Molecular typing, serotyping and cytotoxicity testing of Campylobacter jejuni strains isolated from commercial broilers in Puerto Rico.

AIMS: Thirty Campylobacter jejuni strains isolated from fecal samples (n = 94; 32%) from 13 positive farms (n = 17; 76%) from commercial broiler chickens in Puerto Rico were analysed by molecular methods. METHODS AND RESULTS: Isolates were identified with multiplex polymerase chain reaction assays, tested for their antimicrobial susceptibility and characterized with pulsed-field gel electrophoresis (PFGE), multilocus sequence typing (MLST), serotyping and bacterial cytotoxicity in mammalian cells. Isolates exhibited high resistance to vancomycin (minimum inhibitory concentration, MIC of >256 microg ml(-1)) and trimethoprim (MIC of >32 microg ml(-1)); few were resistant to clindamycin (MIC(90) 4 microg ml(-1)), erythromycin (MIC(90) 8 microg ml(-1)) and tetracycline (MIC(90) 8 microg ml(-1)); but none was resistant to azithromycin (MIC(90) 4 microg ml(-1)), ciprofloxacin (MIC(90) 1 microg ml(-1)) or gentamycin (MIC(90) 4 microg ml(-1)). Most strains restricted with SmaI, but a combination of SmaI-KpnI digestion was more discriminatory. MLST analysis yielded four sequence types (ST), and ST-2624 was the predominant one. Phylogenetic analysis revealed a high degree of recombination for glnA and pgm genes. The predominant serotypes were O:3 and O:5. Most strains had lowest cytotoxicity potential with Caco-2 cells, medium cytotoxicity with INT-407 and Hep-2 cells and high cytotoxicity with CHO cells. CONCLUSION: A low degree of antimicrobial resistance, 13 PFGE profiles, 4 ST and a large variability in cytotoxicity assays were found for these strains. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first characterization of C. jejuni strains isolated from broilers in Puerto Rico. The genetic diversity of these strains suggests that several techniques are needed for strain characterization.

Oxidative stress in laboratory-incubated double-crested cormorant eggs collected from the Great Lakes.

Double-crested cormorant (Phalacrocorax auritus) eggs were collected in 1998 from three sites on Lakes Huron and Superior and either analyzed for 2, 3, 7, 8-tetrachlorodibenzo-p-dioxin (TCDD)-like residues or artifically incubated. Some of the incubated eggs were injected with vitamin E (antioxidant) or piperonyl butoxide (CYPIA blocker) to examine the role of CYPIA and oxidative stress in normal bird development. Embryos (day 23) were analyzed for hepatic ethoxyresorufin O-deethylase (EROD) activity and different measures of oxidative stress. Glutathione-related parameters were also measured in brain. In contrast to the historical data, there were no statistically significant differences in concentrations of chlorinated dioxins, furans, dioxin-like PCBs, or total TCDD-equivalents (TEQs) in eggs among sites. Survival and incidence of abnormalities were comparable at all study sites. Slight differences in liver, heart, and egg weight were observed among sites. A greater incidence of eye deformities was observed in embryos treated with vitamin E. Treatment with the CYPIA blocker, piperonyl butoxide, decreased the body weights of embryos. EROD activities were similar at all locations, but measures of oxidative stress varied among locations. There were greater levels of oxidized glutathione and oxidative DNA damage at Little Charity Island in Saginaw Bay. There was relatively great interindividual variation in biochemical responses and significant interrelation of the parameters of oxidative stress. While exposure to PCDD/DF and PCBs does not seem to explain the observed oxidative stress, the potential of these compounds to cause the observed effects can not be completely excluded.

Dietary inulin suppresses azoxymethane-induced aberrant crypt foci and colon tumors at the promotion stage in young Fisher 344 rats.

