Physiological Route Research Articles

Morning Engagement of Hepatic Insulin Receptors Improves Afternoon Hepatic Glucose Disposal and Storage.

Glucose tolerance improves significantly upon consuming a second, identical meal later in the day (second meal phenomenon). We previously established that morning hyperinsulinemia primes the liver for increased afternoon hepatic glucose uptake (HGU). Although the route of insulin delivery is an important determinant of the mechanisms by which insulin regulates liver glucose metabolism (direct hepatic vs indirect insulin action), it is not known if insulin's delivery route affects the second meal response. To determine whether morning peripheral insulin delivery (as occurs clinically (subcutaneous)) can enhance afternoon HGU, conscious dogs were treated in the morning with insulin delivered via the portal vein, or peripherally (leg vein), while glucose was infused to maintain euglycemia. Consequently, arterial insulin levels increased similarly in both groups, but relative hepatic insulin deficiency occurred when insulin was delivered peripherally. In the afternoon, all animals were challenged with the same hyperinsulinemic-hyperglycemic clamp to simulate identical postprandial-like conditions. The substantial enhancement of HGU in the afternoon caused by morning portal vein insulin delivery was lost when insulin was delivered peripherally. This indicates that morning insulin does not cause the second meal phenomenon via its indirect actions on the liver, but rather through direct activation of hepatic insulin signaling.

Neuronal expression of herpes simplex virus-1 VP16 protein induces pseudorabies virus escape from silencing and reactivation.

Alpha herpesvirus (α-HV) particles enter their hosts from mucosal surfaces and efficiently maintain fast transport in peripheral nervous system (PNS) axons to establish infections in the peripheral ganglia. The path from axons to distant neuronal nuclei is challenging to dissect due to the difficulty of monitoring early events in a dispersed neuron culture model. We have established well-controlled, reproducible, and reactivateable latent infections in compartmented rodent neurons by infecting physically isolated axons with a small number of viral particles. This system not only recapitulates the physiological infection route but also facilitates independent treatment of isolated cell bodies or axons. Consequently, this system enables study not only of the stimuli that promote reactivation but also the factors that regulate the initial switch from productive to latent infection. Adeno-associated virus (AAV)-mediated expression of herpes simplex-1 (HSV-1) VP16 alone in neuronal cell bodies enabled the escape from silencing of incoming pseudorabies virus (PRV) genomes. Furthermore, the expression of HSV VP16 alone reactivated a latent PRV infection in this system. Surprisingly, the expression of PRV VP16 protein supported neither PRV escape from silencing nor reactivation. We compared transcription transactivation activity of both VP16 proteins in primary neurons by RNA sequencing and found that these homolog viral proteins produce different gene expression profiles. AAV-transduced HSV VP16 specifically induced the expression of proto-oncogenes including c-Jun and Pim2. In addition, HSV VP16 induces phosphorylation of c-Jun in neurons, and when this activity is inhibited, escape of PRV silencing is dramatically reduced.IMPORTANCEDuring latency, alpha herpesvirus genomes are silenced yet retain the capacity to reactivate. Currently, host and viral protein interactions that determine the establishment of latency, induce escape from genome silencing or reactivation are not completely understood. By using a compartmented neuronal culture model of latency, we investigated the effect of the viral transcriptional activator, VP16 on pseudorabies virus (PRV) escape from genome silencing. This model recapitulates the physiological infection route and enables the study of the stimuli that regulate the initial switch from a latent to productive infection. We investigated the neuronal transcriptional activation profiles of two homolog VP16 proteins (encoded by HSV-1 or PRV) and found distinct gene activation signatures leading to diverse infection outcomes. This study contributes to understanding of how alpha herpesvirus proteins modulate neuronal gene expression leading to the initiation of a productive or a latent infection.

