Search Results (25)

Background and Clinical Significance: Mucopolysaccharidosis type VI is a rare lysosomal storage disorder due to arylsulfatase B enzyme deficiency, leading to progressive multisystem disease and complex airway. Acute respiratory infections can precipitate airway embarrassment. A structured treatment guideline is currently lacking. We present a 7-year-old MPS VI male with respiratory distress, highlighting the challenges in management. Case Presentation: Case review focusing on clinical presentation, imaging findings, and multidisciplinary decision-making during acute deterioration. A child diagnosed with MPS VI at the age of 3.5 years old, due to low arylsulfatase B enzyme activity and homozygous for c.275C>A p.(Thr92Lys) variant in the ARSB gene. At 7 years of age, he showed the signs of dyspnoea, increased respiratory effort with bilateral crepitations, and noisy breathing. Initial management included facemask oxygen, nebulised adrenaline, corticosteroids, and bronchodilators. Computer tomography scan of the neck and chest showed a complex upper airway, multiple tracheal narrowing, tortuosity, and an extra loop of truncus brachiocephalicus from the arch of the aorta. Potential interventions carried substantial risks due to abnormal airway and multisystem disease. Following extensive multidisciplinary discussion after careful consideration of the significant risks associated with invasive airway interventions, a shared decision was reached with the family to adopt a comfort-focused palliative care approach. Despite the best supportive care, the child unfortunately passed away after 3 months. The family was involved in every decision process and was fully supported. Conclusions: MPS VI is associated with complex airways and multisystem disease. Multidisciplinary decision-making with family is critical to safe and appropriate care. The rarity of the disease, lack of guidelines, complex airways, and multiple comorbidities make management challenging. Full article

The diversification of cover crops grown in soils with granulometric variability may potentially enhance microbial community and enzyme activities. Thus, the main goal was to evaluate the effect of autumn/winter cover crop sequences and cash crops in spring/summer on soil microbial biomass and enzyme activities. The experiment was conducted in open-field microplots (10 m × 1 m × 0.7 m), containing soils from B horizon of five Oxisols with granulometric variability and clay content ranging from 17 to 80 dag kg⁻¹. The treatments were three cover crops and a winter fallow with a completely randomized experimental design with three replicates. Soil samples from the 0–10 cm layer were collected to analyze soil microbial biomass of carbon and nitrogen, enzyme activities of the acid phosphatase, arylsulfatase, urease, and fluorescein diacetate hydrolysis. The number of nodules in soybean roots was average 63 ± 14.42 nodules per plant and dry mass of nodules was 169 ± 13.74 mg plant⁻¹. Soybean nodulation and N uptake ensured the supply of nitrogen to the soybean plants with 331 ± 82 mg plant⁻¹. Overall, diversified autumn/winter cover crop sequences provide plant residue, containing nutrients, and different carbon/nitrogen, which alters microbial biomass, the ratio of C_mic/N_mic, basal respiration, and soil enzyme activities within each Oxisol with different particle size distribution. Full article

The low fertility of tropical Oxisols challenges sustainable agriculture. While biochar-based granular fertilizers (BBGFs) offer a solution, the influence of different organic binders is unclear. This study investigated how BBGFs formulated with bio-oil (BO), pyroligneous extract (PE), and cassava wastewater (CW) impact soil enzyme activities and nutrient dynamics over time. Eucalyptus biochar (B) and natural phosphate (NP) were granulated with three binders at four doses. These treatments, plus controls (unfertilized soil, NP, B with NP, and B alone), were incubated in an Oxisol, assessing soil samples after 10 and 40 days of incubation. All BBGFs significantly enhanced β-glucosidase, acid phosphatase, and arylsulfatase activities over controls, with increases exceeding 8%. While the BBGFs-BO formulation sustained the highest enzymatic activity, BBGFs-PE at 125% maximized acid phosphatase at 10 days, with a subsequent decline, and inhibited arylsulfatase at the 150% dose. BBGFs-CW was most effective for increasing P availability (up to 24.0 mg kg⁻¹). BBGFs-BO and BBGFs-PE also enhanced soil organic carbon and cation exchange capacity by up to 430% and 163%, respectively. The BBGFs-BO at 150% dose is the most effective and stable formulation to enhance nutrient cycling and soil health, offering a viable pathway to convert agricultural residues into high-value fertilizers. Full article

