Effects of Cryptosporidium parvum infection on lymphocyte phenotype and reactivity in calves

Cryptosporidium parvum is a protozoan parasite now recognized as a significant cause of neonatal diarrhea in calves, and infection is also widespread in both immunocompetent and immunocompromised humans. No effective treatment or preventive measures against C. parvum infection are available, owing largely to the lack of understanding of immunologic mechanisms of resistance to and recovery from this parasite. In the present study, we compared phenotypes of lymphocytes from peripheral blood, spleen, mesenteric, and prescapular lymph nodes of calves infected or not infected with C. parvum. We also compared reactivity of these lymphocytes to mitogens and C. parvum antigen in vitro. There were more non-T, non-B (null) lymphocytes in all tissues of infected compared with control calves. The percent of CD8+ lymphocytes was significantly increased in spleens of infected compared with control calves, and there were markedly less CD4+ than CD8+ cells in spleens of both groups (i.e. low CD4/CD8 ratios). Splenic lymphocytes showed significantly decreased in vitro proliferation to pokeweed mitogen and C. parvum antigen stimulation compared with lymphocytes from other tissues. These findings suggest that null lymphocytes and CD8+ lymphocytes may be important in the expression and regulation of bovine immune responses to C. parvum in vivo.     

Prophylactic effect of bovine anti-Cryptosporidium hyperimmune colostrum immunoglobulin in healthy volunteers challenged with Cryptosporidium parvum

Bovine hyperimmune anti-Cryptosporidium colostrum immunoglobulin (BACI) decreases the intensity of Cryptosporidium parvum infection in vitro. We investigated the prophylactic effect of BACI in healthy adults challenged with C. parvum. After we established an oocyst dose that resulted in 100% infection in four volunteers (baseline group), 16 volunteers were randomized to receive (1) BACI prior to C. parvum challenge (BACI group) and a nonfat milk placebo 30 minutes later, (2) BACI prior to and 30 minutes after challenge (reinforced BACI group), or (3) nonfat milk placebo prior to and 30 minutes after challenge. Subjects received BACI (10 g) or nonfat milk placebo three times a day for a total of 5 days and were followed for clinical symptoms and oocyst excretion for 30 days. A trend toward less diarrhea (P = .08) was observed for subjects receiving BACI in comparison with occurrences in placebo recipients. Subjects receiving BACI or nonfat milk placebo had a 100-fold reduction in oocyst excretion as compared with excretion in the baseline group.     

Prophylactic ganciclovir treatment reduces fungal as well as cytomegalovirus infections after heart transplantation

Cytomegalovirus (CMV) infection is associated with an increased incidence of other opportunistic infections in organ transplant recipients. Whether this is related to immunomodulating effects of CMV or independent of CMV but associated with a host risk factor common to both infections is unclear. The purpose of this study was to determine whether the reduction in CMV infections seen with prophylactic ganciclovir treatment after heart transplantation is associated with a reduced incidence of other opportunistic infections. Of 149 patients prospectively enrolled in a multicenter, randomized, double-blind, placebo-controlled trial of ganciclovir to prevent CMV disease, 74 patients enrolled at this center (33 control and 41 ganciclovir-treated) were retrospectively identified. All received prophylactic OKT-3 and standard 3 drug maintenance immunosuppressive therapy. Actuarial survival and rejection rates and incidence of opportunistic infections (bacterial, fungal, and protozoal) for the 2 treatment groups were determined and compared using Cox-Mantel analysis. CMV disease occurred 2.5 times more frequently in the control group. There were no significant differences in survival or rejection rates nor in bacterial or protozoal infection incidence between the 2 groups. Bacterial infections occurred in 54% of control and 39% of ganciclovir-treated patients (P = 0.18). There were significantly fewer fungal infections in the ganciclovir-treated group (7% vs. 27%, P = 0.0071). CMV and fungal infections were both significantly reduced in patients who received ganciclovir prophylaxis. This suggests that active CMV disease may be causally associated with the development of opportunistic fungal infections.     

