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Retortamonas spp. has been reported as an intestinal parasite among various host organisms, including humans; however, its intra-genus molecular diversity has not yet been elucidated. Haplotypes of the 18S small subunit ribosomal RNA locus (1836–1899 bp) of Retortamonas spp. from humans (n = 8), pigs (n = 6), dogs (n = 1), goats (n = 16), water buffalos (n = 23), cattle (n = 7), rats (n = 3), and chickens (n = 5) were analyzed with references isolated from non-human mammals, amphibians, and insects. Phylogenetic and network analyses revealed a statistically supported three cluster formation among the vertebrate-isolated haplotypes, while insect-isolated haplotypes were independently clustered with Chilomastix. In the clade of vertebrate isolates, assemblage A (amphibian genotype), which included the amphibian references, was addressed as an out-group of the other clusters. Assemblage B (mammalian and chicken genotype) included most haplotypes from various mammals including humans with the haplotypes isolated from a chicken. Human isolates were all classified into this assemblage, thus assemblage B might correspond to R. intestinalis. Assemblage C (bovine genotype), which included specific haplotypes from water buffalos and cattle, was addressed as a sister lineage of assemblage B. Among the diversified haplotypes of assemblage B, a specific haplotype, which was identified from multiple host mammals (humans, dogs, pigs, cattle, water buffalos, elks, goats, and rats), indicates the potential zoonotic transmission of the Retortamonas among them. The genotyping classification of retortamonads could contribute to a better understanding of its molecular epidemiology, especially among humans and related host organisms.

Blastocystis sp. is a common intestinal protist found worldwide in a variety of animals, including humans. Currently, 17 subtypes (STs) of Blastocystis isolates from mammalian and avian host species have been reported based on the small subunit ribosomal RNA gene (SSU rDNA). Among these, human Blastocystis were only identified among STs 1-9. Except ST9, all other STs comprised isolates from humans and other animal species. Entire sequence data of the SSU rDNA of nine Blastocystis isolates from laboratory rats or guinea pigs previously showed ST4, whereas Blastocystis isolates from wild rodents have not been addressed genetically. In this study, Blastocystis infection in wild rodents was surveyed in Indonesia and Japan, and 11 and 12 rodent Blastocystis parasites were obtained from Rattus exulans and R. novercious, respectively. All new Blastocystis isolates from wild rodents were identified as ST4 based on the SSU rDNA sequences. The best tree inferred with the entire sequences of the SSU rDNA of all ST4 isolates including 17 data registered in GenBank clearly showed monophyletic ST4A and ST4B clades. Although ST4 isolates from laboratory rats were separated into these two clades, all Blastocystis isolates from wild rodents in the present study were positioned into the clade ST4A and further separated into two sub-clusters within the clade ST4A according to the location of the host species. Considering the fact that laboratory rats were susceptible to both ST4A and ST4B, separation of the monophyletic sub-clusters of Blastocystis isolates from Indonesian Polynesian rats and Japanese brown rats may indicate the presence of geographical variations rather than a host-specific separation. In either way, the robust host preference to rodent species of ST4 Blastocystis was also confirmed.

The genus Blastocystis is one of the most genetically diverse parasites. Blastocystis isolates from humans and animals have been classified into subtypes (STs) based on the phylogeny of the small subunit rRNA gene (SSU rDNA). Although human Blastocystis isolates are limited to STs 1-9, the identification of all 9 STs remains challenging due to the lack of specific primers for several STs. The sequencing of partial SSU rDNA is therefore essential for the identification of several STs. In this study, we developed 9 sets of PCR primers to detect each of the 9 kinds of ST in humans. When these ST-specific primer pairs were examined reference Blastocystis for the 9 STs, all 9 amplified only the target ST even in a DNA mixture of all 9 STs. The specificities of the 9 primer sets were tested against several intestinal parasites and fungi found in human stool samples. No amplification with these common human intestinal eukaryotes was observed using the primer pairs for 8 STs, while the ST5 primer set gave only faint bands with some parasites. Since genomic DNA levels of these parasites extracted from Blastocystis-positive cultures are expected to be markedly lower than the pure or highly concentrated DNA samples tested, the cross-amplifications with these organisms are unlikely to be detected when DNA samples are extracted from Blastocystis-positive cultures. The PCR conditions for all 9 primer sets were the same, hence a one-step analysis by PCR amplification, followed by electrophoresis has potential as a simple tool for the subtyping of human Blastocystis isolates.

Blastocystis is a common unicellular eukaryotic parasite found not only in humans, but also in various kinds of animal species worldwide. Since Blastocystis isolates are morphologically indistinguishable, many molecular biological approaches have been applied to classify these isolates. The complete or partial sequences of the small subunit rRNA gene (SSU rDNA) are mainly used for comparisons and phylogenetic analyses among Blastocystis isolates. However, various lengths of the partial SSU rDNA sequence have been used for phylogenetic inference among genetically different isolates. Based on the complete SSU rDNA sequences, consensus terminology of nine subtypes (STs) of Blastocystis sp. that were supported by phylogenetically monophyletic nine clades was proposed in 2007. Thereafter, eight additional kinds of STs comprising non-human mammalian Blastocystis isolates have been reported based on the phylogeny of SSU rDNA sequences, while STs 11 and 12 were only proposed on the base of partial sequences. Although many sequence data from mammalian and avian Blastocystis are registered in GenBank, only limited data on SSU rDNA are available for poikilotherm-derived Blastocystis isolates. Therefore, the phylogenetic positions of the reptilian/amphibian Blastocystis clades are unstable. The phylogenetic inference of various STs comprising mammalian and/or avian Blastocystis isolates was verified herein based on comparisons between partial and complete SSU rDNA sequences, and the phylogenetic positions of reptilian and amphibian Blastocystis isolates were also investigated using 14 new Blastocystis isolates from reptiles with all known isolates from other reptilians, amphibians, and insects registered in GenBank.

