Periodic acid-Schiff Positive Granules Research Articles

Anatomical, histological, and histochemical alterations in portio vaginalis uteri with an evaluation of the vaginal artery vascularity during the luteal and early pregnant stages in domestic buffaloes (Bubalus bubalis)

The portio-vaginalis uteri (PVU) and its mucus secretion have shown an essential role in conception. A significant endeavour to improve buffaloes' reproductive efficiency is the investigation of their basic reproductive pattern, which provides a reference for applications in breeding and pregnancy. The present study aimed to evaluate the anatomical and histological alterations in PVU regarding to the vaginal artery (VA) hemodynamic at luteal and early pregnant stages in buffalos. Egyptian live buffaloes (n = 16) and fresh genitals (n = 25) of mature buffalo were used. Different luteal and early pregnant stages were macroscopically identified with the shape and mucosal colouration with discharges of the PVU. Histological examination showed a significant difference in area % of alcian blue and periodic acid Schiff positive granules which considered an indication for presence of acidic and neutral mucins respectively in the epithelial cells of PVU mucosa which increased in pregnant stage than in other luteal stages. VA assessment demonstrated an increase in luminal diameter and thickness of tunica muscularis in pregnant stage than other stages (P < 0.05). Middle uterine (MUA) and VA arteries peak velocity point (PSV mm/sec) were elevated (P < 0.05) in pregnant stage, with a marked reduction in both resistance and pulsatility indices (RI and PI), and ratio of systolic /diastolic (S/D). Positive correlation was detected between VA. PSV and, MUA. PSV (r = 0.87), but a negative relation was detected with VA. S/D (r = -0.77), VA.PI (r = -0.89), VA. RI (r = -0.97), MUA. S/D (r = -0.94), MUA. PI (r = -0.85), and MUA. RI (r = -0.88). Doppler indices were negatively corrected with the VA. PSV (r = -0.68). It was concluded that there was a significant alterations in histological features of the cervical PVU at different physiological stages (luteal and early pregnant) in buffalos in relation to the MUA and VA hemodynamic pattern and that hypotheses can be established regarding the female cyclicity that affected by both arteries hemodynamics change.

Effects of citronella oil (Cymbopogon winterianus Jowitt ex Bor) on Spodoptera frugiperda (J. E. Smith) midgut and fat body

The armyworm, Spodoptera frugiperda, is the principal pest of corn in Brazil. Control is achieved primarily by synthetic insecticides, which cause problems for the agro-ecosystem. Alternative methods of control are under investigation and citronella (Cymbopogon winterianus) essential oil appears to be a promising agent. We investigated the effects of citronella oil using histological, histochemical and immunohistochemical methods. The midgut of larvae treated with citronella exhibited altered epithelium including cytoplasmic protrusions, columnar cell extrusion, pyknotic nuclei, and increased periodic acid-Schiff positive granules. Regenerative cells in the epithelium of the midgut increased in number, which facilitated subsequent regeneration of this tissue. After exposure to citronella, trophocytes, the principal cell type of the fat body, possessed enlarged vacuoles and mitotic bodies, and contained reduced amounts of glycogen, lipid, and protein. Citronella oil caused morphological changes of the midgut and reduction of stored resources in the fat body, which may adversely affect insect reproduction and survival.

P.215 - Sensitivity and specificity of the PAS positive lymphocyte vacuoles in the diagnostic approach to late onset Pompe disease

The autosomal recessive Pompe disease, which is caused by deficiency of the lysosomal enzyme acid α-glucosidase (GAA), manifests in the very severe classic-infantile and in milder course late-onset forms (LOPD). Since an effective Enzyme Replacement Therapy is available, a cheap, rapid, sensitive and specific screening test is crucial to early identify homozygous affected and heterozygous carriers. Aim of this study is to verify sensitivity and specificity of the quantitative assessment of PAS-positive granules in lymphocytes to recognize homozygous and heterozygous GAA mutated individuals. Number of lymphocytes containing PAS positive granules as well as the number of granules in each lymphocyte were assessed on periodic acid- Schiff (PAS) stained blood smears from 72 adult patients with persistent hyperckemia and/or muscle weakness, 7 patients with genetically confirmed LOPD, and 13 patients with heterozygous GAA mutations. Percentage of affected lymphocytes and mean number of granula per lymphocyte were calculated for all patients. Accuracy of the test was evaluated by ROC curve. Eleven out of 72 patients with hyperckemia and/or muscular weakness showed more than 4 PAS-positive lymphocytes (6 new diagnosis of LOPD, 4 Glycogen storage myopathies different from Pompe, and 1 caveolinopathy). All the 22 LOPD patients and, surprisingly, also all the 13 GAA-mutation carriers presented number of PAS-positive granules higher than in other myopathies. PAS-positive lymphocytes analysis disclosed a 97%sensitivity and a 94% specificity in recognizing homozygous and heterozygous GAA mutations. Quantification of PAS-positive lymphocytes in blood smears should be routinely used as first level screening test for Pompe disease.

