Enteric Opportunistic Infection and the Impact of Antiretroviral Therapy among HIV/AIDS Patients from Tehran, Iran

Background: Opportunistic parasites have been identified as human pathogens, especially in immunodeficient patients. Microsporidian and coccidian infections cause chronic diarrhea as common clinical manifestation in HIV positive patients. In this study, the frequency of opportunistic infections, including microsporidian and coccidian infections, was evaluated in HIV/AIDS patients from Tehran and phylogenic analysis was performed for E. bieneusi isolates from these patients. Methods: One hundred and two stool samples were collected from confirmed HIV/AIDS patients, referred to Consult Center of Behavior Diseases, West Health Center, Iran University of Medical Sciences in Tehran, Iran. The samples were transferred to Research Center of Pediatric Infectious Diseases, Institute of Immunology and Infectious Diseases, Iran University of Medical Sciences from Jan 2016 to Dec 2016. After conventional formalin-ether concentration, aniline blue staining method and acid-fast staining technique were used for detection of microsporidian spores and Cyclospora oocysts. DNA was extracted and nested PCR was performed. Results: Two (1.96%) cases were found to be positive for intestinal microsporidia infection using aniline blue staining method and were confirmed as E. bieneusi by nested PCR. One patient was found with Cyclospora cayetanensis infection by acid-fast staining method and PCR. Giardia lamblia and Blastocystis hominis were detected as non-opportunistic parasites in 1/102 (0.98%) and 2/102 (1.96%) of the HIV positive patients, respectively. Conclusion: With respect to the use of antiretroviral therapy (ART) in HIV positive patients, we found a low frequency of infection.

Intestinal Protozoa in Domestic Cats (Carnivora: Felidae, Felis catus) in Northwestern Iran: A Cross-Sectional Study with Prev-alent of Microsporidian and Coccidian Parasites

Background: In this study, some microsporidial and coccidian parasites were isolated from 103 domestic cats in the Meshkin Shahr area, northwestern Iran during the Jun 2014 to Jun 2015, and their genera were identified using parasitological methods with emphasis on their zoonotic importance. Methods: One hundred and three fecal samples of domestic cats were collected and preserved in formalin (10%) and conserved in phosphate buffer saline solution, finally examined by microscopy after formalin-ether concentration and specific staining. Preservation in dichromate potassium (2.5%) was performed for all coccidian positive samples and then sporulated coccidian oocysts were investigated. Results: The detected parasites were Isospora spp. 6/103(5.8%). Microsporidian spores were identified in 46/103 (44.6%) of all samples post-stained by the aniline blue staining method. Conclusion: Microsporidial infections were more prevalent in domestic cats. Further studies are needed in the identification of microsporidial spores isolated from infected cats.

Two histological methods for recognition and study of cortical microinfarcts in thick sections

Cortical microinfarcts are the most widespread form of brain infarction but frequently remain undetected by standard neuroimaging protocols. Moreover, microinfarcts are only partially detectable in hematoxylin-eosin-stained (H and E) 4-10 µm paraffin sections at routine neuropathological examination. In this short report, we provide two staining protocols for visualizing cortical microinfarcts in 100-300 µm sections. For low-power microscopy, the first protocol combines aldehyde fuchsine staining for detection of lipofuscin granules and macrophages with Darrow red counterstaining for Nissl material. The second protocol combines collagen IV immunohistochemistry with aldehyde fuchsine/Darrow red or with erythrosin-phosphotungstic acid-aniline blue staining for detailed study of the capillary network. In the first protocol, microinfarcts are recognizable as radially-oriented funnel-like accumulations of aldehyde fuchsine-positive macrophages. The second protocol recognizes microinfarcts and alterations of the capillary network, at whose center accumulations of dead neurons and aldehyde fuchsine-positive macrophages cluster. In addition, the second protocol permits visualization of abnormalities within the capillary network associated with more recent microinfarcts. Both protocols can be useful for comparing MRI datasets with cortical microinfarcts in corresponding whole brain sections of 100-300 µm thickness.