Hemodialysis (HD) patients with critical limb ischemia (CLI) suffer chronic inflammation and repeated infection, require intervention, and may have a protracted hospital stay. Therefore, early prediction is particularly important for management of CLI in patients with suspected peripheral artery disease. The purpose of this study is to develop a simple score for predicting the incidence of CLI in HD patients with suspected peripheral artery disease. The subjects were 139 asymptomatic patients receiving maintenance HD and with ABI

Trichomonad species inhabit a variety of vertebrate hosts; however, their potential zoonotic transmission has not been clearly addressed, especially with regard to human infection. Twenty-one strains of trichomonads isolated from humans (5 isolates), pigs (6 isolates), rodents (6 isolates), a water buffalo (1 isolate), a cow (1 isolate), a goat (1 isolate), and a dog (1 isolate) were collected in Indonesia and molecularly characterized. The DNA sequences of the partial 18S small subunit ribosomal RNA (rRNA) gene or 5.8S rRNA gene locus with its flanking regions (internal transcribed spacer region, ITS1 and ITS2) were identified in various trichomonads; Simplicimonas sp., Hexamastix mitis, and Hypotrichomonas sp. from rodents, and Tetratrichomonas sp. and Trichomonas sp. from pigs. All of these species were not detected in humans, whereas Pentatrichomonas hominis was identified in humans, pigs, the dog, the water buffalo, the cow, and the goat. Even when using the high-resolution gene locus of the ITS regions, all P. hominis strains were genetically identical; thus zoonotic transmission between humans and these closely related mammals may be occurring in the area investigated. The detection of Simplicimonas sp. in rodents (Rattus exulans) and P. hominis in water buffalo in this study revealed newly recognized host adaptations and suggested the existence of remaining unrevealed ranges of hosts in the trichomonad species.

It is uncertain whether dynamic variation in the diameter of the aortic annulus occurs during the cardiac cycle in humans. The purpose of this study was to analyze cyclic changes of the aortic annulus using speckle-tracking trans-esophageal echocardiography. The subjects were 40 patients with aortic stenosis and 40 controls. Absolute and relative changes in the diameter of the aortic annulus and the times at which the maximum and minimum diameters occurred during the cardiac cycle were determined using speckle-tracking trans-esophageal echocardiography. The maximum and minimum diameters were 22.9 ± 2.7 and 20.0 ± 2.9 mm, respectively, in controls. The change in diameter of the aortic annulus was 2.9 ± 0.7 mm, and the relative change was 12.9 ± 3.5%. The maximum aortic annulus diameter was reached at the onset of aortic valve opening, and the minimum diameter occurred in the rapid filling phase. The change in diameter of the aortic annulus was significantly smaller (2.2 ± 0.6 mm vs. 2.9 ± 0.7 mm, p < 0.0001), and the time to reach the maximum diameter was significantly longer (98.5 ± 17.5 ms vs. 83.4 ± 18.2 ms, p = 0.0004), in the aortic stenosis group than in the control group. The study found that dynamic changes of the aortic annulus occur in the cardiac cycle and can be measured using speckle-tracking trans-esophageal echocardiography. We also found that aortic stenosis has an effect on the extent and timing of these changes. This suggests that accurate assessment of aortic annulus diameter requires consideration of the timing of the cardiac cycle.

Objectives: Cilostazol, a type III phosphodiesterase inhibitor, is an antiplatelet agent with vasodilating properties. Positive inotropic and chronotropic effects are frequently observed with cilostazol, but there are few reports on the influence of cilostazol on left ventricular function. The aim of this study was to assess this effect using tissue Doppler imaging (TDI) and two-dimensional speckle-tracking echocardiography (2D-STE). Methods: Thirty-five patients with normal left ventricular ejection fraction were enrolled in the study. Left ventricular cardiac function was assessed by TDI and 2D-STE before and after oral administration of cilostazol. Peak strain was defined using the peak radial strain (PRS), peak circumferential strain (PCS) and peak longitudinal strain (PLS). Time to peak strain was defined based on the times to PRS, PCS, and PLS, as T-PRS, T-PCS, and T-PLS, respectively. Results: After cilostazol administration, there were significant decreases in the left ventricular end-diastolic and end-systolic diameters (47.3 ± 5.2 vs. 43.3 ± 4.9 mm, P < 0.0001; 29.3 ± 6.4 vs. 26.0 ± 5.5 mm, P < 0.0001, respectively), and significant increases in the left ventricular ejection fraction (70.6 ± 9.5 vs. 72.7 ± 7.8%, P = 0.0381) and peak systolic annular velocity (7.9 ± 1.7 vs. 9.5 ± 3.1 cm/sec, P < 0.0001). PRS, PCS, and PLS all increased significantly and T-PRS, T-PCS, and T-PLS all decreased significantly after cilostazol administration. Conclusion: Positive inotropic and chronotropic effects of cilostazol were found based on assessment by TDI and 2D-STE. We suggest that periodic echocardiographic assessment should be performed before and after oral administration of cilostazol. (Echocardiography 2011;28:431-437)

A 29-year-old man presented with complaints of fever and pain and itching of his left neck with atopic dermatitis and abrasion. These symptoms had persisted for two days and dullness and fever developed, but the patient did not consult a physician. On the following day, he had a fever of 40°C and redness and swelling of the left neck. He visited a local clinic and was hospitalized with suspected cellulitis. A thrombus was detected in the internal jugular vein on MRI, and he was referred to our hospital. Neck ultrasonography showed the presence of an immovable thrombus in the area from the left internal jugular vein to the left brachiocephalic peripheral vein. Blood analysis indicated a major inflammatory response and juvenile idiopathic thrombophlebitis was suspected. A filter was carefully inserted into the superior vena cava, and anticoagulant therapy and medication with antibiotics led to remission. We present this case as a rare example of a condition mimicking Lemierre syndrome that was caused by dermal infection, and we include a review of the literature.