Clinical evaluation of postoperative adjuvant arterial infusion chemotherapy in resected hepatoma patients

Eighty surgically treated patients with advanced hepatocellular carcinoma (HCC) were divided into two groups. In group I, twenty patients whose mean diameter of tumors was 56 mm, prophylactically underwent hepatic arterial infusion chemotherapy after liver resection. Chemotherapeutic agents (5-FU, ADM, MMC, CDDP, Lipiodol) were administered 4 times a year via Infuse A port. The remaining 60 patients, whose mean diameter of tumors was 57 mm, served as the control without prophylactic infusion (group II). The 2-year cumulative survival rate was higher in the prophylactic group (71%) than the control (48%, p = 0.040). The two-year disease-free survival rate was improved in group I (38%) compared with that in group II (27%, p = 0.021). Intrahepatic multiple recurrence within 1 year after surgery was recognized in four out of 18 patients of group I (22%) and in thirty-three out of 60 patients of group II (55%, p = 0.029). In group I, two cases who died of hepatic failure with no recurrence, had lower functional reverse and a larger amount of Lipiodol than the remaining 18 patients. Adjuvant arterial infusion chemotherapy can thus be be efficacious in alleviating hepatoma recurrence after liver resection. For patients with poor liver function, a smaller volume of chemotherapeutic agents might be feasible.     

Patient outcomes after reoperation on the lumbar spine

A prospective, randomized trial was conducted to compare the effectiveness and safety of warfarin given in two regimens in prevention of venous thrombosis after total knee replacement. Adult patients scheduled for primary or revision total knee replacement were randomly assigned to receive either a "two-step" warfarin regimen beginning 10-14 days pre-operatively or, alternatively, to begin warfarin the night before surgery. Post-operatively, the dose was adjusted in both groups to achieve a target International Normalized Ratio (INR) of 2.2 and prophylaxis was continued until venography on post-operative days five through nine. Bleeding was assessed by surgical blood loss, transfusion requirements, changes in hematocrit, and clinically identified bleeding complications. The occurrence of deep vein thrombosis was nearly the same in the two treatment groups, 39% in patients randomized to the two-step regimen as compared to 38% in those beginning the night before surgery. The occurrence of proximal vein thrombosis was also similar, 5% versus 7% (p = NS). Patients in the two-step group received 1.33 +/- 1.26 transfusions compared to 0.95 +/- 1.22 in the night before group (p < 0.05) and also had a lower nadir post-operative hematocrit of 26.7 +/- 3.1 as compared to 28.5 +/- 3.2 (p < 0.0001). Major bleeding complications were associated with excessively prolonged INRs and occurred in five patients in the two-step group and two in the night before group. Patients in both groups who developed thrombosis had a significantly lower INR on post-operative days two and three compared to those without thrombosis. We conclude that a prophylactic warfarin regimen for prevention of deep vein thrombosis after total knee replacement beginning the night before surgery is more convenient and may be associated with less bleeding than a regimen beginning warfarin 10-14 days pre-operatively. Careful control of anticoagulant intensity is needed to achieve maximum effectiveness and avoidance of bleeding complications.     

The effect of vaccination against Mycoplasma hypopneumoniae in pig herds with a continuous production system

An inactivated Mycoplasma hyopneumoniae vaccine was evaluated in five pig herds clinically infected with enzootic pneumonia and practising a continuous production system in the growing/finishing unit. In each herd, a vaccinated and control group of approximately 47 pigs each were individually monitored from birth until slaughter. Vaccinated pigs received the first dose at about 1 week of age and the second approximately 3 weeks later. During all production stages, an equal number of vaccinated and control pigs was present in the same pen. Both groups were compared with respect to zootechnical parameters (major variables) and by means of serological, pathological, and bacteriological parameters (ancillary variables). Daily weight gain was improved by 14 gr/day during the period from 8 days of age until slaughter (P = 0.0486) and by 25 gr/day during the growing/finishing period (P = 0.0067). Mortality rate, and the costs for curative medication were not significantly improved by vaccination. The results of the ancillary variables are presented and discussed.     