Silica Wort Supplementation as an Alternative for Yeast Stress Relief on Corn Ethanol Production with Cell Recycling

In very high gravity (VHG) fermentation, yeast cells are subjected to a multitude of challenging conditions, including the osmotic pressure exerted by the high sugar content of the wort and the stress factors associated with the high ethanol concentrations present at the end of the fermentation cycle. The response of this biological system to abiotic stresses may be enhanced through biochemical and physiological routes. Silica may play a significant role in regulating the cellular homeostasis of yeast. Alternatively, it is expected that this outcome may be achieved through biochemical responses from the effects of vitamins on yeast cells and the physiological yeast route changing by the culture medium aeration. The objective of this study was to investigate the effects of adding 500 mg L−1 of silica on corn ethanol wort medium and the possibility of supplementing the same wort with vitamins alongside aeration (0.2 v v−1 min−1) as an alternative resource to sustain the fermentation yield rather than adding silica in a fed-batch fermentation cycle with yeast recycling. Upon completion of the five fermentation cycles, yeast samples subjected to the treatment with the addition of silica exhibited a 3.1% higher fermentation yield in comparison to the results observed in the vitamins plus aeration medium bath. Even though greater biomass production (19.1 g L−1) was observed through aerobic yeast behavior in vitaminized supplemented corn medium, the provided silica had a more beneficial effect on yeast stress relief for very high gravity fermentation in a corn hydrolyzed wort with cell recycling.

Use of Corn Steep Liquor as a Biostimulant in Agriculture

Biostimulants are substances or microorganisms that are applied to plants, soil, or seeds, to improve the growth, development, performance, and quality of crops. Their application is mainly directed towards improving the resistance of crops against abiotic and biotic stresses. These compounds are formulated from a great variety of compounds: humic substances, complex organic materials (sewage sludge extracts, composts, and manure), chemical elements (Al, Co, Na, Se, and Si), inorganic salts including phosphite, seaweed extracts (brown, red, and green), amino acids, etc. As of today, it has been observed that corn steep liquor (CSL), which is obtained from the industrial process of corn transformation, may be a good ingredient for the formulation of biostimulant products. CSL contains a large amount of different chemical compounds with biological activity for the plants and soil. The use of CSL industrial waste, previously formulated, could have a direct or indirect effect on the physiological processes and metabolic routes of plants related to the adaptation to abiotic and biotic stresses, as their compounds are part of these metabolic pathways, act as elicitor compounds, and/or have their own biological activity in the plants. There is evidence that the application of CSL could protect plants from specific abiotic and biotic stresses, such as an excess of light or temperature, nutritional limitations, salinity, drought, or pathogens. In addition, it can improve the growth of the plant when these are grown in hydroponic systems, and can improve the health of soils. The present article is focused on describing the most relevant scientific aspects of CSL when used as an ingredient to formulate biostimulants for agriculture. It will discuss its chemical composition, the analytical techniques utilized to elucidate and quantify its compounds, its uses in agriculture, and mode of action in the plants.

A novel TASK channel antagonist nasal spray reduces sleep apnea severity in physiological responders: a randomized, blinded, trial.

Preclinical and human physiological studies indicate that topical, selective TASK 1/3 K+ channel antagonism increases upper airway dilator muscle activity and reduces pharyngeal collapsibility during anesthesia and nasal breathing during sleep. The primary aim of this study was to determine the effects of BAY2586116 nasal spray on obstructive sleep apnea (OSA) severity and whether individual responses vary according to differences in physiological responses and route of breathing. Ten people (5 females) with OSA [apnea-hypopnea index (AHI) = 47 ± 26 events/h (means ± SD)] who completed previous sleep physiology studies with BAY2586116 were invited to return for three polysomnography studies to quantify OSA severity. In random order, participants received either placebo nasal spray (saline), BAY2586116 nasal spray (160 µg), or BAY2586116 nasal spray (160 µg) restricted to nasal breathing (chinstrap or mouth tape). Physiological responders were defined a priori as those who had improved upper airway collapsibility (critical closing pressure ≥2 cmH2O) with BAY2586116 nasal spray (NCT04236440). There was no systematic change in apnea-hypopnea index (AHI3) from placebo versus BAY2586116 with either unrestricted or nasal-only breathing versus placebo (47 ± 26 vs. 43 ± 27 vs. 53 ± 33 events/h, P = 0.15). However, BAY2586116 (unrestricted breathing) reduced OSA severity in physiological responders compared with placebo (e.g., AHI3 = 28 ± 11 vs. 36 ± 12 events/h, P = 0.03 and ODI3 = 18 ± 10 vs. 28 ± 12 events/h, P = 0.02). Morning blood pressure was also lower in physiological responders after BAY2586116 versus placebo (e.g., systolic blood pressure = 137 ± 24 vs. 147 ± 21 mmHg, P < 0.01). In conclusion, BAY2586116 reduces OSA severity during sleep in people who demonstrate physiological improvement in upper airway collapsibility. These findings highlight the therapeutic potential of this novel pharmacotherapy target in selected individuals.NEW & NOTEWORTHY Preclinical findings in pigs and humans indicate that blocking potassium channels in the upper airway with topical nasal application increases pharyngeal dilator muscle activity and reduces upper airway collapsibility. In this study, BAY2586116 nasal spray (potassium channel blocker) reduced sleep apnea severity in those who had physiological improvement in upper airway collapsibility. BAY2586116 lowered the next morning's blood pressure. These findings highlight the potential for this novel therapeutic approach to improve sleep apnea in certain people.