This study was designed to determine the effect of 10% dietary long-chain inulin on the azoxymethane (AOM)-induced colonic preneoplastic aberrant crypt foci (ACF) and small intestinal and colon tumors at the initiation (I), promotion (P) and I + P stages (20 rats per treatment) in Fisher 344 male weanling rats. After an acclimatization period of 1 wk, groups of Fisher 344 male weanling rats were assigned to consume AIN 93G diet (control) or AIN 93G diet containing 10% inulin. All the rats received 16 mg/kg body AOM dissolved in saline subcutaneously at 7 wk of age followed by a second injection at 8 wk of age. An additional group of five rats received only saline and consumed the control diet. The rats received the assigned diets until asphyxiation by CO(2) at 16 wk of age for the ACF experiment and 45 wk for the end-point tumor experiment. Feed intake, weight gain, diarrheal index, cecal weight, cecal pH, ACF and tumors in the colon were determined. Rats fed inulin had diarrhea after 2 wk of feeding and recovered by approximately 4 wk. Cecal weight was greater in rats fed inulin and cecal pH was lower. The inulin group had more than 66% fewer aberrant crypts and 60% fewer ACF compared with the control group. Tumor incidences in the small intestine and colon of rats in the control, I, P and I + P groups were: 78, 31, 0 and 11% and 90, 73, 69 and 50%, respectively. The corresponding values for the distal portion of the colon were 87, 63, 45 and 33%, respectively. Colon tumors per tumor-bearing rat were 4.2, 3.09, 1.36 and 1.2 for the control, I, P and I + P groups, respectively. All groups differed, P < 0.05. The results of this study indicate that dietary long-chain inulin suppresses AOM-induced ACF formation, an early preneoplastic marker of colon tumorigenesis in rats, and colon tumors, particularly at the promotion stage.

Accumulation of 2,3,7,8-tetrachlorodibenzo-p-dioxin by rainbow trout (Onchorhynchus mykiss) at environmentally relevant dietary concentrations.

Rainbow trout were fed a diet containing 1.8, 18, or 90 pg/g 3H-2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) for up to 320 d. Concentrations of TCDD were determined in muscle, liver, and ovaries at 100, 150, 200, and 250 d. Concentrations of TCDD reached an apparent steady-state concentration in liver after 100 d of exposure, whereas concentrations in other tissues continued to increase until 150 d of exposure. The greatest portion of the total mass of TCDD was present in the muscle tissue with lesser proportions in other organs. As the ovaries developed before spawning, an increase occurred in the total mass of TCDD present in this tissue. The assimilation rate of TCDD during the initial 100 d of the exposure was determined to be between 10 and 30%. This is somewhat less than estimates derived based on both uptake and elimination constants determined during shorter exposures. Biomagnification factors (BMFs) were estimated for all tissues and exposure concentrations, and at all exposure periods. Lipid-normalized BMFs for muscle ranged from 0.38 to 1.51, which is consistent with the value of 1.0 predicted from fugacity theory. Uptake and depuration rate constants were determined and used to predict individual organ TCDD concentrations. Comparison with observed values indicated that the model could be used to predict tissue concentrations from the known concentrations of TCDD in food. This model will allow more refined risk assessments by predicting TCDD concentrations in sensitive tissues such as developing eggs.

Polychlorinated naphthalenes, -biphenyls, -dibenzo-p-dioxins, and -dibenzofurans in double-crested cormorants and herring gulls from Michigan waters of the Great Lakes.

