Using Constellation Pharmacology to Characterize a Novel α-Conotoxin from Conus ateralbus.

The venom of cone snails has been proven to be a rich source of bioactive peptides that target a variety of ion channels and receptors. α-Conotoxins (αCtx) interact with nicotinic acetylcholine receptors (nAChRs) and are powerful tools for investigating the structure and function of the various nAChR subtypes. By studying how conotoxins interact with nAChRs, we can improve our understanding of these receptors, leading to new insights into neurological diseases associated with nAChRs. Here, we describe the discovery and characterization of a novel conotoxin from Conus ateralbus, αCtx-AtIA, which has an amino acid sequence homologous to the well-described αCtx-PeIA, but with a different selectivity profile towards nAChRs. We tested the synthetic αCtx-AtIA using the calcium imaging-based Constellation Pharmacology assay on mouse DRG neurons and found that αCtx-AtIA significantly inhibited ACh-induced calcium influx in the presence of an α7 positive allosteric modulator, PNU-120596 (PNU). However, αCtx-AtIA did not display any activity in the absence of PNU. These findings were further validated using two-electrode voltage clamp electrophysiology performed on oocytes overexpressing mouse α3ß4, α6/α3ß4 and α7 nAChRs subtypes. We observed that αCtx-AtIA displayed no or low potency in blocking α3ß4 and α6/α3ß4 receptors, respectively, but improved potency and selectivity to block α7 nAChRs when compared with αCtx-PeIA. Through the synthesis of two additional analogs of αCtx-AtIA and subsequent characterization using Constellation Pharmacology, we were able to identify residue Trp18 as a major contributor to the activity of the peptide.

Unveiling the paths of COVID-19 in a large city based on public transportation data.

Human mobility plays a key role in the dissemination of infectious diseases around the world. However, the complexity introduced by commuting patterns in the daily life of cities makes such a role unclear, especially at the intracity scale. Here, we propose a multiplex network fed with 9 months of mobility data with more than 107 million public bus validations in order to understand the relation between urban mobility and the spreading of COVID-19 within a large city, namely, Fortaleza in the northeast of Brazil. Our results suggest that the shortest bus rides in Fortaleza, measured in the number of daily rides among all neighborhoods, decreased [Formula: see text]% more than the longest ones after an epidemic wave. Such a result is the opposite of what has been observed at the intercity scale. We also find that mobility changes among the neighborhoods are synchronous and geographically homogeneous. Furthermore, we find that the most central neighborhoods in mobility are the first targets for infectious disease outbreaks, which is quantified here in terms of the positive linear relation between the disease arrival time and the average of the closeness centrality ranking. These central neighborhoods are also the top neighborhoods in the number of reported cases at the end of an epidemic wave as indicated by the exponential decay behavior of the disease arrival time in relation to the number of accumulated reported cases with decay constant [Formula: see text] days. We believe that these results can help in the development of new strategies to impose restriction measures in the cities guiding decision-makers with smart actions in public health policies, as well as supporting future research on urban mobility and epidemiology.

Renal amyloidosis: a new time for a complete diagnosis.

Amyloidoses are a group of disorders in which soluble proteins aggregate and deposit extracellularly in tissues as insoluble fibrils, causing organ dysfunction. Clinical management depends on the subtype of the protein deposited and the affected organs. Systemic amyloidosis may stem from anomalous proteins, such as immunoglobulin light chains or serum amyloid proteins in chronic inflammation or may arise from hereditary disorders. Hereditary amyloidosis consists of a group of rare conditions that do not respond to chemotherapy, hence the identification of the amyloid subtype is essential for diagnosis, prognosis, and treatment. The kidney is the organ most frequently involved in systemic amyloidosis. Renal amyloidosis is characterized by acellular pathologic Congo red-positive deposition of amyloid fibrils in glomeruli, vessels, and/or interstitium. This disease manifests with heavy proteinuria, nephrotic syndrome, and progression to end-stage kidney failure. In some situations, it is not possible to identify the amyloid subtype using immunodetection methods, so the diagnosis remains indeterminate. In cases where hereditary amyloidosis is suspected or cannot be excluded, genetic testing should be considered. Of note, laser microdissection/mass spectrometry is currently the gold standard for accurate diagnosis of amyloidosis, especially in inconclusive cases. This article reviews the clinical manifestations and the current diagnostic landscape of renal amyloidosis.

Nephrotic syndrome in the elderly: epidemiological aspects, clinical data, and renal biopsy findings.