Snake venoms contain numerous toxic proteins, but low-abundance proteins often remain uncharacterized due to identification challenges. This study employs a bioinformatics approach to identify and structurally model low-abundance proteins from the venom gland transcriptomes of Bothrops asper and Bothrops jararaca. Using tools such as tblastn, Jalview, and CHIMERA, we analyzed sequences and structural features of proteins including arylsulfatase, CRISP (Cysteine-Rich Secretory Protein), von Willebrand factor type D (vWFD), and dihydroorotate dehydrogenase (DHODH), and identified potential new isoforms of SVMP-PIIIb (Ba_1) and botrocetin in B. asper. Protein models were generated with AlphaFold2, compared with crystallized structures from the Protein Data Bank (PDB), and validated using Procheck, ERRAT, and Verify3D. Conserved motifs and domains were annotated through Pfam and InterPro, revealing structural elements that suggest possible roles in venom physiology and toxicity. These findings emphasize the potential of computational biology to characterize structurally relevant but experimentally inaccessible venom proteins, and to lay the groundwork for future functional validation. Full article

Defective CFTR (cystic fibrosis transmembrane conductance regulator) is pathognomonic for cystic fibrosis (CF), which is characterized by an accumulation of tenacious secretions in pulmonary airways, as well as by abnormal ductal secretions in other organs, including the pancreas and prostate. The advent of CFTR modulating therapies has markedly improved the clinical status and survival of CF patients, primarily attributable to improved lung function. Previous publications reported that a decline in CFTR function was associated with a decline in activity and expression of the enzyme N-acetylgalactosamine-4-sulfatase (Arylsulfatase B; ARSB). ARSB removes 4-sulfate groups from N-acetylgalactosamine 4-sulfate residues and is required for the degradation of chondroitin 4-sulfate (chondroitin sulfate A) and dermatan sulfate, two sulfated glycosaminoglycans which accumulate in cystic fibrosis. Declines in both ARSB and in CFTR have been associated with the development of malignancies, including prostate malignancy. The experiments in this report show that similar effects on invasiveness are present when either CFTR or ARSB is inhibited in human prostate epithelial cells, and these effects resemble findings detected in malignant prostate tissue. The effects of CFTR inhibition are reversed by treatment with recombinant human ARSB in prostate cells. These results suggest that treatment by rhARSB may benefit patients with cystic fibrosis and prostate cancer. Full article

Microbial inoculants are vital for promoting plant growth and facilitating the ecological restoration of degraded forested regions near abandoned mine sites. However, the direct application of liquid microbial inoculants is often challenging due to low microbial activities and poor transport efficiencies, which limit their effectiveness in complex soil environments. To tackle these challenges, this study utilized immobilized microbial technology to evaluate the effectiveness of solid microbial inoculants sourced from peat (P), biochar (BC), and spent mushroom substrates (SMSs) in enhancing the soil’s multifunctionality and promoting plant growth. Specifically, this research sought to assess the effectiveness of solid microbial inoculants derived from peat (P), biochar (B), and spent mushroom substrates (SMSs) in enhancing soil multifunctionality and promoting plant growth in nutrient-deficient soils that were affected by abandoned mine sites. We aimed to evaluate the performance of different solid microbial inoculants in improving the soil’s nutrient content and enzyme activities. A 24-week pot experiment was conducted using Medicago sativa L. in nutrient-poor soil. The results demonstrated that, in contrast to peat and biochar, SMSs effectively interacted with microbial inoculants and significantly improved the nutrient content and enzyme activities of nutrient-deficient soil. It was noted that β-1,4-glucosidase (BG), invertase, β-1,4-N-acetylglucosaminidase (NAG), urease, and soil available phosphorus increased by 204%, 405%, 118%, 198%, and 297%, respectively. The soil’s multifunctionality improved by 320% compared with the CK, and the plant biomass also increased significantly. Further, our random forest analysis indicated that the soil available phosphorus, ammonium nitrogen, total nitrogen, total carbon content, arylsulfatase, pH, total phosphorus, NAG, and BG were key environmental factors that induced changes in plant biomass. These findings highlighted the potential of SMSs as an effective carrier for immobilized microbial inoculants, which provides a sustainable approach for the restoration of forest soils surrounding abandoned mine sites, as well as a promising avenue for the valorization of agricultural waste. Full article