Concentrations of polychlorinated dibenzo-p-dioxins (PCDDs), dibenzofurans (PCDFs), naphthalenes (PCNs), and biphenyls (PCBs) were measured in eggs of double-crested cormorants and herring gulls collected from Michigan waters of the Great Lakes. Concentrations of PCNs in eggs of double-crested cormorants and herring gulls were in the ranges of 380-2400 and 83-1300 pg/g, wet wt, respectively. Concentrations of 2,3,7,8-substituted PCDDs and PCDFs were 10-200 times less than those of PCNs in eggs whereas those of total PCBs (380-7900 ng/g, wet wt) were 3-4 orders of magnitude greater. While the profile of PCB isomers and congeners between double-crested cormorants and herring gulls was similar, the PCN isomer profile differed markedly between these two species. PCN congeners 66/67 (1,2,3,4,6,7/1,2,3,5,6,7) accounted for greater than 90% of the total PCN concentrations in herring gulls, whereas their contribution to total PCN concentrations in double-crested cormorants ranged from 18 to 40% (mean, 31%). The ratios of concentrations of PCDDs to PCDFs were greater in herring gulls than in double-crested cormorants collected from the same locations, suggesting the ability of the former to metabolize PCDF congeners relatively rapidly. 2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD) equivalents (TEQs) contributed by PCNs in double-crested cormorant and herring gull eggs were 2-3% of the sum TEQs of PCBs, PCDDs, PCDFs, and PCNs. PCB congener 126 (3,3',4,4',5-PeCB) accounted for 57-72% of the total TEQs in double-crested cormorant and herring gull eggs.

Mixed prognosis: equity, access, and emerging issues within British Columbia's health care system.

The author, an American, examines British Columbia's health care system as it struggles to retain equity and access as two of its implacable tenets. Some sources argue that realities fall far short of the abstract ideals. Certainly implementation and funding problems are growing as B.C.'s policy makers struggle with large-scale economic pressures and management priorities. Despite the system's shortcomings, however, U.S. readers would be wise to consider the philosophical and rhetorical impact of these normative concepts. Their presence and power encapsulates the breadth of the contrast between the Canadian and American systems.

Effect of altitude training on serum creatine kinase activity and serum cortisol concentration in triathletes.

In this investigation we evaluated the effect of a 5-week training program at 1860 m on serum creatine kinase (CK) activity and serum cortisol concentration in national-caliber triathletes for the purpose of monitoring the response to training in a hypobaric hypoxic environment. Subjects included 16 junior-level female (n = 8) and male (n = 8) triathletes who were training for the International Triathlon Union (ITU) World Championships. After an initial acclimatization period, training intensity and/or volume were increased progressively during the 5-week altitude training camp. Resting venous blood samples were drawn at 0700 hours following a 12-h overnight fast and were analyzed for serum CK activity and serum cortisol concentration. Subjects were evaluated before [7-10 days pre-altitude (SL 1)] and after [7-10 days post-altitude (SL 2)] the 5-week training camp at 1860 m. At altitude, subjects were evaluated within 24-36 h after arrival (ALT 1), 7 days after arrival (ALT 2), 18 days after arrival (ALT 3), and 24-36 h prior to leaving the altitude training camp (ALT 4). A repeated-measures analysis of variance was used to evaluate differences over time from SL 1 to SL 2. Compared to SL 1, serum CK activity increased approximately threefold (P < 0.05) within the initial 24-36 h at altitude (ALT 1), and increased by an additional 70% (P < 0.05) after the 1st week of altitude training (ALT 2). Serum CK activity remained significantly elevated over the duration of the experimental period compared to pre-altitude baseline levels. Serum cortisol concentration was increased (P < 0.05) at the end of the 5-week altitude training period (ALT 4) relative to SL 1, ALT 1 and ALT 3. These data suggest that: (1) the initial increase in serum CK activity observed in the first 24-36 h at altitude was due primarily to acute altitude exposure and was independent of increased training intensity and/or training volume, (2) the subsequent increases in serum CK activity observed over the duration of the 5-week altitude camp were probably due to the combined effects of altitude exposure and increased training load, and (3) the increase in serum cortisol concentration observed at the end of the altitude training camp reflects the additive effect of 5 weeks of altitude exposure in combination with a progressively increased training intensity and/or volume.

Polychlorinated dibenzo-p-dioxins (PCDDs) and dibenzofurans (PCDFs) in muscle and eggs of salmonid fishes from the Great Lakes.