Nephrotic syndrome is the most common clinical presentation of glomerular disease in elderly patients, and renal biopsy is an important diagnostic resource. The aim of this study was to describe nephrotic syndrome among elderly patients in Brazil, focusing on tubulointerstitial and vascular involvement. This was a retrospective study of patients over 65 years of age with nephrotic syndrome who underwent renal biopsy between January 2012 and December 2019. Of the 123 renal biopsies that occurred during the study period, 44 (35.8%) were performed for the investigation of nephrotic syndrome. Among those 44 cases, the main etiologies were membranous nephropathy in 13 cases (29.5%), amyloidosis in ten (22.7%), non-collapsing focal segmental glomerulosclerosis (FSGS) in four (9.1%), and collapsing FSGS in four (9.1%). Patients with minimal change disease (MCD) had the lowest degree of interstitial fibrosis compared with the other glomerulopathies, and histological signs of acute tubular necrosis (ATN) were less common among those with amyloidosis than among those with membranous nephropathy, FSGS, or MCD (P=0.0077). Of the patients with ATN, the frequency of acute kidney injury (AKI) was highest in those with MCD (P<0.001). All patients had some degree of vascular involvement, regardless of the type of glomerulopathy. In conclusion, the second most common cause of nephrotic syndrome in this population was amyloidosis, and acute interstitial tubule involvement was more marked in MCD. Vascular involvement is something that cannot be dissociated from the age of the patient and is not only due to the underlying glomerulopathy.

Renal amyloidosis: a new time for a complete diagnosis

Amyloidoses are a group of disorders in which soluble proteins aggregate and deposit extracellularly in tissues as insoluble fibrils, causing organ dysfunction. Clinical management depends on the subtype of the protein deposited and the affected organs. Systemic amyloidosis may stem from anomalous proteins, such as immunoglobulin light chains or serum amyloid proteins in chronic inflammation or may arise from hereditary disorders. Hereditary amyloidosis consists of a group of rare conditions that do not respond to chemotherapy, hence the identification of the amyloid subtype is essential for diagnosis, prognosis, and treatment. The kidney is the organ most frequently involved in systemic amyloidosis. Renal amyloidosis is characterized by acellular pathologic Congo red-positive deposition of amyloid fibrils in glomeruli, vessels, and/or interstitium. This disease manifests with heavy proteinuria, nephrotic syndrome, and progression to end-stage kidney failure. In some situations, it is not possible to identify the amyloid subtype using immunodetection methods, so the diagnosis remains indeterminate. In cases where hereditary amyloidosis is suspected or cannot be excluded, genetic testing should be considered. Of note, laser microdissection/mass spectrometry is currently the gold standard for accurate diagnosis of amyloidosis, especially in inconclusive cases. This article reviews the clinical manifestations and the current diagnostic landscape of renal amyloidosis.

Nephrotic syndrome in the elderly: epidemiological aspects, clinical data, and renal biopsy findings

Nephrotic syndrome is the most common clinical presentation of glomerular disease in elderly patients, and renal biopsy is an important diagnostic resource. The aim of this study was to describe nephrotic syndrome among elderly patients in Brazil, focusing on tubulointerstitial and vascular involvement. This was a retrospective study of patients over 65 years of age with nephrotic syndrome who underwent renal biopsy between January 2012 and December 2019. Of the 123 renal biopsies that occurred during the study period, 44 (35.8%) were performed for the investigation of nephrotic syndrome. Among those 44 cases, the main etiologies were membranous nephropathy in 13 cases (29.5%), amyloidosis in ten (22.7%), non-collapsing focal segmental glomerulosclerosis (FSGS) in four (9.1%), and collapsing FSGS in four (9.1%). Patients with minimal change disease (MCD) had the lowest degree of interstitial fibrosis compared with the other glomerulopathies, and histological signs of acute tubular necrosis (ATN) were less common among those with amyloidosis than among those with membranous nephropathy, FSGS, or MCD (P=0.0077). Of the patients with ATN, the frequency of acute kidney injury (AKI) was highest in those with MCD (P<0.001). All patients had some degree of vascular involvement, regardless of the type of glomerulopathy. In conclusion, the second most common cause of nephrotic syndrome in this population was amyloidosis, and acute interstitial tubule involvement was more marked in MCD. Vascular involvement is something that cannot be dissociated from the age of the patient and is not only due to the underlying glomerulopathy.

Non-Peptidic Small Molecule Components from Cone Snail Venoms.

