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Biomedical subjects

M A Suckow

Publications and source records attributed to M A Suckow.

At least 19 recordsLinked to original sources

Use of porcine renal capsule matrix as a full-thickness dermal wound-healing material in rats.

OBJECTIVE: To compare the utility of porcine renal capsule matrix (RCM) with porcine small intestinal mucosa (SIS) in a rat full-thickness skin wound model. METHOD: Groups of rats had surgically-created wounds filled with either SIS or RCM. On each rat a contralateral wound was left unfilled (RCM-U or SIS-U). Wound diameter was measured 3, 7, 12, 17, 26 and 30 days after creation. Wound sites sampled 3, 7, 14, 28, 42 and 56 days after wound creation were numerically graded for degree of histologic change and for collagen content, based on intensity of trichrome staining. RESULTS: Wounds in all groups rapidly contracted to less than 50% of the original diameter within 12 days. There were no differences in wound diameter between RCM- and SIS-treated wounds at any time point, but these wounds had significantly greater (p < 0.001) diameters than their unfilled counterparts on days 7, 12 and 17. There were no differences in histologic scores or trichrome-staining scores between RCM- and SIS-treated wounds and their unfilled counterparts at any time point, except for a greater (p < 0.05) histologic score in SIS-treated wounds compared with unfilled controls on day 14. In both treatment groups an acute inflammatory response at the wound site was soon replaced by an influx of macrophages and fibroblasts. CONCLUSION: The results show that RCM is equivalent to SIS for the treatment of full-thickness wounds and that these materials may enhance wound healing in terms of wound-tissue collagenisation and maturation. These materials therefore merit further study in other wound-care models.

Animals↗

Sebaceous adenocarcinoma of the external auditory canal in a New Zealand white rabbit.

A rare sebaceous gland carcinoma of the external auditory canal in a rabbit is described. The tumour was characterized histologically by foci and cords of markedly pleomorphic cells with abundant cytoplasm and variable numbers of vacuoles. A single pulmonary mass had similar histological characteristics. This is the first such tumour reported in a rabbit.

Adenocarcinoma, Sebaceous↗

The characterization of mice with a targeted combined deficiency of protein c and factor XI.

Activated protein C functions directly as an anticoagulant and indirectly as a profibrinolytic enzyme. To determine whether the fibrin deposition previously observed in PC(-/-) murine embryos and neonates was mediated through the FXI pathway, PC(+/-)/FXI(-/-) mice were generated and crossbred to produce double-deficient progeny (PC(-/-)/FXI(-/-)). PC(-/-)/FXI(-/-) mice survived the early lethality observed in the PC(-/-)/FXI(+/+) neonates, with the oldest PC(-/-)/FXI(-/-) animal living to 3 months of age. However, the majority of these animals was sedentary and significantly growth-retarded. On sacrifice or natural death, all of these PC(-/-)/FXI(-/-) mice demonstrated massive systemic fibrin deposition with concomitant hemorrhage and fibrosis, as confirmed through histological analyses. Several of these animals also presented with enlarged lymph nodes and extensive lymphatic fluid in the thoracic cavity. Thus, although a number of the PC(-/-)/FXI(-/-) mice survived the lethal perinatal coagulopathy seen in the PC(-/-) neonates, they nonetheless succumbed to overwhelming thrombotic disease in later life. This combined deficiency state provided the first clear indication that the course of a severe thrombotic disorder could be manipulated by blocking the intrinsic pathway and provided the first opportunity to study a total protein C deficiency in an adult animal.

Animals↗

Intranasal vaccination of rabbits with Pasteurella multocida A:3 outer membranes that express iron-regulated proteins.

