Secondary Accumulation Research Articles

Pigmented ependymoma, a tumor with predilection for the middle-aged adult: case report with methylation classification and review of 16 literature cases.

Ependymomas have rarely been described to contain pigment other than melanin, neuromelanin, lipofuscin or a combination. In this case report, we present a pigmented ependymoma in the fourth ventricle of an adult patient and review 16 additional cases of pigmented ependymoma from the literature. A 46-year-old female showed up with hearing loss, headaches, and nausea. Magnetic resonance imaging revealed a 2.5 cm contrast-enhancing cystic mass in the fourth ventricle, which was resected. Intraoperatively, the tumor appeared grey-brown, cystic, and was adherent to the brainstem. Routine histology revealed a tumor with true rosettes, perivascular pseudorosettes and ependymal canals consistent with ependymoma, but also showed chronic inflammation and abundant distended pigmented tumor cells that mimicked macrophages in frozen and permanent sections. The pigmented cells were positive for GFAP and negative for CD163 consonant with glial tumor cells. The pigment was negative for Fontana-Masson, positive for Periodic-acid Schiff and autofluorescent, which coincide with characteristics of lipofuscin. Proliferation indices were low and H3K27me3 showed partial loss. H3K27me 3 is an epigenetic modification to the DNA packaging protein Histone H3 that indicates the tri-methylation of lysine 27 on histone H3 protein. This methylation classification was compatible with a posterior fossa group B ependymoma (EPN_PFB). The patient was clinically well without recurrence at three-month post-operative follow-up appointment. Our analysis of all 17 cases, including the one presented, shows that pigmented ependymomas are most common in the middle-aged with a median age of 42 years and most have a favorable outcome. However, one patient that also developed secondary leptomeningeal melanin accumulations died. Most (58.8%) arise in the 4th ventricle, while spinal cord (17.6%) and supratentorial locations (17.6%) were less common. The age of presentation and generally good prognosis raise the question of whether most other posterior fossa pigmented ependymomas may also fall into the EPN_PFB group, but additional study is required to address that question.

Litho‐mineralogy, geochemistry, and chronology for the genesis of the Kamust sandstone‐hosted uranium deposit, Junggar Basin, NW China

The Kamust sandstone‐hosted uranium deposit was recently discovered within the Middle Jurassic Toutunhe Formation of the eastern Junggar Basin. In this study, polarizing microscope and scanning electron microscope observation, electron microprobe analysis, and laser ablation inductively coupled plasma mass spectrometry measurement were carried out to define the petrology, mineralogy, and geochemistry of these sandstone‐hosted uranium ores. In addition, sulphur isotope signatures for the associated ore‐stage pyrites and the ore‐forming age were determined. The results show that the mineralized sandstones, which exhibit excellent reducibility with abundant organic matter and pyrites, are characterized by the secondary accumulation of uranium and are slightly enriched in rare earth elements (REE) and light REE (LREE). The uranium deposits are mainly composed of a large amount of uraninite, followed by some coffinites and uranium‐bearing titanium minerals and very few sorptive uranium‐bearing materials. Uranium minerals primarily exist within the interior and edge of clastic particles in colloidal, dispersed, and vein form, and some are closely interweaved with the colloidal and framboidal pyrites and carbonaceous detritus. Both uraninite and coffinite are strongly enriched in REE and LREE, as determined by in situ trace element measurements. Moreover, the δ34S values of the associated ore‐stage pyrites mostly range from −41.66 to −19.33‰, indicating biological origin, and light 32S demonstrates the relatively low‐temperature and open‐system mineralization processes related to the microbial anaerobic action on the organic matter. One of the δ34S values (6.88‰) is possibly caused by Rayleigh isotope fractionation due to the utilization of residual heavy 34S in the relatively later closed‐like mineralization system. The ore‐reforming ages of 27 ± 3.2 Ma, 22.4 ± 3.9 Ma, and 21.3 ± 2.3 Ma were obtained by U–Pb isotopic dating combined with U–Ra balance correction, corresponding to the rapid uplift event caused by the remote effect of the collision between the Indian and Eurasian plates at the end of the Palaeogene. In conjunction with the tectonic–sedimentary evolution of the study region, a four‐stage ore‐forming model for the Middle Jurassic Toutunhe Formation is preliminarily constructed: ① Development of ore‐bearing strata, ② initiation of the mineralization stage from the Late Jurassic to the Early Cretaceous, ③ deep burial by lacustrine and flood mudstone from the Cretaceous to the Palaeogene, and ④ the superimposed ore‐forming stage during the Neogene. These research results provide significant benefits for further uranium prospecting and in situ leach process mining in this basin.

