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Multiple epidemics of chronic kidney disease of an unknown etiology (CKDu), primarily in young healthy agricultural workers, have emerged in agricultural communities around the world. We have hypothesized that the harvest and burning of sugarcane leading to inhalation of sugarcane ash may contribute to development of CKDu. Sugarcane stalks consist of ~80% amorphous silica and we have demonstrated that following burning of sugarcane, nano-sized silica particles (~200nm) are generated. To determine what effect such exposures have on kidney cells, a human proximal convoluted tubule (PCT) cell line (HK-2) was subjected to treatments ranging in concentration from 0.025µg/mL to 25µg/mL of sugarcane ash, desilicated sugarcane ash, sugarcane ash-derived silica nanoparticles (SAD SiNPs), or manufactured pristine 200nm silica nanoparticles. Following 6 to 48 hours of exposure, mitochondrial activity and viability were found to be significantly reduced when exposed to SAD particles at concentrations 2.5µg/mL or higher. Oxygen consumption rate (OCR) and pH were investigated with a Seahorse XF Analyzer, revealing significant changes to cellular metabolism across treatments as early as 6 hours following exposure. While most treatments slightly increased respiration, SAD SiNPs were found to inhibit mitochondrial function, reduce ATP generation, increase reliance on glycolysis, and reduce glycolytic reserve. Metabolomic analysis following 24 hours of exposure found several cellular energetics pathways (e.g., fatty acid metabolism, glycolysis, and TCA cycle) significantly altered across ash-based treatments. Such changes indicate that exposure to sugarcane ash and its derivatives can promote mitochondrial dysfunction and disrupt metabolic activity of human PCT cells.

We have hypothesized that sugarcane burning in developing countries contributes to an endemic kidney pathology in agricultural workers called chronic kidney disease of an unknown etiology (CKDu). Elemental analysis shows that sugarcane stalks contain a high percentage of amorphous silica (SiO2). Therefore, we hypothesized that burning of sugarcane generates silica nanoparticles (SiNPs) that present an inhalation hazard to these agricultural workers. To determine if SiNPs are present in sugarcane ash and kidney biopsies from agricultural workers, we utilized single particle inductively coupled plasma mass spectrometry and found SiNPs, which ranged in size from 190-212 nm, in sugarcane ash. In a small cohort of CKDu patients from El Salvador and Mexico, we found a significant increase in the number of SiNPs in their kidney biopsies within a similar size range as observed in sugarcane ash. Additionally, Wistar rats exposed to sugarcane ash intranasally for 24 weeks developed focal injury in the tubulointerstitium and glomeruli with deposition of SiNPs in the kidney. To further determine the cellular effects of SiNPs, we exposed human proximal convoluted tubule (PCT) cells to sugarcane ash, de-silicated ash, or sugarcane ash derived SiNPs. Despite not being directly cytotoxic to PCT cells at 24 hrs, SiNPs were taken up and generated reactive oxygen species. Vimentin staining confirmed PCT cells underwent epithelial-mesenchymal transition (EMT) following treatment with ash or SiNPs but not with de-silicated ash. These findings suggest SiNPs present in sugarcane ash are capable of driving EMT in PCT cells potentially contributing to a CKDu phenotype.

Climate change should be of special concern for the nephrologist, as the kidney has a critical role in protecting the host from dehydration, but it is also a favorite target of heat stress and dehydration. Here we discuss how rising temperatures and extreme heat events may affect the kidney. The most severe presentation of heat stress is heat stroke, which can result in severe electrolyte disturbance and both acute and chronic kidney disease (CKD). However, lesser levels of heat stress also have multiple effects, including exacerbating kidney disease and precipitating cardiovascular events in subjects with established kidney disease. Heat stress can also increase the risk for kidney stones, cause multiple electrolyte abnormalities and induce both acute and chronic kidney disease. Recently there have been multiple epidemics of CKD of uncertain etiology in various regions of the world, including Mesoamerica, Sri Lanka, India and Thailand. There is increasing evidence that climate change and heat stress may play a contributory role in these conditions, although other causes, including toxins, could also be involved. As climate change worsens, the nephrologist should prepare for an increase in diseases associated with heat stress and dehydration.