OBJECTIVE: To determine efficacy of intranasal vaccination of rabbits with Pasteurella multocida A:3 outer membrane proteins (OMP) expressing iron-regulated OMP (IROMP) in conferring protection against experimental challenge exposure. ANIMALS: 52 male New Zealand White rabbits. PROCEDURE: Rabbits were vaccinated intranasally on days 0, 7, and 14; some vaccines included cholera toxin (CT) as an adjuvant. Concentrations of intranasal IgA and serum IgG antibodies against P multocida OMP were determined. In experiment A, rabbits were vaccinated with either phospate-buffered saline solution (PBSS), PBSS-CT, OMP-CT, or IROMP-CT, challenge-exposed intranasally on day 16, and euthanatized and necropsied on day 28. Rabbits were also vaccinated with OMP or IROMP without CT and were not challenge-exposed. In experiment B, rabbits were vaccinated with PBSS, PBSS-CT, IROMP, or IROMP-CT. On day 17, rabbits were challenge-exposed intranasally. Nasal bacteria and antibodies were determined on day 24. RESULTS: In experiment A, OMP-CT vaccination stimulated mucosal and systemic antibody responses to the bacterium and enhanced resistance against challenge exposure. Intranasal bacterial counts were not significantly reduced. Vaccination with IROMP-CT stimulated mucosal and systemic antibodies, enhanced resistance to challenge exposure, and significantly reduced nasal bacterial counts. In experiment B, natural infection was detected in several rabbits at challenge exposure; however, IROMP-CT-vaccinated rabbits had significantly higher serum and nasal antibody responses, compared with other rabbits IROMP-CT-vaccinated rabbits had significantly lower nasal bacterial counts compared to control rabbits. CONCLUSIONS AND CLINICAL RELEVANCE: Intranasal vaccination of rabbits with P multocida outer membranes containing IROMP and CT stimulated immunity against experimental pneumonic pasteurellosis.

Adjuvants, Immunologic↗

Immunogenicity of antigens in boiled alginate microspheres.

Vaccine efficacy can be enhanced by delivery of antigens in synthetic microspheres. The process of antigen incorporation into microspheres can expose fragile antigens to damaging conditions, such as high temperatures, and to bacterial contamination. Maintenance of immunogenicity of several antigens and reduction of bacterial load in alginate microspheres following boiling was evaluated. Mice were immunized subcutaneously, initially and again 21 days later, with either non-boiled or boiled microspheres containing ovalbumin (OVA), a culture supernatant vaccine of Pasteurella haemolytica (PHV), or a potassium thiocyanate extract of P. multocida (PTE). Serum samples were obtained prior to immunization and at the time of euthanasia 28 days later. Culture of microspheres showed that boiling completely eliminated aerobic bacterial growth for OVA-containing microspheres, and reduced growth by a factor of 10(4) for PTE microspheres. More bacteria were cultured after boiling than before for PHV microspheres. ELISA performed on serum and intestinal lamina propria explant supernatants showed that immunogenicity of PHV microspheres was not altered by boiling. Boiled OVA microspheres were still able to stimulate a significant serum IgG anti-OVA titer in mice, but boiled PTE microspheres completely lacked immunogenicity. Elispot assays of spleens showed that only PHV microspheres were able to retain immunogenicity after boiling. Results indicate that boiling is not an effective means for reducing the bacterial load of alginate microspheres and that the process is associated with a diminution of vaccine immunogenicity.

Alginates↗

Immunization of rabbits against Pasteurella multocida using a commercial swine vaccine.

Elaboration of heat-labile toxin (PMT) is an important virulence factor in some isolates of Pasteurella multocida from rabbits. Previously, we reported that immunization with inactivated PMT (IPMT) stimulated protective immunity to challenges from PMT. To test the hypothesis that immunization with a commercial swine vaccine containing IPMT stimulates similar protective immunity, groups of five rabbits were inoculated twice intramuscularly (i.m.), 10 days apart, with 0.5 ml of sterile saline or a commercial swine P. multocida bacterin-toxoid (BT). In addition, a group was inoculated intranasally with 5 microg of IPMT. Serum and nasal lavage samples were taken on days 0, 7, 14 and 21 after initial immunization and assayed by ELISA for anti-PMT antibody. Serum IgG and nasal lavage IgA were detectable by day 14 in BT and IPMT-immunized rabbits, but not in the saline controls. Groups of similarly inoculated rabbits were then challenged intranasally with 28 microg of PMT 21 days after initial immunization, and necropsied 7 days later, along with control challenged and non-challenged rabbits. Histological lesion severity was graded on a numerical scale. Non-immunized and saline, challenged controls developed more severe pneumonia, pleuritis, nasal turbinate atrophy and testicular atrophy than IPMT and BT-immunized rabbits. The results confirm the hypothesis that immunization with a commercial swine P. multocida BT confers protective immunity in rabbits against challenges from PMT.