Concentrations of polychlorinated dibenzo-p-dioxins (PCDDs) and dibenzofurans (PCDFs) were measured in dorsal muscle and eggs of coho salmon, lake trout, and chinook salmon collected from Lakes Superior, Michigan, and Huron (Michigan waters). Absolute and relative concentrations of PCDDs and PCDFs varied among sampling locations (inter- and intralake) and fish species. Fish collected from Bay City (Saginaw Bay) contained the greatest concentrations of PCDDs and PCDFs both in muscle and eggs. Among the three fish species, chinook salmon accumulated greater concentrations than did coho salmon or lake trout. Concentrations of PCDFs were greater than those of PCDDs in all fishes. OCDF and TCDF were the predominant congeners of PCDF, whereas OCDD and TCDD concentrations predominated in PCDDs. Homolog compositions of PCDDs and PCDFs suggested the existence of multiple local sources in various locations. Concentrations of PCDDs and PCDFs in eggs of fishes were significantly correlated with those in muscle.

Who are they with and what are they doing? Delinquent behavior, substance use, and early adolescents' after-school time.

The relationship among after-school time, parental monitoring, and problem behavior was examined in a sample of 1,170 early adolescents. Those spending unsupervised time with peers reported higher levels of aggression, delinquency, substance use, and susceptibility to peer pressure, and lower levels of parental monitoring, than did adolescents at home with parents. Adolescents home alone after school were similar to those who spent time with adults or in school activities.

Thiotepa and cyclophosphamide with stem cell rescue for consolidation therapy for children with high-risk neuroblastoma: a phase I/II study of the Pediatric Blood and Marrow Transplant Consortium.

PURPOSE: We report the results of a phase I/II stem cell rescue trial for patients with high risk neuroblastoma. PATIENTS AND METHODS: Fifty-one patients with a median age of 2.3 years (range 1 to 20) who were in their first complete remission (CR) (n = 8), very good partial remission (VGPR) (n = 23), partial remission (PR) (n = 5), or subsequent CR/PR (n = 7) after receiving a platinum-based induction regimen were consolidated with high dose chemotherapy and stem cell rescue. They received an ablative regimen of thiotepa (300 mg/m2/day for 3 days) and cyclophosphamide (1500 mg/m2/day for 4 days) followed by either purged marrow (n = 16), unpurged bone marrow (BM) (n = 23), or peripheral blood stem cell (PBSC) rescue (n = 13). The median nucleated cell doses administered were 2.7 x 10(8)/kg for unpurged marrow (range 1.1 to 13), 1.7 x 10(8)/kg for purged marrow (range 0.8 to 6.4), and 2.1 x 10(8)/kg for the PBSC (range 1.1 to 13). RESULTS: Engraftment was achieved for all patients. The time to achieve an absolute neutrophil count (ANC) >500 x 10(9)/l was 19 days for patients who received purged BM (range 13 to 18), 17.5 days for patients who received unpurged BM (range 9 to 38), and 13 days for patients who received PBSC (range 9 to 25). An unsustained platelet count >20 x 10(9)/l was attained in 33.5 days by patients who received purged BM (range 13 to 100), 35 days for patients who received unpurged BM (range 14 to 128), and 20 days for patients who received PBSC (range 11 to 64). There was one infectious death in the unpurged marrow group caused by aspergillosis pneumonia, but none in the other two groups. Progressive disease (PD) developed in 21 patients at a median of 271 days (range 31 to 1230). The remaining 29 patients are progression-free at a median follow-up of 1190 days (range 530 to 2383). CONCLUSION: We conclude that this regimen is well tolerated, and that progression-free survival (PFS) with this chemotherapy-only regimen compares favorably with regimens containing total body irradiation (TBI).

2-Chlorodeoxyadenosine (2-CDA) for the treatment of refractory or recurrent Langerhans cell histiocytosis (LCH) in pediatric patients.

