Cryptosporidium wrairi sp. n. from the Guinea Pig Cavia porcellus, with an Emendation of the Genus

SYNOPSIS. Cryptosporidium wrairi sp. n. is described from the laboratory guinea pig Cavia porcellus. The life cycle is given insofar as it is known. Two schizogonous generations are described; the 1st with 8 merozoites, the 2nd with 4 merozoites. The latter generation was previously referred to as the sporulated oocyst, but evidence is presented to show that it is a schizont. Micro- and macrogametogony are also described. No oocysts were found. Cross-transmission to mice, chickens, turkeys and rabbits was unsuccessful. The generic character of oocysts with 4 naked sporozoites is discarded and the presence of endogenous stages in the striated border of epithelial cells is used as the emended generic character. A listing of valid and non-valid species is given.     

Comparison of the Host Ranges and Antigenicity of Cryptosporidium parvum and Cryptosporidium wrairi from Guinea Pigs

ABSTRACT. Oocysts of a Cryptosporidium isolate from guinea pigs were not infectious for adult mice, but were infectious for two of three newborn calves and for suckling mice. However, oocysts isolated from calves or mice infected with guinea pig Cryptosporidium were not infectious for guinea pigs. Four isolates of C. parvum from calves were incapable of infecting weanling guinea pigs. Microscopic examination of tissue from the colon and cecum of suckling guinea pigs inoculated with C. parvum revealed sparse infection of some pups. These host range studies and previously described differences in ¹²⁵I-labeled oocyst surface protein profiles between Cryptosporidium sp. from guinea pigs and C. parvum suggest they are distinct species. We propose the name Cryptosporidium wrairi be retained. Studies with monoclonal antibodies indicate that C. wrairi and C. parvum are antigenically related.     

Comparison of the host ranges and antigenicity of Cryptosporidium parvum and Cryptosporidium wrairi from guinea pigs.

Oocysts of a Cryptosporidium isolate from guinea pigs were not infectious for adult mice, but were infectious for two of three newborn calves and for suckling mice. However, oocysts isolated from calves or mice infected with guinea pig Cryptosporidium were not infectious for guinea pigs. Four isolates of C. parvum from calves were incapable of infecting weanling guinea pigs. Microscopic examination of tissue from the colon and cecum of suckling guinea pigs inoculated with C. parvum revealed sparse infection of some pups. These host range studies and previously described differences in 125I-labeled oocyst surface protein profiles between Cryptosporidium sp. from guinea pigs and C. parvum suggest they are distinct species. We propose the name Cryptosporidium wrairi be retained. Studies with monoclonal antibodies indicate that C. wrairi and C. parvum are antigenically related.     

Isolation of Neuropeptide-Containing Vesicles from the Guinea Pig Ileum

Three distinct vesicle fractions enriched 40-60 times in the neuropeptides substance P, somatostatin, and vasoactive intestinal peptide (VIP) were prepared from the myenteric plexus of guinea pig ileum by density gradient centrifugation in a small zonal rotor. Mean densities (in g X ml-1) and diameters (in nm) of the three classes of vesicles were: substance P, 1.123, 65; somatostatin, 1.138, 37; VIP, 1.148, 110; standard deviations were about 5%. These peaks were distinct from the peak of acetylcholine-containing vesicles of density 1.066 g X ml-1 and diameter 61 nm. When a relatively mild method of homogenization was used a second peak of acetylcholine appeared in the same region of the gradient as VIP and the VIP was larger. This may represent a class of vesicles containing both acetylcholine and VIP, though cosedimentation of two classes of vesicles of almost the same density and similar fragility, one containing VIP and the other acetylcholine, cannot be entirely excluded on present evidence.     

Effect of hyperimmune bovine colostrum raised against Cryptosporidium parvum on infection of guinea pigs by Cryptosporidium wrairi.

Oocysts shedding was markedly reduced in guinea pigs inoculated intraintestinally with Cryptosporidium wrairi sporozoites that had been incubated with hyperimmune bovine colostrum raised to C. parvum when compared with shedding in guinea pigs inoculated with sporozoites incubated in either non-immune bovine colostrum or buffered saline. However oocyst shedding was apparently not reduced in guinea pigs inoculated by gavage with oocysts of C. wrairi and subsequently treated twice daily per os with hyperimmune bovine colostrum. Similarly, oocyst shedding was apparently not reduced by oral treatment with hyperimmune bovine colostrum when treatment was begun simultaneously with inoculation of C. wrairi oocysts.     

Quantitation of Cholinergic Synaptosomes from Guinea Pig Brain

An antiserum raised to nerve terminal sacs derived from the electric organ and Torpedo marmorata was used to lyse guinea pig brain synaptosomes in the presence of complement. From the release of the cytoplasmic enzymes choline acetyltransferase, lactate dehydrogenase, tyrosine hydroxylase and glutamate decarboxylase it appears that the antiserum binds specifically to cholinergic terminals. The amount of lactate dehydrogenase released was used to estimate the proportion of cholinergic nerve terminals in different synaptosome preparations.     