Animals↗

Intranasal vaccination of New Zealand white rabbits against pasteurellosis, using alginate-encapsulated Pasteurella multocida toxin and potassium thiocyanate extract.

OBJECTIVE: We evaluated the efficacy of intranasal administration of Pasteurella multocida toxin (PMT) and a potassium thiocyanate extract of P. multocida (CN) encapsulated in alginate microspheres, compared with unencapsulated PMT and CN antigens, in protection of rabbits against pasteurellosis. METHODS: New Zealand male rabbits (n=24) were allotted randomly into four intranasally administered vaccine groups: 1, PMT/CN; 2, microencapsulated PMT/CN with or; 3, without subcutaneous priming; and 4, empty microspheres (control). Blood samples and nasal wash specimens were collected before vaccination and one week after each vaccination (days 7, 21, 35, and 49). Rabbits were primed subcutaneously with either unencapsulated PMT/CN or aluminum hydroxide (control) (day 0), vaccinated intranasally (days 14, 28, and 42), challenged intranasally with live P. multocida (day 56), and necropsied (day 60). RESULTS: Compared with controls, PMT/CN-immunized rabbits had significantly higher concentrations of serum IgG and IgM, nasal IgG, and bronchoalveolar lavage fluid IgA and IgG against CN. Immunized rabbits had 100% survival rate and low numbers of bacteria in liver and lungs; the control group had 50% survival rate and higher numbers of bacteria (> 4x) per gram of tissue in liver and lungs. CONCLUSION: The PMT/CN microspheres stimulated systemic and mucosal immune responses similar in effectiveness (protection) to those in response to unencapsulated PMT/CN administration.

Administration, Intranasal↗

Evaluation of hypothermia-induced analgesia and influence of opioid antagonists in leopard frogs (Rana pipiens).

Hypothermia results in diminished voluntary muscle activity, and is frequently used as a means of providing deep anesthesia to ectotherms and some mammals. In ectotherms, however, it is unclear if hypothermia produces true pain insensation. A needle-probe thermometer was used to demonstrate in frogs (Rana pipiens) that local hypothermia (9 degrees C) could be induced by placement of a tourniqueted leg into ice water (6 degrees C) for 10 min in contrast to the contralateral nontourniqueted leg (21.8 degrees C) kept out of ice water. Analgesia was tested by placement of dilutions of acetic acid on the rear leg. Further tests using groups of 10 frogs demonstrated that frogs with local hypothermia tolerated greater concentrations of acetic acid (mean acetic acid test score = 11) than morphine (10 mg/kg)-treated (9.6) or nontreated (5.8) frogs. Additional studies showed that morphine analgesia was blocked with naloxone doses as low as 0.01 mg/kg and hypothermia-induced analgesia at 10 mg/kg. Naltrexone blocked morphine analgesia at dosages as low as 0.01 mg/kg and hypothermia-induced analgesia at 0.10 mg/kg. In summary, this study demonstrates that hypothermia induces significant analgesia in an amphibian, and that this analgesia is partially blocked by naloxone and naltrexone, suggesting that the effect is mediated at least partially by opioid receptors.

Analgesia↗

Induction of protective immunity in rabbits by coadministration of inactivated Pasteurella multocida toxin and potassium thiocyanate extract.

Pasteurella multocida is a bacterial pathogen that causes rhinitis (snuffles), pneumonia, otitis media, septicemia, metritis, and death in domestic rabbits. Currently, there are no effective vaccines to prevent infection by this organism. Subcutaneous (s.c.) immunization with either exotoxin or thiocyanate extracts of P. multocida induces partial protection in rabbits. Since disease begins at mucosal sites, induction of local immunity may be important in preventing systemic disease. Little is known concerning the efficacy of intranasal (i.n. ) administration of these antigens in inducing protective mucosal immunity to P. multocida in rabbits. The purpose of this study was twofold: (i) to investigate the effectiveness of vaccination with purified P. multocida toxin (PMT) and a potassium thiocyanate extract of P. multocida (CN) in combination and (ii) to evaluate the efficacy of administration of these antigens i.n. versus s.c. Forty-eight rabbits were randomly divided into eight different treatment groups. Rabbits received either one or both antigens by either s.c. or i.n. administration. Following vaccination, each group received an i.n. challenge of P. multocida. Rabbits vaccinated with both antigens i.n. or s.c. had a 100% survival rate, few or no bacteria in the liver and lungs, high serum immunoglobulin G (IgG) and IgM antibody titers, and significant numbers of IgG antibody-secreting cells (ASC) in the spleen and tracheobronchial lymph node. Rabbits vaccinated i.n. had significant nasal and bronchoalveolar lavage IgA antibody levels. Rabbits vaccinated with only one antigen, either PMT or CN, had lower antibody titers, moderate to severe liver and lung infections, and fewer ASC compared to rabbits receiving both antigens. Rabbits in the control groups had moderate to severe liver and lung infections. This study indicates that i.n. immunization with both PMT and CN induces an effective response against homologous P. multocida challenge.

