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Animal model studies of genital chlamydial infections. Immunity to re-infection with guinea-pig inclusion conjunctivitis agent in the urethra and eye of male guinea-pigs.

A previous report demonstrated that male guinea-pigs could be infected in the urethra with guinea-pig inclusion conjunctivitis (GPIC) agent and that the infection was transmitted during mating from infected males to females. In the experiments reported here, inoculation of male guinea-pigs in the urethra with GPIC organisms resulted in infection which subsided spontaneously in about 2 weeks. Males were demonstrated to be completely resistant to urethral challenge with 10(3)ID50 when tested 6 weeks after urethral infection. These guinea-pigs, immune to re-infection of the urethra, remained susceptible to infection of the eye, but this ocular infection was shorter in duration than that in previously uninfected control animals. Infection in the eye resulted in immunity to both ocular and urethral infection when animals were challenged 6 weeks after the ocular infection.

Animals↗

Strain related infectivity of Neisseria gonorrhoeae for the guinea-pig subcutaneous chamber and the variability of the immune resistance in different breeds of guinea-pig.

The ability of gonococci to infect the subcutaneous chamber in guinea-pigs is strictly strain related. This ability was usually present in prototrophic strains and auxotrophs requiring proline only which were obtained from patients with uncomplicated gonorrhoea, but it was invariably absent in Neisseria gonorrhoeae auxotypes requiring arginine, uracil, and hypoxanthine, or arginine only for growth. All the strains obtained from disseminated gonococcal infections (all dependent upon arginine, uracil, and hypoxanthine) were unable to infect the guinea-pig chamber. Hence, the high invasiveness of N. gonorrhoeae for man and its infectivity for guinea-pig chambers appear to be unrelated properties. Although guinea-pigs of the same origin (Dunkin-Hartley) were used throughout, the degree of immune resistance was found to differ between the lines supplied by various breeders--that is, after a standard immunisation schedule using whole cell gonococcal vaccines, the homologous immune resistance to challenge varied from weak or non-existent in some lines, to highly resistant in others.

Animals↗

Molecular cloning of the guinea pig GRO gene and its rapid expression in the tissues of lipopolysaccharide-injected guinea pigs.

BACKGROUND: CXC chemokines, IL-8 and GRO, play a role in the recruitment of neutrophils in the human. The functional orthologues in the rat and mouse are CINC/KC and MIP-2. The lack of IL-8 made these animals less useful to study the role of IL-8 and GRO. METHODS: Guinea pig (gp) cDNA libraries were screened for GRO and IL-1beta. A gp genomic library was screened with a gpGRO cDNA probe. Expression of gpIL-8, gpGRO, gpTNFalpha, and gpIL-1beta was investigated by Northern analysis and/or by in situ hybridization. RESULTS: Two gpGRO cDNAs, a 3.0-kb gpGRO genomic DNA, and a gpIL-1beta cDNA were cloned. gpGRO and gpIL-8 mRNA were detected in different tissues including lungs 1 h after intraperitoneal injection of lipopolysaccharide (LPS) into guinea pigs. gpGRO, gpIL-8, gpTNFalpha, and gpIL-1beta expression peaked at 3 h in the lungs. Both gpGRO and gpIL-8 mRNA were detected in the cells in alveolar spaces and bronchial epithelial cells. However, gpGRO mRNA, but not gpIL-8, was also expressed in endothelial cells and vascular smooth muscle cells. CONCLUSIONS: gpGRO and gpIL-8 mRNA rapidly accumulated in the lungs of guinea pigs after LPS injection. Expression of gpIL-8 and gpGRO mRNA appeared to be independent from TNFalpha- or IL-1beta-stimulation in this model. A high level expression of gpGRO in vascular cells suggest an important role of GRO in the sequestration of neutrophils and multi-organ injuries induced by LPS. The guinea pig will provide an excellent model to study the roles of IL-8 and GRO, important inflammatory mediators in the human.

