Cerebral cystic echinococcosis in Mongolian children caused by Echinococcus canadensis

Recent molecular re-evaluation of Echinococcus granulosus, which causes cystic echinococcosis (CE), has revealed that it is not a single species, but instead consists of 5 cryptic species. Among them, E. granulosus (dog-sheep strain) is predominant (75%) followed by Echinococcus canadensis (22%). The major affected organs, in humans, are the liver (88%) and lungs (11%). Primary cerebral CE comprises less than 1% of all cases. As cerebral CE cases are rare, there are few reports with molecular confirmation of the causative species. This study reports mitochondrial gene analysis from 4 Mongolian pediatric cerebral CE cases. Molecular confirmation was obtained for 3 of the 4 cases, with all 3 cases determined to be due to E. canadensis (G6/G7) infection. None of the cases had other organ involvement. This is only the third report on the molecular identification of the Echinococcus species responsible for cerebral CE, and only the second report of E. canadensis (G6/G7) being the causative agent of cerebral CE.     

First assessment of the population structure of the Asiatic wild ass in Mongolia

The Mongolian Gobi is the most important stronghold of the Asiatic wild ass (Equus hemionus), housing >?75% of the remaining global population. However, even in this remote refuge, poaching and changes in land use are challenging the species’ conservation. Whereas progress has been made in monitoring population size, little data are available on population structure and dynamics. To fill this gap, we determined the age and sex of 440 skulls collected in two regions of the Mongolian Gobi. Foals and yearlings were underrepresented in our skull sample with 3 and 7.3% only, but the rest of the age pyramid was well balanced. The mean age was 7.7 years, the maximum age 29 years, and the sex ratio was not different from even. Mortality risk analysis revealed low annual mortality rates of about 15% in the most productive age classes of 5–10 years, followed by a slow increase with age until about 17 years and a likely faster increase thereafter. As the large majority of carcasses suggested a poaching-related mortality which appears random, our dataset provides the first insight into the structure of the largest remaining Asiatic wild ass population and can be used as a benchmark for future monitoring and population viability modeling.     

Changes in fatigue,autonomic functions,and blood biomarkers due to sitting isometric yoga in patients with chronic fatigue syndrome

Background

In a previous randomized controlled trial, we found that sitting isometric yoga improves fatigue in patients with chronic fatigue syndrome (CFS) who are resistant to conventional therapy. The aim of this study was to investigate possible mechanisms behind this finding, focusing on the short-term fatigue-relieving effect, by comparing autonomic nervous function and blood biomarkers before and after a session of isometric yoga.

Methods

Fifteen patients with CFS who remained symptomatic despite at least 6 months of conventional therapy practiced sitting isometric yoga (biweekly 20 min practice with a yoga instructor and daily home practice) for eight weeks. Acute effects of sitting isometric yoga on fatigue, autonomic function, and blood biomarkers were investigated after the final session with an instructor. The effect of a single session of sitting isometric yoga on fatigue was assessed by the Profile of Mood Status (POMS) questionnaire immediately before and after the session. Autonomic nervous function (heart rate (HR) variability) and blood biomarkers (cortisol, DHEA-S, TNF-α, IL-6, IFN-γ, IFN-α, prolactin, carnitine, TGF-β1, BDNF, MHPG, and HVA) were compared before and after the session.

Results

Sitting isometric yoga significantly reduced the POMS fatigue score (p?<?0.01) and increased the vigor score (p?<?0.01). It also reduced HR (p?<?0.05) and increased the high frequency power (p?<?0.05) of HR variability. Sitting isometric yoga increased serum levels of DHEA-S (p?<?0.05), reduced levels of cortisol (p?<?0.05) and TNF-α (p?<?0.05), and had a tendency to reduce serum levels of prolactin (p?<?0.1). Decreases in fatigue scores correlated with changes in plasma levels of TGF-β1 and BDNF. In contrast, increased vigor positively correlated with HVA.

Conclusions

A single session of sitting isometric yoga reduced fatigue and increased vigor in patients with CFS. Yoga also increased vagal nerve function and changed blood biomarkers in a pattern that suggested anti-stress and anti-inflammatory effects. These changes appear to be related to the short-term fatigue-relieving effect of sitting isometric yoga in patients with CFS. Furthermore, dopaminergic nervous system activation might account for sitting isometric yoga-induced increases in energy in this patient population.

Trial registration

University Hospital Medical Information Network (UMIN CTR) UMIN000009646. Registered Dec 27, 2012.