The purpose of this study is to determine the normative Doppler values and gender differences in left ventricular (LV) diastolic function in healthy subjects at each decade of life. Four hundred sixty-seven male and 455 female healthy subjects, aged 20–90 years, underwent standard echocardiography and tissue Doppler imaging. We divided the subjects into seven groups, based on the decade of their age (i.e., 20s, 30s, 40s, 50s, 60s, 70s, and 80s), and measured standard echocardiographic parameters and Doppler indices. Females younger than 50 years had significantly higher mitral E velocity, and females older than 40 years had significantly higher mitral A velocity, compared with their male counterparts. There was no difference in mitral E to A velocity ratio (E/A) at any age. The tissue Doppler method showed significantly lower early diastole velocity of the mitral annulus (E a) in females—especially females older than 50 years—than in males (10.7 ± 3.7 versus 11.2 ± 3.7 cm/s, p

The stool samples obtained from 94 patients with gastrointestinal symptoms and 109 asymptomatic individuals, who checked in due to other reasons, admitted at a major hospital in Ankara, Turkey were examined with native Lugol's iodine, trichrome, and Kinyoun's acid-fast stainings for parasitology examinations and with in vitro culture method for detection of Blastocystis. In a total of 203 stool samples tested, native Lugol's iodine and trichrome stainings could detect 12 (5.9%) and 20 (9.9%) positive samples for Blastocystis, respectively. Conversely, culture method could detect 66 (32.5%) positive samples, and this method was more sensitive compared to the both microscopic examinations (p < 0.001). Among 66 positive samples for Blastocystis, 27 were from symptomatic patients and 39 were from asymptomatic group. Subtypes (STs) were determined by PCR using seven different sequence-tagged site primers. ST3 was the most dominant in both symptomatic and asymptomatic groups and followed by ST1 or ST2. There were mixed infections with STs 1 and 2 or STs 1 and 3 in nine isolates. There was no statistical significance of the distribution of Blastocystis sp. subtypes between symptomatic and asymptomatic individuals (p > 0.05).

SummaryBackgroundThe disadvantageous effect of kidney dysfunction on left ventricular (LV) diastolic function is still unknown.MethodsForty non-chronic kidney disease (CKD) patients and 202 CKD patients, aged 40–89, were examined by standard echocardiography and the new modality of tissue Doppler imaging. All subjects were divided into 5 groups depending on their estimated glomerular filtration rate (GFR: ml/min/BSA). Classifications by GFR were defined as follows: group 1 (more than 90: normal subjects), group 2 (60–89), group 3 (30–59), group 4 (15–29) and group 5 (less than 15).ResultsThere were no significant differences in LV systolic function among the groups. Mitral E velocity was significantly lower in groups 1–4 (p

To investigate the possible transmission of Blastocystis organisms between local rhesus monkeys and children in Kathmandu, Nepal, we compared the subtype (ST) and sequence of Blastocystis isolates from children with gastrointestinal symptoms and local rhesus monkeys. Twenty and 10 Blastocystis isolates were established from 82 and 10 fecal samples obtained from children and monkeys, respectively. Subtype analysis with seven sequence-tagged site (STS) primers indicated that the prevalence of Blastocystis sp. ST1, ST2 and ST3 was 20%, 20% and 60% in the child isolates, respectively. In contrast to human isolates, ST3 was not found in monkey isolates and the prevalence of ST1 and ST2 was 50% and 70%, respectively, including three mixed STs1 and 2 and one isolate not amplified by any STS primers, respectively. Since Blastocystis sp. ST2 has been reported as the most dominant genotype in the survey of Blastocystis infection among the various monkey species, sequence comparison of the 150bp variable region of the small subunit rRNA (SSU rRNA) gene was conducted among ST2 isolates of humans and monkeys. Sequence alignment of 24 clones developed from ST2 isolates of 4 humans and 4 monkeys showed three distinct subgroups, defined as ST2A, ST2B and ST2C. These three subgroups were shared between the child and monkey isolates. These results suggest that the local rhesus monkeys are a possible source of Blastocystis sp. ST2 infection of humans in Kathmandu.

Blastocystis hominis is one of the most common eukaryotic organisms in the intestinal tract of humans, while its pathogenic potential is still controversial. A total of 286 stool samples obtained from adult and pediatric patients with or without gastrointestinal symptoms in two hospitals in Manisa, Turkey, were cultured to detect B. hominis infection. Forty-one and 51 isolates were obtained from the adults and children, respectively, and these isolates were subjected to subtyping by polymerase chain reaction (PCR) with the known sequence-tagged site primers. The correlation between the genotype and the symptoms was evaluated. PCR subtyping indicated that subtype 3 was the most common genotype in both symptomatic and asymptomatic groups, and the second common genotype was subtypes 1 and 2 in symptomatic and asymptomatic groups, respectively. A significant correlation between subtype 2 and the asymptomatic groups was found among both in pediatric and adult patients (chi(2) (cal) = 4.38, df = 1, p = 0.044). However, there were no significant differences between the other genotypes and the symptomatic or asymptomatic groups, as well as both the age and sex of the patients. The present study suggests that subtype 2 is a non-pathogenic genotype of B. hominis.