Molecular Genetic Analysis of Anaplastic Pleomorphic Xanthoastrocytoma

A case of pleomorphic xanthoastrocytoma in a 10-year-old Malay boy is reported. The patient presented with headache and epilepsy. On computed tomography, a ring-enhancing low-density lesion was observed in the left fronto-temporal area. During surgery, a cystic tumour containing serous fluid was found and almost totally removed. Histologically, the tumour exhibited marked pleomorphism of oval and spindle-shaped cells intermixed with uni- and multinucleated giant cells, and xanthomatous cells with foamy cytoplasm. The tumour displayed pericellular reticulin and periodic acid-Schiff positive granules. Focally, six mitotic characters per 10 high-power fields were seen, and necrosis was confined only to the inner lining of the cyst. Mutational analysis showed that a frameshift mutation (a 4-bp deletion) in the p53 gene had occurred in codons 273 and 274 of exon 8. No mutation was detected in the p16 gene. No allelic loss and/or loss of heterozygosity were observed on chromosome 10 using microsatellite marker D105532. The patient was treated with postoperative radiotherapy because of histological anaplasia and the presence of residual tumour. The patient showed marked neurological recovery after a follow-up period of 2 years.

Immunohistochemical, conventional and immunoelectron microscopical characteristics of periodic acid-Schiff-positive granules in the mouse brain.

Periodic acid-Schiff-positive granules (PGs) appear in the mouse brains in relation to advancing age. The exact location and pathophysiological significance of PGs, however, are not fully understood. The incidence, staining properties, and topographical distributions of PGs in the brains of 17 AKR mice ranging in age from 7 to 18 months were examined histochemically and immunohistochemically using antibody KM279 raised against a polyglucosan. In addition, to define the precise site of PG formation, we investigated the brains of 4 AKR mice of 24 months of age using conventional and immunoelectron microscopy. PGs were seen in all mice examined and the levels were increased with age. The PGs were located predominantly in the hippocampus and, to a lesser extent, in the cerebellum and olfactory bulb. Immunohistochemically, PGs in the hippocampus and cerebellum were labeled uniformly with KM279. On immunoelectron microscopy with this monoclonal antibody, the fibrillar or membranous structures corresponding to PGs seen using light microscopy were labeled specifically with gold particles. With conventional electron microscopy, fibrillar or membranous structures were seen along with synaptic vesicles and dense-core granules. Moreover, around the cells containing PGs, a few synaptic junctions with neighboring cells were observed, indicating that the cells contributing to formation of PGs were neuronal cells. The positive immunoreactivity of AKR mouse PGs for the antibody KM279 suggests that the PGs and similar structures in other species may share a common antigenicity. Thus, it is assumed that PGs in AKR mice might result from some abnormalities in glucose metabolism.

Systemic Mastocytosis in a Goat

Systemic mastocytosis was diagnosed in a 4-year-old, female Nubian goat. Clinically, the animal was depressed and had severe macrocytic hypochromic anemia and leukopenia. Postmortem examination revealed neoplastic mast cells invading the heart, lung, liver, spleen, lymph nodes, and bone marrow. Eosinophils were frequently admixed with infiltrating mast cells in all organs. Using routine light microscopy, histochemistry, and transmission electron microscopy, metachromatic and periodic acid-Schiff-positive granules were identified within the cytoplasm of neoplastic mast cells. Erythrophagocytosis was observed in some neoplastic cells, although its contribution to the anemia was not clear. This report represents the first description of mast cell neoplasia in the goat.

A monoclonal antibody, 4H12, recognizes a surface antigen found on granulated metrial gland cells in the murine decidua

A monoclonal antibody (MAb), designated 4H12, was selected for reactivity to a surface antigen on PYS-2 teratocarcinoma cells. 4H12 was the product of a fusion of lymphoid cells of a non-immunized pregnant C57BL/6 mouse to NS-1 myeloma cells. Initial studies utilizing immunohistochemistry revealed that MAb 4H12 bound to an antigen found on cells in the decidua basalis of 7-, 8- and 10-day pregnant mice. Antigen-positive cells of 11–19-day pregnant mice were also found predominantly in the decidua. A few antigen-positive cells were found in the labyrinth of the placenta and up against Reichert's membrane. Antigen-positive cells were morphologically and spatially distinct, oval to round with large periodic acid Schiff positive granules. Indirect immunofluorescent (IIF) labeling of decidual cultures showed antigen on the surface of cells that were small, oval to round and adherent. The antigen recognized by MAb 4H12 was removed from tissue sections with trypsin and protease and therefore is suggested to be a protein. We conclude that MAb 4H12 recognizes a surface antigen found on cells historically described as granulated metrial gland (GMG) cells. This MAb should greatly facilitate the further analysis of the life history and function of GMG cells during pregnancy.

Inheritance of the Chediak-Higashi syndrome in cats

The phenotypes with respect to the Chediak-Higashi syndrome (CHS) of 245 cats from 67 matings were analyzed. It was determined that the gene for feline CHS, like that in other species with CHS, is inherited in an autosomal recessive manner with complete penetrance. We propose the symbol ch for the gene for CHS in cats. Additional studies revealed enlarged periodic acid-Schiff positive granules in renal tubule cells in CHS cats, which provides further evidence of the homologous nature of CHS among the various species in which it occurs.