白血病肿瘤疫苗联合1-甲基色氨酸免疫治疗荷瘤小鼠疗效观察

目的 探讨白血病肿瘤疫苗(简称瘤苗)主动免疫治疗及联合吲哚2,3双加氧酶(IDO)的抑制剂1-甲基色氨酸(1-MT),在白血病荷瘤小鼠治疗中的作用.方法 采用FBL-3细胞皮下注射建立荷瘤白血病小鼠模型;实验分为5组:正常对照组、PBS对照组、环磷酰胺(CTX)化疗组、单用瘤苗治疗组和瘤苗联合1-MT治疗组;观察各组小鼠的一般状况、肿瘤缓解率、肿瘤大小、转移情况及生存期.结果 PBS对照组小鼠活动迟缓,体质量(含瘤结节质量)比其余各组均高;单用瘤苗组和瘤苗联合1-MT组小鼠活动、进食正常,体质量与正常小鼠筹异不大;化疗组体质量明显减轻,出现脱毛、弓背、活动减少等,差异有统计学意义(F=57.71,P=000);单用瘤苗组和瘤苗联合1-MT组治疗相关死亡率明显低于化疗组(0,0,40%).瘤苗联合1-MT组完全缓解率与单用瘤苗组(61.1%、70.0%)比较,差异无统计学意义(χ2=0.221,P>0.05),但瘤苗联合1-MT组的复发率低于单用瘤苗组(0,36.36%);复发小鼠再应用1-MT,能明显抑制瘤结节的生长.单用瘤苗组和瘤苗联合1-MT组小鼠中位存活期明显高于化疗组和PBS对照组(χ2=52.13,P<0.01).各组小鼠整体瘤结节的变化比较差异有统计学意义(F=89.966,P=0.000).结论 白血病瘤苗在动物实验具有肯定的疗效,能明显抑制肿瘤的生长,延长小鼠生存时间,且副作用小.免疫治疗联合1-MT对白血病进行治疗,可以显著减少肿瘤的复发率;而免疫治疗有效后复发时应用1-MT,可以显著抑制肿瘤的生长.     

DNA vaccination using expression vectors carrying FIV structural genes induces immune response against feline immunodeficiency virus

Following inactivated virus vaccination trials, the surface glycoprotein gp120 of the feline immunodeficiency virus (FIV) was considered as one of the determinants for protection. However, several vaccination trials using recombinant Env protein or some peptides failed to induce protection. To understand the role of the gp120 protein in vivo, we vaccinated cats with naked DNA coding for FIV structural proteins gp120 and p10. We analyzed the ability of these vaccinations to induce immune protection and to influence the onset of infection. Injection in cat muscles of expression vectors coding for the FIV gp120 protein induced a humoral response. Cats immunized twice with the gp120 gene showed different patterns after challenge. Two cats were, like the control cats, infected from the second week after infection onwards. The two others maintained a low proviral load with no modification of their antibody pattern. The immune response induced by gp120 DNA injection could control the level of viral replication. This protective-like immune response was not correlated to the humoral response. All the cats immunized with the gp120 gene followed by the p10 gene were infected, like the control cats, from the second week but they developed a complete humoral response against viral proteins after challenge. Furthermore, they showed a sudden but transient drop of the proviral load at 4 weeks after infection. Under these conditions, one injection of the p10 gene after one injection of the gp120 gene was not sufficient to stimulate protection. On the contrary, after a period, it seems to facilitate virus replication.     

Clinical and antiinflammatory effects of intranasal budesonide aqueous pump spray in the treatment of perennial allergic rhinitis

BACKGROUND: Neutrophils are of great importance for the host's defense against invading organisms. Granulocyte colony-stimulating factor (G-CSF) has been used to augment both the neutrophil number and function, and its prophylactic administration has proved beneficial in animal models of sepsis. However, pretreatment with G-CSF is not practical under clinical conditions. We therefore investigated the effect of recombinant human (rh)G-CSF, administered only after infection, on the survival rate as well as the hemodynamic and cytokine response of the animals. METHODS: Chronically catheterized conscious pigs were challenged with Pseudomonas aeruginosa (8 x 10(7) colony-forming units kg(-1) x h(-1) for 120 h (control group, n = 10). Animals in the G-CSF group (n = 7) also received rhG-CSF (5 microg kg(-1) x day(-1)), the first dose being given 3 h after beginning bacterial infusion. RESULTS: The mortality rate was 50% (5/10) and 29% (2/7) in the control and G-CSF groups, respectively (p = NS, control vs. G-CSF group). Fever, severe pulmonary hypertension, and a hyperdynamic response were recorded in all of the animals. In spite of a prompt and significant recovery from the initial leukopenia (p < .05 vs. control group), the animals of the G-CSF group showed no significant differences in the parameters investigated from those of the controls. Compared with the survivors, the interleukin-1 receptor antagonist was markedly elevated in all nonsurvivors after 6 h of sepsis (p < .05). CONCLUSIONS: These data suggest that treatment with rhG-CSF after the onset of bacterial sepsis might not significantly improve the chances of survival for non-neutropenic patients.     

