Lyme disease in an experimental cat model

This report concerns the susceptibility of the domestic cat to Lyme disease. Three strains ofBorrelia burgdorferi used in the study were administered intradermally (10⁶ live cells) to three groups of cats, five cats in each group. At biweekly intervals, over a 23-week period, blood samples were obtained for hematologic and serologic analysis from each cat. Each 4 weeks during the study, one cat from each group was necropsied for gross- and histopathologic examination. Seroconversion was observed during weeks 3 and 4 in all inoculated animals. Cyclic changes, characterized by decrease in percentage of neutrophils accompanied by increase in percentage of lymphocytes and eosinophils, were observed 11 weeks after initial exposure to the agent and at 2–4-week intervals afterwards for the following 12 weeks of the study. Similar cyclic changes in the IgG levels corresponding to those of the white blood cells were also observed. Histopathologic changes were observed in the stifle joints and in all major organs of infected cats 16 weeks after initial exposure toB. burgdorferi. Overt clinical signs were minimal, but included mild pyrexia and lameness first observed 4–6 weeks following exposure to the agent.Presented at the 35th World Congress, International College of Angiology, Copenhagen, Denmark, July 1993     

Characterization of bronchoalveolar lymphocytes during a specific antibody-forming cell response in the lungs of mice

The goal of this study was to characterize the total numbers and phenotypes of lymphocyte subpopulations recovered by bronchoalveolar lavage during an experimental immune response in the lung parenchyma. Inbred mice (C57BL/6) were primed systemically and then challenged intratracheally with sheep red blood cells, a T-cell-dependent antigen. At various days later, we performed differential cell counts, measured the concentrations of specific antibody-forming cells, and determined lymphocyte phenotypes in bronchoalveolar lavage fluid by flow cytometry, distinguishing lymphocytes by light scatter parameters. We found that the numbers of lymphocytes recovered by bronchoalveolar lavage increased significantly in primed mice challenged with the priming antigen but not in three control groups: unprimed mice challenged intratracheally with the same dose of sheep red blood cells, primed mice challenged with hydrochloric acid, and primed mice challenged with a non-cross-reacting erythrocyte. At all times tested the concentrations of antibody-forming cells in bronchoalveolar lavages were identical to those of cells from minced lungs. Helper T-cells (L3T4-positive) increased earliest and constituted the majority of lymphocytes in bronchoalveolar lavage fluid throughout the immune response. We conclude: first, that there is a major influx of lymphocytes into the lungs during the development of a specific pulmonary immune response; second, that this lymphocyte influx occurs only in the presence of an antigen-driven response; third, that lymphocytes specific for the challenging antigen are in equilibrium between the bronchoalveolar and interstitial compartments; fourth, flow cytometry can be used to determine surface phenotypes of bronchoalveolar lymphocytes in mice.     

Familial hypercholesterolaemia commonly presents with Achilles tenosynovitis

BACKGROUND: Patients with heterozygous familial hypercholesterolaemia (HeFH) develop tendon xanthomata (TX), most commonly in their Achilles tendons. Even before tendons are chronically enlarged, tenosynovitis may occur and medical advice be sought. Untreated HeFH carries a high risk of premature coronary heart disease, which can be ameliorated by early diagnosis. OBJECTIVE: To determine the prevalence of episodes of Achilles tendon pain in HeFH before its diagnosis. METHODS: Patients with definite HeFH (Simon Broome criteria) attending a lipid clinic were identified. They completed a questionnaire asking about symptoms relating to their Achilles tendons. Unaffected spouses or cohabiting partners served as controls. RESULTS: 133 patients (47% men) and 87 controls (51% men) participated. TX had been recognised by the referring physicians in <5% of cases. However, 62 (46.6% (95% confidence interval (CI) 38.1 to 55.1)) patients had experienced one or more episodes of pain in one or both Achilles tendons lasting >3 days, whereas only 6 (6.9% (1.6 to 12.2)) controls had done so (difference p<0.001; likelihood ratio 6.75). Typically, in the patients with HeFH the pain lasted 4 days (median). It was described as severe or very severe in 24/62 (38.7% (30.4 to 47.0)) patients with HeFH, but never more than moderate in controls. 35 (26.3% (18.8 to 33.8)) patients with HeFH had consulted a doctor about Achilles tendon pain, but in no case had this led to a diagnosis of HeFH. None of the controls had consulted a doctor. CONCLUSIONS: Measurement of serum cholesterol in patients presenting with painful Achilles tendon could lead to early diagnosis of HeFH.     