Interspecific Hybridization Processes between Michelia yunnanensis and M. crassipes and Embryogenesis of the Heterozygote

The investigation of hybridization processes and embryogenesis of heterozygote is an effective approach for early hybrids’ identification, which could provide reliable information for successful crossbreeding. In this study, we reported the whole hybridization processes of the direct cross and reciprocal cross between Michelia yunnanensis Franch. ex Finet et Gagnep. and Michelia crassipes Law using fluorescence microscopy after aniline blue staining, with the pollen germination on stigmas, pollen tube growth in styles, and subsequent extension into the embryo sac as well as the double fertilization processes are documented in detail. The M. yunnanensis × M. crassipes combination displayed considerable cross-compatibility, and the heterozygote embryogenesis was further observed with an approach of modified cryosectioning technique. Besides, the whole formation processes of hybrid seeds from artificial pollination to maturation were successfully observed. However, in the reciprocal cross, we found incompatibility between pollen grains of M. yunnanensis and stigmas of M. crassipes for the reason of hysteretic identification, as well as the abnormal callose deposition which belongs to the prefertilization barriers. This is the first study in which the complete and clear hybridization processes in Michelia were reported. We inferred that unilateral incompatibility of M. crassipes detected in this study may also exist in some other Michelia species. In artificial hybridization practices, we suggest some special treatments for overcoming prefertilization barrier should be taken when treating M. crassipes as the maternal parent.

Female sporogenesis in the native Antarctic grass Deschampsia antarctica Desv.

The development of megasporocytes and the functional megaspore formation in Deschampsia antarctica were analyzed with the use of microscopic methods. A single archesporial cell was formed directly under the epidermis in the micropylar region of the ovule without producing a parietal cell. In successive stages of development, the meiocyte was transformed into a megaspore tetrad after meiosis. Most megaspores were arranged in a linear fashion, but some tetrads were T-shaped. Only one of the 60 analyzed ovules contained a cell in the direct proximity of the megasporocyte, which could be an aposporous initial. Most of the evaluated D. antarctica ovules featured monosporic embryo sacs of the Polygonum type. Approximately 30% of ovules contained numerous megaspores that were enlarged. The megaspores were located at chalazal and micropylar poles, and some ovules featured two megaspores – terminal and medial – in the chalazal region, or even three megaspores at the chalazal pole. In those cases, the micropylar megaspore was significantly smaller than the remaining megaspores, and it did not have the characteristic features of functional megaspores. Meiocytes and megaspores of D. antarctica contained polysaccharides that were detectable by PAS-reaction and aniline blue staining. Starch granules and cell walls of megasporocytes, megaspores and nucellar cells were PAS-positive. Fluorescent callose deposits were identified in the micropylar end of the megasporocytes. During meiosis and after its completion, thick callose deposits were also visible in the periclinal walls and in a small amount in the anticlinal walls of megaspores forming linear and T-shaped tetrads. Callose deposits fluorescence was not observed in the walls of the nucellar cells.

Identification of Chrysanthemum (Chrysanthemum morifolium) Self-Incompatibility

There has been a heated argument over self-incompatibilityof chrysanthemum (Chrysanthemum morifolium) among chrysanthemum breeders. In order to solve the argument, we investigated pistil receptivity, seed set, and compatible index of 24 chrysanthemum cultivars. It was found that the 24 cultivars averagely had 3.7–36.3 pollen grains germinating on stigmas at 24 hours after self-pollination through the fluorescence microscope using aniline blue staining method. However, only 10 of them produced self-pollinated seeds, and their seed sets and compatible indexes were 0.03–56.50% and 0.04–87.50, respectively. The cultivar “Q10-33-1” had the highest seed set (56.50%) and compatible index (87.50), but ten of its progeny had a wide range of separation in seed set (0–37.23%) and compatible index (0–68.65). The results indicated that most of chrysanthemum cultivars were self-incompatible, while a small proportion of cultivars were self-compatible. In addition, there is a comprehensive separation of self-incompatibility among progeny from the same self-pollinated self-compatible chrysanthemum cultivar. Therefore, it is better to emasculate inflorescences during chrysanthemum hybridization breeding when no information concerning its self-incompatibility characteristics is available. However, if it is self-incompatible and propagated by vegetative methods, it is unnecessary to carry out emasculation when it is used as a female plant during hybridization breeding.