MR1-restricted mucosal-associated invariant T (MAIT) cells respond to mycobacterial vaccination and infection in nonhuman primates.

Studies on mucosal-associated invariant T cells (MAITs) in nonhuman primates (NHP), a physiologically relevant model of human immunity, are handicapped due to a lack of macaque MAIT-specific reagents. Here we show that while MR1 ligand-contact residues are conserved between human and multiple NHP species, three T-cell receptor contact-residue mutations in NHP MR1 diminish binding of human MR1 tetramers to macaque MAITs. Construction of naturally loaded macaque MR1 tetramers facilitated identification and characterization of macaque MR1-binding ligands and MAITs, both of which mirrored their human counterparts. Using the macaque MR1 tetramer we show that NHP MAITs activated in vivo in response to both Bacillus Calmette-Guerin vaccination and Mycobacterium tuberculosis infection. These results demonstrate that NHP and human MR1 and MAITs function analogously, and establish a preclinical animal model to test MAIT-targeted vaccines and therapeutics for human infectious and autoimmune disease.

Delayed cellulitis associated with conservative therapy for breast cancer.

BACKGROUND AND OBJECTIVES: Delayed breast cellulitis is an infrequently reported entity after conservation therapy for breast cancer. We describe our experience with this entity at Naval Medical Center, San Diego. METHODS: Eight patients who presented with delayed cellulitis after wide local excision/axillary dissection and breast radiotherapy (RT) are presented. Their clinical characteristics and therapy are described and possible causative factors are analyzed. RESULTS: The latency of breast cellulitis is variable after breast conservation therapy, although most cases in our experience and in the literature occur within a year post-RT. These infections are frequently refractory to a single course of antibiotics (n = 4 cases in our experience). Some patients suffer multiple episodes separated by months. CONCLUSIONS: Breast cancer patients are at risk for delayed cellulitis after conservative surgery and RT. The mechanism of such events probably involves lymph stasis, however, therapy is no different from the more frequently occurring cases of cellulitis presenting perioperatively.

Evaluating the readability of informed consent forms used in contraceptive clinical trials.

The readability level of informed consent forms used in clinical trials on contraceptives was determined. Three different formulas for measuring readability were used. Some forms received relatively high scores by all three methods. The most common problems associated with high readability scores were the use of 'unfamiliar' words, long words and long sentences. At present all forms used must be readable, using the SMOG formula, at a grade 6 level or less.

The effect of rate limitation by cytochrome c on the redox state of the ubiquinone pool in reconstituted NADH: cytochrome c reductase.

The kinetic model of Ragan & Cottingham [(1985) Biochim. Biophys. Acta 811, 13-31] for electron transport through a mobile pool of quinone predicts that, under certain conditions, the normal linear dependence of electron flow on the degree of reduction (or oxidation) of the quinone should no longer be found. These conditions can be met by reconstituted NADH: cytochrome c reductase (Complex I-III from bovine heart) when electron flow is rate-limited by a low concentration of cytochrome c. We show that, in such a system, the dependence of activity (varied by inhibition with rotenone) on the steady-state level of quinone reduction is indeed non-linear and very closely accounted for by the theory.

Comparison of two testing methods to determine hepatitis B surface antibody response to hepatitis B vaccine among health-care workers.

Because of variations in reported seroconversion rates and to compare enzyme-linked immunoassay (EIA) and radioimmunoassay (RIA) methods to assess hepatitis B vaccine response, hepatitis B surface antibody (anti-HBs) was tested in 116 of 174 high-risk hospital employees enrolled in a hepatitis B vaccine program. All individuals were vaccinated with three injections of Heptavax-B, 1.0 ml containing 20 micrograms of HBsAg intramuscularly in the deltoid. The same lot of appropriately stored vaccine was used. Of the 41 individuals tested within zero to six months postvaccination, 35 (85%) and 38 (93%) were positive by EIA and RIA, respectively. Of the 75 individuals tested 7-24 months postvaccination, 61 (81%) and 71 (95%) were positive by EIA and RIA, respectively. Results of EIA tests performed at two laboratories were similar. Of 109 individuals positive by RIA, 13 did not have protective antibody levels. In contrast, of 96 individuals positive by EIA, only one did not have a protective antibody level. RIA may be a more sensitive test for anti-HBs, but a positive EIA result correlates better with protective antibody levels.