BACKGROUND: Pediatric patients with Langerhans cell histiocytosis (LCH) may become refractory to conventional therapy or present with repeated recurrences over several years. Current therapeutic options such as prednisone, vinblastine, etoposide, and cyclosporine are associated with significant acute toxicities and late effects. Recent reports suggested that 2-chlorodeoxyadenosine (2-CDA) may be an effective agent in adults with LCH. The purpose of this study was to determine the safety and efficacy of 2-CDA in children with LCH. METHODS: This report presents the data collected from the first three patients that have completed this trial. Patients were enrolled in a prospective study after informed consent was obtained. Patients had a confirmed diagnosis of LCH that had recurred several times or not responded to standard therapy. Patients were given a starting dose of 5 mg/M2 of daily continuous infusion for three days duration. Two patients had their dose increased to 6.5 mg/M2/ day. A total of 4-6 courses were given, and courses were repeated every 3-4 weeks. Thirteen of fifteen courses were given as outpatients at home. RESULTS: Each patient completed therapy with myelosuppression the primary toxicity. Pt. 1 initially received a higher dose of 2-CDA and developed sepsis. The dose was reduced to current study levels and no other incidence of infection, fever, and neutropenia, or blood product transfusion was required. All three patients are free of active disease 10-18 months after completing 2-CDA. CONCLUSION: Three patients with LCH refractory to standard therapy had CR to 2-CDA, given at 5-6.5 mg/M2/day for 3 days, without significant toxicity.

Outpatient therapy with ceftriaxone and oral cefixime for selected febrile children with sickle cell disease.

PURPOSE: Children with sickle cell disease are at increased risk for bacterial sepsis and, when febrile, are usually hospitalized for intravenous antibiotic therapy pending results of blood cultures. In this study, we prospectively identified a group of febrile patients with sickle cell disease who were at low risk for sepsis and treated them with outpatient therapy. PATIENTS AND METHODS: Children identified as low risk for sepsis were treated with an initial dose of intravenous ceftriaxone, followed by outpatient therapy with oral cefixime, and were monitored for 14 days after the initial visit. Compliance was assessed by phone calls to parents and by analysis of urine samples. RESULTS: In 107 eligible febrile episodes (80 patients) over a 21-month period, no patient developed sepsis. One child developed bacteremia 3 days after completing the course of cefixime, and one had splenic sequestration on the fourth study day. Both patients did well. Side effects of cefixime were modest, and overall compliance was excellent (approximately 95%), although urine samples were returned by only 56% of parents. CONCLUSION: We conclude that outpatient therapy is safe and effective in febrile patients with sickle cell disease who meet the criteria for a low risk of sepsis.

Cure of X-linked lymphoproliferative disease (XLP) with allogeneic hematopoietic stem cell transplantation (HSCT): report from the XLP registry.

Seven male patients in the David T Purtilo International X-linked Lymphoproliferative Disease (XLP) Registry have undergone allogeneic hematopoietic stem cell transplantation (HSCT). All patients received HSCT from HLA-identical donors: sibling BM, five; unrelated BM, one; and sibling umbilical cord blood, one. Ages at time of HSCT ranged from 5 to 30 years. Pre-HSCT clinical course varied, but four boys had a significant history of chronic and/or serious infections. Conditioning regimens varied: TBI containing regimens, four, chemotherapy only, three. All patients engrafted. Six developed grade I-II acute GVHD but no chronic GVHD. Four are alive and well with normal immune function greater than 3 years following HSCT. Three died within 100 days: disseminated adenovirus, one; polymicrobial sepsis, one; and multiple organ system failure and bleeding diathesis, one. No EBV-associated post-transplant complications were observed, even though all donors except the umbilical cord blood were EBV-seropositive. Unsuccessful HSCT was associated with age at HSCT (> 15 years), TBI-containing regimen and significant history for pre-HSCT infections. These results provide evidence that HSCT performed during childhood with HLA-identical sibling donors, regardless of EBV serostatus, offers the only curative therapy for XLP.

Polychlorinated biphenyls and 2,3,7,8-tetrachlorodibenzo-p-dioxin equivalents in eggs of double-crested cormorants from a colony near Green Bay, Wisconsin, USA.