Amino Acid Sequence↗

Density and kinetics of I(Kr) and I(Ks) in guinea pig and rabbit ventricular myocytes explain different efficacy of I(Ks) blockade at high heart rate in guinea pig and rabbit: implications for arrhythmogenesis in humans.

BACKGROUND: Class III antiarrhythmic agents commonly exhibit reverse frequency-dependent prolongation of the action potential duration (APD). This is undesirable because of the danger of bradycardia-related arrhythmias and the limited protection against ventricular tachyarrhythmias. The effects of blockade of separate components of delayed rectifier K(+) current (I(K)) may help to develop agents effective at high heart rate. METHODS AND RESULTS: We assessed the density and kinetics of the 2 components of the delayed rectifier K(+) current, I(Kr) and I(Ks), in rabbit and guinea pig ventricular myocytes. The effects of their specific blockers (chromanol 293B for I(Ks) and E-4031 for I(Kr)) on the action potential was studied at different heart rates by use of whole-cell patch-clamp techniques. In guinea pig ventricular myocytes only, blockade of I(Ks) causes APD prolongation in a frequency-independent manner, whereas blockade of I(Ks) in rabbit ventricular myocytes shows reverse frequency dependence, as does blockade of I(Kr) in both species. This result can be explained primarily by the higher density of I(Ks) in guinea pig ventricle and by its slow deactivation kinetics, which allows I(Ks) to accumulate at high heart rate because little time is available for complete deactivation of it during diastole. CONCLUSIONS: Density and kinetics of components of I(K) explain why blockade of I(Ks) is more effective at high heart rate in the guinea pig ventricle than in the rabbit ventricle, without adverse effects at low heart rate.

Action Potentials↗

Immunohistochemical identification and localization of actin and fimbrin in vestibular hair cells in the normal guinea pig and in a strain of the waltzing guinea pig.

Using immunohistochemical methods, actin and fimbrin were identified and localized in vestibular epithelia iin the normal guinea pig and in the waltzing guinea pig. In the normal guinea pig, actin was found in the stereocilia and in the cuticular plate. Fimbrin was detected in the stereocilia but surprisingly not in the cuticular plate. As fimbrin was found in the cuticular plates in cochlear hair cells it is suggested that the hair cells in the two organs have different mechanoreceptor properties which can demand different cuticular plate stability. In the waltzing guinea pig, actin was found in the stereocilia, in the rod and n the cuticular plate. Fimbrin was seen in the stereocilia and in the rod, but could not be detected in the cuticular plate. These results emphasize the resemblance between the rod and the stereocilia. It is suggested that the rod is a genetically induced pathological intracellular type of stereocilium which grows in an uncontrolled manner.

Actins↗

Antibodies to guinea pig lymphokines. II. Suppression of delayed hypersensitivity reactions by a "second generation" goat antibody against guinea pig lymphokines.

A "second generation" antibody to a highly purified lymphocyte product was raised in a goat against material eluted from a rabbit anti-guinea pig lymphokine immunoadsorbent column. This anti-lymphokine serum, in constrast to anti-lymphocyte serum (ALS) did not appear to contain cytotoxic antibodies directed against membrane antigens on guinea pig lymph node lymphocytes. Furthermore, the anti-lymphokine serum did not inhibit the formation of spontaneous T rosettes nor significantly depress lymphocyte response to mitogens. The anti-lymphokine serum totally suppressed the delayed skin reactivity to PPD and contact sensitivity to DNCB when injected intradermally around the site of antigen challenge. By contrast, intradermally injected ALS did not appear to suppress the PPD response in sensitized guinea pigs. Intravenously and i.p. administered anti-lymphokine serum was somewhat less effective in suppressing the delayed skin response to PPD. The intradermal injection of the antiserum had no effect on nonspecific inflammation evoked by turpentine-olive oil or on the extravasation of circulating Evans blue evoked by intradermally injected histamine. Histologic examination of 24-hr DNCB-induced skin lesions from sensitized guinea pigs treated with intradermally injected anti-lymphokine serum showed marked reduction of mononuclear infiltration of the dermis and of epidermal lesions, as compared with skin sites taken from sensitized animals pretreated with normal goat serum. The anti-lymphokine serum injected i.v. also markedly reduced the perivascular infiltration of the dermis and subcutis in skin reaction sites from sensitized animals challenged with PPD. Intravenous treatment with ALS for 3 consecutive days caused extensive depletion of the paracortical areas of peripheral lymph nodes whereas treatment with normal serum and anti-lymphokine serum caused no such depletion. It is proposed that the anti-lymphokine serum is directed against activated lymphocyte products, one of them being MIF. These products are involved in the mediation of delayed hypersensitivity reactions. This is in marked contrast to ALS, the suppressive action of which appears to be central rather than peripheral.