Most Blastocystis hominis isolates from humans are believed to be potentially zoonotic. This is because B. hominis isolates found in a variety of other host species have been found to have identical or relatively similar genotypes to those found in human isolates. However, the transmission of human B. hominis isolates to other animals has not been confirmed experimentally. In this study, the infectivity associated with several unique human Blastocystis genotypes (subtypes 2, 3, 4 and 7) was therefore investigated by infecting chickens and rats with two isolates of each subtype experimentally. The results showed that one isolate of subtype 4 and one isolate of subtype 7 was capable of infecting both chickens and rats, while two isolates of subtype 2, another isolate of subtype 4, and another isolate of subtype 7 could only infect chickens. Conversely, two isolates of subtype 3 failed to infect either of the animals. These results confirmed that several genotypes from human isolates could infect chickens and/or rats, indicating that chickens and rats are suitable experimental animal models for studying the zoonotic potential of human Blastocystis isolates.

Blastocystis sp. is a common parasite found in human and animal fecal samples. Currently, human Blastocystis isolates are classified into nine subtypes (STs) based on the phylogeny of their small subunit ribosomal RNA genes (SSU rDNAs). Since eight of the nine STs, except for ST9, have been reported in both humans and animals, these parasites are considered to be potentially zoonotic STs. To evaluate whether zoonotic transmissions play a main role in the lifecycle of Blastocystis, STs derived from humans, domestic pigs, domestic chickens, and wild rodents in a community with poor hygiene in Sumba Island, Indonesia were surveyed. Although fecal cross-contaminations between humans and animals were likely common at the investigation site, the confirmed major Blastocystis STs, which were detected as intense bands on gels following PCR targeting of the SSU rDNA, were different in each host species. STs 1-3 were found in resident children, while ST5, ST7, and ST4 were found in domestic pigs and chickens, and in wild rodents, respectively. Faint bands of STs 1, 2, and 7 were detected in samples from pigs, while no minor STs were observed in samples from the other host species. The distinct distributions of the major STs among the host animals examined, including humans, indicate host specificity in the lifecycle of Blastocystis. Considering the coprophagous nature of pigs, the presence of minor STs observed only in pigs could be explained by the mechanical passage of contaminated fecal materials.

Hemodialysis (HD) patients with critical limb ischemia (CLI) suffer chronic inflammation and repeated infection, require intervention, and may have a protracted hospital stay. Therefore, early prediction is particularly important for management of CLI in patients with suspected peripheral artery disease. The purpose of this study is to develop a simple score for predicting the incidence of CLI in HD patients with suspected peripheral artery disease. The subjects were 139 asymptomatic patients receiving maintenance HD and with ABI1.0. Multivariate logistic regression analysis was used to identify factors associated with development of CLI. These factors were subsequently weighted and integrated into a scoring system for the prediction of onset of CLI. Twenty-five patients had onset of CLI. Five factors selected from the multivariate model were weighted proportionally using their respective odds ratio (OR) for incidence of CLI (history of cerebral vascular disease, OR 6.42 [3 points]; diabetes, OR 3.92 [2 points]; hypoesthesia, OR 4.21 [2 points]; left ventricular ejection fraction50%, OR 3.89 [2 points]; serum albumin3.5 g/dL, OR 4.39 [2 points]). Three strata of risk were defined (low risk, 0 to 3 points; intermediate risk, 4 to 6 points; and high risk 7 to 11 points) with excellent prognostic accuracy for progression to CLI using the Kaplan-Meier method. Five factors were identified that increased the risk of progression to CLI in HD patients with suspected peripheral artery disease. A combination of those factors permitted establishment of three risk strata for accurate prediction of onset of CLI.

The infectivity of two Blastocystis hominis strains, RN94-9 and NIH:1295:1, was examined in 3-week-old SPF Wistar rats. The NIH:1295:1 strain, originally isolated from a guinea pig, was only able to infect rats via intracecal inoculation of the cultured organisms, while the RN94-9 strain, originally isolated from a laboratory rat, was able to infect rats by oral inoculation of the cultures due to the presence of a cystic form in the in vitro culture. Since many cysts were discharged in the feces of the infected rats, the infectivity of the concentrated cysts was compared between the two strains. Successful oral infection was observed in rats inoculated with 1 x 10(2)-1 x 10(6) cysts of the RN94-9 and NIH:1295:1 strains. The infectivity of the ten cysts varied in the three experiments of ten rats, being 20-100% and 30-100% in the RN94-9 and NIH:1295:1 strains, respectively. When an uninfected normal rat was housed with five experimentally inoculated rats, the normal rat became infected, demonstrating the fecal-oral transmission of the cyst form of this parasite. These results show that the Wistar rat is an ideal host for the propagation of strains RN94-9 and NIH:1295:1 of B. hominis, and demonstrate that the cyst form is the only transmissible form of this parasite.

Blastocystis infection in amphibians was surveyed in three species of anuran and one species of urodele amphibians captured at two distinct locations in Japan. All three species of frogs were highly infected with Blastocystis, while 69 individual urodele newts, Cynopus pyrrhogaster, were negative for infection. Eleven Blastocystis isolates (47.8%) were recovered from 23 Rana nigromaculata leopard frogs. Twenty-three (92%) of 25 Rana catesbeiana bullfrogs and all (100%) of 24 Bufo japonicus japonicus toads were positive for Blastocystis. Two distinct populations of the toad and bullfrog showed a high prevalence (100 or 84.6%) of Blastocystis infection, while in two populations of the leopard frog only one population was positive for Blastocystis (84.6%). Three Blastocystis isolates from different species of the frogs were established. Since none of the three isolates could survive at 37 degrees C, a temperature tolerance assay was performed to assess the optimal growth temperature and to determine the range of non-lethal temperatures. During the exponential growth phase of 3- or 4-day cultures at 25 degrees C, three isolates were exposed to 4, 28, 31, or 34 degrees C for 3 days and then returned to 25 degrees C to monitor the cell growth. Based on the optimal growth temperatures and different ranges of temperature tolerance among the three new isolates from frogs and two known species, Blastocystis hominis and Blastocystis lapemi, it was established that the three isolates recovered from different species of frogs had different physiological features from B. hominis and B. lapemi.