A Synaptosomal Preparation from the Guinea Pig Ileum Myenteric Plexus

Abstract: Our interest in investigating the presynaptic modulation of acetylcholine release led to the development of a synaptosomal preparation from the guinea pig ileum myenteric plexus-longitudinal muscle. A crude synaptosomal fraction (P₂) was obtained by homogenization and differential centrifugation. The preparation exhibited a specific uptake system for choline and for nor-adrenaline (NA), but not for 5-hydroxytryptamine (5-HT). Synaptosomes were isolated from this P₂ fraction by an isoosmotic density gradient prepared from sucrose and metrizamide. The resultant synaptosomal fraction was enriched about sevenfold in both choline uptake and in choline acetyltransferase (ChAT). Choline was transported by a high-affinity system with a K_m of 6.5 × 10⁻⁷ M and a V_max of 41 pmol/mg protein/min. Electron microscopy confirmed the synaptosomal nature of the gradient fraction. Some synaptosomal profiles contained only small, translucent vesicles whereas others also contained large (approx. 100 nm diameter) electron-opaque vesicles. The crude synaptosomal fraction synthesized acetylcholine (ACh) from exogenous choline and it released the synthesized ACh in a calcium-dependent manner.     

Properties of a CDP-Diglyceride Hydrolase from Guinea Pig Brain

Enzymatic hydrolysis of the pyrophosphate bond of CDP-diglyceride (CDP-DG), previously shown to occur in bacteria, is demonstrable in mammalian tissues. Activity was enriched in a lysosomal fraction obtained from guinea pig cerebral cortex and was purified 92-fold relative to the homogenate by a combination of XM-300 ultrafiltration and DEAE-cellulose column chromatography. When incubated with CDP-dipalmitin, the purified enzyme produced stoichiometric amounts of CMP and phosphatidate. dCDP-DG served as a substrate, while ADP-DG was an inhibitor, as were 5'-AMP and 5'-dAMP. CDP-DG hydrolysis was not affected by the presence of excess amounts of CDP-choline, CDP-glycerol, sodium pyrophosphate, or cyclic 3',5'-AMP.     

Acetylcholine Releases ATP from Varicosities Isolated from Guinea Pig Myenteric Plexus

The effects of cholinergic agonists and antagonists on the release of ATP from isolated myenteric varicosities were studied using a firefly luciferin-luciferase technique. In a previous study, acetylcholine and nicotine released ATP from isolated myenteric varicosities, whereas the muscarinic agonist bethanechol did not. In the present study, release of ATP by acetylcholine was shown to be Ca2+ dependent. d-Tubocurarine competitively antagonized the release of ATP by either acetylcholine or nicotine. Maximal release of ATP by acetylcholine (10(-3) M) was approximately 24% that observed with the depolarizing drug veratridine (5 X 10(-5) M), suggesting either that not all of the varicosities capable of releasing ATP possess nicotinic receptors or that acetylcholine does not depolarize the varicosities to the degree that veratridine does. Tetrodotoxin slightly but significantly reduced ATP release induced by acetylcholine or nicotine, indicating some involvement of Na+ channels in the release process. Finally, 6-hydroxydopamine pretreatment produced a 48% reduction in the acetylcholine-evoked release of ATP, suggesting that much, but possibly not all, of the ATP release occurs from noradrenergic varicosities present in the preparation.     

Sequence of Guinea Pig Myelin Basic Protein

This paper proposes a tentative amino acid sequence of guinea pig myelin basic protein obtained by comparison of peptide fragments of the guinea pig and bovine proteins. Analyses of the tryptic peptides confirmed the known sequence differences in the NH2-terminal half of the molecule and showed that in the COOH-terminal half of the guinea pig protein Ser131 was missing, Ala136 - His137 was deleted, Leu140 was replaced by Phe, and an extra Ala was inserted somewhere within sequence 142-151 (tryptic peptide T23 ). Sequence determination of guinea pig tryptic peptides corresponding to residues 130-134 ( T20 ), 135-138 ( T21 ), and 142-151 ( T23 ) of the bovine protein confirmed the above sequence changes and placed the extra Ala between Gly142 and His143 . The sequence of the region corresponding to bovine residues 130-143 is thus Ala-Asp-Tyr-Lys-Ser-Lys-Gly-Phe-Lys-Gly-Ala-His. No species differences were observed in the amino acid compositions of the remaining tryptic peptides obtained from the COOH-terminal half of the molecule. Based upon these results, the guinea pig basic protein contains 167 amino acid residues and has a molecular weight of 18,256.     