Resection as elective treatment of hilar cholangiocarcinoma (Klatskin tumor)

A retrospective analysis of our experience in the treatment of hiliary cholangiocarcinoma or Klatskin tumor was performed with the aim of evaluating the morbi-mortality and prognosis of its treatment to thereby determine the usefulness of the different therapeutic options. From 1989 to 1997, 51 patients diagnosed with hiliary cholangiocarcinoma were treated in our hospital. Surgery was indicated in 16 with curative aims (group I) while palliative treatment with percutaneous biliary drainage was indicated in 35 (group II). Biliary resection was carried out in 8 patients being associated with hepatic resection in 4 (group IA) and in 8 patients undergoing liver transplantation (group IB). Clinico-epidemiologic data and hospital stay were similar in all the groups. The frequency of complications was similar in groups I and II although the frequency of cholangitis (49%) in group II was noticeable. The percentage of readmissions was also greater in group II (12 vs 46%, respectively; p = 0.03) with prosthesis obstruction being the most frequent cause. Accumulated survival at 1, 2, and 3 years in group I was 84, 64 and 48% with a median survival of 33 months, while in group II the median survival was of 6 months with no patient surviving more than 2 years (p = 0.0001). When groups IA and IB were compared, greater frequency of complications in groups IA (100 vs 37%; p = 0.002), similar frequency of readmissions (87 vs 75%; p = NS), median survival greater in group IB (12.5 months vs 48 months) and significantly higher actuarial survival in group IB (48% in 2 years vs 83% to 2 years; p = 0.02) was observed. In conclusion, surgery is the treatment of choice in hiliary cholangiocarcinoma whenever possible, given the greater survival without a significant increase in morbimortality. Likewise, we consider that liver transplantation is a useful option in the treatment of patients with cholangiocarcinoma type IV of Bismuth.     

Prevention of preeclampsia with heparin and antiplatelet drugs in women with renal disease

In a retrospective cohort study of women with renal disease in pregnancy we investigated if: 1. low dose aspirin reduced the prevalence of preeclampsia and improved fetal outcome compared to no anticoagulant therapy. 2. heparin plus low dose aspirin and/or dipyridamole reduced the prevalence of preeclampsia and improved fetal outcome compared to i. no treatment ii. low dose aspirin alone. Women with renal disease were allocated into 3 groups according to the treatment received during their pregnancies: I. no prophylactic heparin or antiplatelet drugs, n = 76 II. prophylactic low-dose aspirin 75(50-150)mg, n = 27 III. prophylactic subcutaneous heparin 10,000 (5000-12,500) IU b.d. combined with low-dose aspirin 50 (50-150)mg and/or dipyridamole 400 (200-400)mg, n = 44. Preeclampsia and fetal outcome was analysed according to treatment group. Preeclampsia was less common in the heparin group (2.3%) compared with 27.6% in the no treatment group [O.R. 0.06 (0.01-0.30)] and 25.9% in the aspirin group [O.R. 0.07 (0.01-0.38)]. Women on aspirin, who developed preeclampsia, delivered later in pregnancy [35.4 (33-38.2) weeks] than preeclamptic women on no treatment [29 (22-38) weeks], p = 0.04. There was a trend to reduced perinatal deaths in the heparin + antiplatelet drug group, [2.3%; O.R., 0.17 (0.02-1.4)] and in the aspirin group [0%, O.R., 0.13 (0.01-2.3)] compared with 11.7% perinatal deaths in the no treatment group. Heparin with anti-platelet drugs may prevent preeclampsia in high risk women with renal disease. Further investigation in a randomized trial is indicated.     

Vitreoretinal surgery after inadvertent globe penetration during local ocular anesthesia

BACKGROUND: The efficacy of prophylactic epsilon-aminocaproic acid and tranexamic acid to reduce transfusions after primary myocardial revascularization was evaluated in a teaching hospital context. METHODS: Patients (n = 134) received either epsilon-aminocaproic acid (15-g bolus + infusion of 1 g/h), high-dose tranexamic acid (10-g bolus + placebo infusion), or normal saline solution in a double-blind fashion. Anticoagulation and conduct of cardiopulmonary bypass were standardized. RESULTS: Tranexamic acid and epsilon-aminocaproic acid produced a significant reduction in postoperative blood loss compared with placebo (median loss, 438 mL, 538 mL, and 700 mL, respectively). Transfusion of red cells was similar in all three groups. Nonetheless, the percentage of patients receiving hemostatic blood products was significantly decreased in the epsilon-aminocaproic acid group compared with the placebo group (20% versus 43%; p = 0.03). Both tranexamic acid and epsilon-aminocaproic acid significantly decreased total exposure to allogeneic blood products compared with placebo (p = 0.01 and p = 0.05, respectively), and this reduction was clinically important (median exposure, 2, 2, and 7.5 units, respectively). Fibrinolysis was inhibited significantly in both treatment groups. CONCLUSIONS: We conclude that either high-dose tranexamic acid or epsilon-aminocaproic acid effectively reduces transfusions in patients undergoing primary, elective myocardial revascularization.     

