Modified Trichrome Stain Research Articles

Abstract A71: Isolation of aglycons and glycosides from Scutellaria baicalensis and exploration of responsible constituents on human breast cancer chemoprevention

Abstract Numerous effective anticancer drugs have been developed from botanical sources. Although the anticancer properties of Scutellaria baicalensis extract (SbE) have been reported, in some of our studies, SbE did not inhibit the growth of breast cancer cells significantly. In certain concentrations, it increased cancer cell growth. To identify the active anticancer constituents in SbE, we isolated a baicalin-knockout fraction (SbF1) and a baicalin-fraction (SbF3). We evaluated the anti-breast cancer activities of the fractions and their constituents on human breast cancer MCF-7 cells. Using a Waters 2960 high performance liquid chromatography (HPLC) instrument and a Phenomenex Prodigy ODS(2) column, we determined the contents in the extract and fractions: scutellarin, baicalin, baicalein and wogonin. The antiproliferative effects were assayed using the modified trichrome stain (MTS) method. The active properties of the fraction/constituents on the cancer cell cycle and on apoptosis were evaluated using a BD FACScan flow cytometer. A one-way ANOVA was used to measure statistical significance. HPLC data showed that the content of scutellarin, baicalin, baicalein and wogonin in SbE were 0.03, 15.67, 5.19 and 1.42%, respectively. The major constituents of SbF1 are the aglycons, baicalein (40.54%) and wogonin (12.35%). SbF3 contains only the glycoside baicalin (16.68%). SbF1 showed a significant antiproliferative effect. Treatment with 100 g/ml of SbF1 for 72 h inhibited MCF-7 cell growth by 81.6% (P<0.01); at the same treatment concentration, SbF3 increased cell growth by 22.6% (P<0.01). Therefore, SbF1 is the active fraction of SbE. Compared to control (48% of G1-phase, 33% of M-phase, and 10% of G2/M-phase), after treatment with 50 g/mL of SbF1 for 72 h, 27% of cells were in G-phase, 41% of cells were in M-phase, and 23% of cells were in G2/M-phase. SbF1 increased the S- and G2/M-phases in MCF-7 cells. After treatment for 48 h, the percentage of early apoptotic cells induced by 50 g/mL of SbF1 was 12.4%; by 100 g/mL, the percentage was 24.2%. SbF1 significantly increased induction of cell apoptosis compared to control, 6.7%. Studying the effects of four flavonoids in SbE, two glycosides (scutellarin and baicalin) and two aglycons (baicalein and wogonin), on MCF-7 cell proliferation showed that baicalein and wogonin significantly inhibited MCF-7 cell growth at 10–100 µM. In contrast, at certain concentrations (5–50 µM), the glycosides scutellarin and baicalin increased cancer cell growth. These phytochemical and biological data suggest that the active anticancer constituents in SbE are flavonoid aglycons. Since the two glycosides promoted MCF-7 cells at certain concentrations, these compounds should be removed from the S. baicalensis extract to ensure chemopreventive activities. Citation Information: Cancer Prev Res 2010;3(1 Suppl):A71.

Apport de la PCR dans la recherche et l’identification des microsporidies intestinales chez les sujets infectés par le VIH

AimIntestinal microsporidiosis are among the most frequent opportunistic diseases in immunocompromised subjects. This study aimed to evaluate the contribution of PCR for a better detection and species identification of microsporidia in stool specimens of HIV-infected patients. Patients and methodsStool samples obtained from 119 HIV-infected Tunisian subjects were screened for intestinal microsporidiosis by light microscopy using Weber's modified Trichrome stain and by a PCR method using universal primers V1/PMP2 which amplified a common fragment of the small subunit rRNA gene of microsporidia. The obtained PCR products were then sequenced using an ABI PRISM 377 DNA sequencer. ResultsThe results showed a better sensitivity of PCR in the detection of microsporidia with an infection rate of 14.3% significantly higher than that of 6.7% obtained by light microscopy (p=0.03). As previously described, intestinal microsporidiosis was associated with low CD4 cell counts; 23.9% infection rate in patients having CD4 cell count under 200/mm3 against 5.6% in patients with higher CD4 cell count (p=0.008). The sequencing of 15 out of the 17 positive PCR products has confirmed in all cases the species identified based on the PCR fragment size i.e., 250pb for Enterocytozoon bieneusi (seven cases) and about 270pb for Encephalitozoon intestinalis (nine cases); one case revealed a double infection. ConclusionPCR proved to be more effective than classical Trichrome stain for the diagnosis of intestinal microsporidiosis. Moreover, the ability of PCR to identify the species involved could also be useful for cases management.