A preliminary study of the innate immune memory of Kupffer cells induced by PEGylated nanoemulsions

The accelerated blood clearance (ABC) phenomenon describes a dilemma of polyethylene glycol (PEG) applied in drug delivery system (DDS) caused by its immunogenicity, that results in the enhanced blood clearance rate and increased hepatic and splenic accumulation after secondary injection of PEGylated nanocarriers. However, the ABC index, as the judgement of ABC phenomenon, only describes the accelerated blood clearance rate, but ignores the enhanced hepatic and splenic accumulation. Therefore, we proposed the hepatic accumulation (HA) index and the splenic accumulation (SA) index as supplements for assessing the ABC phenomenon, to emphasize the contribution of liver and spleen, especially the liver, possessing the most population of tissue resident macrophages. By altering the first injection site from the tail vein to the liver portal vein, there was no impact on anti-PEG IgM production, and the secondary hepatic accumulation of PEGylated nanoemulsions (PE) was observed to be proportionate to the first PE stimulation strength on the liver. We also determined that Kupffer cells (KCs) were the main contributor to this enhancement. On this basis, we revealed a definite phenomenon that PE could induce innate immune memory in KCs, by enhancing the phagocytosis of KCs toward PE during the secondary stimulation. The PE-stimulated KCs could carry this memory to the naïve rats through adoptive transfer, resulting in increased hepatic accumulation in the recipient rats without antibody production. Studies examining the phagocytosis of KCs in vivo, ex vivo and in vitro revealed that the memory of KCs against PE triggered by first-stimulated PE could be maintained independently of other cells or components until 21 days after the first stimulation, and possessing specificity to PEG, which was invalid to long-circulating GE (GM1 modified nanoemulsions). The discovery of immune memory in KCs induced by PE highlights the importance of focusing on the relationship between the innate immune system and PEGylated nanocarriers during the development of DDS to improve medication safety in the clinic.

Anaerobic digestion of sulphate wastewater mediated by biochar

ABSTRACT In this paper, the influences of biochar on the anaerobic digestion of sulphate wastewater, including the COD removal rate, methane yield, intermediate products and the change of microbial community structure, were investigated. The results showed that sulphate could promote the anaerobic digestion with the SO4 2−/COD ratio increasing from 0 to 0.1, while the activity of MPB was inhibited, which led to the decrease of COD removal rate and methane yield with the SO4 2−/COD ratio increasing from 0.1 to 2. At 1 g biochar loading, 344.97 mL CH4/gCODremoval was obtained compared with the control group (220.70 CH4/gCODremoval) at 2 of SO4 2−/COD. Biochar could also reduce the secondary accumulation of NH4 +–N and TVFA. Meanwhile, methanogenic microorganisms were selectively enriched especially for methanobacterium, methanosaeta and methanolinea, while the growth of SRB was inhibited with biochar addition.