Venomous molluscs (Superfamily Conoidea) comprise a substantial fraction of tropical marine biodiversity (>15,000 species). Prior characterization of cone snail venoms established that bioactive venom components used to capture prey, defend against predators and for competitive interactions were relatively small, structured peptides (10-35 amino acids), most with multiple disulfide crosslinks. These venom components ("conotoxins, conopeptides") have been widely studied in many laboratories, leading to pharmaceutical agents and probes. In this review, we describe how it has recently become clear that to varying degrees, cone snail venoms also contain bioactive non-peptidic small molecule components. Since the initial discovery of genuanine as the first bioactive venom small molecule with an unprecedented structure, a broad set of cone snail venoms have been examined for non-peptidic bioactive components. In particular, a basal clade of cone snails (Stephanoconus) that prey on polychaetes produce genuanine and many other small molecules in their venoms, suggesting that this lineage may be a rich source of non-peptidic cone snail venom natural products. In contrast to standing dogma in the field that peptide and proteins are predominantly used for prey capture in cone snails, these small molecules also contribute to prey capture and push the molecular diversity of cone snails beyond peptides. The compounds so far characterized are active on neurons and thus may potentially serve as leads for neuronal diseases. Thus, in analogy to the incredible pharmacopeia resulting from studying venom peptides, these small molecules may provide a new resource of pharmacological agents.

Characterization of the First Conotoxin from Conus ateralbus, a Vermivorous Cone Snail from the Cabo Verde Archipelago.

Conus ateralbus is a cone snail endemic to the west side of the island of Sal, in the Cabo Verde Archipelago off West Africa. We describe the isolation and characterization of the first bioactive peptide from the venom of this species. This 30AA venom peptide is named conotoxin AtVIA (δ-conotoxin-like). An excitatory activity was manifested by the peptide on a majority of mouse lumbar dorsal root ganglion neurons. An analog of AtVIA with conservative changes on three amino acid residues at the C-terminal region was synthesized and this analog produced an identical effect on the mouse neurons. AtVIA has homology with δ-conotoxins from other worm-hunters, which include conserved sequence elements that are shared with δ-conotoxins from fish-hunting Conus. In contrast, there is no comparable sequence similarity with δ-conotoxins from the venoms of molluscivorous Conus species. A rationale for the potential presence of δ-conotoxins, that are potent in vertebrate systems in two different lineages of worm-hunting cone snails, is discussed.

Worse renal outcome of subclass IV-G lupus nephritis patients over IV-S.

Background International Society of Nephrology/ Renal Pathology Society (ISN/RPS) consensus on the classification of lupus nephritis (LN) subdivided class IV into diffuse segmental (IV-S) and diffuse global (IV-G). Nephrologists and nephropathologists believe that this subclassification would be clinically relevant based on hypothetical distinct immunopathogenesis of those subclasses guiding therapy as well as judging prognosis. Methods All adult patients with a renal biopsy-confirmed diagnosis of LN class IV undergoing regular follow-up in the Nephrology Division between January 2004 and December 2014 were enrolled excluding those with diabetes, hepatitis B, hepatitis C, HIV as well as those with insufficient clinical and hystopathological data. Biopsies were reviewed and reclassified according to ISN/RPS 2003 classification by two experienced pathologists and were examined by light microscopy and direct immunofluorescence. Results On baseline subclass IV-G compared to IV-S showed higher frequency of males and histologically higher activity (7.5 ± 2.8 vs 5.1 ± 2.3, p = 0.004) and chronicity index (3.4 ± 1.6 vs 2.4 ± 1.8, p = 0.016) as well as a higher percentage of epithelial crescents (12.9 vs 5.1, p = 0.0001) and vessel abnormalities (72% vs 42%, p = 0.017). Although renal function on baseline was not different between subclasses, IV-G showed lower levels, although not significant, of estimated glomerular filtration based on CKD-EPI formula (91.0 ± 34.8 vs 64.4 ± 44.5, p = 0.059) at the end of follow-up. In addition, we observed a higher rate of patients reaching CKD-EPI under 60 mL/min/1.73 m2 in subclass IV-G over IV-S on last follow-up. Conclusion Subclasses IV-S and IV-G patients show some clinical and pathological differences that might represent distinct stages of the same disease and they should thus be treated the same.

Small Molecules in the Cone Snail Arsenal.

Cone snails are renowned for producing peptide-based venom, containing conopeptides and conotoxins, to capture their prey. A novel small-molecule guanine derivative with unprecedented features, genuanine, was isolated from the venom of two cone snail species. Genuanine causes paralysis in mice, indicating that small molecules and not just polypeptides may contribute to the activity of cone snail venom.

Lupus nephritis is more severe in children and adolescents than in older adults.