Twenty two Blastocystis isolates from cattle and pigs were genotyped by PCR using diagnostic primers, and the homology among isolates was then confirmed by restriction fragment length polymorphism (RFLP) analysis of the small subunit ribosomal RNA gene. Two distinct genotypes, subtypes 1 and 3, were detected in one and two of the ten isolates from cattle, and three and one of the 12 isolates from pigs, respectively. The RFLP profiles of all isolates designated as subtype 1 and subtype 3 were identical to those of known ribodemes 1 and 2, respectively. We found that 31.8% (7/22) of isolates examined were zoonotic genotypes of Bastocystis hominis. However, 68.2% (15/22) of isolates were not amplified with any of the diagnostic primers, and the profiles of these isolates were different from any ribodemes of B. hominis reported previously. This study shows that Blastocystis organisms in cattle and pigs are a potential source of human infection.

After cultured cysts are osmotically shocked by treating with distilled water, there is an exponential increase in the cyst form of Blastocystis pythoni; this was demonstrated by an immunofluorescence antibody assay against the culture organisms. In 11-day-old cultures of B. pythoni, 68.8% of the organisms (= 2.2 x 10(8) cysts/ml) were in the cyst form. Examination of thin sections of cysts revealed many similarities to the cyst forms of Blastocystis obtained from fecal samples in previous investigations. Freeze-fracture images of the plasma membrane of non-cyst cells also revealed a similar distribution of the intramembrane particles (IMP) when compared to non-cysts of B. hominis, while the plasma membrane of the cyst form showed practically no IMP. The size and morphology of particle-rich small depressions and smooth small protrusions observed on the P face and E face of non-cyst cells, respectively, were similar to endocytic sites reported for B. hominis. In the present study glycogen was cytochemically demonstrated at the ultrastructural level by an alkaline bismuth staining method in both cyst and non-cyst cells.

Introduction: Takotsubo cardiomyopathy (TTC) is characterized by a transient systolic dysfunction of the left ventricle. A few case was reported about left ventricular apical hypertrophy (APH) during recovery from TTC. The purpose of this study was to investigate the incidence of transient APH and the differences in clinical characteristics in TTC patients. Methods: Forty seven TTC patients were enrolled in this study. Patients were divided into two groups as N-APH group which did not show APH like findings in the process of wall motion recovery and T-APH group which showed transient APH findings. Cardiac complications were defined as cardiac death, pump failure, sustained ventricular tachycardia or ventricular fibrillation and advanced atrioventricular block. Results: Twelve of 47 (25.5%) TTC patients demonstrate typical APH findings during recovery from wall motion abnormality. Finally, this APH findings disappeared and left ventricular morphology recovered to normal structure. There is no significant difference between two groups in clinical characteristics (table). Incidence of cardiac complication showed significantly lower in T-APH group than that in N-APH group (P< 0.005). Conclusions: Transient left ventricular APH were observed in 25.5% in TCC patient by serial echocardiographic observation. Incidence of cardiac complications in T-APH group is significantly lower than that in N-APH group. Detection of T-APH is useful as a prognostic indicator in patients with TTC.

Introduction: It is reported that aortic valve reconstruction (AVrC) by using autologous pericardium is useful in patients with various aortic valve diseases. This method may be more effective in preserving the natural motion of the aortic valve annulus. Significant dynamic changes in the aortic annulus have previously been reported in animal models and clinical studies. The purpose of this study is to analize the cyclic changes of the aortic annulus using speckle-tracking trans-esophageal echocardiography in patients who underwent AVrC surgery. Methods: The subjects were 35 patients underwent AVrC surgery for aortic regurgitation or aortic stenosis and 40 controls. Cyclic aortic annulus diameter of the maximum and minimum diameters were determined from a speckle-tracking displacement curve (figure). These data was used to calculate the absolute change in the diameter of the aortic annulus, which was defined as the difference in diameters throughout the cardiac cycle. Results: There was significant difference between maximum and minimum aortic annulus diameter in AVrC group (18.7±3.0 mm vs 16.1±2.9 mm, p Conclusions: Our study suggests that dynamic changes of the aortic annulus occur in the cardiac cycle and can be measured using speckle-tracking trans-esophageal echocardiography. AVrC may preserve the coordination of the left ventricle, aortic annulus, sinus of Valsalva and aorta compared with conventional aortic valve replacement.

Genomic polymorphisms among nine strains of Blastocystis hominis were examined by random amplified polymorphic DNA (RAPD) using four different arbitrary polymerase chain reaction (PCR) primers. Based on the RAPD patterns, nine strains were classified into three groups. The specific primers designed from the unique bands yielded a single band from within each same group, but did not amplify between all the groups examined. Specificity of these diagnostic primers was tested against several common intestinal parasites and a yeast, and no amplification was confirmed. Since the current criteria indicates that Blastocystis organisms isolated from humans are designated as B. hominis, the authors propose to classify several subtypes among B. hominis groups based on the difference of genomic DNA using three diagnostic primers.