Mannose-binding lectin promotes blood-brain barrier breakdown and exacerbates axonal damage after traumatic brain injury in mice

Leukocyte infiltration and blood-brain barrier breakdown contribute to secondary brain damage after traumatic brain injury (TBI). TBI induces neuroimmune responses triggering pathogenic complement activation through different pathways, including the lectin pathway. We investigated mechanisms underlying mannose-binding lectin (MBL)-mediated brain damage focusing on neutrophil infiltration and blood-brain barrier breakdown in a TBI mouse model. Wild type mice and MBL−/− null mice were subjected to controlled cortical impact. We studied neutrophil infiltration and regional localization by confocal microscopy 1, 4 and 15 days post-trauma, and investigated neutrophil extracellular trap (NET) formation. By immunofluorescence and/or Western blotting in various brain regions we studied the presence of fibrin(ogen), pentraxin-3, albumin and immunoglobulin G. Finally, we studied neurofilament proteins, synaptophysin, and αII-spectrin, and assessed white matter content in the injured tissue. TBI triggered an acute wave of neutrophil infiltration at day 1 followed by a more discrete persistence of neutrophils in the injured tissue at least until day 15. We detected the presence of NETs and pentraxin-3 in the injured tissue, as well as accumulation of fibrin(ogen), increased blood-brain barrier permeability, and neurofilament, synaptophysin and white matter loss, and calpain-mediated αII spectrin breakdown. MBL−/− mice showed reduced number of Ly6G+ neutrophils 4 days after TBI, lower accumulation of pentraxin-3 and fibrin(ogen) in the injured tissue, reduced global plasma protein extravasation, and better preservation of axonal and white matter integrity. These results show that MBL participates in secondary neutrophil accumulation and blood-brain barrier breakdown, and promotes axonal and white matter damage after TBI in mice.

Transcriptome analysis and validation of key genes involved in biosynthesis of iridoids in Gentiana lhassica

As the main chemical constituents, iridoids are widely distributed within Gentiana, Gentianaceae, with promising bioactivities. Based on the previous work, the transcriptome of G. lhassica, an original plant of Tibetan herb "Jieji Nabao", was sequenced and analyzed in this study, and the transcriptome databases of roots, stems, leaves, and flowers were constructed so as to explore unigenes that may encode the key enzymes in the biosynthetic pathway of iridoids. Then, qRT-PCR was used to validate the relative expression levels of 11 genes named AACT, DXS, MCS, HDS, IDI, GPPS, GES, G10H, 7-DLNGT, 7-DLGT, and SLS in roots, stems, leaves, and flowers. Also, the total contents of gentiopicroside and loganic acid were determined by HPLC, respectively. The results are as follows:(1)a total of 76 486 unigenes with an average length of 852 bp were obtained;(2)335 unigenes were involved in 19 stan-dard secondary metabolism pathways in KEGG database, with phenylpropanoid biosynthesis having the maximum number(75 unigenes), and no isoflavone biosynthetic pathway was annotated;(3)171 unigenes participatedin 27 key enzymes encoding in the biosynthetic pathway of iridoids, and 1-deoxy-D-xylulose-5-phosphate reductoisomerase(DXR) gene was highly expressed;(4)qRT-PCR results were approximately consistent with RNA-Seq data and the relative expression levels of the 11 genes were higher in the aboveground parts(stem, leaf, and flower) than in the underground part(root);(5)the total contents of gentiopicroside and loganic acid were higher in the aboveground parts(stem, leaf, and flower) than in the underground part(root), and the difference was significant. This study provides basic scientific data for accurate species identification, evaluation of germplasm resources, research on secondary pro-duct accumulation of medicinal plants within Gentianaceae, and protection of endangered alpine species.

Decoupling of Plant Growth and Accumulation of Biologically Active Compounds in Leaves, Roots, and Root Exudates of Hypericum perforatum L. by the Combination of Jasmonate and Far-Red Lighting.

The plant hormone jasmonic acid (JA) fine tunes the growth–defense dilemma by inhibiting plant growth and stimulating the accumulation of secondary compounds. We investigated the interactions between JA and phytochrome B signaling on growth and the accumulation of selected secondary metabolites in Hypericum perforatum L., a medically important plant, by spraying plants with methyl jasmonate (MeJA) and by adding far-red (FR) lighting. MeJA inhibited plant growth, decreased fructose concentration, and enhanced the accumulation of most secondary metabolites. FR enhanced plant growth and starch accumulation and did not decrease the accumulation of most secondary metabolites. MeJA and FR acted mostly independently with no observable interactions on plant growth or secondary metabolite levels. The accumulation of different compounds (e.g., hypericin, flavonols, flavan-3-ols, and phenolic acid) in shoots, roots, and root exudates showed different responses to the two treatments. These findings indicate that the relationship between growth and secondary compound accumulation is specific and depends on the classes of compounds and/or their organ location. The combined application of MeJA and FR enhanced the accumulation of most secondary compounds without compromising plant growth. Thus, the negative correlations between biomass and the content of secondary compounds predicted by the growth-defense dilemma were overcome.