Iron ore extraction can lead to significant environmental degradation, particularly due to the generation of tailings during the beneficiation process. This issue was highlighted by the B1 dam collapse in Brumadinho, Brazil, in 2019. Therefore, the study and monitoring of affected areas is essential to assess soil quality throughout the rehabilitation process, whether through natural recovery or active rehabilitation practices. Microbial indicators can serve as valuable tools to track the recovery of these areas, given their high sensitivity and rapid response to environmental changes. The aim of this study was to evaluate soil microbial indicators, such as enzyme activity, microbial biomass carbon, microbial basal respiration and microbial diversity, and to select microbial approaches for monitoring the area affected by mining tailings in Brumadinho. The results indicated that the reference area initially outperformed the affected area on all evaluated bioindicators, highlighting environmental stress in the affected zone. Over the course of the study, the two areas began to show greater similarity, suggesting a natural recovery of the soil together with the return of natural vegetation. Indicators such as microbial carbon biomass went from values close to 50 mg of C Kg of soil⁻¹ in the affected area, to around 200, statistically equal to the reference. qCO₂ also varied in the affected area to values statistically equal to those of the reference over time, variated in the first collection to 0.25 mg of C-CO₂ mg of C⁻¹ h⁻¹ in the affected area against 0.1 in the reference area; in the last collection, both areas presented values close to 0.2. Enzymatic activity had superior values in the reference area about the affected area, being urease, and arylsulfatase more sensitive to show differences between areas over time. The metataxonomic data again revealed indicator species for each environment, including genera such as Bacillus, Mycobacterium, Acidibacter, and Burkholderia representative of the reference, and the genera Ramlibacter, Sinomonas, Psedarthrobacter, and Knoellia indicators of the affected area. By the end of this study, the applicability of microbial indicators for monitoring soil microbiota and its ecosystem services was successfully demonstrated. In addition, specific microbial indicators were proposed for monitoring areas affected by iron mining tailings. Full article

Previous proteomic studies of viperid venom revealed that it is mainly composed of metalloproteinases (SVMPs), serine proteinases (SVSPs), phospholipase A2 (PLA2), and C-type lectins (CTLs). However, other proteins appear in minor amounts that affect prey and need to be identified. This study aimed to identify novel toxic proteins in the venom gland transcriptome of Bothrops asper and Bothrops jararaca, using data from NCBI. Bioinformatics tools were used to assemble, identify, and compare potentially novel proteins in both species, and we performed functional annotation with BLASTX against the NR database. While previous assemblies have been performed for B. jararaca, this is the first assembly of the B. asper venom gland transcriptome. Proteins with potentially novel functions were identified, including arylsulfatase and dihydroorotate dehydrogenase, among others, that could have implications for venom toxicity. These results suggest that the identified proteins may contribute to venom toxic variation and provide new opportunities for antivenom research. The study improves the understanding of the protein composition of Bothrops venom and suggests new possibilities for the development of treatments and antivenoms. Full article

Phosphatases are enzymes that catalyze the hydrolysis of phosphate esters. They play critical roles in diverse biological processes such as extracellular nucleotide homeostasis, transport of molecules across membranes, intracellular signaling pathways, or vertebrate mineralization. Among them, tissue-nonspecific alkaline phosphatase (TNAP) is today increasingly studied, due to its ubiquitous expression and its ability to dephosphorylate a very broad range of substrates and participate in several different biological functions. For instance, TNAP hydrolyzes inorganic pyrophosphate (PP_i) to allow skeletal and dental mineralization. Additionally, TNAP hydrolyzes pyridoxal phosphate to allow cellular pyridoxal uptake, and stimulate vitamin B6-dependent reactions. Furthermore, TNAP has been identified as a key enzyme in non-shivering adaptive thermogenesis, by dephosphorylating phosphocreatine in the mitochondrial creatine futile cycle. This latter recent discovery and others suggest that the list of substrates and functions of TNAP may be much longer than previously thought. In the present review, we sought to examine TNAP within the alkaline phosphatase (AP) superfamily, comparing its sequence, structure, and evolutionary trajectory. The AP superfamily, characterized by a conserved central folding motif of a mixed beta-sheet flanked by alpha-helices, includes six subfamilies: AP, arylsulfatases (ARS), ectonucleotide pyrophosphatases/phosphodiesterases (ENPP), phosphoglycerate mutases (PGM), phosphonoacetate hydrolases, and phosphopentomutases. Interestingly, TNAP and several ENPP family members appear to participate in the same metabolic pathways and functions. For instance, extra-skeletal mineralization in vertebrates is inhibited by ENPP1-mediated ATP hydrolysis into the mineralization inhibitor PP_i, which is hydrolyzed by TNAP expressed in the skeleton. Better understanding how TNAP and other AP family members differ structurally will be very useful to clarify their complementary functions. Structurally, TNAP shares the conserved catalytic core with other AP superfamily members but has unique features affecting substrate specificity and activity. The review also aims to highlight the importance of oligomerization in enzyme stability and function, and the role of conserved metal ion coordination, particularly magnesium, in APs. By exploring the structural and functional diversity within the AP superfamily, and discussing to which extent its members exert redundant, complementary, or specific functions, this review illuminates the evolutionary pressures shaping these enzymes and their broad physiological roles, offering insights into TNAP’s multifunctionality and its implications for health and disease. Full article