Selective fraction of Scutellaria baicalensis and its chemopreventive effects on MCF-7 human breast cancer cells

Based on our previous observation, the whole Scutellaria baicalensis extract (SbE) did not show significant breast cancer cell inhibitory effect. In this study, we isolated a baicalin-deprived-fraction (SbF1) of Scutellaria baicalensis, and baicalin-fraction (SbF3), and evaluated their anti-breast cancer properties using MCF-7 cells. The content of four flavonoids in extract/fractions were determined using high performance liquid chromatography. Analytical data showed that in SbF1, the major constituents are baicalein and wogonin, while SbF3 only contains baicalin. The antiproliferative effects of fractions and SbE were assayed using modified trichrome stain method. SbF1 showed significant antiproliferative effect. Treated with 100mug/ml of SbF1 for 72h inhibited MCF-7 cell growth by 81.6%, while in the same treatment concentration, SbF3 increased cell growth by 22.6%. SbF1 was recognized as an active fraction of SbE. The effects of four flavonoids in SbE, scutellarin, baicalin, baicalein and wogonin, were determined, and data showed that baicalein and wogonin significantly inhibited MCF-7 cell growth. In contrast, in certain concentrations, scutellarin and baicalin increased cancer cell growth. The effects of SbF1 on cell cycle and apoptosis were assayed using flow cytometry. SbF1 arrested MCF-7 cells in S- and G2/M-phases, and significantly increased induction of cell apoptosis. These combined phytochemical and biological data provide evidence for further chemopreventive studies of the baicalin-deprived SbE on breast cancer.

Emerging Prevalence of Microsporidial Keratitis in Singapore: Epidemiology, Clinical Features, and Management

To investigate the incidence and epidemiologic factors involved in the development of microsporidial keratitis. The association of host immune status and clinical pattern, clinical features, and the role of fluoroquinolone monotherapy in treatment are also examined. Retrospective, noncomparative case series. All cases (124 patients, 134 eyes) of microsporidial keratitis confirmed with modified trichrome stain positive of corneal scrape over a 4-year period. Epidemiologic factors were observed. Host immune status with human immunodeficiency virus (HIV) serology and CD4/CD8 analysis was performed when consent was obtained. Visual acuity (VA) and slit-lamp examination throughout the course of keratitis was recorded. Treatment used included topical fluoroquinolones (ciprofloxacin 0.3%, moxifloxacin 0.5%, gatifloxacin 0.5%, levofloxacin 0.5%, or norfloxacin 0.3%) as monotherapy or in combination with topical fumagillin and/or systemic albendazole. Where corneal edema developed, ultrasound corneal pachymetry was recorded. Demographic features and epidemiologic factors, including host immune status. Clinical features and disease course, including the response to different therapeutic regimes. Patients ranged in age from 11 to 68 years (mean, 31.9; median, 30) with a male:female ratio of 8:1 (females n = 17 [13.7%]). We performed HIV serology and CD4/CD8 in 45.9% of cases (n = 57); all the cases tested were negative with normal T-cell indices. Epidemiologic factors included soil exposure (50%), contact lens wear (21.1%), and topical steroid treatment (17.1%). The VA on presentation ranged from 20/20 to 20/100 (median, 20/30) with no loss in lines of VA on resolution. Common features were follicular papillary conjunctivitis and coarse punctate epithelial lesions in 3 patterns--diffuse, peripheral, and paracentral--evolving into nummular keratitis before resolution. Resolution occurred in 99% of cases on topical fluoroquinolone monotherapy. Four patients had recurrent disease that resolved with repeat fluoroquinolone or fluoroquinolone/oral albendazole combination. Two new clinical features were identified--diffuse endotheliitis (19.4%) with corneal edema and limbitis. This study identifies an increasing incidence of microsporidial keratitis in Singapore with a strong correlation with prior soil exposure. Diffuse endotheliitis and limbitis have not been described and resolves with topical steroid therapy. Topical fluoroquinolone monotherapy is a valid treatment option.