Acid Sphingomyelinase, a Lysosomal and Secretory Phospholipase C, Is Key for Cellular Phospholipid Catabolism.

Here, we present the main features of human acid sphingomyelinase (ASM), its biosynthesis, processing and intracellular trafficking, its structure, its broad substrate specificity, and the proposed mode of action at the surface of the phospholipid substrate carrying intraendolysosomal luminal vesicles. In addition, we discuss the complex regulation of its phospholipid cleaving activity by membrane lipids and lipid-binding proteins. The majority of the literature implies that ASM hydrolyses solely sphingomyelin to generate ceramide and ignores its ability to degrade further substrates. Indeed, more than twenty different phospholipids are cleaved by ASM in vitro, including some minor but functionally important phospholipids such as the growth factor ceramide-1-phosphate and the unique lysosomal lysolipid bis(monoacylglycero)phosphate. The inherited ASM deficiency, Niemann-Pick disease type A and B, impairs mainly, but not only, cellular sphingomyelin catabolism, causing a progressive sphingomyelin accumulation, which furthermore triggers a secondary accumulation of lipids (cholesterol, glucosylceramide, GM2) by inhibiting their turnover in late endosomes and lysosomes. However, ASM appears to be involved in a variety of major cellular functions with a regulatory significance for an increasing number of metabolic disorders. The biochemical characteristics of ASM, their potential effect on cellular lipid turnover, as well as a potential impact on physiological processes will be discussed.

Disease modeling for Mucopolysaccharidosis type IIIB using patient derived induced pluripotent stem cells

Mucopolysaccharidosis type IIIB (MPS IIIB) is a lysosomal disease caused by mutations in the NAGLU gene encoding α-N-acetylglucosaminidase (NAGLU) which degrades heparan sulfate in lysosomes. Deficiency in NAGLU results in lysosomal accumulation of glycosaminoglycans (GAGs) and neurological symptoms. Currently, there is no effective treatment or cure for this disease. In this study, induced pluripotent stem cell lines were established from two MPS IIIB patient fibroblast lines and differentiated into neural stem cells and neurons. MPS IIIB neural stem cells exhibited NAGLU deficiency accompanied with GAG accumulation, as well as lysosomal enlargement and secondary lipid accumulation. Treatments with recombinant NAGLU, δ-tocopherol, and 2-hydroxypropyl-b-cyclodextrin significantly reduced the disease phenotypes in these cells. These results indicate the MPS IIIB neural stem cells and neurons have the disease relevant phenotype and can be used as a cell-based disease model system for evaluation of drug efficacy and compound screening for drug development.

Phytopharmacological Assessment of Some Medicinal Plants of Thal Desert of Pakistan

The presence of secondary metabolites and various ions in a plant determines its phytophamacological potential. Desert plants are adapted to stressful environmet by synthesizing secondary metabolites and ions accumulation as osmoticum.The present study was conducted to evaluate the pharmacological potential of Thal desert plants in term of their metabolites and nutrient ions concentrations. Five specimens of seasonally available herbs and three of trees of Thal desert plants were colected. After collection specimens were analysed for alkaloids, terpenoids, tannins, sugar and ion contents. The data were analyzed statistically and means were compared by Duncan’s Multiple Range Test. Among the herbs Panicum antidotale root showed highest terpenoid, K+ ion and Ca+ ion contents. The herb Aerva javanica stem showed lowest alkaloid, tannin, soluble sugar, phosphorus, potassium and calcium contents. Among trees specimens, Tamarix aphylla leaves showed highest soluble sugar, phosphorous, potassium and calcium contents. Stem of Acacia modesta showed the lowest alkaloid, terpenoid, phosphorous and potassium contents.