(1) Background: Colorectal cancer (CRC) remains a significant cause of morbidity and mortality worldwide, with its prognosis influenced by genetic and clinicopathological factors. This study investigates the associations between the gene expressions of Arylsulfatase B (ARSB), TP53, and Maspin, alongside traditional clinicopathological features, and their impact on CRC survival outcomes. (2) Methods: 70 consecutive CRC cases were analyzed for ARSB, TP53, and Maspin gene expression using RT-qPCR, and their protein levels were assessed through immunohistochemistry. Clinicopathological parameters—age, gender, tumor localization, macroscopic and microscopic aspects, lymph node ratio, pT stage, and tumor budding—were evaluated for their prognostic significance. Kaplan–Meier survival analysis with Cox proportional hazards regression was used to determine their impact on overall survival. (3) Results: No significant survival differences were observed based on age, gender, tumor localization, and macroscopic aspect. The microscopic aspect and pT stage showed significant associations with survival, with poorer outcomes in G3 and pT3/pT4 stages, respectively. Immunohistochemical positivity for ARSB and Maspin indicated a longer survival, while TP53 protein expression alone did not significantly impact the prognosis. Dual high gene expression (ARSB + TP53, TP53 + Maspin) and triple high gene expression (ARSB + TP53 + Maspin) were significantly associated with better survival outcomes. (4) Conclusions: The combined gene expression profile of ARSB, TP53, and Maspin presents a novel prognostic marker in CRC, offering insights into the molecular dynamics of cancer cells and potential therapeutic targets. These findings emphasize the importance of integrating molecular markers with traditional clinicopathological factors for a more accurate prognostication and personalized treatment approach in CRC. Full article

The effects of the enzyme N-acetylgalactosamine-4-sulfatase (Arylsulfatase B, ARSB), which removes the 4-sulfate group at the non-reducing end of chondroitin 4-sulfate, on the expression of PD-L1 were determined, and the underlying mechanism of PD-L1 expression was elucidated. Initial experiments in human melanoma cells (A375) showed that PD-L1 expression increased from 357 ± 31 to 796 ± 50 pg/mg protein (p < 10⁻¹¹) when ARSB was silenced in A375 cells. In subcutaneous B16F10 murine melanomas, PD-L1 declined from 1227 ± 189 to 583 ± 110 pg/mg protein (p = 1.67 × 10⁻⁷), a decline of 52%, following treatment with exogenous, bioactive recombinant ARSB. This decline occurred in association with reduced tumor growth and prolongation of survival, as previously reported. The mechanism of regulation of PD-L1 expression by ARSB is attributed to ARSB-mediated alteration in chondroitin 4-sulfation, leading to changes in free galectin-3, c-Jun nuclear localization, HDAC3 expression, and effects of acetyl-H3 on the PD-L1 promoter. These findings indicate that changes in ARSB contribute to the expression of PD-L1 in melanoma and can thereby affect the immune checkpoint response. Exogenous ARSB acted on melanoma cells and normal melanocytes through the IGF2 receptor. The decline in PD-L1 expression by exogenous ARSB may contribute to the impact of ARSB on melanoma progression. Full article