The patient was a 42-year-old male, who visited the hospital after he became aware of exertional dyspnoea. There were no abnormal physical findings. Transthoracic echocardiography indicated the presence of an 84 × 75 mm mass with homogeneous echogenicity, which was adjacent to the left atrium ( Panel A : apical four-chamber view, Supplementary data online, …

Basic proteins ofBlastocystis hominis were detected by the ammoniacal silver and ethanolic phosphotungstic acid techniques using electron microscopy. The central vacuole showed many silver grains when treated with ammoniacal silver and an increased electron density when treated with phosphotungstic acid. The intensity of positive reactions correlated with the electron density of the central vacuole, because cells having an electron-lucent central vacuole showed no silver grain deposits. Since it is known that the concentration of electron-dense materials in the central vacuole increases during log phase of growth, and then decreases in stationary phase, this organelle must accumulate basic proteins during cell growth.

Morphological changes in the central vacuole during the growth in in vitro culture ofBlastocystis hominis were investigated by light and electron microscopy. Most cells in log phase and an early stationary phase showed a positive staining reaction in the central vacuole with PAS or Sudan black B stain, whereas cells in late stationary phase showed few positive reactions. Electron microscopic observations revealed that 95% ofB. hominis cells in log phase and 50% of cells in early stationary phase, had a substantial accumulation of electron-dense material in the central vacuole. In contrast, only 25% of the organisms in late stationary phase had an electron-dense central vacuole, while more than 50% of cells had an electron-lucent central vacuole. These results indicate thatB. hominis accumulated carbohydrates and lipids in the central vacuole during cell growth and that the organism probably consumed these metabolic substances during stationary growth. Therefore, it is strongly suggested that the central vacuole is an important organelle for storage of metabolic substances, such as carbohydrates and lipids, required for cell growth.

Blastocystis is a common unicellular anaerobic eukaryote that inhabits the large intestine of many animals worldwide, including humans. The finding of Blastocystis in faeces in mammals and birds has led to proposals of zoonotic potential and that these hosts may be the source of many human infections. Blastocystis is, however, a genetically diverse complex of many distinct organisms (termed subtypes; STs), and sampling to date has been limited, both geographically and in the range of hosts studied. In order to expand our understanding of host specificity of Blastocystis STs, 557 samples were examined from various non-primate animal hosts and from a variety of different countries in Africa, Asia and Europe. STs were identified using 'barcoding' of the small subunit rRNA gene using DNA extracted either from culture or directly from faeces. The host and geographic range of several STs has thereby been greatly expanded and the evidence suggests that livestock is not a major contributor to human infection. Two new STs were detected among the barcode sequences obtained; for these, and for three others where the data were incomplete, the corresponding genes were fully sequenced and phylogenetic analysis was undertaken.

Background In patients with left ventricular hypertrophy (LVH), LV midwall fractional shortening (FS) is used as a measure of LV systolic performance that is more physiologically appropriate than conventional FS. For evaluation of LV volume and ejection fraction (EF), 2-dimensional (2D) echocardiography is more accurate than M-mode echocardiography. The purpose of this study was to assess systolic performance by midwall EF using 2D speckle tracking echocardiography (STE). Methods Sixty patients were enrolled in the study. Patients were divided into two groups with LVH (n = 30) and without LVH (control group, n = 30). LV systolic function was compared between the two groups and the relationships of left ventricular mass index (LVMI) with LV systolic parameters, including midwall EF, were investigated. Results Midwall EF in the LVH group was significantly lower than that in the control group (42.8±4.4% vs. 48.1±4.1%, p Conclusions These results show that midwall EF can be determined using 2D STE. Midwall EF can be used to monitor LV systolic dysfunction, which is not possible with conventional EF. Evaluation of midwall EF may allow assessment of new parameters of LV systolic function in patients with LV geometric variability.

The present study is aimed to identify the prevalence of Blastocystis subtypes isolated from patients in a major hospital in northeastern Thailand. A total of 562 stool samples were examined by culture technique, and 56 Blastocystis-positive samples were analyzed further by the combination of restriction fragment length polymorphism (RFLP) followed by polymerase chain reaction with sequence-tagged site primers (PCR-STS). By RFLP profiles, Blastocystis genotypes were categorized into four groups: group A (12, 21.4%), group B (32, 57.1%), group C (10, 17.9%), and group D (2, 3.6%). By PCR-STS, only four subtypes were identified. All 12 (21.4%) isolates in group A were identified as subtype 1. Similarly, all 32 (57.1%) isolates in group B were subtype 3. In group C, 10 (17.9%) samples were all subtype 7, and two samples (3.6%) in group D were both subtype 6. Of 56 Blastocystis-positive patients, 31 (55.4%) were asymptomatic and 22 (39.4%) have gastrointestinal symptoms. No significant association was observed between the Blastocystis subtypes and the clinical features. Among the Blastocystis-positive patients, the most characteristic stool samples were loose (78.6%) and soft (17.9%). In conclusion, the most common Blastocystis spp. in northeastern Thailand was subtype 3 followed by subtype 1. Relatively minor subtypes, subtype 6 and subtype 7 which are considered as avian subtypes, were found for the first time in humans in Thailand.

This report describes an obese 39-year-old man who experienced ST-segment elevation myocardial infarction with total thrombotic occlusion of the right coronary artery. Culprit vessel flow was improved by aspiration. Data suggested that myocardial infarction had resulted from paradoxical embolus via a patent foramen ovale triggered by the Mueller maneuver, which had induced negative intrathoracic pressure following an acute increase of right-heart volume in the context of obesity and sleep-disordered breathing (SDB). Obesity is increasing among younger populations and it represents a risk for SDB and thrombosis. Thus, this mechanism should be included within the differential diagnosis for myocardial infarction in young patients.