As the world braces to enter its third year in the coronavirus disease 2019 (COVID-19) pandemic, the need for accessible and effective antiviral therapeutics continues to be felt globally. The recent surge of Omicron variant cases has demonstrated that vaccination and prevention alone cannot quell the spread of highly transmissible variants. A safe and nontoxic therapeutic with an adaptable design to respond to the emergence of new variants is critical for transitioning to treatment of COVID-19 as an endemic disease. Here, we present a novel compound, called SBCoV202, that specifically and tightly binds the translation initiation site of RNA-dependent RNA polymerase within the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) genome, inhibiting viral replication. SBCoV202 is a Nanoligomer,™ a molecule that includes peptide nucleic acid sequences capable of binding viral RNA with single-base-pair specificity to accurately target the viral genome. The compound has been shown to be safe and nontoxic in mice, with favorable biodistribution, and has shown efficacy against SARS-CoV-2 in vitro. Safety and biodistribution were assessed after three separate administration methods, namely intranasal, intravenous, and intraperitoneal. Safety studies showed the Nanoligomer caused no outward distress, immunogenicity, or organ tissue damage, measured through observation of behavior and body weight, serum levels of cytokines, and histopathology of fixed tissue, respectively. SBCoV202 was evenly biodistributed throughout the body, with most tissues measuring Nanoligomer concentrations well above the compound KD of 3.37 nM. In addition to favorable availability to organs such as the lungs, lymph nodes, liver, and spleen, the compound circulated through the blood and was rapidly cleared through the renal and urinary systems. The favorable biodistribution and lack of immunogenicity and toxicity set Nanoligomers apart from other antisense therapies, while the adaptability of the nucleic acid sequence of Nanoligomers provides a defense against future emergence of drug resistance, making these molecules an attractive potential treatment for COVID-19.

Electronic cigarettes (EC) and other electronic nicotine delivery systems (ENDS) have recently become popular choices for nicotine consumption due to the lower perceived harm compared to conventional tobacco products. Currently, only nicotine levels in EC fluids are regulated by the FDA. Besides nicotine and solvents such as propylene glycol and glycerin, the heating of EC fluids may produce thermal degradants that could impact a user's health. We proposed to use direct sample introduction (DSI) GC-MS/MS as a fast and reliable instrumental method to analyze EC fluid components and their thermal degradants generated in a simulated ENDS-like environment within the DSI enclosure. DSI GC-MS/MS separates and detects the target analytes even in the presence of a complex, viscous, and often “dirty” sample matrix. DSI utilizes a programmable-temperature vaporization (PTV) injector in conjunction with a ChromatoProbe accessory, in which a solid or liquid sample in a disposable glass vial can be heated and vapors introduced into the GC column for separation. DSI was used to mimic the heating behavior of an EC atomizer and introduce the EC vapors and degradants into GC-MS/MS in almost real-time. Subsequently, through tandem mass spectrometry, signature ion fragments for thermal breakdown components (e.g. aldehydes) were detected and quantified. Relative peak ratios of those thermal breakdown products and an internal standard were employed to study the effects of temperature ramp rate, maximum heating temperature, and maximum temperature hold time on the outcomes of thermal degradation.

We present an in vivo model for specific protection of normal hepatocytes from damage by the highly specific hepatotoxin galactosamine. The idea is based on the fact that normal, unlike malignant, hepatocytes possess unique cell-surface receptors that can bind and internalize galactose terminal (asialo)glycoproteins by receptor-mediated endocytosis. A targetable carrier-antagonist conjugate was formed by coupling asialofetuin to the galactosamine antagonist uridine monophosphate. Intravenous injection of the antagonist conjugate resulted in specific uptake by the liver. Rats treated with carrier-antagonist conjugate together with a toxic dose of galactosamine developed significantly less hepatotoxicity than did controls. We conclude that a galactosamine antagonist can be targeted to liver, resulting in specific protection of hepatocytes from galactosamine toxicity in vivo. Because hepatoma cells lack asialoglycoprotein receptor activity, this "targeted rescue" may be of value in the differential protection of normal cells in the treatment of hepatocellular carcinoma.

Interspecific hybridization between the viviparous fishes Poeciliopsis monacha and P. lucida has produced two all-female forms: P. monacha-lucida, which is diploid, and P. monacha-2 lucida, a triploid. To reproduce, both must be inseminated by males of P. lucida. Expansion of unisexual populations is limited by the ability to attract males, which discriminate against hybrid forms. Unisexual clones in nature differ in their proportions of pregnant females, and laboratory studies comparing reproductive behavior and pregnancy rates demonstrate that some clones are more successful than others at acquiring sperm. When clones are at equal frequencies in a population, some attract males more readily than others. Learning by males of P. lucida to discriminate against one unisexual clone does not generalize completely to new clones; thus, unfamiliar clones are more likely to be courted than familiar clones (Keegan-Rogers 1984). Although all members of a triploid clone are genetically identical, members of a diploi...