Adjuvant-independent airway sensitization and infection mouse models leading to allergic asthma.

Asthma is a chronic respiratory disease of global importance. Mouse models of allergic asthma have been instrumental in advancing research and novel therapeutic strategies for patients. The application of relevant allergens and physiological routes of exposure in such models has led to valuable insights into the complexities of asthma onset and development as well as key disease mechanisms. Furthermore, environmental microbial exposures and infections have been shown to play a fundamental part in asthma pathogenesis and alter disease outcome. In this review, we delve into physiological mouse models of allergic asthma and explore literature reports on most significant interplays between microbial infections and asthma development with relevance to human disease.

Distribution of APP in endo‐lysosomal system induced by Aβ is modulated by Gβγ signaling

AbstractBackgroundAlzheimer´s disease is characterized by the deposition of aggregated species of amyloid beta (Aβ) in the brain, which leads to progressive cognitive deficits and dementia. We recently found that Aβ assemblies, oligomers and fibrils, increase APP and BACE1 interaction in recycling endosomes and accumulation of intracellular Aβ42 in human neurons derived from iPSCs by a mechanism dependent of APP/Go/Gβγ signaling (Antonino et al 2022, https://doi.org/10.3389/fcell.2022.852738). Now, we are interested on deepen in the APP trafficking in the endo‐lysosomal pathway in order to understand the mechanism underlying such effect.MethodN2a and human neurons derived from iPSc cultures were transfected with fluorescently tagged proteins (APP‐YFP, BACE‐CH, Rab11‐RFP, LAMP1‐CH, Rab7‐CH, APP‐VN, BACE‐VC). Cultures were treated with vehicle or the Gβγ inhibitor gallein and 30 minutes later with vehicle or 1 µM of Aβ oligomeric or fibrillar. After 24 h of treatment quantitative colocalization analysis and bimolecular fluorescence complementation assays were performed.ResultWe found that Aβ induced an enrichment of APP in recycling endosomes at the expense of a decrement of its levels in lysosomes. This change in APP intracellular distribution is drive by Gβγ signaling. Moreover, we found that the changes on APP distribution correlate with an increase in its interaction with BACE1 and with an increase in the number of recycling endosomes, both events modulated by Gβγ signaling (Figure 1,2).ConclusionTogether, these results suggest that Aβ pathological assemblies induce a re‐direction of APP from its physiological route to lysosomes, to recycling endosomes which favors its encounter with BACE1 and its amyloidogenic processing. These finding elucidate the intracellular process which sustain the feed‐forward mechanisms implicated in the amyloidogenesis induced by pathological assemblies of Aβ.