Animals↗

Methoxyphenamine inhibits histamine-induced bronchoconstriction in anaesthetized guinea-pigs and histamine-induced contractions of guinea-pig ileum in vitro.

Measurements were made of the effects of methoxyphenamine hydrochloride on histamine-, acetylcholine- and U46619-induced bronchoconstriction in pentobarbitone-anaesthetized guinea-pigs, and on histamine- and acetylcholine-induced contractions of guinea-pig ileum in vitro. Methoxyphenamine (20 mg/kg, i.v.) did not affect bronchoconstriction induced by acetylcholine or the thromboxane A2-mimetic U46619. However, it produced a parallel rightward shift [2.94-(1.79, 4.41) fold, 95% confidence limits in parentheses] of the curve relating bronchoconstrictor responses to log-dose of histamine at a total dose of 13 mg/kg, i.v., which was not significantly different from the shift [3.30-(1.93, 5.56) fold] produced by 3 micrograms/kg, i.v., of the histamine antagonist mepyramine maleate. Histamine-induced contractions of the isolated guinea-pig ileum were antagonized by methoxyphenamine (10(-5) to 10(-3) M). The histamine log-concentration-response curve was shifted to the right in a parallel manner by methoxyphenamine (10(-5) to 10(-4) M), without depression of maximum responses. However, at higher concentrations, maximum responses were reduced. The slope of the Schild plot was significantly different from -1. The degree of the rightward shift of the concentration-response curves to histamine, produced by 10(-5) M of methoxyphenamine [3.90-(2.83, 4.97) fold], was not significantly different from that produced by 3 x 10(-9) M of mepyramine [4.60-(2.86, 6.52) fold]. Methoxyphenamine, at concentrations of 10(-5) to 3 x 10(-4) M, had no significant effect on responses of guinea-pig ilea to acetylcholine (10(-9) to 10(-5) M). These results indicate that methoxyphenamine antagonizes the effects of histamine both in vivo and in vitro. In vitro studies indicate a noncompetitive antagonism.

15-Hydroxy-11 alpha,9 alpha-(epoxymethano)prosta-5↗

[The effect of rotating magnetic fields on the growth of Deal's guinea pig sarcoma transplanted subcutaneously in guinea pigs].

The effect of rotating magnetic field (RMF) was studied on the growth of Deals' guinea pig sarcoma (DGPS) transplanted subcutaneously into the inguen of guinea pigs. The 0.2 ml of transplanted suspension contained 1.5-2.0 x 10(6) of vital DGPS cells. Over a period of 24 days the animals were daily two times exposed to RMF for 3 hours. The animals were sacrificed in chloroform anesthesia, the inguinal tumors, lungs and hearts were removed and weighed. The following findings were recorded: 1. RMF inhibited the growth of DGPS cells in experimental guinea pigs compared to control animals (p < 0.001); 2. In the experimental animals DGPS cells did not metastasize into the lung parenchyma; 3. The pathological characteristics of the tumors in the experimental and control group of guinea pigs tended to be different. (Tab. 3, Ref. 14.)