Effects of water deficit and high N fertilization on wheat storage protein synthesis, gluten secondary structure, and breadmaking quality

The content and composition of wheat storage proteins are the major determinants of dough rheological properties and breadmaking quality and are influenced by cultivation conditions. This study aimed to investigate the effects of water deficit and high N-fertilizer application on wheat storage protein synthesis, gluten secondary structure, and breadmaking quality. Reverse-phase ultrahigh-performance liquid chromatography analysis showed that storage protein and gluten macropolymer accumulation was promoted under both independent applications and a combination of water-deficit and high N-fertilizer treatments. Fourier-transform infrared spectroscopy showed that water deficit and high N-fertilizer treatments generally improved protein secondary structure formation and lipid accumulation, and reduced flour moisture. In particular, high N-fertilizer application increased β-sheet content by 10.4% and the combination of water-deficit and high N-fertilizer treatments increased random coil content by 7.6%. These changes in gluten content and secondary structure led to improved dough rheological properties and breadmaking quality, including superior loaf internal structure, volume, and score. Our results demonstrate that moderately high N-fertilizer application under drought conditions can improve gluten accumulation, gluten secondary structure formation, and baking quality.

The localization of the photosensitizer determines the dynamics of the secondary production of hydrogen peroxide in cell cytoplasm and mitochondria

Photodynamic therapy (PDT) is based on the production of the cytotoxic reactive oxygen species (ROS) by light irradiation of a photosensitizer dye in the presence of molecular oxygen. Along with photochemical ROS production, it becomes evident that PDT induces massive secondary production of ROS which is registered long after the irradiation is completed. We created cell lines of human epidermoid carcinoma with the cytoplasmic and mitochondrial localization of protein sensor HyPer sensitive to hydrogen peroxide to compare its concentration in two cellular compartments. The lag-period between irradiation and accumulation of hydrogen peroxide in cells was registered; its duration was dose-dependent and increased up to 80 min when lowering the exposition dose from 50 to 15 J/cm2. We have shown that localization of the photosensitizer determines the spatiotemporal pattern of the cell response to PDT: secondary hydrogen peroxide accumulation in cell cytoplasm induced by photodynamic treatment with lysosome-localized phtalocyianine Photosens occurs several minutes prior to that in mitochondria; on the contrary, membranotropic arylcyanoporphyrazine dye leads to massive mitochondrial hydrogen peroxide production followed by its cytoplasmic accumulation. We hypothesize that photosensitizers with various physicochemical properties and intracellular localization can trigger different patterns not only of primary but also secondary ROS production leading to different cell fate outcomes.

Provenance, and environment context of pedogenic carbonates formation from MIS 3 to MIS 1 in the Teotihuacan Valley, Mexico

Pedogenic carbonates are considered important indicators of past climatic changes. Their formation is due to calcite accumulation through evaporation processes in arid or semi-arid regions. Over time, secondary carbonate accumulation results in the development of calcic and petrocalcic horizons in soil profiles. In this study, we conducted an analysis of pedogenic carbonates as records of paleoenvironmental characteristics in the Teotihuacan Valley, Basin of Mexico. Micromorphological, mineralogical, and geochemical characteristics of secondary carbonates from three paleosol profiles (Cerro Gordo 1, Cerro Gordo 2, and Maseca 1) were analyzed. Pedogenic carbonates were characterized by irregular and variable carbonate crystal sizes, inclusions of soil particles, and dark or light micrite and microsparite laminations. Micromorphology and scanning electron microscopy revealed weathered plagioclase minerals as a potential source of calcium for carbonate precipitation. The 87Sr/86Sr ratio of carbonate laminar crusts and nodules from the analyses pedons in the study area also indicated that rocks of the Transmexican Volcanic Belt were the principal source of Ca and Sr for carbonates, while contribution of a far-distance eolian input was minimal. The δ13C isotopic composition evidenced a mixture of C3 and C4 plants during the Pleistocene and a transition to a higher proportion of C4 vegetal cover during the Holocene. This agreed with an increase in modeled paleo-temperature from 10.9 °C to 13.7 °C based on 18O-temperature equations. Our results support those determined for the regional paleoenvironment during the Pleistocene, the Pleistocene-Holocene transition and Mid-Late Holocene.

