Septata intestinalis frequently isolated from stool of AIDS patients with a new cultivation method

SummaryTwo species of microsporidia, Enterocytozoon bieneusi and Septata intestinalis have been reported as intestinal parasites of AIDS patients. In attempts to establish E. bieneusi in vitro, spores were concentrated from stool samples from 4 AIDS patients with biopsy-proven E. bieneusi infections. After sterilization of the concentrate in antibiotic solution, the spores were added to monolayers of RK13 cells grown on the membranes of Transwells. Cultures were established from 7 stool samples from the 4 patients but in every case the species established was S. intestinalis not E. bieneusi. On retrospective examination of the stools, a very small number of spores of a size comparable to that of S. intestinalis was found but this species was not detected in biopsies. Typical septate vacuoles containing Type I tubules were observed in vitro but in contrast to the original description, meronts were intravacuolar and sporogony was mainly disporoblastic. The cultivation system, used for the first time for microsporidia, revealed the presence of unsuspected S. intestinalis infections and indicates that this species may be much more common than hitherto suspected. S. intestinalis has not previously been cultured.

Download Full-text

Strategies to Obtain Encapsulation and Controlled Release of Pentamidine in Mesoporous Silica Nanoparticles

Pharmaceutics ◽

10.3390/pharmaceutics10040195 ◽

2018 ◽

Vol 10 (4) ◽

pp. 195 ◽

Cited By ~ 8

Author(s):

Enrico Peretti ◽

Ivana Miletto ◽

Barbara Stella ◽

Flavio Rocco ◽

Gloria Berlier ◽

...

Keyword(s):

Controlled Release ◽

Mesoporous Silica ◽

Silica Nanoparticles ◽

Mesoporous Silica Nanoparticles ◽

Type I ◽

Pentamidine Isethionate ◽

Kinetic Release ◽

First Time ◽

Severe Limit

Pentamidine (PTM), an antiprotozoal agent used in clinics as pentamidine isethionate salt (PTM-S), recently showed high potential also for the treatment of cancer and myotonic dystrophy type I. However, a severe limit to the systemic administration of PTM is represented by its nephrotoxicity, leading to the need for a system able to achieve a controlled release of the drug. In this study, mesoporous silica nanoparticles (MSNs) were employed for the first time to encapsulate PTM. PTM-S was first used for loading experiments into bare (MSN-OH) and aminopropyl, cyanopropyl and carboxypropyl-functionalized MSNs (MSN-NH2, MSN-CN and MSN-COOH respectively) but it was not adequately loaded in any MSNs. The free base of PTM (PTM-B) was then obtained from PTM-S and successfully loaded into MSNs. Specifically, MSN-COOH exhibited the highest loading capacity. In vitro evaluation of PTM-B kinetic release from the different MSNs was carried out. An influence of the functional groups in slowing the release of the drug, when compared to bare MSNs was observed. Altogether, these results demonstrate that MSN-COOH could be a promising system to achieve a controlled release of PTM.

Download Full-text

Intestinal parasites, including Cryptosporidium species, in Iraqi patients with sickle-cell anaemia

Eastern Mediterranean Health Journal ◽

10.26719/2002.8.2-3.345 ◽

2002 ◽

Vol 8 (2-3) ◽

pp. 345-349

Author(s):

N. K. Mahdi ◽

N. H. Ali

Keyword(s):

Sickle Cell ◽

Sickle Cell Anaemia ◽

Intestinal Parasites ◽

Cryptosporidium Species ◽

Parasitic Infections ◽

Blastocystis Hominis ◽

Significant Difference ◽

Stool Samples ◽

First Time ◽

Apparently Healthy

Stool samples were obtained from individuals admitted to three hospitals in Basra during November 1997-May 1998. Of 40 patients with sickle-cell anaemia, 25 [62.5%] had parasitic infections. In the apparently healthy comparison group, 26 of 175 individuals [14.8%] had intestinal parasitic infections, a statistically significant difference. The most common intestinal parasites isolated in the sickle-cell patients were Blastocystis hominis [36%] and Giardia lamblia [28%]. The isolation rate of Cryptosporidium species in sickle-cell patients [5%] was not significantly different from that in apparently healthy individuals [1.14%]. We report for the first time the isolation of Isospora belli from a sickle-cell patient in Iraq and the Mediterranean region.

Download Full-text

Molecular diversity of Giardia duodenalis, Cryptosporidium spp. and Blastocystis sp. in asymptomatic school children in Leganés, Madrid (Spain)

10.21203/rs.2.19429/v1 ◽

2019 ◽

Author(s):

Aly Salimo Muadica ◽

Pamela C Köster ◽

Alejandro Dashti ◽

Begoña Bailo ◽

Marta Heernández de Mingo ◽

...

