A questionnaire study for 128 patients with Gaucher disease.

Gaucher disease is an uncommon autosomal recessive disorder characterized by lysosomal storage of glucosyl ceramide, a material released during cell degradation. Patients with Gaucher disease often have significant hematologic, bone structural, and visceral problems which sometimes greatly affect their health and life style. Based on some extraordinary scientific discoveries over the past 45 years, a treatment system has evolved which consists of administration of an enzyme, which destroys the lysosome-stored material and to some extent restores the patients to good health. There are still some problems for these patients; however, and the purpose of the study is to define some of the clinical, sociologic, and psychologic problems with a specially designed questionnaire. Questionnaire data was collected for 128 patients from two institutions with complete anonymity, and the information compared against data from a National Health Interview Survey. The results show that many of the patients still have fairly extensive problems, which could possibly be helped by some alterations in treatment protocols.

Hyperbaric oxygen as an adjunct in zygomycosis: randomized controlled trial in a murine model.

Zygomycosis was induced by injecting CD-1 mice with 5 mg of intraperitoneal deferoxamine and then 10(6) CFU of intravenous and intrasinus Rhizopus arrhizus. The addition of hyperbaric oxygen (2.0 atm absolute twice daily) to amphotericin B did not improve survival over that achieved with amphotericin B and placebo air treatments.

Biomechanical analysis of a chondrocyte-based repair model of articular cartilage.

The objective of this study was to evaluate the biomechanical properties of newly formed cartilaginous tissue synthesized from isolated chondrocytes. Cartilage from articular joints of lambs was either digested in collagenase to isolated chondrocytes or cut into discs that were devitalized by multiple freeze-thaw cycles. Isolated cells were incubated in suspension culture in the presence of devitalized cartilage matrix for 3 weeks. Multiple chondrocyte/matrix constructs were assembled with fibrin glue and implanted subcutaneously in nude mice for up to 6 weeks. Testing methods were devised to quantify integration of cartilage pieces and mechanical properties of constructs. These studies showed monotonic increase with time in tensile strength, fracture strain, fracture energy, and tensile modulus to values 5-10% of normal articular cartilage by 6 weeks in vivo. Histological analysis indicated that chondrocytes grown on dead cartilage matrix produced new matrix that integrated individual cartilage pieces with mechanically functional tissue.

Cartilage degeneration in relation to repetitive pressure: case study of a unilateral hip hemiarthroplasty patient.

In vivo acetabular contact pressures were measured over 32 months in an elderly man with a pressure instrumented hemiarthroplasty. After death, left (hemiarthroplasty) and right (control) acetabula were explanted. Cartilage thickness and degeneration were quantified from magnetic resonance imaging and histological analysis. Highest repetitive in vivo contact pressures during gait (4.5 to 6.5 MPa) were measured in the superior dome of the acetabulum and decreased at a rate of approximately 1 MPa per year after implant (R2 = 0.48, P < .001). Contact pressure magnitudes measured during gait correlated positively with regional histology score (R2 = 0.34, P < .0001) and negatively with cartilage thickness (R2 = 0.35, P < .0001). Although histology scores were typical of early osteoarthritis (histological grade of 4-6), there were no significant differences in overall histology score for the left and right acetabula (P = .23). We conclude that acetabular cartilage degeneration was explained, in part, by repetitive stress, but the degeneration did not appear to be mediated solely by articulation with the metallic endoprosthesis.

The porcine model for intratemporal facial nerve trauma studies.

Facial nerve anatomy was compared in the rat, rabbit, cat, and pig in an effort to develop a model for facial nerve trauma within the temporal bone. The porcine model was found to have the most suitable anatomy. Landmarks for the nerve were excellent. The pig had a definite facial nerve mastoid segment that was vertical, as in the human, and long enough to allow for performance of sequential procedures on the nerve. It was also large enough for grafting and electrical testing. A detailed description of the advantages of the pig model and the anatomy of the surgical approach to the facial nerve in the porcine model is presented.

Efficacy of a commercial bacterin in protecting strain 13 guineapigs against Bordetella bronchiseptica pneumonia.