OBJECTIVE: To evaluate clinicopathological features and treatment response in patients with lupus nephritis (LN), comparing the childhood- and late-onset forms of the disease. METHODS: We retrospectively analyzed clinical presentation, treatment and evolution in patients diagnosed with LN by renal biopsy between 1999 and 2008. Patients were grouped by age-≤18 years (n = 23); and ≥50 years (n = 13)-and were followed for the first year of treatment. RESULTS: The baseline features of the childhood- and late-onset groups, respectively, were as follows: mean age, 15 ± 2 and 54 ± 5 years; female gender, 87% and 92%; hypertension, 87% and 77%; Systemic Lupus Erythematosus Disease Activity Index, 29 ± 9 and 17 ± 7 (p = 0.002); estimated glomerular filtration rate (eGFR), 86 ± 66 and 70 ± 18 ml/min; concurrent SLE/LN diagnosis, 90% and 15% (p < 0.001); crescents on biopsy, 74% and 30% (p = 0.02); activity index on biopsy, 4.8 ± 2.6 and 3.3 ± 1.9 (p = 0.10); and interstitial fibrosis (>10%), 39% and 61% (p = 0.08). Treatment consisted mainly of methylprednisolone, prednisone and intravenous cyclophosphamide, average cumulative doses being similar between the groups. After 12 months of treatment, the eGFR in the younger and older patients was 116 ± 62 and 78 ± 20 ml/min, respectively (p = 0.005). Three of the younger patients progressed to dialysis at 12 months, compared with none of the older patients. CONCLUSION: Childhood-onset LN seems to be more severe than is late-onset LN.

Cultivation of Arthrospira (spirulina) platensis in desalinator wastewater and salinated synthetic medium: protein content and amino-acid profile

Arthrospira (Spirulina) platensis was cultivated in laboratory under controlled conditions (30ºC, photoperiod of 12 hours light/dark provided by fluorescent lamps at a light intensity of 140 µmol photons.m-2.s-1 and constant bubbling air) in three different culture media: (1) Paoletti medium (control), (2) Paoletti supplemented with 1 g.L-1 NaCl (salinated water) and (3) Paoletti medium prepared with desalinator wastewater. The effects of these treatments on growth, protein content and amino acid profile were measured. Maximum cell concentrations observed in Paoletti medium, Paoletti supplemented with salinated water or with desalinator wastewater were 2.587, 3.545 and 4.954 g.L-1, respectively. Biomass in medium 3 presented the highest protein content (56.17 percent), while biomass in medium 2 presented 48.59 percent protein. All essential amino acids, except lysine and tryptophan, were found in concentrations higher than those requiried by FAO.

Arthrospira (Spirulina) platensis foi cultivada em laboratório sob condições controladas (30ºC, intensidade luminosa de 140 µmol fótons.m-2.s-1, 12 horas claro/escuro e insuflação constante de ar atmosférico), em três meios de cultivo: (1) meio de Paoletti (controle), (2) meio de Paoletti suplementado com 1,0 g.L-1 de NaCl (água salinizada) e (3) meio de Paoletti preparado com rejeito de dessalinizador. Foi verificado o efeito destes tratamentos no crescimento, teor de proteínas e aminoácidos. As concentrações celulares máximas obtidas foram de 2,587; 3,545 e 4,954 g.L-1 no meio controle, meio de Paoletti suplementado com água salinizada ou com rejeito de dessalinizador, respectivamente. Com relação às concentrações protéicas, estas foram maiores na biomassa cultivada no meio 3, com 56,17 por cento, enquanto que a biomassa cultivada no meio 2 apresentou 48,59 por cento. A maioria dos aminoácidos essenciais encontrou-se acima dos limites requeridos pela FAO, com exceção apenas de lisina e triptofano.

Cultivation of Arthrospira (spirulina) platensis in desalinator wastewater and salinated synthetic medium: protein content and amino-acid profile.

Arthrospira (Spirulina) platensis was cultivated in laboratory under controlled conditions (30°C, photoperiod of 12 hours light/dark provided by fluorescent lamps at a light intensity of 140 µmol photons.m(-2).s(-1) and constant bubbling air) in three different culture media: (1) Paoletti medium (control), (2) Paoletti supplemented with 1 g.L(-1) NaCl (salinated water) and (3) Paoletti medium prepared with desalinator wastewater. The effects of these treatments on growth, protein content and amino acid profile were measured. Maximum cell concentrations observed in Paoletti medium, Paoletti supplemented with salinated water or with desalinator wastewater were 2.587, 3.545 and 4.954 g.L(-1), respectively. Biomass in medium 3 presented the highest protein content (56.17%), while biomass in medium 2 presented 48.59% protein. All essential amino acids, except lysine and tryptophan, were found in concentrations higher than those requiried by FAO.