Prevalence of microsporidia in an indigenous Orang Asli community in Pahang, Malaysia

Microsporidia are ubiquitous parasites thought to be closely related to fungi. Their presence in the environment means that humans are frequently exposed to infection. Stool samples were collected from 151 indigenous villagers from the eastern state of Pahang in 2005. The samples were concentrated with water-ether sedimentation, stained with modified trichrome stain and examined under oil-immersion microscopy. Thirty-two specimens (21.2%) were positive for microsporidia. Microsporidia were observed as ovoid or rounded ovoid shapes measuring approximately 1mum, with a bright pink outline containing a central or posterior vacuole. PCR amplification with specific primers on microscopy-positive specimens amplified Encephalitozoon intestinalis DNA from five of the ten specimens used.

Microsporidiose et toxoplasmose disséminées chez une patiente présentant une leucémie prolymphocytaire T

We report a case of microsporidiosis in a 72-year-old woman presenting with prolymphocytic leukemia. The underlying conditions 7 months after leukemia was diagnosed were pancytopenia and immunosuppression due to alemtuzumab and pentostatin. The patient's status had worsened and she presented with dysuria. Urine cultures for bacteria were repeatedly negative. She was first empirically treated with broad-spectrum antibiotics. Three months later, urinary symptoms were persisting. Her blood lymphocyte count was 90/μl. Urine examination was positive for microsporidia using modified trichrome staining and Uvitex® 2B fluorescence. Microsporidia were also detected in stools. The patient was cured by albendazole. This was consistent with an infection due to Encephalitozoon sp. Concurrently, disseminated toxoplasmosis was diagnosed. Toxoplasma gondii was detected in bone marrow, broncho-alveolar lavage and cerebrospinal fluid. She was successfully treated with sulfadiazine–pyrimethamine. Four cases of microsporidiosis in myeloid leukemic patients have been already described. The present case in a patient with lymphoid leukemia is the first to be reported.

Incidence of Microsporidia in Cancer Patients

Microsporidia are considered opportunistic pathogens as evidenced by the significant detection in immunocompromised HIV/AIDS population. Cancer patients receiving chemotherapy are considered to be immunosuppressed. Stool samples were collected from 311 cancer patients in the Klang Valley. Each sample underwent water-ether concentration and staining with modified trichrome stain. Sixty-eight samples were positive by oil immersion examination. Polymerase chain reaction amplification with specific primers on those samples amplified Encephalitozoon intestinalis from two of the samples and Encephalitozoon hellem from one sample.

Comparison of weber green and ryan blue modified trichrome staining for the diagnosis of microsporidial spores from stool samples of HIV-positive patients with diarrhoea

Comparison of weber green and ryan blue modified trichrome staining for the diagnosis of microsporidial spores from stool samples of HIV-positive patients with diarrhoea

Preliminary studies on histological changes after repairing the facial nerve defect with acellular facial nerve

To investigate the morphological changes after chemically extracted acellular nerve allografts transplant. Seventy-two rabbits were divided into four groups. Acellular allografts of facial nerve were used in experimental group, and facial nerve autografts, acellular peroneal nerve allografts and peroneal nerve autografts respectively used in three control groups. The morphological changes after transplant were evaluated by modified trichrome staining, immunohistological staining and transmission electron microscope. The two facial nerve grafts showed numerous regenerated nerve fibers, vessels and as well as a spindle schwann cells arranged longitudinally. No significant difference was observed in the fiber number and myelin thickness between the two groups,while the two peroneal nerve groups showed poor regeneration 6 months after operation. The facial nerve allografts showed more neurite regeneration six months after transplant, and the regenerated axons passed through the distal stoma and there were well revascularized and proliferated schwann cells in the grafts.