Enhanced human T-cell lymphotropic virus type I expression following induction of the cellular stress response

Human T-cell lymphotropic virus type I (HTLV-I) infection is typically associated with long incubation periods between virus exposure and disease manifestation. Although viral protein expression is considered to play an important role in the pathogenesis of HTLV-I-associated diseases, limited information is known regarding host cell mechanisms that control viral gene expression. This study was designed to evaluate modulation of HTLV-I gene expression following induction of the cellular stress response in HTLV-I-infected lymphocytes. The cellular stress response was elicited by treatment with either Na arsenite or thermal stress and was monitored by demonstrating increased expression of the 72-kDa heat shock protein. Induction of the cellular stress response in HTLV-I-infected lymphocytes resulted in significantly increased HTLV-I-mediated syncytia formation due to enhanced HTLV-I envelope (gp46) expression. Intracellular viral proteins and released p24 capsid protein were increased in stressed infected lymphocytes as compared to nonstressed infected lymphocytes. Furthermore, HTLV-I-LTR reporter gene constructs had increased activity (three- to sixfold) in a transiently transfected, uninfected lymphocyte cell line following induction of the cellular stress response. Quantitation of HTLV-I RNA expression by slot blot analysis of infected lymphocytes suggested variable increases in RNA accumulation. Northern blot analysis demonstrated no qualitative changes in expression of RNA species. These data suggest a relationship between modulation of viral replication and a basic cellular response to stress and have important implications for understanding host cell control mechanisms of HTLV-I expression.     

Characterisation and cloning of a calmodulin-like domain protein kinase from Chlamydomonas moewusii (Gerloff)

Traditional herbal medicines with anti-herpes simplex virus type 1 (HSV-1) activity in vivo were examined for their prophylactic effects on recurrent HSV-1 infection in mice. Mice were intradermally infected with HSV-1 in the pinna and recurrent HSV-1 disease was induced by ultraviolet irradiation. Herbal extracts arrested the progression of recurrent HSV-1 disease, reduced the incidence of severe erythema and/or vesicles in the pinna, and/or shortened the period of severe recurrent lesions compared with water-administered mice (P < 0.01 or 0.05). Similarly, the prophylactic treatment of herbal extracts limited the development of recurrent skin lesions induced by stripping with cellophane tape physically. The prophylactic efficacy on recurrence was confirmed by the absence of HSV DNA in the skin lesions. HSV-1 genome was revealed to exist in the trigeminal ganglia but not in the pinna of latently infected mice before stimuli by a nested-polymerase chain reaction assay. After stimuli, HSV-1 genome was detected in both pinna and trigeminal ganglia of latently infected mice administered with water. However, prophylactic treatment decreased the rate of detection of HSV-1 genome in the stimulated pinna. Thus, the herbal extracts exhibited prophylactic efficacy against recurrent HSV-1 disease in mice and modulated the recurrent HSV-1 infection.     

Response-acquisition and cognitive self-statement modification approaches to dating-skills training.

Three training programs for girl-shy males were designed using an empirically derived domain of problem situations from shy males and response alternatives from a "competent" population. The effectiveness of a response-acquisition treatment was compared with a cognitive self-statement modification treatment, a combination of these 2 treatments, and a waiting-list control group (no treatment). Two enhanced-treatment groups were used to control for the longer time of the combined-treatment group. 61 college men replying to the program announcement were randomly assigned to 1 of the 6 groups. Assessment included in vivo measures made by women phoned by the Ss, questionnaire measures, and ratings of role-play performance in taped, laboratory, problem situations. A 6-mo follow-up assessment was also included. Results indicate that Ss trained in cognitive self-statement modification showed significantly better performance in role-play situations for which they were not trained, made significantly more phone calls, and made a significantly better impression on the women than Ss in other groups. These effects were generally maintained at follow-up, and the cognitive self-statement groups' performance on the role-play measures improved from posttreatment to follow-up. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Role of immunoglobulin A monoclonal antibodies against P23 in controlling murine Cryptosporidium parvum infection

