC was greater at all ages in males than in females. We also compared os calcis BMC with spinal bone mineral density (BMD), measured by quantitative computed tomographic (CT) scanning, in 85 subjects: 33 were normal controls, 19 had osteoporosis defined by the presence of one or more pathological fractures, and in the remainder the CT examination was performed at the patient's request. Os calcis BMC correlated with spinal BMD in both females (r = 0.69, p less than 0.001) and males (r = 0.84, p less than 0.001). However, the os calcis BMC did not reliably predict spine values around the CT "fracture threshold" of 90-100 mg/cm3 and did not correlate with osteoporotic fracture as well as did spinal BMD. It is concluded that measurement of the os calcis BMC is of limited clinical usefulness for the early diagnosis of osteoporosis.

Six patients with angiodysplasias of the small intestine were diagnosed preoperatively with selective angiography during the past eight years. Enteroscopy was used to identify these vascular lesions intraoperatively. Lesions were ulcerated in four patients, had a bleeding vessel tip in one patient and appeared as a nonulcerated reddish patch in the other. Exact localization of these lesions allowed limited segmental resection of the small intestine to be performed. With complete enteroscopy, we were confident that no bleeding lesion was left undetected. The feeding artery and vein to the resected intestinal segment were separately cannulated, and the resected specimen was prepared and injected with hot barium-gelatine. The lesions were confirmed histopathologically to be angiodysplasias in all six patients. There was no recurrence of gastrointestinal bleeding at a median follow-up study of 34 months. There were no operative deaths, and no complications arose from enteroscopy.