Red American Ginseng: Ginsenoside Constituents and Antiproliferative Activities of Heat-Processed Panax quinquefolius Roots

Red Asian ginseng ( Panax ginseng C. A. Meyer, Araliaceae) is used in many Oriental countries. In this study, the saponin constituents and anticancer activities of steamed American ginseng ( Panax quinquefolius L.) roots were evaluated. The contents of 12 ginsenosides in the roots were determined using high performance liquid chromatography (HPLC). After the steaming treatment (100 - 120 degrees C for 1 h and 120 degrees C for 0.5 - 4 h), the quantity of 7 ginsenosides decreased and that of 5 others increased. The content of ginsenoside Rg3, a previously recognized anticancer compound, increased significantly when the root was steamed at 120 degrees C for 0.5 - 3 h. The antiproliferative effects of unsteamed and steamed (120 degrees C for 1 h and 2 h) American ginseng root extracts were assayed by the modified trichrome stain (MTS) method using three cancer cell lines (SW-480, HT-29, NSCLC). Heat-processing increased the antiproliferative effect of American ginseng significantly, and the activity of the extract from roots steamed for 2 h was greater than that of roots steamed for 1 h. Chemical constituents and antiproliferative activities of white and red Asian ginseng have also been evaluated. Five representative ginsenosides, Rb1, Rd, Re, Rg2 and Rg3, were studied. Ginsenoside Rg3 had the most potent effect. The antiproliferative activities of red American ginseng are augmented when ginsenoside Rg3 is increased.

Therapeutic effect of passive mobilization exercise on improvement of muscle regeneration and prevention of fibrosis after laceration injury of rat

OBJECTIVE: To evaluate the muscle healing effect of passive mobilization exercises after a laceration injury.DESIGN: Randomized‐controlled trial.SETTING: Basic science laboratory.ANIMALS: Male Sprague–Dawley rats (N=36), age ranging from 8 to 10 weeks and weight ranging from 300 to 400 g.INTERVENTION: The bilateral gastrocnemius muscles were lacerated. The left‐leg muscles were used as the study groups and the right side was used as the control (lacerated muscles without any treatment, n=8). In the exercise group (n=24), passive mobilization exercise (15 min/day) was performed for 5 days starting from different time points (2, 7, and 14 days postlaceration). The decorin group (n=8) was injected with decorin (50 μg at 14 days postlaceration), which is a well‐known antifibrotic agent. Four animals were used as the normal controls, in which only the muscle strength was evaluated. All the animals were killed 4 weeks after the laceration.MAIN OUTCOME MEASURES: The histologic characterization of muscle regeneration (hematoxylin and eosin staining, number and diameter of the centronucleated, regenerating myofibers), muscle fibrosis (vimentin‐positive area, Masson modified trichrome staining positive area), and muscle strength (analysis of fast twitch strength).RESULTS: The level of fibrosis was more than 50% lower in the exercise and decorin groups than in the control (P<0.05). The decorin group showed the highest number of regenerated, new myofibers, and the highest muscle strength. All of the exercise groups, regardless of the starting time of exercise, also showed significant improvement in regeneration and strength. However, the exercise group starting 14 days after the laceration showed the best results.CONCLUSIONS: Stretching exercises after a muscle laceration injury has a strong antifibrotic effect, as much as a well‐known antifibrotic agent, decorin. According to the results, the best time to begin stretching exercises is 14 days after laceration for antifibrosis and muscle regeneration.