Keyword(s):

Molecular Diversity ◽

Parasite Species ◽

Intestinal Parasites ◽

Giardia Duodenalis ◽

Small Subunit ◽

Enterocytozoon Bieneusi ◽

Published Data ◽

Cryptosporidium Hominis ◽

Additional Sequence ◽

Stool Samples

Abstract Background: The protozoa Giardia duodenalis, Cryptosporidium spp., the stramenopile Blastocystis sp. and the microsporidia Enterocytozoon bieneusi are among the most frequent diarrheal pathogens affecting humans globally. This molecular epidemiological study assesses the frequency and molecular diversity of these intestinal parasites in schoolchildren in central Spain, complementing previously published data on risk and protective factors associated with parasite infection (Reh et al., Euro Surveill. 2019;24).Methods: Stool samples were collected from voluntary asymptomatic schoolchildren (4‒14 years) and their siblings (1‒16 years) attending 12 primary and secondary schools in Leganés (Madrid). Initial detection of pathogens was conducted by PCR-based methods targeting the small subunit (ssu) ribosomal RNA or the internal transcribed spacer (ITS) genes of these parasite species. Genotyping of G. duodenalis-positive samples was carried out by PCR and Sanger sequencing of appropriate markers including the glutamate dehydrogenase (gdh), the ß-giardin (bg), and triose phosphate isomerase (tpi) loci. For C. hominis/C. parvum-positive samples the 60-kDa glycoprotein (gp60) locus was used.Results: A total of 1,512 stool samples were analysed. Giardia duodenalis was the most prevalent pathogen (17.4%, 95% CI: 15.5‒19.4%), followed by Blastocystis sp. (13.0%, 95% CI: 11.4‒14.8%), and Cryptosporidium spp. (0.9%, 95% CI: 0.5%‒1.5%). Enterocytozoon bieneusi was not detected. Sequence analyses of the 24 G. duodenalis isolates genotyped at the gdh, bg, and/or tpi loci revealed the presence of sub-assemblages AII (16.6%, 4/24) and BIV (79.2%, 19/24). An additional sequence (4.2%, 1/24) represented an ambiguous BIII/BIV result. Analyses of the 14 Cryptosporidium sequences generated at the ssu rRNA allowed the identification of C. hominis (71.4%; 10/14) and C. parvum (21.4%; 3/14). An additional sequence (7.2%, 1/14) was only identified at the genus level. A total of 162 Blastocystis sp. isolates were successfully genotyped, revealing the presence of five subtypes including ST1 (22.8%; 37/162), ST2 (36.4%; 59/162), ST3 (21.6%; 35/162), ST4 (18.6%; 30/162), and ST8 (0.6%; 1/162).Conclusions: Giardia duodenalis sub-assemblage BIV, Cryptosporidium hominis and Blastocystis ST2 were the genetic variants of these parasite species more prevalent in the asymptomatic schoolchildren population investigated. These findings are very similar to those previously reported in clinical, symptomatic populations in Spain. Enterocytozoon bieneusi was absent in apparently healthy schoolchildren.

Download Full-text

Structures of Arenaviral Nucleoproteins with Triphosphate dsRNA Reveal a Unique Mechanism of Immune Suppression

Journal of Biological Chemistry ◽

10.1074/jbc.m112.420521 ◽

2013 ◽

Vol 288 (23) ◽

pp. 16949-16959 ◽

Cited By ~ 48

Author(s):

Xue Jiang ◽

Qinfeng Huang ◽

Wenjian Wang ◽

Haohao Dong ◽

Hinh Ly ◽

...

Keyword(s):

Immune Suppression ◽

Lassa Fever ◽

Lassa Virus ◽

Type I ◽

Molecular Pattern ◽

Catalytically Active ◽

Suppression Mechanism ◽

First Time ◽

Unique Mechanism

A hallmark of severe Lassa fever is the generalized immune suppression, the mechanism of which is poorly understood. Lassa virus (LASV) nucleoprotein (NP) is the only known 3′-5′ exoribonuclease that can suppress type I interferon (IFN) production possibly by degrading immune-stimulatory RNAs. How this unique enzymatic activity of LASV NP recognizes and processes RNA substrates is unknown. We provide an atomic view of a catalytically active exoribonuclease domain of LASV NP (LASV NP-C) in the process of degrading a 5′ triphosphate double-stranded (ds) RNA substrate, a typical pathogen-associated molecular pattern molecule, to induce type I IFN production. Additionally, we provide for the first time a high-resolution crystal structure of an active exoribonuclease domain of Tacaribe arenavirus (TCRV) NP. Coupled with the in vitro enzymatic and cell-based interferon suppression assays, these structural analyses strongly support a unified model of an exoribonuclease-dependent IFN suppression mechanism shared by all known arenaviruses. New knowledge learned from these studies should aid the development of therapeutics against pathogenic arenaviruses that can infect hundreds of thousands of individuals and kill thousands annually.