Bordetella bronchiseptica is known to be endemic in many guineapig (Cavia porcellus) colonies, and periodically is the aetiological agent of fatal epizootics of bronchopneumonia. A commercial, non-adjuvant B. bronchiseptica bacterin, which is approved for use in canines, was evaluated for induction of a protective immune response in Strain 13/N guineapigs against an airborne challenge of virulent B. bronchispeptica in small-particle aerosol. Seronegative animals were vaccinated on days 0 and 21 with intramuscular injections of 0.2 ml of bacterin. Humoral antibody titres of the vaccinated animals, as determined by ELISA, ranged from 128-1024 on day 49. On day 30 following the second dose of bacterin (study day 51), 12 vaccinated and 12 PBS sham-vaccinated animals were exposed to an inhaled dose of 4.3 x 10(5) CFU of B. bronchiseptica (325 LD50). Vaccinated, challenged animals remained clinically normal, although each guineapig did develop a localized upper respiratory infection. The rate of weight gain as well as rectal temperature of these animals were analogous to those exhibited by the control groups. Examination of 4 of the vaccinated, challenged animals on day 7 after exposure showed bacteria present in moderate to high numbers in the larynx and trachea but only minimally detectable in the lungs; by 30 days after exposure, the numbers of bacteria in the larynx and trachea were diminished, with none being detected in the lungs. Pathological alterations induced by B. bronchiseptica were not detected at either day 7 or day 30 after challenge in any of the vaccinated, challenged animals. Protection induced in Strain 13/N guineapigs by the commercial canine bacterin was sufficient to preclude the development of pulmonary disease, even in animals presented with a massive challenge of virulent bacteria in a small-particle aerosol.

Kinetic and biochemical heterogeneity in vertebrate chondroepiphyseal regions during development.

The purpose of this study was to see if kinetic and biochemical heterogeneity could be documented in vertebrate chondroepiphyseal regions as they develop from mesenchymal condensations to cartilage. The kinetics of developing proximal and distal femoral chondroepiphyseal regions were studied from early limb bud stage to newborn animals in chicks, mice, and rabbits with thymidine autoradiography. Proteoglycan synthesis in the proximal femoral chondroepiphyseal region of the rabbit was studied with radioactive sulfate incorporation at 28 days of gestation and at 1 and 4 days after birth. The results indicated that these kinetic and biochemical characteristics of the developing chondroepiphyseal regions became heterogeneous very early in development. This early programming of populations of cells for division and for different biochemical functions existed during the fetal period when heterogeneity has been described histologically but has not been well documented.

Ribavirin prophylaxis and therapy for experimental argentine hemorrhagic fever.

Junin virus-infected rhesus macaques received prophylactic and therapeutic ribavirin to assess the potential of this drug for treating humans with Argentine hemorrhagic fever. When ribavirin was administered intramuscularly at the time of experimental infection with the lethal P3790 strain of Junin virus, all animals were protected from clinical disease. A delay in the initiation of therapy until after the onset of illness resulted in improvement and resolution of systemic signs of disease; however, survivors subsequently developed a late-onset central nervous system infection which was fatal in two of three animals. Side effects of ribavirin included thrombocytosis and severe anemia, both of which resolved promptly on withdrawal of drug therapy. Results of this study suggest that ribavirin may prove useful in treating humans with Argentine hemorrhagic fever.

Epiphyseal replacement using developing tissue donors in a murine model: a combined histologic and radiographic study.

Epiphyseal transplantation has long been a goal of orthopaedic surgeons. While microvascular surgery has raised hopes that this goal could be achieved, factors other than blood supply also appear capable of affecting the function of the epiphysis. Basic research into the biology of the epiphysis appears to be required. This would be facilitated with a model of epiphyseal transplantation using a small mammal. The purpose of this experiment was to develop such a model in the mouse. Developing CD1 mouse or Lewis rat limb tissue was used to replace knee tissue that had been resected from CD1 postnatal mouse hosts. Donor tissue ranged from 14-day embryonic mouse to 9-day postnatal mouse or 18- and 19-day fetal rat, which has a gestation similar to the mouse. The murine tissue is known to be avascular prior to the sixth postnatal day. The limbs were analyzed radiographically and histologically. The results show that epiphyseal replacement could be studied using developing tissue donors in a murine model. The results suggest that donor tissue prior to vascularization and tissue combinations with the least developmental time mismatch (the least heterochronicity) produced relatively the best, although still abnormal epiphyses.

Airborne-induced experimental Bordetella bronchiseptica pneumonia in strain 13 guineapigs.

To evaluate the efficacy of a commercial bacterial vaccine in protecting Strain 13 guineapigs against fatal Bordetella bronchiseptica pneumonia, it was necessary to establish the infectivity and disease pathogenesis induced by virulent organisms. When guineapigs were exposed to small-particle aerosols of varying concentrations of virulent B. bronchiseptica, a spectrum of disease was produced that ranged from inapparent illness to fulminant bronchopneumonia. Clinical signs began by day 4 after exposure, and were evidenced by anorexia, weight loss, respiratory distress and serous to purulent nasal discharge. Pathological alterations were limited to the respiratory system. Moribund animals exhibited a suppurative necrotizing bronchopneumonia and necrotizing tracheitis. In animals that survived the challenge, the bacteria were eliminated from the lungs by day 28 but continued to persist in the laryngeal area and the trachea. The median infectious dose and the median lethal dose were estimated to be 4 colony-forming units (CFU) and 1314 CFU respectively. These data suggest that the guineapig will be a valuable model system in which to study interactions between Bordetella species and host cells as well as to evaluate potential B. bronchiseptica immunogens.