GCase and LIMP2 Abnormalities in the Liver of Niemann Pick Type C Mice.

The lysosomal storage disease Niemann–Pick type C (NPC) is caused by impaired cholesterol efflux from lysosomes, which is accompanied by secondary lysosomal accumulation of sphingomyelin and glucosylceramide (GlcCer). Similar to Gaucher disease (GD), patients deficient in glucocerebrosidase (GCase) degrading GlcCer, NPC patients show an elevated glucosylsphingosine and glucosylated cholesterol. In livers of mice lacking the lysosomal cholesterol efflux transporter NPC1, we investigated the expression of established biomarkers of lipid-laden macrophages of GD patients, their GCase status, and content on the cytosol facing glucosylceramidase GBA2 and lysosomal integral membrane protein type B (LIMP2), a transporter of newly formed GCase to lysosomes. Livers of 80-week-old Npc1−/− mice showed a partially reduced GCase protein and enzymatic activity. In contrast, GBA2 levels tended to be reciprocally increased with the GCase deficiency. In Npc1−/− liver, increased expression of lysosomal enzymes (cathepsin D, acid ceramidase) was observed as well as increased markers of lipid-stressed macrophages (GPNMB and galectin-3). Immunohistochemistry showed that the latter markers are expressed by lipid laden Kupffer cells. Earlier reported increase of LIMP2 in Npc1−/− liver was confirmed. Unexpectedly, immunohistochemistry showed that LIMP2 is particularly overexpressed in the hepatocytes of the Npc1−/− liver. LIMP2 in these hepatocytes seems not to only localize to (endo)lysosomes. The recent recognition that LIMP2 harbors a cholesterol channel prompts the speculation that LIMP2 in Npc1−/− hepatocytes might mediate export of cholesterol into the bile and thus protects the hepatocytes.

5-lipoxygenase- and Glucocorticoid-dependent eosinophilia in a novel surgical model in mice

BackgroundSubcutaneous implants of heat–coagulated egg white (egg white implants, EWI) induce intense local eosinophilia and prime for hyperreactivity following airway ovalbumin challenge. The roles of allergen sensitization, surgical trauma–induced glucocorticoids, and the 5-lipoxygenase (5-LO) pathway were hitherto unexplored in this model, in which quantitative recovery and large-scale purification of the eosinophils from the inflammatory site for functional and immunopharmacological studies are difficult to achieve. MethodsWe overcame this limitation by shifting the implantation site to the peritoneal cavity (EWIp), thereby enabling quantitative leukocyte retrieval. ResultsBy day 7 post–surgery, eosinophil counts reached ~ 30% of all leukocytes recovered. Eosinophilia was prevented by: a) induction of allergen–specific oral tolerance to ovalbumin, the main allergen in egg white; b) inactivation of the 5–lipoxygenase pathway; c) blockade of endogenous glucocorticoid signaling by pretreatment with metirapone plus mifepristone before surgery. Highly purified eosinophils (~99% pure) could be obtained from the peritoneal exudate of EWIp–carrier mice in 2 simple, antibody–free steps. Preparative-scale yields, suitable for most biochemical, pharmacological, and molecular applications, were routinely obtained, and could be further enhanced through addition of pre–or post–surgery immunization steps (active or adoptive). The recovered eosinophils were fully functional in vivo, as demonstrated by the transfer of purified eosinophils into eosinophil–deficient Δdbl–GATA–1-KO mice, which upon subsequent challenge with eotaxin-1 present secondary accumulation of neutrophils, but not of mononuclear phagocytes. ConclusionThese findings document glucocorticoid−, allergen– and 5–lipoxygenase–dependent eosinophilia, which makes EWIp carriers an abundant source of pure, nontransgenic eosinophils for immunopharmacological studies.

