We studied two patients from a nonconsanguineous family with life-long abnormal liver function, hepatomegaly and abnormal fatty acid profiles. Abnormal liver function, hypoglycemia and muscle weakness are observed in various genetic diseases, including medium-chain acyl-CoA dehydrogenase (MCAD) deficiency and glycogen storage diseases. The proband showed increased free fatty acids, mainly C8 and C10, resembling fatty acid oxidation disorder. However, no mutation was found in ACADM and ACADL gene. Sequencing of theamylo-alpha-1, 6-glucosidase, 4-alpha-glucanotransferase (AGL) gene showed that both patients were compound heterozygotes for c.118C>T (p.Gln40X) and c.753_756 del CAGA (p.Asp251Glufsx29), whereas their parents were each heterozygous for one of these mutations. The AGL protein was undetectable in EBV-B cells from the two patients. Transcriptome analysis demonstrated a significant different pattern of gene expression in both of patients’ cells, including genes involving in the PPAR signaling pathway, fatty acid biosynthesis, lipid synthesis and visceral fat deposition and metabolic syndrome. This unique gene expression pattern is probably due to the absence of AGL, which potentially accounts for the observed clinical phenotypes of hyperlipidemia and hepatocyte steatosis in glycogen storage disease type IIIa.

Tomato (Lycopersicumesculentum) is one of the important horticultural commodity unfortunately the production is still low in quantity or quality. One of the important problems for the low productivity is the bacterial wilt disease caused by Ralstonia solanacearum. Some control methods have been developed but the disease still caused damages. One alternative control which can be prospective to be developed is the use of biocontrol agents. Among them the endophytic bacteria and Plant Growth Promoting Rhizobacteria (PGPR). The experiment was conducted to evaluate the viability of biocontrol agents, i.e. S.epidermidis BC4 and P. fluorescens RH4003 in the formula during storage and 3 the effectiveness of biopesticides formulation to control the bacterial wilt disease on tomato. Biopesticides were applied to the root during transplanting. Disease incidence and the height of the plant were calculated every week. Population of the bacteria in the formula was calculated at 0, 2, 4, and 8 weeks during storage. P. fluorescens RH4003 in granule formula applied into the soil caused disease suppression index up to 46.15%, which significantly lower compared with control, while S. epidermidis BC4 in liquid formulation with pH 5 caused disease suppression index up to 57.69%. Formula containing both of the bacteria did not affect the height growth of tomato plant. Population of the bacteria in the formula grew well at pH 5, 6, 7 and it was stable until 8 weeks of storage.

Fabry disease is a monogenic X-linked lysosomal storage disease caused by α-galactosidase A (αGalA) deficiency. Enzyme replacement therapy through administration of the missing αGalA is currently the only accepted therapeutic option. However, this treatment is connected to high costs, has ill-defined indication criteria and its efficacy is controversially discussed. Our aim was to explore the possibility of a novel targeted substrate reduction therapy for Fabry disease. Owing to the fact that αGalA-deficient humans and mice accumulate the same glycosphingolipids (i.e. globosides, galabiosylceramide and isoglobosides), αGalA-deficient mice were crossed with mice deficient in enzymes synthesizing these classes of glycosphingolipids (i.e. globotrihexosylceramide and isoglobotrihexosylceramide synthase, respectively). Functional heart and kidney tests were performed together with an extensive biochemical analysis of urine and serum in aged mice. Lysosomal storage was assessed by thin layer chromatography and electron microscopy. We showed that depletion of globosides was sufficient to fully abolish the storage of glycosphingolipids in heart, kidney and liver and was paralleled by a complete restoration of lysosomal morphology in these organs. In contrast, in dorsal root ganglia, a depletion of both globosides and isoglobosides was necessary to fully counteract the lysosomal storage. The deficiency in globosides and/or isoglobosides did not cause any adverse effects. We conclude that substrate reduction therapy through inhibition of the synthesis of globosides and isoglobosides represents a valuable therapeutic option for Fabry disease, all the more as globosides and isoglobosides seem to be dispensable.