Heavy Metal Content in Fish of the Barguzin River (Eastern Cisbaikalia) and Assessment of Potential Risks to Human Health

In this paper, the levels of nine heavy metals in the organs and tissues of six commercially important fish species from the Barguzin River (Eastern Cisbaikalia, Russia), bream Abramis brama, roach Rutilus rutilus, crucian carp Carassius carassius, common carp Cyprinus carpio, perch Perca fluviatilis and northern pike Esox lucius, were investigated. The distribution of HMs in the fish organs and tissues was unequal and was determined by both their physiological functions and routes of entry. The study also characterized the environmental habitats, including water and sediments, and conducted an assessment of potential risks to human health associated with fish consumption. The results showed that the levels of Fe, Zn, Cu, Ni, Cd, Pb, Cr and Hg in the muscle tissue of fish from the Barguzin River were generally low and did not exceed the maximum allowable concentrations established in Russia and worldwide. However, Mn levels in a number of samples exceeded the corresponding maximum allowable concentrations, which may be attributed to its elevated presence in the habitat. The calculated fish condition factor K showed good health status of fish from the Barguzin River. The highest Metal Pollution Index values were recorded for northern pike scales (5.9), which, however, corresponded to a low level of contamination. The other metal pollution index values for fish organs and tissues were all below five, indicating either no or very low contamination. Risk to human health was assessed using target hazard quotients and hazard index. None of these indices, both for each fish species studied and for each of the nine heavy metals, exceeded one. This indicated that consumption of the fish species studied did not pose an excessive risk of non-carcinogenic health effects to humans. The data on the content of nine heavy metals in the main commercially caught fish species from the Barguzin River indicated a low level of tissue and organ contamination by heavy metals and the absence of health risks to humans associated with their consumption as food.

Pathogenesis of the crosstalk between reproductive function and stress in animals-part 1: Hypothalamo-pituitary-adrenal axis, sympatho-adrenomedullary system and kisspeptin.

Stress is defined as a disruption of the body homeostasis in response to modest as well as perceived challenge. Two main physiological routes, the hypothalamic-pituitary-adrenal system (HPA) and the sympatho-adrenomedullary system (SAM), aim to maintain or restore homeostasis by mutual interaction. SAM is quickly-reacting as it primarily works through the nervous system-the sympathetic nervous system. In response to stress, signals are sent to activate the adrenal medulla which releases catecholamines (primarily adrenaline and norepinephrine). The catecholamines have a momentary effect on the body's organs that are prepared for a fight situation. At the same time, the stressor activates the HPA axis by signals from the brain causing secretion of the pituitary hormone adrenocorticotropic hormone (ACTH). ACTH acts on the adrenal cortex, which secretes glucocorticoids, including cortisol. Since HPA primarily works through hormones, the system is slightly slower than SAM and gives rise to a metabolic effect. While short-term stress response is an adaptive and beneficial process, chronic or excessive stress can lead to a range of negative health outcomes including reproductive disorders and infertility. Several mechanisms have been proposed to explain the link between stress and reproduction. This includes in particular kisspeptin, which is closely related to reproduction, as it is a powerful stimulator of the Hypothalamic-pituitary-gonadal (HPG) system. The present review, through current knowledge in various male and female species, deals with the role of the SAM and the HPA, including the major action of kisspeptin and glucocorticoids that trigger the consequences of psychological or physiological stress on reproductive function.

OMNI®, the new surgical option in the treatment of glaucoma

AbstractPurpose: The mechanisms of elevation of intraocular pressure (IOP) in eyes with open‐angle glaucoma are complex and probably include problems related to alterations in the trabecular network, Schlemm channel and distal collecting channels. Traditional surgeries, such as trabeculectomy and the implantation of a shunt microtube, avoid these structures and divert the aqueous humour to the subconjunctival space. Recently, a growing family of minimally invasive glaucoma (MIGS) surgeries has evolved to achieve the reduction of IOP without blistering by restoring flow through the conventional physiological route of exit. One of the devices in this family is the OMNI® surgical system, which was designed to address the three sources of resistance to output flow: trabecular network, Schlemm channel and distal collectors. Thanks to this device it is possible to perform two procedures with a single surgery and overcome three points of flow resistance.The relative novelty of this device makes it important to monitor and analyse long‐term results in normal clinical practice.Methods: We present 6 clinical cases of chronic open‐angle glaucoma that were subjected to surgery with the OMNI® device, associating cataract phacoemulsification in 5 of them.Results: The reduction in IOP that we have observed at 3 and 6 months is around 30%, which is very similar to what has been published in the literature.Conclusions: In our experience, the OMNI® surgical system has provided a clinically relevant decrease in IOP and the number of active ingredients needed, with a good safety profile, although a greater volume of patients and follow‐up time are required to draw more precise conclusions.