Therapeutic Effect of Passive Mobilization Exercise on Improvement of Muscle Regeneration and Prevention of Fibrosis After Laceration Injury of Rat

Hwang JH, Ra Y-J, Lee KM, Lee JY, Ghil SH. Therapeutic effect of passive mobilization exercise on improvement of muscle regeneration and prevention of fibrosis after laceration injury of rat. Objective To evaluate the muscle healing effect of passive mobilization exercises after a laceration injury. Design Randomized controlled trial. Setting Basic science laboratory. Animals Male Sprague-Dawley rats (N=36), age ranging from 8 to 10 weeks and weight ranging from 300 to 400g. Intervention The bilateral gastrocnemius muscles were lacerated. The left leg muscles were used as the study groups and the right side was used as the control (lacerated muscles without any treatment, n=8). In the exercise group (n=24), passive mobilization exercise (15min/d) was performed for 5 days starting from different time points (2, 7, and 14d postlaceration). The decorin group (n=8) was injected with decorin (50μg at 14d postlaceration), which is a well-known antifibrotic agent. Four animals were used as the normal controls, in which only the muscle strength was evaluated. All the animals were killed 4 weeks after the laceration. Main Outcome Measures The histologic characterization of muscle regeneration (hematoxylin and eosin staining, number and diameter of the centronucleated, regenerating myofibers), muscle fibrosis (vimentin-positive area, Masson modified trichrome staining positive area), and muscle strength (analysis of fast twitch strength). Results The level of fibrosis was more than 50% lower in the exercise and decorin groups than in the control ( P<.05). The decorin group showed the highest number of regenerated, new myofibers and the highest muscle strength. All of the exercise groups, regardless of the starting time of exercise, also showed significant improvement in regeneration and strength. However, the exercise group starting 14 days after the laceration showed the best results. Conclusions Stretching exercises after a muscle laceration injury has a strong antifibrotic effect, as much as a well-known antifibrotic agent, decorin. According to the results, the best time to begin stretching exercises is 14 days after laceration for antifibrosis and muscle regeneration.

Sensitivity and Specificity of a Monoclonal Antibody-Based Fluorescence Assay for Detecting Enterocytozoon bieneusi Spores in Feces of Simian Immunodeficiency Virus-Infected Macaques

Enterocytozoon bieneusi is clinically the most significant among the microsporidia causing chronic diarrhea, wasting, and cholangitis in individuals with human immunodeficiency virus/AIDS. Microscopy with either calcofluor or modified trichrome stains is the standard diagnostic test for microsporidiosis and does not allow species identification. Detection of E. bieneusi infection based on PCR is limited to a few reference laboratories, and thus it is not the standard diagnostic assay. We have recently reported the development and characterization of a panel of monoclonal antibodies against E. bieneusi, and in this publication we evaluated the specificity and sensitivity of an immunofluorescence assay (IFA), compared with PCR, in simian immunodeficiency virus-infected macaques. The IFA, which correlated with the primary PCR method, with a detection limit of 1.5 x 10(5) spores per gram of feces, will simplify considerably the detection of E. bieneusi spores in clinical and environmental specimens and in laboratory and epidemiological investigations.

Detection of Cryptosporidium - specific coproantigen in human immunodeficiency virus/acquired immunodeficiency syndrome patients by using a commercially available immunoenzymatic assay

The aim of this study was to verify the occurrence of Cryptosporidium infection in 52 human immunodeficiency virus (HIV)/acquired immunodeficiency syndrome (AIDS) patients (group 1) and 38 clinically healthy individuals (group 2) by using enzyme immunoassay (EIA). All fecal samples collected were submitted to the Baermann, Lutz and Ritchie methods, the Safranin/Methylene Blue, and Weber's chromotrope modified Trichrome staining techniques, and EIA. In group 1, parasitological staining techniques and EIA were both positive for Cryptosporidium sp. infection in 3/52 (5.8%) samples and both negative in 45/52 (86.5%) samples, while 4/52 (7.7%) samples were positive in EIA and negative in parasitological staining techniques. Concerning group 2, all samples were negative by EIA and microscopy for Cryptosporidium infection. In conclusion, EIA may be an alternative method for detecting Cryptosporidium-specific coproantigen in HIV/AIDS patients.