Characterization of Bromodeoxyuridine-Induced Endogenous Guinea Pig Virus

An endogenous virus (GPV) was induced after 5-bromodeoxyuridine treatment of cultured guinea pig cells. Compared to Gross murine leukemia virus (G-MuLV) GPV has a reproducibly heterogenous density of about 1.16 to 1.18 g/ml. The virion-associated RNA is slightly larger than that in G-MuLV. Sodium dodecyl sulfate-polyacrylamide gel electrophoretic analysis of dissociated GPV resolved five major structural proteins: I (molecular weight 70,000), II (molecular weight 36,000), III (molecular weight 24,000), IV (molecular weight 18,000), and V (molecular weight 16,000) which are similar to but distinct from G-MuLV proteins. Proteins I and II were demonstrated to be glycoproteins by incorporation of [(3)H]glucosamine. GPV and G-MuLV did not have any appreciable genetic homology or any common group-specific antigens when analyzed by immunodiffusion, radioimmunoassay, and indirect immunofluorescence. Morphogenesis of GPV also differed from that of a typical type C oncornavirus and proceeded via two pathways: (i) a majority of virus particles were formed in cytoplasmic vacuoles and were released after cellular disruption; and (ii) a minor population of particles were assembled in the cytoplasmic matrix and then migrated to the plasma membrane where they budded into the extracellular space. To date, GPV has been unable to initiate or maintain a productive replication in any cell line tested.     

豚鼠生长激素受体cDNA的克隆

报道了豚鼠肝生长激素受体（ＧＨＲ）的ｃＲＮＡ克隆和编码区序列。它由１８９９ｂｐ组成,编码６１０个氨基酸。此外,还报道豚鼠ＧＨＲ的结构特征和同源性比较的结果。     

Formation of Cyclohexylamine and Cyclohexanone from Cyclamate by Microorganisms Isolated from the Feces of Guinea Pig

The assimilation of sodium cyclamate (CHS-Na) by microorganisms was studied. Fifteen strains of cyclamate-assimilating bacteria were isolated from the feces of guinea pig excreting cyclohexylamine (CHA) in urine. The majority of the strain isolated seems to belong to the genera Pseudomonas and Corynebacterium. All strains were able to assimilate CHS-Na as a sole source of carbon and nitrogen, and accumulated clearly CHA in the culture medium. It was confirmed with the cell-free extract that the strains possessed the enzyme system which formed cyclohexanone (CHnone) from CHS-Na via CHA. It seems that the desulfation of CHS-Na to CHA is catalyzed by hydrolase, and that the deamination of CHA to CHnone is catalyzed by amine oxidase depending on oxygen.     

Preparation of Neurofilament Protein from Guinea Pig Peripheral Nerve and Spinal Cord

Abstract: A simple and rapid method for preparation of enriched neurofilament protein from mammalian peripheral nerve or spinal cord is described. Tissue extracts from guinea pig nerve or spinal cord are fractionated by ammonium sulfate fractionation, chromatography on Sepharose 4B, and precipitation with ethanol. Molecular exclusion chromatography on Sepharose 4B, in which the neurofilament protein elutes quantitatively in the exclusion volume of the column, with little contamination by other proteins, is found to be a highly effective purification step. The protein is found to precipitate in ammonium sulfate fractions over a wide range of salt concentration, from 20 to 80% saturation. It is found to be quantitatively precipitated in 40% v/v ethanol-water. The preparative method described yields 0.25 mg of neurofilament protein per gram of nerve or spinal cord, with a purity of approximately 50%. The three principal neurofilament polypeptides, which have molecular weights by SDS-polyacrylamide gel electrophoresis of 200K, 145K, and 68K, are found to be present in the preparation in a molar ratio of 1:2:6. A variant form of neurofilament protein occurring in approximately 20% of Hartley strain guinea pigs is described, which has the polypeptide composition: 200K, 192K, 145K, 68K.     

Putative Cholinergic-Specific Gangliosides in Guinea Pig Forebrain

The nature of the cholinergic-specific antigen Chol-1 recognized by an antiserum raised against Torpedo cholinergic electromotor synaptosomal plasma membranes was investigated in guinea pig forebrain to establish whether it has a gangliosidic nature in guinea pig as in Torpedo. Gangliosides extracted from guinea pig forebrain and extensively purified to eliminate peptide contaminants were effective in inhibiting the selective lysis of the cholinergic subpopulation of cortical synaptosomes induced by the antiserum. Neuraminidase, protease, alkali, and heat treatment did not impair the inhibitory activity of gangliosides. Whereas the antiserum recognized many gangliosides from Torpedo electric organ, the immunostaining of guinea pig forebrain gangliosides separated on TLC showed only two immunopositive bands migrating close to GT1b and GQ. After affinity purification on Torpedo electric organ gangliosides the activity of the antiserum in inducing complement-mediated lysis was increased and it still recognized the two ganglioside bands on TLC. These results strongly suggest the existence of two polysialogangliosides bearing antigenic determinants specific for the cholinergic neurons.