In the spring of 2013 a five year old bear kept in Kalvene Zoo (Nature Park of Riga Zoo) developed sudden motor dysfunction, loss of coordination and difficulty to eat. During summer neurological deficits progressed and bear was killed in September of 2013. Postmortem necropsy performed at FVM showed no gross lesions in the brain, spinal cord or muscles. Microscopic examination revealed accumulation of large amount of fine eosinophilic-yellow vacuoles in nearly neurons of all levels of brain and spinal cord. Neurons were often fragmented, necrotic or lost. Vacuoles were intensively positive by LFB stain and moderately positive for PAS stain confirming lysosomal storage disease. The appearance of vacuoles and staining properties suggested neurodegenerative storage disease -- neuronal ceroid lipofuscinosis. Neuronal ceroid lipofuscinosis (NCL) is a lysosomal storage disease characterized by accumulation of lipopigment within central nervous system and peripheral tissues. This occurs due to mutations in genes that code for a variety of proteins including lysosomal enzymes and membranes of various compartments. Accumulation of lipopigment is progressive and cumulative, resulting in disruption of neuronal integrity and impairing neuronal functions. The bear affected by NCL was offspring of close sibling / parent matings several generations in a row which likely facilitated expression of this recessively inherited disease. NCL is rare disease best characterized in humans but also affecting domestic animals – most commonly dogs, cats, sheep and cattle.

Gaucher disease (GD) and Fabry disease (FD) are two relatively common inherited glycosphingolipidoses caused by deficiencies in the lysosomal glycosidases glucocerebrosidase and alpha-galactosidase A, respectively. For both diseases enzyme supplementation is presently used as therapy. Cells and tissues of GD and FD patients are uniformly deficient in enzyme activity, but the two diseases markedly differ in cell types showing lysosomal accumulation of the glycosphingolipid substrates glucosylceramide and globotriaosylceramide, respectively. The clinical manifestation of Gaucher disease and Fabry disease is consequently entirely different and the response to enzyme therapy is only impressive in the case of GD patients. This review compares both glycosphingolipid storage disorders with respect to similarities and differences. Presented is an update on insights regarding pathophysiological mechanisms as well as recently available biochemical markers and diagnostic tools for both disorders. Special attention is paid to sphingoid bases of the primary storage lipids in both diseases. The value of elevated glucosylsphingosine in Gaucher disease and globotriaosylsphingosine in Fabry disease for diagnosis and monitoring of disease is discussed as well as the possible contribution of the sphingoid bases to (patho)physiology. This article is part of a Special Issue entitled New Frontiers in Sphingolipid Biology.

Phacidiopycnis washingtonensis is the cause of speck rot, a recently reported postharvest fruit rot disease of apple. The pathogen is believed to incite infections in the field, and disease symptoms become evident only during storage. To determine the timing of apple fruit infection in relation to development of speck rot in storage, ‘Red Delicious’ and ‘Fuji’ apple fruit were inoculated in the orchard with P. washingtonensis at different times during the growing season, harvested, and monitored for decay development during storage at 0°C. Fruit inoculated in both field and laboratory also were used to identify the infection courts and mode of apple fruit penetration by P. washingtonensis. In all 3 years, stem-end speck rot and calyx-end speck rot developed during cold storage on fruit inoculated during the growing season, regardless of inoculation time; and the incidence of total speck rot in storage increased as the fruit inoculation time approached harvest. On fruit floral parts, the pathogen colonized sepals at higher rates than stamens. Availability of naturally occurring necrotic tissues favored the colonization of the fungus on sepals. Histological studies indicated that infection occurred through micro-cracks on the surfaces of pedicels and sepals of the fruit, and invasion of these tissues was restricted between the cuticle and epidermis. Findings of this study will assist in the development of effective control strategies for speck rot.

Ralstonia solancearum is a pathogen of bacterial wilt disease on tomato. One of the alternatives for disease control is the application of biocontrol agents. Biocontrol agents which have been able to control the disease are Bacillus subtilis AB89 (PGPR) and Staphylococcus epidermidis BC4 (endophyte). This research was conducted to evaluate the viability of those bacteria in formulation and the effectiveness of the formulation to control the bacterial wilt disease of tomato. B. subtilis AB89 (BS) and S. epidermidis BC4 (BC) ware formulated singly in liquid (BS and BC) and powder (TBS and TBC) formulation contained talc powder and/or xanthan gum. The liquid formulation of B. subtilis AB89 (BS) and S. epidermidis BC4 (BC) effectively suppressed the incidence of the disease, with control effectiveness was up to 55.55% and 54.94%, respectively. The liquid formulation BC, BS and the powder formulation TBS were able to promote the plant growth. Populations of S. epidermidis BC4 was stable within powder and liquid formulations during 8 weeks of storing. Population of B. subtilis AB89 in powder and liquid formulations declined on 1 week after storage, but stable until 8 weeks after storage.