Cryptosporidium parvum is an important diarrhea-causing protozoan parasite of immunocompetent and immunocompromised hosts. Immunoglobulin A (IgA) has been implicated in resistance to mucosal infections with bacteria, viruses, and parasites, but little is known about the role of IgA in the control of C. parvum infection. We assessed the role of IgA during C. parvum infection in neonatal mice. IgA-secreting hybridomas were developed by using Peyer's patch lymphocytes from BALB/c mice which had been orally inoculated with viable C. parvum oocysts. Six monoclonal antibodies (MAbs) were selected for further study based on indirect immunofluorescence assay reactivity with sporozoite and merozoite pellicles and the antigen (Ag) deposited on glass substrate by gliding sporozoites. Each MAb was secreted in dimeric form and recognized a 23-kDa sporozoite Ag in Western immunoblots. The Ag recognized comigrated in sodium dodecyl sulfate-polyacrylamide gel electrophoresis with P23, a previously defined neutralization-sensitive zoite pellicle Ag. MAbs were evaluated for prophylactic or therapeutic efficacy against C. parvum, singly and in combinations, in neonatal BALB/c mice. A combination of two MAbs given prophylactically prior to and 12 h following oocyst challenge reduced the number of intestinal parasites scored histologically by 21.1% compared to the numbers in mice given an isotype-matched control MAb (P < 0.01). Individual MAbs given therapeutically in nine doses over a 96-h period following oocyst challenge increased efficacy against C. parvum infection. Four MAbs given therapeutically each reduced intestinal infection 34.4 to 42.2% compared to isotype-matched control MAb-treated mice (P < 0.05). One MAb reduced infection 63.3 and 72. 7% in replicate experiments compared to isotype-matched control MAb-treated mice (P < 0.0001). We conclude that IgA MAbs directed to neutralization-sensitive P23 epitopes may have utility in passive immunization against murine C. parvum infection.     

Cryptosporidium parvum infection of human intestinal xenografts in SCID mice induces production of human tumor necrosis factor alpha and interleukin-8

The protozoan parasite Cryptosporidium parvum invades intestinal epithelial cells and can cause life-threatening diarrhea in immunocompromised individuals. Despite the clinical importance of this organism, much remains to be learned about the pathogenesis of C. parvum-induced diarrhea. To explore the role of the intestinal inflammatory response in C. parvum disease, using C. parvum oocysts we infected human intestinal xenografts in severe combined immunodeficient (SCID) mice. Seven days after infection, we found levels of human tumor necrosis factor alpha and interleukin-8 in C. parvum-infected human intestinal xenografts that were significantly higher than those seen in uninfected control xenografts. These results demonstrate that human intestinal cells produce proinflammatory cytokines in response to C. parvum infection and establish SCID-HU-INT mice as a model system to study the interactions of C. parvum with the human intestine.     

Experimental cancer immunotherapy: comparison of tumor rejection if F344 rats given live Mycobacterium bovis (Strain BCG) and killed Corynebacterium parvum

F344 rats received grafts of syngeneic 13762 mammary adenocarcinoma cells previously admixed with either living BCG of killed Corynebacterium parvum administered sc or intradermally (id). Animals given id transplants of tumor cells admixed with either BCG or killed C. parvum exhibited tumor growth for an average of 10 days, then regression in size and rejection of the tumor nodules. Lesions were found in rats given sc transplants of tumor cells admixed with the killed microorganism for an average of 13 days with the same results. When live BCG was added to the sc transplants, accelerated rates of tumor growth and early death were noted, compared with the group receiving tumor cells alone sc. Suppressed rates of tumor growth and prolonged survival were observed in the groups receiving id inoculations of tumor cells followed by treatment with killed C. parvum administered weekly ip or id 1 cm away and around the growing tumor. On the other hand, weekly treatment of BCG injected either ip or id 1 cm away and around the growing tumor resulted in accelerated rates of tumor growth and early death. Animals exhibiting C. parvum of BCG-mediated tumor rejection displayed tumor-specific protection to sc challenge injections of the cell line initially used, but they died with growing tumors and metastases when challenged with tumor cells of an antigenically different line syngeneic to F344 RATS. Microscopic examination of histologic sections of tumors formed from id inoculations of tumor cells admixed with either BCG or killed C. parvum revealed a nonspecific infiltrate of macrophages and polymorphonuclear leukocytes in the tumor, whereas sections of tumors formed from sc grafts of cells admixed with killed C. parvum revealed a specific organized infiltrate of mostly macrophages around the tumor follicles.     