     

Isometric yoga improves the fatigue and pain of patients with chronic fatigue syndrome who are resistant to conventional therapy: a randomized,controlled trial

Background

Patients with chronic fatigue syndrome (CFS) often complain of persistent fatigue even after conventional therapies such as pharmacotherapy, cognitive behavioral therapy, or graded exercise therapy. The aim of this study was to investigate in a randomized, controlled trial the feasibility and efficacy of isometric yoga in patients with CFS who are resistant to conventional treatments.

Methods

This trial enrolled 30 patients with CFS who did not have satisfactory improvement after receiving conventional therapy for at least six months. They were randomly divided into two groups and were treated with either conventional pharmacotherapy (control group, n?=?15) or conventional therapy together with isometric yoga practice that consisted of biweekly, 20-minute sessions with a yoga instructor and daily in-home sessions (yoga group, n?=?15) for approximately two months. The short-term effect of isometric yoga on fatigue was assessed by administration of the Profile of Mood Status (POMS) questionnaire immediately before and after the final 20-minute session with the instructor. The long-term effect of isometric yoga on fatigue was assessed by administration of the Chalder’s Fatigue Scale (FS) questionnaire to both groups before and after the intervention. Adverse events and changes in subjective symptoms were recorded for subjects in the yoga group.

Results

All subjects completed the intervention. The mean POMS fatigue score decreased significantly (from 21.9?±?7.7 to 13.8?±?6.7, P?<?0.001) after a yoga session. The Chalder’s FS score decreased significantly (from 25.9?±?6.1 to 19.2?±?7.5, P?=?0.002) in the yoga group, but not in the control group. In addition to the improvement of fatigue, two patients with CFS and fibromyalgia syndrome in the yoga group also reported pain relief. Furthermore, many subjects reported that their bodies became warmer and lighter after practicing isometric yoga. Although there were no serious adverse events in the yoga group, two patients complained of tiredness and one of dizziness after the first yoga session with the instructor.

Conclusions

Isometric yoga as an add-on therapy is both feasible and successful at relieving the fatigue and pain of a subset of therapy-resistant patients with CFS.

Trial registration

University Hospital Medical Information Network (UMIN CTR) UMIN 000009646.

     

Analysis of geographic centrality and genetic diversity in the declining grasshopper species Bryodemella tuberculata (Orthoptera: Oedipodinae)

Human-induced ecological and climatic changes have led to the decline and even local extinction of many formerly widely distributed temperate and cold-adapted species. Determining the exact causes of this decline remains difficult. Bryodemella tuberculata was a widely distributed orthopteran species before the mid-19th century. Since then, many European populations have suffered drastic declines and are now considered extinct or critically endangered. We used ecological niche modelling based on a large dataset of extant and extinct occurrence data to investigate whether poor climatic suitability in the periphery of its global range was a possible cause of the local extinction of the European populations of B. tuberculata. We also used population genetics based on the COI marker to estimate and compare the genetic diversity of extant populations. We found that Europe still provides highly suitable habitats close to the climatic optimum, contradicting the assumption of climate change as major driver of this decline. Instead, changes in land-cover and other anthropogenic modifications of the habitats at the local scale seem to be the major reasons for local extinctions. Genetic analysis suggests Central Asia as center of diversity with a stable population size, whereas the effective sizes of the remaining European populations are decreasing. We found European genetic lineages nested within Central Asian lineages, suggesting a Central Asian source distribution area. Our results suggest that the declining European populations represent relics of a formerly wider distribution, which was fragmented by changes in land-use. These relics are now threatened by limited connectivity and small effective population sizes. Specific conservation actions, such as the restoration of former or potential new habitats, and translocation of individuals from extant populations to these restored sites may help slow, stall, or even revert the extinction process.

     

A Comparative Analysis of Constitutive Promoters Located in Adeno-Associated Viral Vectors

The properties of constitutive promoters within adeno-associated viral (AAV) vectors have not yet been fully characterized. In this study, AAV vectors, in which enhanced GFP expression was directed by one of the six constitutive promoters (human β-actin, human elongation factor-1α, chicken β-actin combined with cytomegalovirus early enhancer, cytomegalovirus (CMV), simian virus 40, and herpes simplex virus thymidine kinase), were constructed and introduced into the HCT116, DLD-1, HT-1080, and MCF-10A cell lines. Quantification of GFP signals in infected cells demonstrated that the CMV promoter produced the highest GFP expression in the six promoters and maintained relatively high GFP expression for up to eight weeks after infection of HCT116, DLD-1, and HT-1080. Exogenous human CDKN2A gene expression was also introduced into DLD-1 and MCF-10A in a similar pattern by using AAV vectors bearing the human β-actin and the CMV promoters. The six constitutive promoters were subsequently placed upstream of the neomycin resistance gene within AAV vectors, and HCT116, DLD-1, and HT-1080 were infected with the resulting vectors. Of the six promoters, the CMV promoter produced the largest number of G418-resistant colonies in all three cell lines. Because AAV vectors have been frequently used as a platform to construct targeting vectors that permit gene editing in human cell lines, we lastly infected the three cell lines with AAV-based targeting vectors against the human PIGA gene in which one of the six promoters regulate the neomycin resistance gene. This assay revealed that the CMV promoter led to the lowest PIGA gene targeting efficiency in the investigated promoters. These results provide a clue to the identification of constitutive promoters suitable to express exogenous genes with AAV vectors, as well as those helpful to conduct efficient gene targeting using AAV-based targeting vectors in human cell lines.     