Tetraedron minimum, First Reported Member of Hydrodictyaceae to Accumulate Secondary Carotenoids.

We isolated a novel strain of the microalga Tetraedron minimum in Iceland from a terrestrial habitat. During long-term cultivation, a dish culture turned orange, indicating the presence of secondary pigments. Thus, we characterized T. minimum for growth and possible carotenoid production in different inorganic media. In a lab-scale photobioreactor, we confirmed that nitrogen starvation in combination with salt stress triggered a secondary carotenoid accumulation. The development of the pigment composition and the antioxidant capacity of the extracts was analyzed throughout the cultivations. The final secondary carotenoid composition was, on average, 61.1% astaxanthin and 38.9% adonixanthin. Moreover, the cells accumulated approx. 83.1% unsaturated fatty acids. This work presents the first report of the formation of secondary carotenoids within the family Hydrodictyaceae (Sphaeropleales, Chlorophyta).

Mechanism and restoration strategy of lysosomal abnormalities induced by urinary protein overload in proximal tubule epithelial cells.

Persistent elevated concentrations of urinary protein can destroy proximal tubule epithelial cells (PTECs) by inducing lysosomal abnormalities, thereby aggravating PTEC damage and renal fibrosis. However, the specific mechanisms of these serial biochemical events and methods for treating or preventing PTEC damage upon proteinuria need further investigation. In this study, electron microscopy and dual-labeled immunofluorescence analysis for identifying lysosome type revealed inadequate primary lysosome biogenesis and secondary lysosome accumulation in the PTECs of patients with minimal change nephrotic syndrome or membranous nephropathy who suffered from proteinuria. In vitro studies on HK-2 cells indicated that this abnormality was associated with decreased expression of transcription factor EB (TFEB). In contrast, TFEB overexpressing HK-2 cells under urinary protein overload exhibited significantly reduced accumulation of secondary lysosomes and increased proportion and quantity of primary lysosomes as indicated by dual-labeled immunofluorescence. Further, these cells could upregulate lysosomal degradation functions, as determined using Cathepsin L activity assays and flow cytometry for dye quenched-albumin. These results indicate that abnormal TFEB expression is a key mechanism of lysosomal dyshomeostasis caused by protein overload in PTECs. TFEB is thus a potential therapeutic target for the treatment of urinary protein-related kidney disease.

Geology and genesis of the Shalipayco evaporite-related Mississippi Valley-type Zn–Pb deposit, Central Peru: 3D geological modeling and C–O–S–Sr isotope constraints

The Shalipayco Zn–Pb deposit, in central Peru, is composed of several stratabound orebodies, the largest of which are the Resurgidora and Intermedios, contained in carbonate rocks of the Upper Triassic Chambara Formation, Pucara group. Petrography suggests that a single ore-forming episode formed sphalerite and galena within vugs, open spaces, and fractures. Three-dimensional (3D) geological modeling has allowed division of the Chambara Formation into four members (Chambara I, II, III, and IV) that better define lithological controls on sulfide formation. Diagenetic replacement of evaporite minerals with the organic matter (OM) presence likely generated secondary porosity and H2S accumulation by bacterial sulfate reduction (BSR), providing ground preparation for the later Zn–Pb mineralizing event. The least-altered host rocks have C–O isotope compositions of 1.8 ± 0.1‰ (VPDB) and 29.9 ± 2.1‰ (VSMOW), respectively, within the Triassic marine carbonate ranges. Early dolomite contains lighter C–O composition (1.1 ± 0.9 and 23.8 ± 2.9‰, respectively) consistent with OM decomposition during burial diagenesis. Post-mineralization calcite has still lighter C–O composition (− 5.1 and 13.3‰, respectively), suggesting meteoric water that had migrated through organic-rich strata. The strontium isotopes of Mitu group basalts (0.709654–0.719669) indicate it as a possible, but not the unique source of strontium and probably of other metals. Highly negative sulfide sulfur isotope values (− 23.3 to − 6.2‰ (VCDT)) indicate a major component of the ore sulfur derived ultimately from BSR. However, multiple lines of evidence suggest that preexisting H2S underwent thermochemical redox cycling prior to ore formation. The influx of hot metalliferous brines to dolomitized zones containing trapped H2S is the preferred model for ore deposition at Shalipayco.