Great Lakes colonial waterbirds have experienced poor reproduction and a greater incidence of birth defects than those in remote areas. An egg was collected from each of 1,000 marked cormorant nests at Spider Island (Lake Michigan). Nine pools comprised of three eggs were randomly selected for instrumental quantification of polychlorinated biphenyls (PCB) congeners, calculation of 2,3,7,8-tetrachlorodibenzo-p-dioxin equivalents (TEq) and measurement of equivalents by bioassay (TCDD-EQ). PCB analysis of the nine samples was semi-automated with high performance liquid chromatography (HPLC) columns including a porous graphitic carbon column. TEqs were calculated from concentrations of PCB congeners and bioassay-derived toxic equivalency factors (TEfs), and TCDD-EQ were measured directly with an H4IIE bioassay. Total PCB concentrations ranged from 9.7 to 38 micrograms/g, wet weight (ww). Mean concentrations of PCB 77, 126, and 169 were 2, 7, and 1 ng/g, ww. The mean TEqs and TCDD-EQ were 150 and 350 pg/g, ww, respectively. Thus, PCB congeners contributed less than 50% of the total TCDD-EQs as measured by the bioassay.

Plasma 3-methoxy-4-hydroxyphenylglycol (MHPG) and clinical symptoms in Alzheimer's disease.

Postmortem findings point to significant abnormalities in central noradrenergic function in Alzheimer's disease (AD) which may be associated with changes in peripheral markers. In this study, the relationship between the peripheral noradrenergic marker, plasma 3-methoxy-4-hydroxyphenylglycol (MHPG), and clinical symptoms was examined in 23 patients with probable AD. Basal MHPG levels correlated significantly with increased cognitive impairment (r = .58, p = .005), controlling for age, age at onset, gender, and time interval between plasma MHPG determination and cognitive testing. These results suggest that plasma MHPG increases as cognitive function in AD deteriorates, further supporting preliminary evidence for increases in noradrenergic indices in association with disease severity in AD.

Biased attention in childhood anxiety disorders: a preliminary study.

This study provides preliminary tests of two hypotheses: (1) Anxiety-disordered children show an attentional bias toward emotionally threatening stimuli, and (2) normal controls show an attentional bias away from emotionally threatening stimuli. Twelve children, 9 to 14 years of age, with primary diagnoses of anxiety disorder were compared with 12 normal controls matched for age, gender, vocabulary level, and reading ability. Subjects completed a reaction time task that measured visual attention toward threatening versus neutral words. The anxious group showed the predicted attentional bias toward threat words. However, controls did not show the predicted bias away from threat words. These results are the first showing that biased attentional processing occurs among clinically anxious children. The potential role of such an attentional bias in childhood anxiety disorders and future direction for research are discussed.

Skeletal changes in multiparous and nulliparous mice fed a nutrient-deficient diet containing cadmium.

Female mice were given nutrient-deficient, purified diets containing either 0.25 (environmental), 5, or 50 ppm Cd; the nutrient quality of each was patterned after deficiencies known to be present in food consumed by Japanese women who contracted Itai-Itai disease. One-half of the mice were bred for six consecutive, 42-day rounds of pregnancy/lactation (PL mice); remaining females were non-pregnant, virgin controls (NP mice). PL and NP mice were sacrificed at the end of rounds 1, 2, 3, 5, or 6. PL mice taken during the first three rounds were successively pregnant; those taken in later rounds experienced gestation/lactation either four (round 5) or three (round 6) non-successive times. No consistent round-by-round decreases in diet consumption or body weight occurred among NP mice during the 252 days of cadmium exposure, however a significant decrease in femur calcium content (11-17%) was observed in virgin groups exposed to 50 vs. 0.25 ppm Cd. Similar femur decalcification (14-20%) was observed in PL mice, however calcium loss at 50 ppm Cd paralleled decreases in food consumption (24%) and body weight (9-17%). Significant but smaller decreases in the calcium/dry weight (Ca/DW) ratio were found for NP and PL groups consuming 50 ppm dietary Cd. Over the 6-round experiment, exposure to cadmium was found to effect smaller decreases in both femur Ca content and Ca/DW ratio than either consumption of nutrient-deficient diet or multiparous experience. Demineralization results for PL mice provide evidence that the combination of chronic ingestion of cadmium in a nutrient-deficient diet and multiparous activity likely played a role in the etiology of Itai-Itai disease; results for NP mice additionally suggest that decalcification may have been initiated in human females at a time prior to the multiparous and menopausal stages of life.