Effects of neonatal exposure to isophilic and-or heterophilic antigens on immune responses of adult rats

Abstract. Isophilic immune response was profoundly depressed in rats injected with heterophilic antigen during the first week of life and challenged with sheep red cells 10 to 18 weeks later. Untreated litter mates served as controls. When neonatal treatment was given during the second week of life and/or challenge with sheep red cells was deferred until 20 weeks, the isophilic immune response was inhibited to a lesser degree. Rats injected neonatally with sheep red cells had decreased amounts of isophilic and heterophilic antibodies upon challenge with sheep red cells 12 weeks later, while intervals of 16 to 20 weeks between neonatal treatment and adult immunization with sheep red cells increased heterophilic antibodies without significant changes in isophilic antibodies. Rats exposed neonatally to either heterophilic antigen or sheep red cells, and challenged with heterophilic antigen as adults, formed significantly more heterophilic antibodies than did control litter mates. These observations were interpreted as resulting from imprinting of large numbers of cells with receptors for heterophilic antigen after neonatal exposure to the antigen, and cellular diversion of antigen upon challenge of adult animals with sheep red cells. The possible relationship of this phenomenon to the interference with Rh immunization of ABO incompatibility in man was considered.     

Hand flexor tenosynovitis in rheumatoid arthritis

Tenosynovitis of one or more flexor tendons of the hand (mean 3.1 tendons per patient) was noted in 55% of 100 patients with rheumatoid arthritis (RA) examined periodically during a mean period of 5 years. The third flexor tendon was involved most frequently (71% of patients), followed by the second (62%), fourth (53%), fifth (27%), and first (13%). Patients with flexor tendonitis (FT) had a significantly higher prevalence of rheumatoid nodules (56% vs 33%), carpal tunnel syndrome (47% vs 13%), wrist extensor tenosynovitis (47% vs 9%), and elbow epicondylitis (22% vs 7%) than patients without FT. Dupuytren's contracture, DeQuervain's tenovaginitis, flexor carpi radialis and ulnaris tendonitis, and Achilles tendonitis were found exclusively in patients with FT. A control group of 50 non-RA patients with FT had statistically fewer diseased tendons per patient (mean 1.5) and a different digital distribution, the thumb being affected more frequently (P < 0.05) than in RA patients.     

The immune response of regional lymph nodes during the early stages of Fasciola hepatica infection in cattle

In this study we examined regional immune responses to Fasciola hepatica infection in the natural ruminant host. Naïve cattle and those pre‐exposed to a drug‐abbreviated infection were subsequently challenged and lymph nodes extracted at slaughter. In vitro proliferation and cytokine production by mononuclear cells isolated from hepatic and mesenteric lymph nodes were measured after culture with whole fluke antigen (WFA). Hepatic lymph node cells had a significantly greater response to parasite antigen than mesenteric lymph node cells (P < 0·02), although there was no difference in the magnitude of the proliferative response between naïve and pre‐exposed challenged cattle. Mononuclear cells from hepatic lymph nodes produced interferon gamma, interleukin 2 and interleukin 4 after culture with parasite antigen, indicative of a mixed, T helper type 0, response. Comparison of the hepatic node response to a variety of F. hepatica antigens showed that proliferation was lower after culture with cathepsin‐L, than with a high molecular weight fraction, WFA or excretory–secretory antigen. Cell culture supernatant fluid from unstimulated hepatic lymph node cells showed an IgG1 response to antigens of 48, 52–70, 82, 96 and 120–190 kDa on Western blot in pre‐exposed, but not naïve, challenged animals.     