Regulation of electron transfer by the quinone pool.

Strong evidence for a random collisional mechanism for ubiquinone-mediated electron transfer is provided by the characteristic kinetic properties of respiratory chains originally explored by Kröger, A., and Klingenberg, M. (1973), Eur. J. Biochem. 34, 313-323. A kinetic model which leads to this so-called "simple Q-pool behavior" has been described and we use this in reviewing evidence that electron transfer is diffusion-controlled as well as diffusion-coupled. We also consider mechanisms by which the kinetics of electron transfer might deviate from simple Q-pool behavior and how these might be implicated in the regulation of electron transport.

Ultrastructural study of the liver in argininosuccinase deficiency.

The liver of a 19-year-old woman with argininosuccinase deficiency was studied ultrastructurally under conditions for excellent tissue fixation. Dilated smooth endoplasmic reticulum and mitochondrial crystalloids were the most prominent abnormalities found. These features were compared to fine-structural abnormalities found in other disorders of the Krebs-Henseleit pathway. No change was considered specific or related to subcellular enzyme localization.

When alcoholics drink aftershave: a study of nonbeverage alcohol consumers.

A one-year prospective study was conducted to describe nonbeverage alcohol (NBA) consumption (the use of substitutes for traditional forms of ethanol) among alcoholics. It was found that 11% of inpatient alcoholics at a veterans' hospital and 8.6% of alcoholics at a community hospital had consumed NBA. Substances consumed ranged from toiletries to organic solvents, often in quantities exceeding the theoretical lethal dose. 'Ready availability' was cited as the primary reason for consumption. Social, demographic and psychiatric parameters were then compared between 48 VA alcoholics who used NBA and 48 nonusing alcoholics. No social or demographic differences were found, but NBA drinkers drank more alcohol (P less than 0.0001), had higher global alcoholic severity scores (P less than 0.0001), more severe withdrawal symptoms (P less than 0.0001), and a higher frequency of antisocial personality disorder (P = 0.009) and drug abuse (P = 0.005). When NBA drinkers were subdivided by quantity of NBA consumption and recency of latest ingestion, no social or psychological differences were found between groups, except for more frequent and heavier illicit drug use among 'heavy' NBA consumers (P less than 0.0001).

Alcoholics who drink mouthwash: the spectrum of nonbeverage alcohol use.

Nonbeverage alcohol (NBA), or substitutes for traditional forms of beverage alcohol, includes such substances as mouthwash, aftershave lotion and alcohol-based fuels. Literature pertaining to the prevalence, clinical significance and toxicity of this practice is reviewed, using illustrative cases from a series of 48 NBA consumers. It was found that 10-15% of alcoholics hospitalized in detoxication units have consumed NBA; half of these patients are regular consumers. Addiction to NBA itself may occur. Its use is primarily related to easy accessibility, rather than social or monetary factors. Polydrug misuse and antisocial personality disorder are more frequent in NBA users, but use is not pathognomic of end-stage alcoholism. The 48 NBA users reported surprisingly few toxic symptoms from acute ingestion, perhaps because tolerance to some substances in NBA may occur. Isopropyl alcohol was the exception, reproducibly causing symptoms suggestive of severe gastritis.

Effect of exchange transfusion with an oxygen-carrying resuscitation fluid on the efficacy of penicillin therapy of pneumococcal infection in rats.

The effects of exchange transfusion with Fluosol DA (FDA) or stroma-free hemoglobin on the outcome of pneumococcal infection in rats were determined. Rats were sham transfused or exchange transfused with 25 ml of FDA or stroma-free hemoglobin. They were then challenged intraperitoneally with Streptococcus pneumoniae type 3 and treated with penicillin for 120 h. Only 2 of 15 (13.3%) FDA-transfused rats were alive at 312 h compared with 11 of 15 (73.3%) concurrently studied sham-transfused control rats (P = 0.0016). Of 10 stroma-free hemoglobin-transfused rats and 10 concurrently studied sham-transfused control rats (P = 0.98), 8 from each group (80%) were alive at 312 h. Penicillin therapy only suppressed pneumococcal infection in FDA-transfused rats, and relapse occurred after therapy was stopped. This effect could not be attributed to interference with the bactericidal activity of penicillin against pneumococci, to an alteration in the pneumococcal burden before penicillin therapy or to an alteration of the leukocyte and polymorphonuclear leukocyte response by FDA. In contrast, pneumococcal infection in stroma-free hemoglobin-transfused rats was cured with penicillin therapy. These data showed that FDA altered the ability of rats to respond to pneumococcal infection.