The responses of sulforaphane synthesis and endogenous NO to Penicillium chrysogenum elicitor in Broccoli cells

In this study, we investigated the effects Penicillium chrysogenum elicitor and NO synthesis inhibitor on phenylalanine ammonia lyase (PAL) activity, cell viability, endogenous nitric oxide (NO) and NO synthase (NOS) and nitrate reductase (NR), secondary metabolite sulforaphane accumulation. In addition, the link of between endogenous nitric oxide (NO) and sulforaphane, as well as between NR and NOS pathway was also assessed. The results showed that all the concentrations of Penicillium chrysogenum elicitor induced NO synthesis, and the synthesis reaction time was earlier under high concentrations of Penicillium chrysogenum elicitor, while the released rate of NO was relatively slower under low concentration of Penicillium chrysogenum elicitor. In addition, NO scavenger carboxy-PTIO (c-PTIO) blocked the effect of Penicillium chrysogenum elicitor on the promotion that sulforaphane accumulation and the activation of PAL activity, and decrease cell vitality induced by Penicillium chrysogenum elicitor. The content of sulforaphane was related with NO accumulation, and the accumulation of sulforaphane was increased when endogenous NO release was excited, while sulforaphane accumulation amount decreased when NO synthesis was inhibited. Under the induction of Penicillium chrysogenum elicitor, the release of NO from in vitro cells of broccoli was mainly through NOS and NR pathways. When NR pathway was inhibited by high concentrations of NR inhibitor Tungstate, NOS activity was activated via high concentration of K+ from Tungstate. However, NR pathway could not be activated by high concentration of K+ when the NOS pathway was inhibited by NOS inhibitor L-NAME.

Systemic scAAV9.U1a.hSGSH Delivery Corrects Brain Biochemistry in Mucopolysaccharidosis Type IIIA at Early and Later Stages of Disease.

Mucopolysaccharidosis type IIIA (MPS IIIA, Sanfilippo A syndrome) is a single gene (SGSH) childhood onset neurodegenerative disease for which gene therapy is in clinical trial. Theoretically, the transfer of a working gene should enable functional expression of the defective protein and rescue the phenotype when administered before the onset of irreversible disease. Recombinant adeno-associated virus (AAV) is being used as a vehicle for a number of gene therapy applications and the neurotropism of serotype 9 affords utility for monogenetic neurological disorders. To assess the efficacy of restoring the underlying biochemistry in the MPS IIIA brain, tail vein injections of self-complementary AAV9 human N-sulfoglucosamine sulfohydrolase (scAAV9.U1A.hSGSH) at 3 × 1013 vg/kg were administered to 6- and 16-week-old MPS IIIA mice. Heparan sulfate (HS) and GM2 and GM3 gangliosides were cleared from the cortex, hippocampus and subcortex with residual storage remaining in the brain stem and cerebellum. SGSH activity increased in the brain of the MPS IIIA-treated mice, but remained significantly reduced compared with wild-type. Motor activity as assessed in an open-field arena, and gait length, improved in MPS IIIA mice treated at both 6 and 16 weeks of age. However, functional assessment of cognition in the water cross-maze test, as well as gait width, normalized in mice treated at 6 weeks of age only, with mice treated at 16 weeks performing similar to untreated MPS IIIA mice. Astrogliosis was reduced in mice treated at 6 and 16 weeks of age compared to untreated MPS IIIA mice. These results demonstrate that the gene product is actively clearing primary HS and secondary ganglioside accumulation in MPS IIIA mice, but in older mice, neurocognitive impairments remain. This is likely due to secondary downstream consequences of HS affecting neurological functions that are not reversible upon substrate clearance.