Local immune responses in sensitized sheep following challenge infection with Teladorsagia circumcincta

Sheep were sensitized by weekly infections with Teladorsagia circumcincta over a 9-week period. After a 12-week rest, sheep were divided into four groups and killed without challenge or 3, 5 and 10 days post challenge (DPC) with 50000 L3. Recovery of challenge larvae from abomasal scrapings was highest at 3 DPC while no parasites were recovered by 10 DPC. Abomasal lymph nodes (ALN) of challenged sheep were significantly larger at 5 DPC, coinciding with an increase in the proportion of CD4 T cells and a decrease in CD21+ cells, probably reflecting the loss of CD21 from terminally differentiated antibody secreting cells. A significant increase was observed in gammadelta-TCR+ cells at 3 DPC in the ALN, while their number slightly decreased in the abomasal tissues throughout the challenge period. The number of tissue eosinophils was dramatically increased after challenge compared with the unchallenged controls, with a peak at 3 DPC, coinciding with the peak in larval recovery. CD4+ cells significantly increased in the abomasal tissues at 5 DPC, while no changes in globule leucocytes were observed until 10 DPC. Antibody-secreting cell probes (ASC-probes) generated from the ALN showed highest reactivity against larval antigens at 5 DPC. This reactivity was predominantly directed against regions between 90 and 100 kDa and 30-35 kDa in the L3 preparation and lower molecular weight antigens in the L4. No reactivity was observed against the adult extract. The 30-35 kDa antigen seemed to exist as a high molecular weight complex in L3 homogenate and was not susceptible to protease K treatment, suggesting it may be non-protein in nature.     

Experimental Hypersensitivity Pneumonitis: Location of Transferring Cells

Cultured cells from Micropolyspora faeni-sensitized donors can adoptively transfer murine experimental hypersensitivity pneumonitis (EHP). We sought to determine the location of transferred cells in recipient animals, the influence of the origin of the cultured cells, and the effect of specific intratracheal challenge. We labeled cultured sensitized spleen or lung-associated lymph node (LALN) cells with CFDA-SE, a cytoplasmic stain, before transfer to naive recipients, which were sacrificed 1 h, 1 day, or 4 days thereafter. We also transferred labeled cultured spleen cells to recipients that were challenged with intratracheal M. faeni and sacrificed 4 days later (MF). Controls were recipients of M. faeni-sensitized and cultured cells challenged with intratracheal normal saline (NS) and recipients of ovalbumin (OVA)-sensitized cells cultured with M. faeni and challenged with intratracheal M. faeni (OVA). The number and proportion of cells that were stained were determined in dispersed spleen, peripheral and lung-associated lymph nodes, and lung parenchyma. The extent of the pulmonary inflammatory response was measured by determining the proportion of microscopic fields that were abnormal and the total number of dispersed pulmonary cells. CFDA-SE stained cells uniformly, and stained cells could be detected in recipients for up to 7 days after transfer. CFDA-SE treatment (0.5 μM) did not affect the ability of cells to transfer EHP adoptively. Transferred cells could be detected easily in lung, lung-associated and peripheral lymph nodes, blood, and spleen. Transferred cells localized to the lung at 1 h but then rapidly decreased with no difference between labeled cells from spleen and LALN. After intratracheal M. faeni challenge, there was no difference in the proportion of labeled cells in the lung among any of the groups (MF, NS, or OVA). There was an increase in the number of lung cells in the MF group compared with the control (NS and OVA) groups. We conclude that cells capable of transfer are transiently (1 h) trapped in the lung but are much decreased in the lung by four days after transfer. After intratracheal antigen challenge of recipients, there is a substantial increase in the number of pulmonary cells in animals exhibiting adoptive EHP but not in the control groups. Transferred cells responsible for EHP are increased in the lungs of animals with adoptive EHP. Accepted for publication: 30 June 1997     

A clinical and experimental study of colostomy blood flow and healing after closure

Colostomy blood flow and healing have been studied in both man and rat. Laser Doppler velocimetry (LDV) was used to measure flow and showed a significant difference between colostomy blood flow measured at 1 week and at more than 8 weeks after fashioning (p < 0.002). Hartmann colostomies were constructed in rats and closed at 3 or 6 weeks later. There was a correlation between distal colonic (stump) blood flow and anastomotic bursting pressures 3 days after closure in the 3-week group (r=0.080;p < 0.01). Colostomies closed at 6 weeks were significantly stronger than those closed at 3 weeks (p < 0.02). Collagen concentration in proximal (stoma) colon was higher in the 6-week group compared with the 3-week group prior to anastomosis. There was also a fall in the proximal collagen after anastomosis in both 3-(p < 0.01) and 6-week groups (p < 0.01). Poor blood flow measured by LDV and colonic collagen concentration may predict poor healing after colostomy closure. LDV is a non-invasive technique with clinical application in this field.     