We present evidence that normal hepatocytes can be specifically protected from galactosamine toxicity in vitro by targeting an antagonist to these cells via receptor-mediated endocytosis. The strategy is based upon the following principles: 1) galactosamine is a highly selective hepatotoxin that causes a dose-dependent depletion of uridine intermediates; 2) galactosamine toxicity can be antagonized by supplemental administration of uridine; 3) normal hepatocytes possess unique cell-surface receptors that can internalize galactose terminal (asialo-)glycoproteins with subsequent degradation of the glycoprotein ligand. Based on these facts, we hypothesized that chemical coupling of a galactosamine antagonist to an asialoglycoprotein could result in cell-specific delivery and protection of normal hepatocytes by targeting the antagonist via asialoglycoprotein receptors. Using a model system consisting of freshly isolated rat hepatocytes (receptor (+)) and Morris 7777 rat hepatoma (receptor (-)) cells, sensitivity to galactosamine in vitro was determined and found to be similar for both types of cells. A targetable antagonist was synthesized by coupling uridine monophosphate to asialoorosomucoid in a molar ratio of 5 to 1. Exposure of Morris 7777 cells to the targetable antagonist in the presence of a toxic concentration of galactosamine did not protect these cells as evidenced by a steady decline in the number of viable cells in a fashion identical to cells treated with galactosamine alone. However, normal hepatocytes that received the conjugate in the presence of galactosamine were protected as their viable cell number remained the same as control (untreated) cells. Competition by an excess of asialoglycoprotein inhibited the protective effect of the conjugate, supporting the concept that the asialoglycoprotein component of the conjugate was responsible for the specific delivery of the antagonist to the target cells.

Hybridization between the viviparous fishes Poeciliopsis monacha and P. lucida of northwestern Mexico has resulted in the formation of diploid and triploid all-female ‘species’, P. monacha-lucida and P. monacha -2 lucida . These females reproduce by mating back to P. lucida , and are essentially clonally reproducing sexual parasites superimposed on that species. In a series of behavioural experiments, one diploid clone proved to be significantly more aggressive than one triploid and four other diploid clones. No differences in aggression were exhibited among the other five clones. The aggressiveness of this one clone may explain why only two clones live in the small tributary where it is found but up to 10 diploid and triploid clones occur where it is absent.

We present evidence that normal hepatocytes can be specifically protected from galactosamine toxicity in vitro by targeting an antagonist to these cells via receptor-mediated endocytosis. The strategy is based upon the following principles: 1) galactosamine is a highly selective hepatotoxin that causes a dose-dependent depletion of uridine intermediates; 2) galactosamine toxicity can be antagonized by supplemental administration of uridine; 3) normal hepatocytes possess unique cell-surface receptors that can internalize galactose terminal (asialo-)glycoproteins with subsequent degradation of the glycoprotein ligand. Based on these facts, we hypothesized that chemical coupling of a galactosamine antagonist to an asialoglycoprotein could result in cell-specific delivery and protection of normal hepatocytes by targeting the antagonist via asialoglycoprotein receptors. Using a model system consisting of freshly isolated rat hepatocytes (receptor (+)) and Morris 7777 rat hepatoma (receptor (-)) cells, sensitivity to galactosamine in vitro was determined and found to be similar for both types of cells. A targetable antagonist was synthesized by coupling uridine monophosphate to asialoorosomucoid in a molar ratio of 5 to 1. Exposure of Morris 7777 cells to the targetable antagonist in the presence of a toxic concentration of galactosamine did not protect these cells as evidenced by a steady decline in the number of viable cells in a fashion identical to cells treated with galactosamine alone. However, normal hepatocytes that received the conjugate in the presence of galactosamine were protected as their viable cell number remained the same as control (untreated) cells. Competition by an excess of asialoglycoprotein inhibited the protective effect of the conjugate, supporting the concept that the asialoglycoprotein component of the conjugate was responsible for the specific delivery of the antagonist to the target cells.

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