Download Full-text

Collagen fibrillogenesis in vitro: interaction of types I and V collagen

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100142670 ◽

1986 ◽

Vol 44 ◽

pp. 210-211

Author(s):

Arthur J. Wasserman ◽

Kathy C. Kloos ◽

David E. Birk

Keyword(s):

Type I Collagen ◽

Corneal Stroma ◽

Minor Component ◽

Homogeneous Distribution ◽

Type I ◽

Type V Collagen ◽

Fibril Morphology ◽

Type V ◽

In Vitro Interaction

Type I collagen is the predominant collagen in the cornea with type V collagen being a quantitatively minor component. However, the content of type V collagen (10-20%) in the cornea is high when compared to other tissues containing predominantly type I collagen. The corneal stroma has a homogeneous distribution of these two collagens, however, immunochemical localization of type V collagen requires the disruption of type I collagen structure. This indicates that these collagens may be arranged as heterpolymeric fibrils. This arrangement may be responsible for the control of fibril diameter necessary for corneal transparency. The purpose of this work is to study the in vitro assembly of collagen type V and to determine whether the interactions of these collagens influence fibril morphology.

Download Full-text

In vitro effect of an aqueous extract of Artocarpus lakoocha on the intestinal parasites in cattle

Planta Medica ◽

10.1055/s-0030-1264720 ◽

2010 ◽

Vol 76 (12) ◽

Author(s):

N Saowakon ◽

P Chaichanasak ◽

C Wanichanon ◽

V Reutrakul ◽

P Sobhon

Keyword(s):

Aqueous Extract ◽

Intestinal Parasites ◽

Vitro Effect

Download Full-text

Plasminogen Activation in Diabetes Mellitus: Normalization of Blood Sugar Levels Improves Impaired Enzyme Kinetics In Vitro

Thrombosis and Haemostasis ◽

10.1055/s-0038-1657861 ◽

1985 ◽

Vol 54 (02) ◽

pp. 413-414 ◽

Cited By ~ 10

Author(s):

Margarethe Geiger ◽

Bernd R Binder

Keyword(s):

Plasminogen Activator ◽

Blood Sugar ◽

Metabolic Disorders ◽

Normal Values ◽

Diabetic Patients ◽

Type I ◽

Plasminogen Activation ◽

Lag Period ◽

Sugar Levels

SummaryWe have demonstrated previously that fibrin enhanced plasmin formation by the vascular plasminogen activator was significantly impaired, when components isolated from the plasma of three uncontrolled diabetic patients (type I) were used to study plasminogen activation in vitro. In the present study it can be demonstrated that functional properties of the vascular plasminogen activators as well as of the plasminogens from the same three diabetic patients are significantly improved after normalization of blood sugar levels and improvement of HbAlc values. Most pronounced the Km of diabetic vascular plasminogen activator in the presence of fibrin returned to normal values, and for diabetic plasminogen the prolonged lag period until maximal plasmin formation occurred was shortened to almost control values. From these data we conclude that the observed abnormalities of in vitro fibrinolysis are not primarily associated with the diabetic disease, but might be secondary to metabolic disorders caused by diabetes.

Download Full-text

Type I Interferons promote maintenance and function of follicular T helper cells in vitro

10.1055/s-0038-1677255 ◽

2019 ◽

Author(s):

S Ehrlich ◽

K Wild ◽

M Smits ◽

K Zoldan ◽

M Hofmann ◽

...

Keyword(s):

T Helper Cells ◽

Type I Interferons ◽

Type I ◽

T Helper ◽

Helper Cells ◽

Follicular T Helper Cells ◽

And Function

Download Full-text

Somatic Embryogenesis and In vitro Plant Regeneration in Vigna trilobata (L.) Verd. - A Potential Pasture Legume

Plant Tissue Culture and Biotechnology ◽

10.3329/ptcb.v19i1.4990 ◽

1970 ◽

Vol 19 (1) ◽

pp. 89-99

Author(s):

K. Choudhary ◽

M. Singh ◽

M. S. Rathore ◽

N. S. Shekhawat

Keyword(s):

Somatic Embryogenesis ◽

Growth Regulators ◽

Somatic Embryos ◽

Basal Medium ◽

Long Term Study ◽

Continuous Application ◽

First Time ◽

Pasture Legume

This long term study demonstrates for the first time that it is possible to propagate embryogenic Vigna trilobata and to subsequently initiate the differentiation of embryos into complete plantlets. Initiation of callus was possible on 2,4-D. Somatic embryos differentiated on modified MS basal nutrient medium with 1.0 mg/l of 2,4-D and 0.5 mg/l of Kn. Sustained cell division resulted in globular and heart shape stages of somatic embryos. Transfer of embryos on to a fresh modified MS basal medium with 0.5 mg/l of Kn and 0.5 mg/l of GA3 helped them to attain maturation and germination. However, the propagation of cells, as well as the differentiation of embryos, were inhibited by a continuous application of these growth regulators. For this reason, a long period on medium lacking these growth regulators was necessary before the differentiation of embryos occurred again. The consequences for improving the propagation of embryogenic cultures in Vigna species are discussed. Key words: Pasture legume, Vigna trilobata, Globular, Heart shape, somatic embryogenesis D.O.I. 10.3329/ptcb.v19i1.4990 Plant Tissue Cult. & Biotech. 19(1): 89-99, 2009 (June)

Download Full-text