Animals↗

Derivation of Pasteurella multocida-free rabbit litters by enrofloxacin treatment.

Pasteurella multocida is an important bacterial pathogen of rabbits that is easily transmitted from infected does to their kits prior to weaning. Enrofloxacin, a flouroquinolone antibiotic, is effective at limiting nasal carriage of P. multocida in rabbits. To determine if enrofloxacin treatment of pregnant does infected with P. multocida can be used to produce P. multocida-free litters, groups of 3 rabbits were inoculated intranasally on day 10 of gestation with 1.0 x 10(6) P. multocida CFUs. Beginning on day 14, one group received enrofloxacin IM (5 mg kg-1, BID), and a second group received enrofloxacin in the drinking water (200 mgl-1). IM treatment continued until kindling, while PO treatment continued 1 week after kindling. A third group was infected but received only IM saline, and a fourth group was infected but not treated. In addition, a fifth group was neither infected nor treated. Culture of nasal lavage samples and tissues from does and kits showed that both routes of enrofloxacin treatment failed to completely eliminate P. multocida from does, but all kits from enrofloxacin-treated does were free from P. multocida. These results suggest that treatment does with enrofloxacin during the periparturient period may interrupt transmission of P. multocida from infected does to their kits and that this treatment may be useful for deriving Pasteurella-free rabbits from infected does.

Animals↗

Oral immunization of rabbits against Pasteurella multocida with an alginate microsphere delivery system.

Oral delivery of microencapsulated antigens is a potential means to vaccinate rabbits against Pasteurella multocida, a common bacterial pathogen. Groups of five rabbits were dosed orally on days 0, 7, and 14 with alginate microspheres prepared to contain no added protein, 5 mg of a potassium thiocyanate extract of P. multocida (PTE), or 5 mg of PTE with 200 micrograms of cholera toxin (CT). In addition, groups were dosed orally with 5 mg of soluble PTE with or without 200 micrograms CT, intranasally (IN) with 1 mg of soluble PTE, or with saline. Serum and nasal lavage samples collected prior to initial immunization and 10, 16, and 21 days later were assayed by ELISA for anti-PTE IgG and IgA. Strong nasal lavage IgA and serum IgG activities were found in samples from rabbits immunized with PTE IN or orally when incorporated into microspheres. Addition of CT did not significantly enhance either response. To examine the development of protective immunity, groups were similarly immunized and challenge-exposed IN on day 16 with 10(6) CFU of P. multocida. One week later, rabbits were euthanized, and specimens from the lungs, nasopharynx, liver, and inner ear were cultured for P. multocida. Less severe infections of the lung and nasopharynx developed in rabbits immunized with PTE IN or orally in microspheres, with or without added CT. In addition, culture of liver and tympanic bullae samples from these rabbits yielded growth of P. multocida less frequently compared to other P. multocida-challenged rabbits. Coadministration of CT and PTE did not significantly improve protective immunity to challenge.

Adjuvants, Immunologic↗

Gastric trichobezoar in a banner-tailed kangaroo rat (Dipodomys spectabilis).

A male, wild-caught kangaroo rat developed anorexia and wasting. The animal was euthanized and a gastric trichobezoar found at necropsy. The paucity of information regarding the clinical medicine of this species is a hindrance to those charged with the care of kangaroo rats. Gastric trichobezoar should be considered as a differential diagnosis in cases of anorexia in kangaroo rats.