The patient was a 13-year-old male with chief complaints of exertional chest pain and dyspnea. Cardiac murmur was suspected in a medical checkup at 1 month old, at which time he was diagnosed with subvalvular aortic stenosis. He had subsequently been under follow-up observation at a nearby hospital for subvalvular aortic stenosis. He was admitted to our department for surgery due to aggravation of symptoms that had occurred over the previous year. Transthoracic echocardiography after admission showed an abnormal structure in the subvalvular aortic area, and the maximum pressure gradient between the left ventricle and aortic valve was 84 mmHg. The preoperative valve area was 0.71 cm(2), as measured by the Doppler method. Measurement of valve area by the trace method was difficult. Transesophageal echocardiography (TEE) showed a septum-like structure extending from the ventricular septum in the subvalvular area. On 3D TEE, the valve areas in the systolic and diastolic phases were 0.86 and 0.49 cm(2), respectively. Postoperative echocardiography showed resection of the structure in the subvalvular area, and the postoperative course was favorable.

This report presents the case of a 51-year-old female who was admitted to a local hospital because of a persistent headache. A diagnosis of multiple cerebral infarctions was thereafter made, but there was no evidence of either atherosclerosis or atrial fibrillation. The case was thought to be a cryptogenic stroke, however, Doppler ultrasonography of the lower extremities showed venous insufficiency. Transesophageal echocardiography revealed a secundum atrial septal defect (ASD) with a left to right shunt. Therefore, the final diagnosis was paradoxical brain emboli, and transcatheter ASD closure was successfully performed by cardiologists without any sequelae.

Blastocystis was first reported as a harmless yeast found in human stool samples in 1912. This organism has been classified into the subphylum Sporozoa and then reclassified into the subphylum Sarcodina based on the morphological and physiological characteristics. Molecular sequencing study on the SSU rRNA gene of a human Blastocystis isolate showed that Blastocystis is not monophyletic with yeasts (Saccharomyces), fungi (Neurospora), sarcodines (Naegleria, Acanthamoeba, and Dictyostelium), or sporozoans (Sarcocystis and Toxoplasma) based on the partial sequence of the gene. Phylogenetic studies inferred with the entire sequence of the SSU rRNA gene of a human and a guinea pig isolates of Blastocystis revealed that the organism is a member of the stramenopiles. Furthermore, accumulation of the molecular data of human and animal Blastocystis isolates showed not only extensive genetic diversity among Blastocystis isolates from the same hosts but also genetic similarities were observed among the isolates from different hosts. Therefore, a consensus terminology of Blastocystis sp. is recently proposed for the isolates from mammalian and avian hosts including humans and it classifies all isolates into nine subtypes. Recently, however, several additional novel subtypes have been reported based on the phylogenetic studies of the partial sequences of the SSU rRNA gene. Therefore, the genus Blastocystis may be the most genetically polymorphic organism that parasitizes a wide range of animals.

This chapter represents a list of papers that were considered by the authors of this book as important “breakthroughs” in the knowledge of the different features in the history of the Blastocystis research during the last 101 years. Of course this is a selection, but due to space, others were not considered in this chapter, but belong to the basic knowledge collected in the other chapters of this book and are cited at the end of each chapter.

Blastocystis, an anaerobic unicellular eukaryotic protozoon, is one of the most encountered microorganisms in fecal samples of various host animals including humans. The parasite is generally spherical in shape but shows a great variation in size from 5 to 50 μm. When the isolates from the feces were cultured in vitro, the size of the cell increases and some cells reach up more than 200 μm in diameter. Since a large central vacuole occupies the central part of the organisms, the cytoplasm containing nucleus, mitochondria-like organelle, and Golgi apparatus is pushed to the peripheral rim. The central vacuole possesses storage function and contains various substances such as carbohydrates, lipids, and basic proteins identified under the light and electron microscopy. These substances are accumulated in the vacuole via Golgi apparatus or crathrin-based endocytosis. There are various naming for morphologically different organisms, such as vacuolar (vacuolated), multivacuolar, avacuolar, granular, amoeboid, and cyst forms, which appear in fecal and in vitro samples. The vacuolar and granular forms are predominantly detected in the fresh feces or in vitro cultures, while some other forms, multivacuolar and avacuolar forms are rare and small in size and are mainly characterized by electron microscopy. The amoeboid (ameba) forms are also small in size and are generally more frequently observed in the samples of in vitro culture. Irregular shape of the amoeboid form is difficult to be distinguished from the amoebic forms of other intestinal protozoa, and locomotion of the amoeboid form of Blastocystis has not been recognized. The cyst forms are mainly seen in the fecal samples and rarely in in vitro cultures. The nuclei in vacuolar and granular forms can be clearly seen when the cells are stained with DAPI or Giemsa. In general, small cells show one or two nuclei, while bigger cells show many nuclei. When there were two nuclei in a cell, both the nuclei were usually located at the opposite pole of the cell. The life cycle of Blastocystis is still not clear. The most conflicting mode is the reproductive stages in the proposed life cycle. Binary fission is frequently observed in fresh fecal and cultured samples. However, other reproductive stages such as schizogony, plasmotomy (budding), endodyogeny, and sac-like pouches are not accepted generally for the true modes of Blastocystis. Schizogony-like and budding-like organisms, which possess many nuclei in the peripheral cytoplasm or have many daughter cell-like structures in the central vacuole, are observed especially in in vitro cultures. Many reproductive processes have been proposed for Blastocystis, however, to date, only binary fission, budding, or plasmotomy have been proven. Pseudopodal activity, on a few occasions, is seen in the amoeboid forms, and this form may be proposed as another alternative reproduction process except three identified modes of reproduction, binary fission, plasmotomy, and budding (or schizogony) under ultrastructural insights while locomotion of the amoeboid form has not still been confirmed.