Animals↗

Ectoparasites of guinea fowl (Numida meleagris galeata Pallas) and local domestic chicken (Gallus gallus) in southern Guinea Savanna, Nigeria.

The ectoparasites of poultry in a southern guinea savanna zone were investigated by the examination of guinea fowl and local domestic chickens in the range and guinea fowl under intensive management. The prevalent ectoparasites of range guinea fowl and local chickens include seven species of lice Menacanthus stramineus, Menopon gallinae, Goniodes gigas, Goniocotes gallinae, Lipeurus caponis, Numidilipeurus tropicalis, Damalinia bovis; three mites Bdellonyssus bursa, Megninia cubitalis, Dermanyssus gallinae; two fleas Echidnophaga gallinacea, Ctenocephalides felis and two ticks Argas persicus and Ambylomma variegatum. Under intensive management, infestation by G. gigas, L. caponis and M. gallinae led to clinical signs, feather damage, reduced food intake and death.

Animals↗

Molecular cloning of cDNA for guinea pig CYP1A2 comparison with guinea pig CYP1A1.

Guinea pig CYP1A2 cDNA was isolated by RT-PCR from liver tissue of 3,3'-methylcholanthrene-treated guinea pigs. It shares considerable sequence identity with guinea pig CYP1A1 (nt 77%, aa 65%), but differs in levels of constitutive expression, function, and inducibility. Western blot analysis of protein expressed by full-length cDNA in COS-1 cells identified CYP1A2 (56 kDa) and CYP1A1 (53 kDa) proteins in corun liver microsomes. CYP1A2 transfectants metabolized methoxyresorufin and ethoxyresorufin, while CYP1A1 transfectants metabolized only ethoxyresorufin. Constitutive expression of CYP1A2 mRNA (2.0 kb) and protein was much lower than that of CYP1A1 mRNA (2.6 kb) and protein, but the fold induction of CYP1A2 by 3,3'-methylcholanthrene was greater than that of CYP1A1. Changes in splicing of CYP1A2 pre-mRNA occur upon treatment with 3,3'-methylcholanthrene.

Amino Acid Sequence↗

Two distinct insulin-related molecules in the guinea pig: immunological and biochemical characterization of insulin-like immunoactivity from extrapancreatic tissues of the guinea pig.

In this study we extracted guinea pig brain and testis with; the extract was adsorbed to and eluted from cartridges (the Sep-Pak C18 procedure). We found this procedure superior for recovering crystalline insulin added to buffers or tissues, and for recovering endogenous insulin from plasma, but inferior for recovery of insulin from tissues. However, we did find 'rat/pork' type-insulin in guinea pig brain and testis (5-50 pg/g wet weight tissue). Our results with the Sep-Pak C18 procedures were reproduced by four other laboratories (who found 4-60 pg/g wet weight of tissue) and similar findings were also obtained by an independent investigator. Thus, we conclude that extrapancreatic tissues of guinea pigs have a second type of insulin-related material that is more typical of other mammalian insulins, but that the amount recovered is dependent upon the extraction procedure utilized.

Animals↗

Ultrastructural development and persistence of guinea pig cytomegalovirus in duet cells of guinea pig submaxillary gland.

Salivary glands from Hartley guinea pigs were experimentally infected with guinea pig cytomegalovirus (GPCMV) and examined by light and electron microscopy at different time intervals. Characteristic intranuclear and intracytoplasmic viral inclusions were observed in duct cells of infected animals. Viral inclusion counts and infectivity titers in the salivary gland reached maximum levels by 3 to 4 weeks after infection; infectivity persisted, though at reduced levels, for at least 30 weeks. Electron microscopic examination of viral inclusions revealed several developmental events including nucleocapsid assembly, envelopment of nucleocapsids at the inner nuclear membrane and their enclosure by a thin vacuolar membrane. While contained within cytoplasmic vacuoles, enveloped virions acquired surface spikes. Cytoplasmic vacuoles containing virions subsequently coalesced and discharged mature virions at the cell surface into the lumen of the salivary gland duct. The data indicate that the ultrastructural development of GPCMV in the guinea pig salivary gland shows many similarities to that of human cytomegalovirus in humans. The salivary gland may provide a primary locus for virus shedding and horizontal transmission of cytomegalovirus.