The blood leukocytes of Bufo alvarius: a light, phase-contrast, and histochemical study

Blood leukocytes of Bufo alvarius were studied by light and phase-contrast microscopy and histochemical techniques for the localization of glycogen and several hydrolytic enzymes, i.e., acid and alkaline phosphatases, nonspecific esterase, beta-glucuronidase, aryl-sulfatase, and myeloperoxidase (peroxidase). Neutrophils were the only leukocytes to demonstrate alkaline phosphatase activity, while beta-glucuronidase and aryl-sulfatase were not observed in any leukocytes. Periodic acid – Schiff (PAS) positive granules and granules containing hydrolytic enzymes occurred in varying amounts in leukocytes. In eosinophils, most glycogen was associated with smaller granules, while the larger refractile granules were PAS negative. Small lymphocytes were myeloperoxidase (peroxidase) negative. The present study agrees with previous investigations in mammals which indicate that specific granules in granulocytes may be PAS positive as well as contain one or more hydrolytic enzymes. In small lymphocytes of B. alvarius, PAS positive and acid phosphatase positive granules correspond to neutral red granules seen in supravital films. Furthermore, the appearance and histochemical reactivity of acid phosphatase granules in mature neutrophils, metamyelocytes, and late myelocytes correspond closely with the appearance and number of specific neutrophilic granules seen in Wright–Giemsa preparations and with PAS positive granules.

Isospora felis: Development in Cultured Cells with Some Cytological Observations

Isopora felis undelwent development in monolayer cultures of human cells (embryonic intestine, esophageal epithelium, amnion, lung, and epithelioid carcinoma of the cervix) as well as cells from embryonic bovine trachea, canine kidney, and chick kidney. Pairs of daughter organisms developed by endodyogeny and possibly schizogony. Special stains were used to study the cytological changes that occurred during development. Disappearance of red-stained material from the refractile body in organisms stained with Whipf's polychrome stain suggested utilization or conversion of this material by the sporozoite or change in its ionic nature; its reappearance in mature daughter organisms suggested replacement, reconversion, or return to the previous ionic composition for the next generation. Carbohydrate, identified as periodic acid-Schiff positive granules, also appeared to be utilized by the sporozoites and replaced in the mature daughter organisms. Nucleic acids were identified in the cytoplasm and the nucleus of sporozoites and daughter organisms by staining with gallocyaninchromalum after RNase or DNase digestion. The distribution of nucleic acids in these two stages was similar. Several species of Isospora have developed in cultured cells (Turner and Box, 1970; Fayer, 1972; Fayer and Kocan, 1972; Fayer and Mahrt, 1972). However, previous attempts to obtain development of I. felis have been unsuccessful. Sheffield and Melton (1970) inoculated monkey kidney cell cultures with mixture of Toxoplasma gondii, I. felis, and I. rivolta and found that a typical T. gondii infection resulted. They also inoculated similar cultures with mixture of I. felis and I. rivolta sporozoites and found no intracellular organisms. Shibalova and Petrenko (1972) inoculated cultures of chick fibroblasts, quail fibroblasts, monkey kidney, human embryo, rabbit, sheep embryo, green marmoset, and human amniotic cells with sporozoites of I. bigemina, I. felis, and I. revolta. Each of the three species entered cells, but only I. bigemina appeared to have undergone development. The present study represents further attempt to obtain development of I. felis by employing some of the same cell types used by previous investigators as well as several different cell types. This study also represents the first investigation of cytochemical changes of an isosporan in cell culture. Received for publication 5 July 1973. *Animal Parasitology Institute, Beltsville Agricultural Research Center, ARS, Beltsville, Maryland 20705. MATERIALS AND METHODS

Malacoplakia

A case of malacoplakia of the bladder and intestinal tract is reported in which bacilliform organisms are demonstrated within the malacoplakic macrophages by light and electron microscopy. On electron microscopy, two types of cytoplasmic inclusions are found, those corresponding to the periodic acid Schiff-positive granules which are phagolysosomes, and the Michaelis-Gutmann bodies. Bacteria, probably coliforms, in various stages of degradation, are found within phagocytic vacuoles and phagolysosomes, sometimes in close apposition to phospholipid membranes. The Michaelis-Gutmann bodies develop within the phagolysosomes. It is postulated that the following sequence of events occurs in the pathogenesis of Michaelis-Gutmann bodies. Bacteria are phagocytosed by the malacoplakic macrophages, incorporated into phagolysosomes, and killed, but are incompletely digested. They persist as dense amorphous aggregates and phospholipid membranes which later become encrusted with calcium phosphate crystals to form the laminated Michaelis-Gutmann bodies. Why the macrophages respond in this unusual fashion to a common infectious organism is not clear. It may be due to an immunological abnormality affecting intracellular digestion. Other factors, such as infection with an unusual strain of coliform or intrinsically abnormal macrophages, are discussed. The world literature of intestinal malacoplakia is reviewed and the aggressive nature of intestinal involvement is stressed.