The genus Blastocystis is one of the most frequently encountered protozoans in human and animal fecal samples. In general, prevalence of Blastocystis infection in humans is higher in developing countries than in developed countries. However, detectability of the parasite is closely related to the diagnostic methods. Namely, routine parasitological diagnostic methods including direct fecal smear with or without staining and formalin–ether concentration methods are known to be considerably lower detection or almost undetectable. Therefore, suitable methods should be used for diagnosis of this parasite. The actual mode of transmission of Blastocystis between human-to-human, human-to-animal, and animal-to-animal is not conclusively demonstrated. However, the direct or indirect transmission of the parasite via fecal–oral route has been demonstrated between individuals in the closed human communities and among experimental animals in the same gages. In addition, only cyst form is known to be responsible for transmission. Therefore, contamination of fecal materials in drinking water or foods seems to be the most possible route for infection. Indeed, viable Blastocystis cysts have been found in the source of drinking water. Although zoonotic potential of the parasite has been proposed based on the genetic similarities among Blastocystis isolates from humans and animals, positive infections of some human isolates in rats and chickens indicate the zoonotic feature of this parasite. However, there are variability in the infectivity of the isolates in relation to genetic diversity of Blastocystis.

Blastocystis sp. is now recognized as one of the most common intestinal parasite in human fecal examinations. Recently, PCR-based diagnostic methods of Blastocystis infection using direct DNA extraction from fresh fecal samples with commercially available kits are reported. Several kits have been developed, but little has been done in comparing the detective sensitivity between PCR methods using the commercial kits. In this study, we compared the detective sensitivity among five commercially available kits (MagNA Pure LC DNA Isolation Kit I, Roche; QuickGene SP Kit DNA, FujiFilm; NucleoSpin Plant II, Macherey-Nagel; QIAamp DNA Stool Mini Kit, Qiagen; ZR Fecal DNA Kit, Zymo Research) and fecal culture method. In a preliminary test, the DNA isolated with two kits (FujiFilm and Macherey-Nagel) showed negative PCR, while the other three kits showed positive PCR. Then, DNA from 50 clinical samples that was Blastocystis-positive in the examination of fecal culture method were isolated with the three kits and 1.1 kbp SSU rRNA gene was detected with PCR. The positive rates of the three kits (Roche, Qiagen, and Zymo Research) were 10, 48 and 94%, respectively. The present study indicated that there is different detective sensitivity among the commercial kits, and fecal culture method is superior in detection rate and cost performance than DNA-elution kits for diagnosis of Blastocystis sp. subtypes.

The mode and organization of the attachment site of Cryptosporidium muris to gastric glands of stomach were investigated by the freeze-fracture method. Cryptosporidium muris was enveloped by a double membrane, of host plasma membrane origin, which formed the parasitophorous vacuole. The outer membrane of the double membrane was continuous with host plasma membrane, while the inner membrane was connected with the anterior part of the parasite plasma membrane at the annular ring. The density of intramembranous particles (IMP) was severely altered at the above two junctures. The parasitophorous outer membrane showed low IMP-density when compared to the host plasma membrane, although both membranes were continuous at the dense band. The inner membrane had few IMP, whereas the parasite plasma membrane showed numerous IMP, although both membranes were continuous at the annular ring. The size of dense band and annular ring was similar in diameter. The feeder organelle was clearly visible as membrane folds in freeze-fracture and some of them were connected with small vesicles of cytoplasm, indicating that the feeder organelle may play an important role for incorporation of nutrients from the host cell.

Blastocystis infection has been reported to be associated with irritable bowel syndrome (IBS), inflammatory bowel disease (IBD) and chronic diarrhoea. The availability of data on the subtypes of Blastocystis found in these patient groups would be of interest in understanding the significance of Blastocystis infection in chronic illness. In this study, we identify Blastocystis subtypes found in patients presenting with IBS, IBD, chronic diarrhoea and asymptomatic patients in Ankara, Turkey. Blastocystis was detected in 11 symptomatic patients by microscopy and 19 by stool culture. Stool culture was more sensitive than microscopy in identifying Blastocystis. Using standard nomenclature adopted in 2007, Blastocystis sp. subtype 3 was the most common in all groups, followed by Blastocystis sp. subtype 2. Identical subtypes of Blastocystis are found in patients with IBS, IBD and chronic diarrhoea. These particular subtypes show low host specificity and are carried by humans and some farm animals. The subtypes of Blastocystis that are commonly found in rodents and certain wild birds were not found in these patients. We suggest a model in which the severity of enteric protozoan infection may be mediated by host factors.

Blastocystis hominis is a zoonotic intestinal protozoan parasite whose pathogenic potential is still controversial. The aim of the present study was to clarify the pathogenicity of Blastocystis parasites in rats. Oral inoculation with 1 x 10(5) cysts of Blastocystis sp. strain RN94-9 in rats resulted in chronic infection in the cecum at least until 4 weeks after infection. Histological examination revealed neither mucosal sloughing nor inflammatory cell infiltration but showed a slight but significant increase in goblet cell numbers in the cecal mucosa 1-3 weeks post-infection. Differential staining of acidic and neutral mucins by the alcian blue-periodic acid-Schiff method showed that the predominantly increased cells were neutral mucin(+) but not acidic mucin(+) goblet cells. Reverse transcription real-time polymerase chain reaction studies demonstrated significant upregulation of the expression of interferon-gamma, interleukin (IL)-12, and tumor necrosis factor alpha, but not IL-6 or granulocyte-macrophage colony-stimulating factor, in the cecal mucosa at 2 and/or 3 weeks post-infection. The induction of local host responses, including mild goblet cell hyperplasia, and significant upregulation of type-1 and proinflammatory cytokines, suggest that Blastocystis sp. strain RN94-9 is a weakly pathogenic organism that could elicit proinflammatory as well as protective responses in local tissues.