Animals↗

Immunochemical characterization of a cytochrome P450 isozyme and a protein purified from liver microsomes of male guinea pigs and their roles in the oxidative metabolism of delta 9-tetrahydrocannabinol by guinea pig liver microsomes.

A protein (designated as protein-B) was purified from liver microsomes of adult male guinea pigs by an affinity chromatography with omega-aminooctyl Sepharose 4B, followed by HPLC using DEAE-5PW and hydroxyapatite columns which had been used to purify a cytochrome P450 (P450) isozyme (P450-A) from the same subcellular fraction (Narimatsu et al., Biochem Biophys Res Commun 172: 607-613, 1990). Protein-B had a molecular mass of 49 kDa in SDS-PAGE, but did not show absorbance at 417 nm for heme. Further, it did not show any oxidative activities towards aniline (AN), d-benzphetamine (d-BP), p-nitroanisole (p-NA) or delta 9-tetrahydrocannabinol (delta 9-THC) in a reconstituted system including dilauroylphosphatidylcholine, NADPH-P450 reductase, and cytochrome b5. However, antiserum against protein-B raised in rabbits suppressed liver microsomal oxidative activities towards d-BP and p-NA dose-dependently. The antibody decreased delta 9-THC oxidative activity most effectively, but did not decrease AN hydroxylation activity. Antiserum against P450-A suppressed all the activities towards these four substrates, especially towards delta 9-THC, in liver microsomes of male guinea pigs. Moreover, reconstitution with hemin made it possible for protein-B to produce some oxidative activity toward delta 9-THC. These results suggest that protein-B is also a cytochrome P450 isozyme which has lost a heme moiety during purification steps. Both P450-A and protein-B could have a role as cytochrome P450 isozymes in the oxidative metabolism of drugs, especially that of delta 9-THC by the liver microsomes of adult male guinea pigs.

Amino Acid Sequence↗

Structural difference between the two forms of guinea-pig beta 2-microglobulin and their occurrence in inbred guinea-pig strains.

The structural difference between two forms (basic and acidic) of guinea-pig beta 2-microglobulin (beta 2m) has been established. Both forms are present in urine from inbred guinea-pig strains. The beta 2m forms were each digested with carboxypeptidase Y and carboxypeptidase A contaminated with carboxypeptidase B. Released amino acids were separated from remaining protein, dansylated and analysed by 2-dimensional TLC on polyamide layer sheets. From the results it was concluded that the basic beta 2m form has lysine and the acidic beta 2m form has asparagine as their respective C-terminal amino acids. The acidic form is also 1 amino acid (lysine) shorter than the basic form, which is supported by electrophoretic studies on sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The presence of the 2 forms of beta 2m in urine from inbred guinea-pig strains 2 and 13, shown by gel filtration and ion exchange chromatography, makes it unlikely that the 2 forms are a result of genetic polymorphism.

Amino Acid Sequence↗

Ethylcholine mustard aziridinium ion (AF64A) impairs cholinergic neuromuscular transmission in the guinea-pig ileum and urinary bladder, and cholinergic neuromodulation in the enteric nervous system of the guinea-pig distal colon.

AF64A was examined for its ability to impair cholinergic neurotransmission in the autonomic nervous system. In the guinea-pig ileum AF64A impaired cholinergic neuromuscular transmission. In the guinea-pig urinary bladder AF64A selectivity impaired the cholinergic but not the purinergic component of neuromuscular transmission. In the guinea-pig colon circular muscle AF64A impaired cholinergic neuromodulation of the inhibitory transmission but not the inhibitory transmission itself. The nature of impairment of cholinergic transmission by AF64A is discussed.