A western Eurasian male is found in 2000‐year‐old elite Xiongnu cemetery in Northeast Mongolia

We analyzed mitochondrial DNA (mtDNA), Y‐chromosome single nucleotide polymorphisms (Y‐SNP), and autosomal short tandem repeats (STR) of three skeletons found in a 2,000‐year‐old Xiongnu elite cemetery in Duurlig Nars of Northeast Mongolia. This study is one of the first reports of the detailed genetic analysis of ancient human remains using the three types of genetic markers. The DNA analyses revealed that one subject was an ancient male skeleton with maternal U2e1 and paternal R1a1 haplogroups. This is the first genetic evidence that a male of distinctive Indo‐European lineages (R1a1) was present in the Xiongnu of Mongolia. This might indicate an Indo‐European migration into Northeast Asia 2,000 years ago. Other specimens are a female with mtDNA haplogroup D4 and a male with Y‐SNP haplogroup C3 and mtDNA haplogroup D4. Those haplogroups are common in Northeast Asia. There was no close kinship among them. The genetic evidence of U2e1 and R1a1 may help to clarify the migration patterns of Indo‐Europeans and ancient East‐West contacts of the Xiongnu Empire. Artifacts in the tombs suggested that the Xiongnu had a system of the social stratification. The West Eurasian male might show the racial tolerance of the Xiongnu Empire and some insight into the Xiongnu society. Am J Phys Anthropol, 2010. © 2010 Wiley‐Liss, Inc.     

Simple Monitoring of Gene Targeting Efficiency in Human Somatic Cell Lines Using the PIGA Gene

Gene targeting in most of human somatic cell lines has been labor-intensive because of low homologous recombination efficiency. The development of an experimental system that permits a facile evaluation of gene targeting efficiency in human somatic cell lines is the first step towards the improvement of this technology and its application to a broad range of cell lines. In this study, we utilized phosphatidylinositol glycan anchor biosynthesis class A (PIGA), a gene essential for the synthesis of glycosylphosphatidyl inositol (GPI) anchors, as a reporter of gene targeting events in human somatic cell lines. Targeted disruption of PIGA was quantitatively detected with FLAER, a reagent that specifically binds to GPI anchors. Using this PIGA-based reporter system, we successfully detected adeno-associated virus (AAV)-mediated gene targeting events both with and without promoter-trap enrichment of gene-targeted cell population. The PIGA-based reporter system was also capable of reproducing previous findings that an AAV-mediated gene targeting achieves a remarkably higher ratio of homologous versus random integration (H/R ratio) of targeting vectors than a plasmid-mediated gene targeting. The PIGA-based system also detected an approximately 2-fold increase in the H/R ratio achieved by a small negative selection cassette introduced at the end of the AAV-based targeting vector with a promoter-trap system. Thus, our PIGA-based system is useful for monitoring AAV-mediated gene targeting and will assist in improving gene targeting technology in human somatic cell lines.     

Technical note: improved ancient DNA purification for PCR using ion-exchange columns

A novel method of ancient DNA (aDNA) purification was developed using ion-exchange columns to improve PCR-amplifiable DNA extraction from ancient bone samples. Thirteen PCR-resistant ancient bone samples aged 500-3,300 years were tested to extract aDNA using a recently reported, silica-based aDNA extraction method and an ion-exchange column method for the further purification. The PCR success rates of the aDNA extracts were evaluated for the amplification ability of the fragments of mitochondrial DNA, a high-copy DNA, and amelogenin, a low-copy DNA. The results demonstrate that the further purification of silica-based aDNA extracts using ion-exchange columns considerably improved PCR amplification. We suggest that the ion-exchange column-based method will be useful for the improvement of PCR-amplifiable aDNA extraction, particularly from the poorly preserved, PCR-resistant, ancient samples.