Microsporidial keratoconjunctivitis in healthy individuals: A case series

Purpose To present a series of 6 cases of microsporidial keratoconjunctivitis in healthy, nonimmunocompromised individuals. Design Retrospective, noncomparative case series. Participants Six individuals with unilateral keratoconjunctivitis. Methods Cornea epithelial scrapings were taken and evaluated by modified trichome staining. Blood was taken for human immunodeficiency virus (HIV) enzyme-linked immunosorbent assay in all cases and for CD4 and CD8 T-lymphocyte counts in 5 cases. Main outcome measures The individuals were evaluated based on symptoms, visual acuity, slit-lamp biomicroscopy, and pathologic examination of the corneal scrapings. Results All cases occurred in men whose ages ranged from 16 to 37 years. Initial symptoms included unilateral pain and redness. All experienced subsequent worsening of symptoms and blurring of vision after using topical steroids prescribed by general practitioners. Slit-lamp biomicroscopy revealed coarse, multifocal, punctate epithelial keratitis in all 6 cases, anterior stromal infiltrates in 2 cases, with accompanying conjunctivitis in all cases. Modified trichrome staining of corneal epithelial scrapes revealed pinkish to red spores characteristic of microsporidia in all cases. Results of an HIV enzyme-linked immunosorbent assay were negative in all cases, and CD4 and CD8 T-lymphocyte counts and ratios were normal in all 5 tested cases. On diagnosis, topical steroid therapy was stopped in all cases. Treatment with topical Fumidil B (bicyclohexylammonium fumagillin; Leiter’s Park Ave Pharmacy, San Jose, CA) together with oral albendazole was given in 3 cases, oral albendazole alone in a single case, and broad-spectrum antibiotic treatment with topical norfloxacin or chloramphenicol in two cases. Two cases had keratic precipitates with mild cellular activity in the anterior chamber and one such case was restarted subsequently on topical steroids. All six cases showed resolution of epithelial keratitis but with residual visually inconsequential subepithelial scars by the end of 1 month of treatment. Conclusions Microsporidial keratoconjunctivitis can occur more commonly than expected in healthy, nonimmunocompromised individuals. Topical steroids seem to contribute to the persistence of this infection and may be a predisposing factor in these cases by creating a localized immunocompromised state. The clinical course is variable and may be self-limiting with cessation of topical steroid use.

An Evaluation of Three Conventional Histological Techniques for Staining the Cerata of Cratena pilata

Fixation and staining methods for different types of tissue in the marine nudibranch Cratena pilata were evaluated. Cratena pilata, a marine snail in the Phylum Mollusca, has the ability to take stinging cells, called nematocysts, from ingested animals belonging to another phylum, the coelenterates, and use them for their own defense. A total of I0 nudibranchs were used; 6 were preserved in 10% formalin and 4 in Helly's fixative. Three different types of stains were compared: hematoxylin and eosin (H&E), Cason's modified trichrome stain, and Bielschowsky's silver stain. The animals fixed in Helly's fixative were stained with Cason's modified trichrome stain and H&E. The formalin fixed animals were stained with either Cason's modified trichrolne stain or Bielschowsky's silver stain. Helly's fixative in conjunction with H&E proved to be the best combination to examine the general morphology of Cratena pilata. Cason's modified trichrome stain was the best stain for differentiating gut tissue, muscle, basement membrane, nematocysts, and the cerebral ganglion. Although Bielschowsky's silver stain stained nerves, Cason's modified trichrome stain proved to be the better stain to differentiate neuronal axons and cell bodies. (The J Histotechnol 24:255, 2001)Submitted: February 2, 2001; Accepted: April 6, 2001

Intestinal microsporidiosis: a current infection in HIV-seropositive patients in Portugal