Induced oral infection of the owl monkey (Aotus trivirgatus) with hepatitis A virus.

Several species of nonhuman primates have served as animal models for hepatitis A virus (HAV) infection and disease. This study was to determine the suitability of Aotus trivirgatus as an orally induced model for HAV infection and to reconfirm the owl monkey's susceptibility to the intravenous route of inoculation. Animals were inoculated, either orally or intravenously, with varying concentrations of PA-33 strain of HAV. Serum enzymes ALT, AST and GGTP levels were monitored and liver biopsies performed when values exceeded three standard deviations above individualized mean baseline values. All animals had postinoculation elevations of serum ALT and AST values, shed virus in their feces, and were seropositive to HAV by 60 days after inoculation. Eight of the ten postinoculation biopsy specimens had histologic lesions compatible with acute viral hepatitis. We conclude that the owl monkey is a useful and valuable model for the study of HAV disease.

Vascular events associated with the appearance of the secondary center of ossification in the murine distal femoral epiphysis.

Although the formation of a secondary center of ossification is often compared with that of the primary center, there are striking differences between these processes. In the formation of the primary center, vascular invasion is always associated with the maturation of chondrocytes, whereas vascularization of the epiphysis can proceed in two different ways. In some species, the epiphysis is vascularized by cartilage canals before the appearance of the secondary center. However, in the mouse, the distal femoral epiphysis is vascularized by peripheral vascular invasion without pre-existing cartilage canals. Histological study of serial sections and studies of vascularization by injection with India ink demonstrated the relationship between hypertrophic chondrocyte formation, vascular invasion, and the formation of the secondary center of ossification in the murine distal femoral epiphysis.

Preparation of a prototype inactivated hepatitis A virus vaccine from infected cell cultures.

Studies were conducted on the preparation, inactivation, safety, and immunogenicity of a prototype hepatitis A virus vaccine prepared from infected cell cultures. BS-C-1 cells maintained in medium 199 without serum were infected with the HM175 strain of hepatitis A virus and harvested after 21-28 days. The harvested virus preparation contained 6.8-7.4 (log 10) cell culture infectious doses/ml. After exposure to 1:4,000 formalin at 35 C, the infectivity titer decreased 10(6)-fold in 30 hr at an exponential rate, although virus was detected in 5.0-ml vaccine samples for up to three days. Three separate vaccine lots elicited antibody in all the guinea pigs given three doses. Owl monkeys given three doses of vaccine did not have any evidence of HAV infection but developed antibodies identifiable by radioimmunoassay and serum neutralization tests. After either oral or intravenous challenge with at least 10(6) monkey infectious doses of a virulent field strain of hepatitis A virus, none of the vaccinated monkeys shed virus in their feces or had elevated serum levels of alanine aminotransferase. The findings suggest that an effective inactivated whole virus hepatitis A vaccine can be prepared from cell culture.

Comparative rates of proteoglycan synthesis and size of proteoglycans in normal and osteoarthritic chondrocytes.

Normal human and osteoarthritic cells were isolated from cartilage with clostridial collagenase. The cells were grown in media as a suspension culture in the presence of 35SO4. Osteoarthritic cartilage of moderate histologic grade (4-8) yielded chondrocytes which incorporated 35SO4 at a rate 3-4 times greater than did normal chondrocytes. The rate of incorporation, however, decreased to normal levels with chondrocytes isolated from mild (grade 0-3) or more advanced (grade 9-13) stages of the disease. These results corroborate those obtained in earlier studies using organ cultures and show that when osteoarthritic cells are isolated from their matrix environment, they continue to synthesize macromolecules at an increased rate. Analysis of the material synthesized by the isolated cells on sizing column demonstrated an inverse relationship between the size of the 35SO4 containing molecules and the severity of the disease.

Quantitative histologic analyses of articular cartilage and subchondral bone from osteoarthritic and normal human hips.

Twelve femoral heads (two normal, four after fracture and six osteo-arthritic) were obtained at surgery or autopsy. Circumferential slices were obtained and five separate areas were analyzed in each for ash content; histological-histochemical grading of the severity of the cartilage changes; and quantitative morphometric analyses to establish the percentage of trabecular area, osteoblastic area and osteoclastic area. Analyses were performed to compare weight-bearing and non-weight-bearing areas of the femoral heads and to determine correlations between the bony and cartilaginous alterations. The data obtained showed wide variations in all parameters in the osteoarthritic specimens but consistently more marked cartilage and bony changes in the weight-bearing areas. Bone structures and metabolic parameters were distinctly increased for the osteoarthrities, differing significantly from both normal and fracture control groups. The bony change correlated directly with the severity of the cartilage lesions, as determined by the histological-histochemical grade.