Primary omental leiomyosarcoma masquerading as hemorrhagic ascites.

An 85-year-old man with unexplained hemorrhagic pseudoascites of four years' duration was discovered to have a primary omental leiomyosarcoma. Neoplasms arising in the greater omentum are rare. To date, only three primary omental leiomyosarcomas have been described. Clinical and pathologic findings of this case are presented and the topic of primary omental neoplasms is reviewed.

Prolongation of the prodrome to acute hepatitis B infection by corticosteroids.

A 56-year-old woman had rash, arthralgia, and lymphadenopathy. Prednisone therapy caused the symptoms to abate but not disappear. Medication was continued for almost eight weeks, during which time the symptoms persisted. While the patient was receiving therapy, serologic evidence of hepatitis B infection was noted. When prednisone therapy was stopped, the patient rapidly passed from the prodrome to typical, acute, icteric hepatitis. Prednisone may have suppressed normal immunologic responses to the hepatitis virus, resulting in persistence of the serum sickness-like state. Corticosteroids are not indicated in the treatment of the prodrome to hepatitis B infection.

Hemodynamic effects on oxygen consumption and bile flow in isolated skate liver.

A technique is described for perfusion of livers from the little skate, Raja erinacea, in the isolated state at 15 degrees C utilizing a recirculating, well-oxygenated elasmobranch Ringer solution. Bile flow and oxygen consumption were optimal and remained relatively constant over a 4- to 5-h perfusion at portal vein pressures of 2.5-5.0 cm Ringer. At these pressures, bile flow rates were comparable to values previously observed in the free-swimming skate. In contrast to mammalian species, bile secretion in the elasmobranch is particularly sensitive to small changes in perfusion pressure. Hepatic clearance of sulfobromophthalein and [14C]sodium taurocholate demonstrated initial fractional disappearance rates that were also similar to values obtained in vivo. Only small amounts of sulfobromophthalein appeared in bile by 5 h, whereas 78.5 +/- 6.5% of [14C]sodium taurocholate was recovered in bile at this time. These experiments establish portal vein perfusion pressures for study of skate livers in the isolated state in which oxygen consumption is maximal and organic anion clearance from perfusate to bile exceeds values obtained in vivo.

Determinants of biliary secretion in isolated perfused skate liver.

In the isolated perfused liver of the little skate, Raja erinacea, bile flow averaged 5.07 +/- 0.58 (mean +/- SE) microliters.h-1.g liver-1 in 21 experiments at a perfusion pressure of 5.0 cm Ringer compared to 3.79 +/- 0.32 in 38 experiments at 2.5 cm (P less than 0.05). [14C]inulin readily entered skate bile. Bile-to-plasma [14C]inulin ratios corrected for delay in transit time, averaged 0.46 +/- 0.07 at 1 h and rose to 0.74 +/- 0.06 by 4 h, although bile flow remained constant. In experiments in which [14C]inulin reached equilibrium between bile and plasma, the bile-to-plasma ratio conformed to the theoretical relationship between bile flow, solvent drag, and inert solute diffusion predicted at extremely low bile flows, but demonstrated that the skate biliary tree is more permeable to inulin than that of the rat. Electron microscopic studies demonstrated that ionic lanthanum could traverse the tight junctions. However, freeze-fracture studies of junction structure did not differ qualitatively from similar studies in the rat. Partial dependence of bile flow on perfusion pressure, high bile-to-plasma inulin ratios, and permeability of the canalicular tight junctions to ionic lanthanum all suggest that the paracellular pathway may be an important component of bile formation in the skate.

Needlestick and puncture wounds: definition of the problem.