Acetylcholine↗

Studies on thymocyte subpopulations in guinea pigs. VII. Characterization of cell populations responsive to guinea pig interleukin 1 and interleukin 2.

The properties of thymocytes responding by proliferation to a mitogenic lectin (PHA), interleukin 1 (IL 1), or interleukin 2 (IL 2) were studied and compared to the properties of cells known to respond to a separate thymocyte growth factor (TGP), which has so far only been studied in guinea pigs. Thymocytes from guinea pigs were separated into subpopulations by density gradient centrifugation with Percoll and by rosette formation with rabbit erythrocytes. PHA-responsive cells were recovered exclusively in a non-rosetting, low-density population designated Ia, RFC-, constituting approximately 4 per cent of all thymocytes. Thus, according to the prevailing view of lymphocyte mitogenesis, IL 1-producing, IL 2-producing as well as IL 2-responding cells are all present in this population. The mitogen-responsive cells could be further stimulated by addition of IL 1 or IL 2, indicating that the magnitude of the mitogenic response was regulated by the production of these factors and was not restricted by the number of IL 2-responding cells. IL 1, to some extent, also enhanced the mitogen response in a non-rosetting, intermediate density population designated Ib, RFC-. None of the factors could affect the lack of mitogen responsiveness in the high-density population II, constituting approximately 85% of the total population and probably including most small cortical thymocytes. We conclude that IL 2-responding thymocytes are present above all in the quantitatively small population Ia, RFC- (shown to contain also the mature, mitogen-responding cells) and to a smaller extent in Ib, RFC- (where a deficit in IL 1-producing cells may explain the poor mitogen responsiveness), but not in the major, high-density population II. From our data, it is also evident that the mitogen, IL 1- and IL 2-reactive cells can be separated from a population of intensely proliferating thymic precursor cells which are stimulated to grow by TGP. Therefore, the growth of these immature cells does not seem to be regulated by IL 1 or IL 2 in the guinea pig.

Animals↗

Usefulness and limitations of various guinea-pig test methods in detecting human skin sensitizers-validation of guinea-pig tests for skin hypersensitivity.

Several guinea-pig predictive test methods were evaluated by comparison of results with those obtained with human predictive tests, using ten compounds that have been used in cosmetics. The method involves the statistical analysis of the frequency with which guinea-pig tests agree with the findings of tests in humans. In addition, the frequencies of false positive and false negative predictive findings are considered and statistically analysed. The results clearly demonstrate the superiority of adjuvant tests (complete Freund's adjuvant) in determining skin sensitizers and the overall superiority of the guinea-pig maximization test in providing results similar to those obtained by human testing. A procedure is suggested for utilizing adjuvant and non-adjuvant test methods for characterizing compounds as of weak, moderate or strong sensitizing potential.

Allergens↗

Effects of genistein, an isoflavone isolated from Genista tridentata, on isolated guinea-pig ileum and guinea-pig ileal myenteric plexus.

The inhibitory action of the major constituent of Genista tridentata L. (Papilionaceae), 4',5,7-trihydroxyisoflavone (genistein), on contractions induced by agonists and electrical field stimulation of smooth muscle was analysed. Genistein inhibited twitches evoked by electrical-stimulation of strips of guinea-pig ileum with an IC50 value of 34 microM. Genistein (34 microM) inhibited contractions of the guinea-pig ileum by several agonists in a non-selective, antispasmodic action and had no effect on inhibition of 3H-ACh release from ileal myenteric plexus. Genistein (34 microM) produces an increase in cAMP levels of guinea-pig ileum which resulted in a smooth muscle relaxation which leads us to think that there must be a blockade of its phosphodiesterase.

Animals↗