Animals↗

Enhancement of respiratory immunity to Pasteurella multocida by cholera toxin in rabbits.

Cholera toxin (CT) is a potent adjuvant for the mucosal immune system. The purpose of this study was to determine if coadministration of CT with a potassium thiocyanate extract of Pasteurella multocida (PTE) leads to enhanced anti-PTE antibody activity and increased protection of rabbits against infection with P. multocida and associated disease. Groups of rabbits were immunized intranasally on days 0, 7, and 14, with phosphate buffered saline (PBS), 200 micrograms of CT, 1.0 mg of PTE, or 1.0 mg PTE with 200 micrograms CT. Nasal lavage and serum samples were collected over 28 days after initial immunization and evaluated by ELISA for specific antibody directed against PTE. Marked increases in serum (IgG) and nasal lavage (IgA) anti-PTE antibody activity were found beginning after day 14 in rabbits immunized with PTE. Rabbits immunized with PTE and CT demonstrated further increases in this activity. Tracheobronchial lavage samples collected at the time of necropsy demonstrated a significant level of anti-PTE IgA activity in animals immunized with PTE, and coadministration with CT stimulated a further significant increase in this activity. Groups of similarly immunized rabbits were challenged 16 days after initial immunization with 5 x 10(7) CFUs of P. multocida. Nasal lavage samples were cultured for P. multocida over the next 10 days. Rabbits were euthanized within 10 days after challenge, tissues cultured for P. multocida, and histopathologic lesion severity graded using a numeric scale. Rabbits immunized with PTE survived longer, had less severe lesions of the lungs, pleura, and liver, and fewer P. multocida CFUs cultured from samples than PBS or CT controls. Coadministration of CT led to further reductions in lesion severity of those tissues and numbers of P. multocida CFUs cultured from samples. Increased nasal turbinate atrophy of rabbits immunized with PTE with or without CT was associated with increased mean survival time. In summary, coadministration of CT with PTE enhanced protective immunity to P. multocida disease and infection in rabbits.

Adjuvants, Immunologic↗

Protective immunity to Pasteurella multocida heat-labile toxin by intranasal immunization in rabbits.

Heat-labile Pasteurella multocida toxin (PMT) is an important virulence factor of some isolates from rabbits. To determine whether protective immunity to PMT could be induced in rabbits by intranasal immunization with heat-inactivated PMT, we immunized groups of rabbits intranasally at days 0, 7, 14, and 21 with inactivated PMT, with or without cholera toxin, an adjuvant for the mucosal immune system. Significant increases in anti-PMT IgA in nasal lavage samples and anti-PMT serum IgG, as determined by enzyme-linked immunosorbent assay, developed within 2 weeks after initial immunization. Coadministration of cholera toxin with inactivated PMT enhanced anti-PMT activity in the samples. Rabbits similarly immunized on days 0, 7, and 14 were challenged with PMT, and tissues were graded histologically on a numeric scale of lesion severity. Immunization conferred partial protection against development of pneumonia, pleuritis, hepatic necrosis, and testicular atrophy in rabbits challenged 16 days after initial immunization. Thus, immunization with inactivated PMT stimulates a protective response to PMT challenge in rabbits that is enhanced by coadministration of cholera toxin.

Administration, Intranasal↗

Stimulation of gastrointestinal antibody to Shiga toxin by orogastric immunization in mice.

Shiga toxin (ST) is a protein toxin of Shigella dysenteriae type 1, a causative agent of severe diarrhoea and dysentery. In this report we describe the gastrointestinal secretory antibody response of mice following orogastric immunization with ST. Gastrointestinal secretions were sampled by a gastrointestinal lavage technique weekly for 5 weeks after initial immunization. Assay of lavage samples by ELISA showed that mice vaccinated orogastrically with various doses of ST developed gastrointestinal antibody to ST in a dose-dependent manner. Serum anti-ST activity developed by 5 weeks after initial immunization. The ability of ST to act as a mucosal immune adjuvant was investigated by coadministration of ST and keyhole limpet haemocyanin. In contrast to cholera toxin, a potent adjuvant, ST did not demonstrate adjuvant activity. The mouse gastrointestinal lavage model could be useful for further analysis of the cellular basis of ST immunogenicity.

Adjuvants, Immunologic↗