During 1979, all hospital personnel who were victims of needlestick or puncture wounds completed an incident questionnaire so that the epidemiology of these events could be determined. The attack rate varied from a high of 20 to a low of 0.8 incident/100 employee years worked (mean 7.5) for different employee groups. Personal carelessness accounted for 55% of the 81 incidents, whereas 35% of the involved personnel were innocent victims. Needles were responsible for 58% of the incidents. Screening for hepatitis B surface antigen (HBsAg) revealed no positive patient or employee associated with needlestick or puncture wounds. Only 2 of 45 employees were hepatitis B surface antibody (anti-HBs) positive. Hepatitis B immune globulin (HBIG) was recommended for 4 employees, immune serum globulin (ISG) for 13, and no globulin for 65. The estimated cost of this surveillance program and globulin therapy was $60 per incident. This data is being used as the basis for in-service training of high-risk personnel in order to decrease the number of incidents.

Bone disease in primary biliary cirrhosis: reversal of osteomalacia with oral 25-hydroxyvitamin D.

Bone disease and the response to oral 25-hydroxy-vitamin D were assessed in 7 women with primary biliary cirrhosis utilizing histomorphometric analysis of undecalcified iliac crest bone biopsies. Low serum concentrations of 25-hydroxyvitamin D rose from 9.2 +/- 6.7 (SD) to 151.0 +/- 103.7 ng/ml on oral 25-hydroxyvitamin D therapy (100--200 micrograms daily). On initial biopsy 5 of 7 patients had osteomalacia. Repeat bone biopsy in 6 patients after 6--8 mo of continued therapy revealed significant improvement in bone mineralization as reflected in fractional osteoid surface and relative osteoid volume. Osteomalacia healed in 4, improved in 1, and remained absent in 1. Trabecular bone volume, an index of mineralized bone mass, was diminished in 6 of 7 patients and did not change significantly over the 6--8-mo treatment period. We conclude that oral 25-hydroxyvitamin D corrects vitamin D deficiency and reverses osteomalacia in primary biliary cirrhosis.

Effect of chlorpromazine on hepatic perfusion and bile secretory function in the isolated perfused rat liver.

The hepatotoxicity of CPZ was studied in the isolated perfused rat liver in order to more closely define possible mechanisms of phenothiazine-induced cholestasis. Perfusate concentrations of CPZ were increased from 5 x 10(-6) M to 5 x 10(-4) M until bile secretion was significantly inhibited. Measurements were then made of determinants of bile secretory function, including the magnitude of lobar distribution of perfusate flow, BAIF, and liver plasma membrane enzyme activity, Na+,K+-ATPase, Mg++-ATPase and 5'-nucleotidase. BAIF diminished significantly from control values of 1.76 +/- 0.07 microliter min-1gm-1 of liver to 1.34 +/- 0.15 and 0.80 +/- 0.09 following 2.5 and 5 x 10(-4) M CPZ, respectively. Perfusate flow also diminished from 5.64 +/- 0.44 to 1.24 +/- 0.12 ml min-1 gm-1 of liver 20 min following 5 x 10(-4) M CPZ and was associated with reduced flow to peripheral areas of the hepatic lobes as demonstrated by Tc-HAM. By 30 min, perfusate flow had returned to baseline values. CPZ also transiently diminished the excretion of bile acids in livers receiving a constant infusion of 40 mumol hr-1 sodium taurocholate. Defects in hepatic perfusion could not account entirely for the impairment in BAIF, since comparable mechanical restriction of perfusate flow in controls only diminished BAIF to 1.49 +/- 0.08 microliter min-1gm-1 of liver. CPZ signofocamt;u rediced tje secofoc actovotu pf Mg++-ATPase and 5'-nucleotidase but did not affect Na+,K+-ATPase in liver plasma membrane isolated 20 min after 5 x 10(-4) M CPZ. CPZ also resulted in a profound shift in the recovery of protein in isolated liver plasma membrane fractions from the light (density = 1.16) to heavier (density = 1.18) fractions. These findings, together with previous observations demonstrating alterations in hepatic ultrastructure, indicate that CPZ interacts in a complex manner with hepatocyte plasma and cytoplasmic membrane components and suggest that these drug-membrane interactions independently result in diminished hepatic perfusion, impairment of bile acid excretion, and inhibition of bile acid-independent bile secretion.