Upper limb pyrophosphate tenosynovitis outside the carpal tunnel.

Three cases of calcium pyrophosphate dihydrate (CPPD) crystal deposits in tendon sheaths outside the carpal tunnel are reported. Crystals were shown by x ray diffraction analysis in one case and by compensated light microscopy in the other two. Surgical excision of the tendon synovial sheath had to be done in two cases (one case with CPPD crystal deposits).     

Treatment of rheumatoid tenosynovitis with cytokine inhibitors

Hand function depends on tendon integrity, but in rheumatoid arthritis tenosynovitis can result in tendon adhesions and rupture. Cytokine inhibitors have proved effective in rheumatoid joint disease; however, their effect on the tenosynovium is not well understood. We investigated the ability of inhibitors of tumour necrosis factor alpha and interleukin 1 to reduce production of collagenolytic matrix metalloproteinases 1 and 13 in tenosynovial tissue obtained from patients with rheumatoid arthritis. Our data show that cytokine blockade can reduce collagenase concentrations in tenosynovial tissue, suggesting cytokine inhibitors could be effective in reduction of tendon damage.     

狂犬病毒抗原刺激小鼠的细胞免疫反应试验

为了研究狂犬病毒抗原刺激机体所产生的细胞介导免疫反应的抗病毒作用 ,我们利用小鼠进行试验。注射环磷酰胺 (Cy)的小鼠再接种狂犬疫苗和稀释液 ,然后将其脾细胞转移到 2 4小时前脑内接种 1或 10 0 0LD50 狂犬病毒标准攻击毒(CVS)的同系小鼠 ,结果表明小鼠的脾细胞不仅可以转递抗狂犬病毒攻击的能力 ,而且可以转移“早期死亡”现象 ;另一方面 ,注射Cy的小鼠接种狂犬病毒抗原 ,两周后用CVS攻击 ,结果发现接种狂犬病毒抗原的小鼠有一定程度的保护作用 ,但中和试验结果阐明这些小鼠体内没有产生中和抗体 ,从而说明狂犬病毒抗原刺激所产生的细胞免疫作用有一定抗狂犬病毒攻击的能力。     

Changes in lysosomal morphology and enzyme activities during the development of adriamycin-induced cardiomyopathy

Morphologic and enzymic changes in heart lysosomes were studied following a chronic treatment of animals with a cumulative dose of 15 mg/kg of adriamycin. Myocardial cell damage due to adriamycin included lysosomal changes, sarcotubular swelling, vacuolization and myofibrillar drop-out. These structural changes were more pronounced in the 6-week treated group as opposed to the 3-week treated group. The number of lysosomes per unit area increased from a control value of 3.6 +/- 1.7 to 17.8 +/- 4.0 in the 3-week treated group and 35.9 +/- 9.2 in the 6-week treated groups, respectively. The scatter in the size distribution of lysosomes was much wider in treated animals. Lysosomal hydrolases in the 3-week and 6-week adriamycin-treated group changed as follows: N acetyl beta-glucosaminidase activity fell in the homogenate (H) and nonsedimentable (NS) and rose in the serum (Ser) fractions; a drop in alpha-mannosidase was seen in the sedimentable (S) and Ser fractions; an increase in beta-galactosidase was noted in the H, S and Ser fractions; acid phosphatase was increased in H, S, NS and Ser fractions. Lanthanum staining, used as a cytochemical probe for normal membrane permeability, revealed intracytoplasmic localization of the tracer only in the 6-week group. Malondialdehyde content was increased significantly in the 3-week and 6-weed treated groups. These results show lysosomal changes in adriamycin-treated hearts which precede as well as accompany nonspecific permeability changes in the sarcolemma.(ABSTRACT TRUNCATED AT 250 WORDS)     