Abnormal responses to cold stress in Charcot-Marie-Tooth I syndrome.

In Charcot-Marie-Tooth syndrome (CMT, Hereditary Motor Sensory Neuropathy), patient complaints of cold intolerance are common but their peripheral responses to cold have not been documented. Using digital plethysmography, a simple test of vascular reactivity with 1 minute cold stress, 20 unrelated adult CMT patients showed a significantly increased average heart rate and decreased average arterial oxygen saturation following cold when compared to 50 age-matched normal controls. There did not appear to be a unique or characteristic CMT vascular reaction to cold stress in CMT patients because their abnormal peripheral vascular responses were variable. Variability in CMT neuropathic responses to cold is consistent with the known irregular segmental demyelination of CMT peripheral nerves as well as abnormal CMT sweating patterns. Though understanding the precise patterns of CMT patient peripheral nerve disturbances remains difficult, awareness of CMT patient's abnormal responses to cold may facilitate CMT patient care and rehabilitation.

XomaZyme-CD5 immunotoxin in conjunction with partial T cell depletion for prevention of graft rejection and graft-versus-host disease after bone marrow transplantation from matched unrelated donors.

Patients who receive bone marrow transplants from unrelated donors have a high incidence of graft-versus-host disease (GVHD). If the donor marrow is first T cell-depleted, the everity of GVHD declines but the risk of rejection rises. In an attempt to prevent both graft rejection and GVHD, we included an anti-T cell antibody-toxin conjugate (CD-5-Ricin; XomaZyme H65) in the transplant conditioning regimen. After receiving a partially T cell-depleted marrow, patients then received a second course of immunotoxin as additional GVHD prophylaxis. Eight recipients of unrelated donor marrow transplants were studied. All engrafted (ANC > 500 x 10(6)/l by day 15, range 13-20 days). One patient had grade II skin GVHD and one developed grade IV disease but the other six patients had no acute GVHD. However, there was high morbidity and mortality from virus infections associated with a sluggish return of CD4 and CD8 T cells into the normal range. Four patients died from virus disease (CMV, n = 2; EBV, n = 1; adenovirus, n = 1) and the remaining patients had frequent documented viral illnesses during the first year. We conclude that improvement in the outcome of unrelated donor marrow transplantation will require strategies which prevent rejection and GVHD coupled with attempts to accelerate immune reconstitution.

Altered membrane fatty acids of cultured human retinal pigment epithelium persistently infected with rubella virus may affect secondary cellular function.

Persistent infection with rubella virus (RV) can alter secondary functions of host cells. Previously we had documented defective phagocytosis of latex beads by cultured human retinal pigment epithelial cells (RPE), persistently infected with M-33 RV (RPE/RV). Here, examining possible mechanisms for altered function, we reported significant differences between the total esterified fatty acids (FA) of RPE and RPE/RV membranes, measured by gas liquid chromatography. RPE/RV contained an increased proportion of saturated FA, particularly palmitic acid, with a presence of unusual chromatographic FA peaks co-eluting with odd-numbered long-chain carbon atom FA not normally found in human cells. Apical membrane microvilli, structures essential to phagocytic activity of RPE and RPE/RV, observed by scanning and transmission electron microscopy, were similar in number and appearance between uninfected RPE and RPE/RV cells before and after latex bead addition. However, RPE/RV microvilli, possibly reflecting altered membrane FA composition, engaged latex beads less effectively than uninfected RPE microvilli. In addition, microvilli remained abnormally distributed on RPE/RV cell surfaces at 48 h after latex addition. Thus, RV persistent infection may affect the cellular membrane fluidity and functional activity of human cells with increased saturated FA proportions and altered FA components of membrane phospholipids. These changes may participate in the defective phagocytosis of RPE/RV.