A role for host phosphoinositide 3-kinase and cytoskeletal remodeling during Cryptosporidium parvum infection

Cryptosporidium parvum preferentially infects epithelial cells lining the intestinal mucosa of mammalian hosts. Parasite development and propagation occurs within a unique intracellular but extracytoplasmic parasitophorous vacuole at the apical surface of infected cells. Parasite-induced host cell signaling events and subsequent cytoskeletal remodeling were investigated by using cultured bovine fallopian tube epithelial (BFTE) cells inoculated with C. parvum sporozoites. Indirect-immunofluorescence microscopy detected host tyrosine phosphorylation within 30 s of inoculation. At >30 min postinoculation, actin aggregates were detected at the site of parasite attachment by fluorescein isothiocyanate-conjugated phalloidin staining as well as by indirect immunolabeling with monoclonal anti-actin. The actin-binding protein villin was also detected in focal aggregates at the site of attachment. Host cytoskeletal rearrangement persisted for the duration of the parasitophorous vacuole and contributed to the formation of long, branched microvilli clustered around the cryptosporidial vacuole. The phosphoinositide 3-kinase inhibitor wortmannin significantly inhibited (P < 0.05) C. parvum infection when BFTE cells were pretreated for 60 min at 37 degreesC prior to inoculation. Similarly, treatment of BFTE cells with the protein kinase inhibitors genistein and staurosporine and the cytoskeletally acting compounds 1-(5-iodonaphthalene-1-sulfonyl)-1H-hexahydro-1,4-diazapine, cytochalasin D, and 2,3-butanedione monoxime significantly inhibited (P < 0.05) in vitro infection at 24 h postinoculation. These findings demonstrate a prominent role for phosphoinositide 3-kinase activity during the early C. parvum infection process and suggest that manipulation of host signaling pathways results in actin rearrangement at the site of sporozoite attachment.     

Cytotoxicity of sera from rats with puromycin aminonucleoside nephrosis

BACKGROUND: Sensitization to allergens has been shown to be a risk factor for adults with acute asthma first seen in the emergency department. OBJECTIVE: The purpose of this study was to evaluate the prevalence of specific IgE to common aeroallergens in children with asthma first seen in the emergency department and in control subjects. METHODS: Fifty-four children, aged 3 to 16 years (mean age, 8.34 years) who visited the emergency department for treatment of acute bronchospasm or other illness, were evaluated. Specific IgE to seven common aeroallergens and four common storage mites was determined. Group I consisted of 29 patients who had acute bronchospasm and histories of recurrent asthma. Group II consisted of 25 control subjects who had no clinical history of atopic disease. Group I and II were compared for differences in the prevalence of positive RAST responses to the 11 allergens tested. Dust samples were collected from 17 homes of subjects in group I and from 13 homes of subjects in group II and were analyzed for levels of Der p 1 and Der f 1. RESULTS: Statistically significant differences in the prevalence of positive RAST results between groups I and II were found in response to: Dermatophagoides pteronyssinus, 89.6% versus 36% (p = 0.0001); Blattella germanica, 45.8% versus 9.5% (p = 0.018); Alternaria tenuis, 44.8% versus 4% (p = 0.001); and the storage mites Aleuroglyphus ovatus, 39.2% versus 4% (p = 0.002); Blomia tropicalis, 42.8% versus 0% (p = 0.0002); Chortoglyphus arcuatus, 46.4% versus 0% (p = 0.0001); and Lepidoglyphus destructor, 32.1% versus 0% (p = 0.0019). Mean specific IgE levels, expressed as percent of the total counts bound, were significantly higher in group I compared with group II only in response to D. pteronyssinus, 21.9% versus 2.1% (mean percent of total counts bound) (p = 0.0001). Analysis of dust samples revealed no significant differences between the two groups, except for a higher concentration of Der f 1 in the sofas of subjects in group II. CONCLUSION: Sensitization to D. pteronyssinus, storage mites, and, to a lesser extent, to A. tenuis and B. germanica is associated with acute childhood asthma that requires emergency treatment in Florida.