Seeking new yeast strains having the ability to protect apple fruits against blue mould for a long time under different storage conditions was the main goal of this work. Based on the in vitro test, yeast strains KKUY0017 and KKUY0051 were selected as the most effective antagonists against Penicillium expansum . Sequencing of 26S rDNA of both yeasts confirmed that the identity of KKUY0017 and KKUY0051 was Cryptococcus albidus and Wickerhamomyces anomalus , respectively. The two strains protected the apple fruits from the blue mould disease under a wide range of temperature (5–30°C); however, W. anomalus KKUY0051 was more effective. At 25°C, W. anomalus KKUY0051 involved in the reduction of disease severity and disease incidence of blue mould by 56.49% and 57.78%, respectively. When either of the two yeasts was applied in concentration of 10 ⁸ or 10 ⁹ cells/mL, the maximum reduction in disease severity and disease incidence was achieved. Under cold storage (5°C), both yeast strains succeeded to protect the apple fruits free from the infection up to 24 days. Electron micrograph showed a fit attachment between the cells of C. albidus KKUY0017 and the fungal hyphae leading to the degrading of the hyphae; however, W. anomalus killed the fungal hyphae without direct attachment to them. Gas chromatography–mass spectrometry analysis of the cell-free extract of W. anomalus KKUY0051 revealed the presence of toxic compounds such as the nitrophenol derivatives. The results support the assumption that the main mode of action of this yeast is by killer toxins. We conclude that application of these yeasts under cold storage condition could keep the apple fruits free from blue mould infection for a long time.

Hemochromatosis, or iron storage disease, has been associated with significant liver disease and mortality in captive Egyptian fruit bats (Rousettus aegyptiacus). The physiologic basis for this susceptibility has not been established. In humans, a deficiency or resistance to the iron regulatory hormone, hepcidin has been implicated in the development of hereditary hemochromatosis. In the present study, we compared the coding sequence of the hepcidin gene in eight species of bats representing three distinct taxonomic families with diverse life histories and dietary preferences. Bat hepcidin mRNA encoded a 23 amino acid signal peptide, a 34 or 35 amino acid pro-region, and a 25 amino acid mature peptide, similar to other mammalian species. Differences in the sequence of the portion of the hepcidin gene that encodes the mature peptide that might account for the increased susceptibility of the Egyptian fruit bat to iron storage disease were not identified. Variability in gene sequence corresponded to the taxonomic relationship amongst species.

National audience | Pompe disease (glycogen storage disease type II) is a lysosomal storage disorder caused by acidalpha- glucosidase (GAA) deficiency leading to progressive accumulation of glycogen in the heart, muscles, and central nervous system (CNS). The disease manifests as a fatal cardiomyopathy in infantile form. Cardiac correction by enzyme replacement therapy (ERT) has recently prolonged the lifespan of these patients, revealing a new natural history. The emergent neurologic phenotype and the poor correction of skeletal muscles in survivors are currently partly attributed to CNS glycogen storage, uncorrected by ERT. We evaluated a gene therapy strategy using the neurotropic and cardiotropic AAV serotype 9 injected intrathecally (ie in the cerebrospinal fluid) to restore GAA activity in the CNS and the heart. GAA-KO mice were injected with AAV9-gaa into the cisterna magna at one month. Their neurologic and motor skills were periodically monitored from three to twelve months by hind limb clasping reflex, brainstem auditory evoked potentials, wire-hang test, and accelerating rotarod; cardiac function was assessed by M-mode echocardiography at 12 months. Glycogen content, GAA activity, and disease-related pathology were assessed in the CNS and heart at end-point. We also used real-time RT-PCR to examine transcriptional markers of cardiomyopathy and denervation atrophy in the heart and muscles respectively. We demonstrate a significant functional neurologic correction in treated animals from 4 months onward, a neuromuscular improvement from 9 months onward, and a correction of the hypertrophic cardiomyopathy at 12 months. The regions most affected by the disease i.e the brainstem, spinal cord, spinal ganglia, and the left cardiac ventricular wall all show enzymatic, biochemical and histological correction. This unprecedented global and long-term CNS and cardiac cure offer new perspectives for the management of patients.