Intestinal microsporidiosis is recognised as an important cause of opportunistic infections in immunocompromised patients, especially those with AIDS. Two species are implicated in diarrhoea and other gastrointestinal disease in HIV-infected patients: Enterocytozoon bieneusi and Encephalitozoon intestinalis. Diagnosis of gastrointestinal microsporidiosis was made by detecting spores of the parasite in stool specimens with Weber’s modified trichrome stain and with some optical brightening agents such as UVITEX 2B or calcofluor white M2R. The identification of microsporidiosis at the species level was made using appropriate primers with PCR. The diagnosis of intestinal microsporidiosis is currently performed in the parasitology laboratory. In a study of 215 HIV-infected patients, conducted from 1996 to 1999 (approximately n = 60/year), we found a prevalence of spores of microsporidia of 51.5% ( n = 31) in 1996, 14.0% ( n = 5) in 1997 and 12.5% ( n = 8) in 1998 and 42.8% ( n = 25) in 1999. Using PCR we found that E. intestinalis was the only species responsible for the gastrointestinal symptoms in 49 patients with microsporidian spores (71%) and E. bieneusi in 29% ( n = 20).

Morphological and molecular characterization of Encephalitozoon hellem in hummingbirds

Microsporidiosis was identified as a cause of enteritis in wild, migratory hummingbirds (Calypte anna). Electron microscopic examinations of parasites showed microsporidian spores with a double spore coat and a polar filament containing four to six coils, compatible with the genus Encephalitozoon. Molecular analysis of ribosomal RNA genes further identified the parasites from droppings and small intestinal segments as Encephalitozoon hellem, genotype I. Microsporidial spores were identified in 19% of droppings from C. anna, Archilochus alexandri and Selasporus sasin using Gram or modified trichrome staining methods. Since E. hellem is an opportunistic pathogen in immunocompromised humans, the pathogenic potential in avian hosts, the zoonotic potential of this parasite, and the role of birds as reservoirs needs to be further explored.

CHRONIC DIARRHEA AS A RESULT OF INTESTINAL MICROSPOSIDIOSIS IN A LIVER TRANSPLANT RECIPIENT

Microsporidia are common pathogens among patients infected with human immunodeficiency virus. They account for a substantial proportion of chronic diarrhea and malabsorption in acquired immune deficiency syndrome, but their appearance after solid organ transplantation has only rarely been reported. Methods. We report what we believe is the first case of documented Enterocytozoon bieneusi infection in a liver transplant recipient. Results. Our patient presented with chronic diarrhea and colicky abdominal pain. Although symptoms were severe, only mild microscopical mucosal changes were found in the intestinal tract. A modified trichrome stain of stool specimens revealed microsporidial spores, and species differentiation by restriction fragment length polymorphism polymerase chain reaction identified Enterocytozoon bieneusi. Albendazole therapy brought symptomatic relief but no microbiological clearance. Enterocytozoon bieneusi may cause chronic diarrhea not only in immunosuppression as a result of human immunodeficiency virus infection but also among patients with therapeutic immunosuppression after organ transplantation. Therefore, microsporidial infection should be considered in immunosuppressed patients with otherwise unexplained diarrhea.

Evaluation of a combined acid-fast-trichrome stain for detection of microsporidia and Cryptosporidium parvum.

M and Cryptosporidium parvum are important agents of enteritis, capable of causing severe chronic disease in immunocompromised individuals.1,2 Currently, detection of these organisms in stool specimens requires at least 2 special stains or procedures; a chemofluorescent agent or a modified trichrome stain is commonly used to detect microsporidia,3–5 and an acid-fast stain, direct immunofluorescent assay (DFA), or enzyme immunoassay is used to detect C parvum.6–8 Recently, Ignatius et al9 described a combination acid-fast–trichrome (AFT) stain, which allows detection of these organisms, as well as Cyclospora cayetanensis and Isospora belli, with a single procedure. We modified this AFT procedure, incorporating the use of commercially available reagents, and compared the results of the modified AFT stain with those of our standard procedures for stool specimens received in the laboratory for a 15-month period.