Analyzing urine is common in drug-facilitated sexual assault cases if the analysis of blood is not optimal. The efficient enzymatic pretreatment of urine is important for cleaving glucuronides and improving the detection of the parent drug. The aim was to investigate the efficiency of three β-glucuronidases on eleven glucuronides relevant to DFSA at different incubation periods and temperatures. Human drug-free urine was fortified with 11 glucuronides, hydrolyzed with either β-glucuronidase/arylsulfatase (Helix Pomatia), recombinant β-glucuronidase B-One™ or recombinant β-glucuronidase BGTurbo™ and incubated for 5, 10, 60 min, 18 h and 24 h at 20 °C/40 °C/55 °C before UHPLC–MS/MS analysis. The stability of 141 drugs and metabolites relevant to DFSA was investigated by incubating fortified urine under the same hydrolysis conditions. B-One™ showed efficient hydrolysis (>90%) of most glucuronides in 5 min at all temperatures, while BGTurbo™ showed a similar efficiency (>90%), but the optimal temperature (20–55 °C) and incubation time (5–60 min) varied among analytes. The β-glucuronidase/arylsulfatase had the lowest efficiency and required the longest incubation (24 h) at 40–55 °C. The stability of 99% of 141 drugs and metabolites was not affected by incubation at 20–55 °C for 24 h. Recombinant enzymes show promising results for the simple and efficient hydrolysis of a broad panel of glucuronides relevant for DFSA. Full article

Mucopolysaccharidoses (MPSs) are rare inherited lysosomal storage diseases (LSDs) caused by deficient activity in one of the enzymes responsible for glycosaminoglycans lysosomal degradation. MPS II is caused by pathogenic mutations in the IDS gene, leading to deficient activity of the enzyme iduronate-2-sulfatase, which causes dermatan and heparan sulfate storage in the lysosomes. In MPS VI, there is dermatan sulfate lysosomal accumulation due to pathogenic mutations in the ARSB gene, leading to arylsulfatase B deficiency. Alterations in the immune system of MPS mouse models have already been described, but data concerning MPSs patients is still scarce. Herein, we study different leukocyte populations in MPS II and VI disease patients. MPS VI, but not MPS II patients, have a decrease percentage of natural killer (NK) cells and monocytes when compared with controls. No alterations were identified in the percentage of T, invariant NKT, and B cells in both groups of MPS disease patients. However, we discovered alterations in the naïve versus memory status of both helper and cytotoxic T cells in MPS VI disease patients compared to control group. Indeed, MPS VI disease patients have a higher frequency of naïve T cells and, consequently, lower memory T cell frequency than control subjects. Altogether, these results reveal MPS VI disease-specific alterations in some leukocyte populations, suggesting that the type of substrate accumulated and/or enzyme deficiency in the lysosome may have a particular effect on the normal cellular composition of the immune system. Full article

Metachromatic leukodystrophy (MLD) is a hereditary neurodegenerative disease characterized by demyelination and motor and cognitive impairments due to deficiencies of the lysosomal enzyme arylsulfatase A (ARSA) or the saposin B activator protein (SapB). Current treatments are limited; however, gene therapy using adeno-associated virus (AAV) vectors for ARSA delivery has shown promising results. The main challenges for MLD gene therapy include optimizing the AAV dosage, selecting the most effective serotype, and determining the best route of administration for ARSA delivery into the central nervous system. This study aims to evaluate the safety and efficacy of AAV serotype 9 encoding ARSA (AAV9-ARSA) gene therapy when administered intravenously or intrathecally in minipigs, a large animal model with anatomical and physiological similarities to humans. By comparing these two administration methods, this study contributes to the understanding of how to improve the effectiveness of MLD gene therapy and offers valuable insights for future clinical applications. Full article

A digestate with amendments provides plants with available nutrients and improves the microbiological properties of treated soil. Modification of a digestate through the addition of a biochar and sulphur source is less well-known. This pot experiment aimed at comparing the short- and long-time fertilization effects of a digestate enriched with biochar, with elemental sulphur, or with a combination of both on soil health and plant biomass. The experiment was carried out with maize, cultivated twice (1st–12th week = pre-cultivation; re-sowing after shoot harvest, 13th–24th = main cultivation) in soil amended with prepared digestate. The digestate used in pre-cultivation was incubated untreated (D) and was then treated with biochar (D + B), with elemental sulphur at a low (LS) and high (HS) dose, or with a combination of both (D + B + LS and D + B + HS). An additional unamended digestate (D) was added to each soil variant before the main cultivation. The application of digestate with a high dose of elemental sulphur and biochar mediated the most significant differences in the soil. The increase (compared to the unamended soil) was of short-term type (+11% and +6% increased total nitrogen and carbon after 12 weeks), then of long-term type (+54% and +30% increased sulphur and arylsulfatase activity after 24 weeks), and later emerged in the 13th to the 24th week of the experiment (+57% and +32% non-inhibited urease, increased N-acetyl-β-D-glucosaminidase and phosphatase). No significant differences in the effect of the applied amendments on dry aboveground plant biomass were observed. Full article