Volunteer Assistance in the Treatment of Chronic Alcoholism*

Chronic alcoholics often have great difficulty in adjusting their lifestyle to accommodate the goals agreed upon in treatment. A 16-week behavioral self-management program, which emphasized creating lifestyle changes in the community, was offered to 193 clients. To assist in the process of treatment delivery, half of the clients were offered the support of trained community volunteers during the treatment program. To give volunteers a time period in which to to meet and start working with their clients, only those clients who completed at least the first 4 weeks of the 16-week program (N = 106) were included in the evaluation. Independent follow-up was conducted over a 12-month period. A significant reduction in alcohol consumption was found for clients of both the volunteer-support (VS; N = 52) and the office-based (OB; N = 54) groups; this reduction was maintained over the 12 months of review, with no major differences found between groups. Volunteers rated high on particular characteristics assisted the client more effectively during treatment than those rated low. The variability in hours spent between volunteer-client pairs appeared to mask any main effects, and also the length of the contact period with the volunteer was too short. It is recommended that the use of volunteers be evaluated further by extending the client contact time into the follow-up period.     

Defective mucosal immunity and normal systemic immunity of Mongolian gerbils,Meriones unguiculatus,to reinfection with Strongyloides venezuelensis

The systemic and local protective activity of Mongolian gerbils was examined after re-infection with Strongyloides venezuelensis. Mongolian gerbils were unable to expel S. venezuelensis adult worms from the intestine for over ten weeks after a primary infection. Therefore, immune animals were prepared by treating with mebendazole four weeks after a primary infection and then they were challenged by different maturation stages of the parasite; subcutaneous inoculation with the infective larvae ( L₃₎ obtained by faecal culture, oral administration of L3 obtained from the lungs of rats three days after a primary infection, or oral implantation of adult worms obtained from the intestines of rats seven days after a primary infection. The results show that, although immune animals were highly resistant against challenge infection by subcutaneous inoculation with cultured L₃, they were unable to expel orally administered lung-recovered L3 nor orally implanted adult worms. Although potentiated mastocytosis was induced by challenge infections with lung-recovered L₃ and adult worms, all mast cells were formalin-resistant, heparin-containing cells and never seen in the epithelial layer. In spite of the defective protective capacity at the intestinal mucosa, circulating antibody production specific to S. venezuelensis adult as well as L3 antigen was positive. Therefore, the inability of Mongolian gerbils to expel S. venezuelensis adult worms from the intestine seems to be due to the defects of effector/regulator cells, presumably mast cells, but not due to immune unresponsiveness to parasite antigen.     

CD5-negative cell mediated experimental hypersensitivity pneumonitis.

Experimental hypersensitivity pneumonitis (EHP) can be transferred to Strain 2 guinea pigs by peripheral lymph node (PLN) cells cultured in vitro with antigen. The phenotype of the active cells has not been delineated. In addition, it is not known if cultured lung-associated lymph node (LALN) cells can transfer EHP. PLN and LALN cells from donor animals were cultured with a soluble extract of Micropolyspora faeni (10 micrograms/ml), and blast cells were isolated by centrifugation over Percoll gradients. Cultured PLN cells were passed through nylon wool columns, and the adherent and nonadherent fractions were tested for their ability to transfer EHP. PLN blast cell populations were depleted of CD5+ cells by treatment with monoclonal anti CD5 antibody (8BE6) and complement. Syngeneic recipients received media or LALN or PLN blast cells treated with antibody plus complement, media, or complement intravenously. Recipients were challenged intratracheally with M. faeni 48 h after the cell transfer, and they were killed 4 days later. The nonadherent PLN cell population was enriched for CD5+ (T) cells and depleted of surface immunoglobulin-positive (SIg+) cells. Treatment of the PLN blast cell population with 8BE6 and complement decreased CD5+ cells from 25 to 4% and increased SIg+ cells from 62 to 80%. All animals were maintained in HEPA-filtered air. Randomly selected microscopic fields of the lung (250/animal) were judged to be normal or abnormal without knowledge of treatment. There was a low level of pulmonary response to an intratracheal challenge of M. faeni in media recipients. There was a substantial increase (p less than 0.01) of the extent of pulmonary abnormalities in the animals receiving cultured PLN cells.(ABSTRACT TRUNCATED AT 250 WORDS)     

CT and MRI evaluation of tenosynovitis of the rheumatoid hindfoot

Summary Thirty-nine patients with rheumatoid arthritis who had presented with tarsitis before, were investigated at the level of the rearfoot. The first 17 patients had CT with previous tenography when it was possible; the following 22 patients had MRI with gadolinium injection. Tendon involvement appeared in 52,9% of the cases on CT, and in 90% of the feet on MRI; therefore, in case of clinical or radiological signs of tarsitis, it appears that tendon involvement must be suspected. With the two procedures the tibialis posterior tendon lesions were very predominant. In the majority of the patients (31/39), there was associated involvement of two or more tendons. If there is a ruptured tendon, the authors think that one must be cautious with surgical tendinous transfer; indeed, the long-term results of this surgical procedure present a strong probability of being compromised in rheumatoid arthritis which is a progressive disease.     

Ultrasound-guided retro-calcaneal bursa corticosteroid injection for refractory Achilles tendinitis in patients with seronegative spondyloarthropathy: efficacy and follow-up study

Ultrasound (US)-guided corticosteroid injection has been shown to be safe and effective for varied causes of plantar fasciitis; however, its use for Achilles tendinitis is controversial. We studied the efficacy and changes in US findings at Achilles enthesitis after corticosteroid injection in patients with spondyloarthropathy (SpA). Patients with SpA with symptomatic Achilles enthesitis, refractory to 6 weeks of full-dose NSAIDs, were offered US-guided local corticosteroid injection. Injected entheses were examined by US (both B mode and power Doppler) at baseline and 6 weeks after injection. Standard OMERACT definitions were used to define enthesitis. Achilles tendon thickness >5.29 mm, 2 cm proximal to insertion in long axis, was considered thickened. Twenty-seven symptomatic Achilles tendons (in 18 patients) were injected with 20 mg methylprednisolone under US guidance baseline, and 6-week follow-up US features were compared. All patients reported improvement in pain (VAS) in the affected tendon after injection (p < 0.0001). Simultaneously, improvement in local inflammatory changes were noted, in the form of significant reduction in tendon thickness (p < 0.0001), vascularity (p < 0.0001), peritendinous oedema (p = 0.001), bursitis and bursal vascularity (p < 0.001 and < 0.0001, respectively). There was no change in bone erosions and enthesophyte. None of the patients had tendon rupture or other injection-related complications at 6 weeks of follow-up. US-guided local corticosteroid injection is an effective and safe modality for refractory Achilles enthesitis in patients with SpA and leads to reversion of acute changes at entheseal site.     

Fibrosing cytolytic liver failure secondary to recurrent hepatitis B after liver transplantation.

Four patients who underwent transplantation for hepatitis B virus-related liver disease developed rapidly progressive liver failure attributable to recurrent hepatitis B disease typified by hyperbilirubinemia and distinctive hepatocyte ballooning and progressive fibrosis consistent with recently reported fibrosing cholestatic hepatitis. Among these four patients, the mean interval from transplantation to redocumentation of hepatitis B surface antigen (HBsAg) and hepatitis B e antigen (HBeAg) was 5 months, to development of malaise and jaundice 6 months, to histological diagnosis 7 months, and to graft failure 8 months. The only patient who underwent retransplantation had accelerated recurrence of the same syndrome with biopsy documentation 1 month later and graft failure 2 months later. Distinctive histological features included confluent hepatocellular ballooning and progressive periportal fibrosis followed by lobular collapse over 4-6 weeks without significant inflammation. Immunohistochemical staining showed marked HBsAg and hepatitis B core antigen (HBcAg) immunoreactivity. The rapid development of cytolytic hepatocellular necrosis and lobular collapse with prominent HBcAg immunoreactivity without significant inflammation suggests a cytolytic rather than immune pathogenesis for this unique and devastating form of recurrent hepatitis B that might better be termed "fibrosing cytolytic hepatitis."