Aβ plaques do not protect against HSV-1 infection in a mouse model of familial Alzheimer's disease, and HSV-1 does not induce Aβ pathology in a model of late onset Alzheimer's disease.

The possibility that the etiology of late onset Alzheimer's disease is linked to viral infections of the CNS has been actively debated in recent years. According to the antiviral protection hypothesis, viral pathogens trigger aggregation of Aβ peptides that are produced as a defense mechanism in response to infection to entrap and neutralize pathogens. To test the causative relationship between viral infection and Aβ aggregation, the current study examined whether Aβ plaques protect the mouse brain against Herpes Simplex Virus 1 (HSV-1) infection introduced via a physiological route and whether HSV-1 infection triggers formation of Aβ plaques in a mouse model of late-onset AD that does not develop Aβ pathology spontaneously. In aged 5XFAD mice infected via eye scarification, high density of Aβ aggregates did not improve survival time or rate when compared with wild type controls. In 5XFADs, viral replication sites were found in brain areas with a high density of extracellular Aβ deposits, however, no association between HSV-1 and Aβ aggregates could be found. To test whether HSV-1 triggers Aβ aggregation in a mouse model that lacks spontaneous Aβ pathology, 13-month-old hAβ/APOE4/Trem2*R47H mice were infected with HSV-1 via eye scarification with the McKrae HSV-1 strain, intracranial inoculation with McKrae, intracranial inoculation after priming with LPS for 6 weeks, or intracranial inoculation with high doses of McKrae or 17syn + strains that represent different degrees of neurovirulence. No signs of Aβ aggregation were found in any of the experimental groups. Instead, extensive infiltration of peripheral leukocytes was observed during the acute stage of HSV-1 infection, and phagocytic activity of myeloid cells was identified as the primary defense mechanism against HSV-1. The current results argue against a direct causative relationship between HSV-1 infection and Aβ pathology.

Intraperitoneal Insulin Delivery: Evidence of a Physiological Route for Artificial Pancreas From Compartmental Modeling.

Intraperitoneal insulin delivery has proven to safely overcome a major limit of subcutaneous delivery-meal announcement-and has been able to optimize glycemic control in adults under controlled experimental conditions. In addition, intraperitoneal delivery avoids peripheral hyperinsulinemia resulting from the subcutaneous route and restores a physiological liver gradient. Relying on a unique data set of intraperitoneal closed-loop insulin delivery obtained with a Model Predictive Controller (MPC), we develop a compartmental model of intraperitoneal insulin kinetics, which, once included in the UVa/Padova T1D simulator, will facilitate the investigation of various control strategies, for example, the simpler Proportional Integral Derivative controller versus MPC. Intraperitoneal insulin kinetics can be described with a 2-compartment model including liver and plasma. Intraperitoneal insulin transit is fast enough to render irrelevant the addition of a peritoneal compartment, proving the peritoneum being a virtual-not actual-transit space for insulin delivery.

Endoscopic third ventriculostomy in an infant with vein of Galen aneurysmal malformation treated by endovascular occlusion: Case report and a review of literature

IntroductionVein of Galen aneurysmal malformations (VGAMs) can, through multiple mechanisms, complicate with hydrocephalus (HCP). It is generally agreed that management strategies in this scenario should focus on endovascular embolizations. Treatment options for non-responders, however, have been only scarcely reported upon. Case presentationWe present a nine-month-old boy with a mural type VGAM complicated by HCP. Despite endovascular occlusion of the sole feeder, the child exhibited hydrocephalus progression prompting an Endoscopic Third Ventriculostomy (ETV). This procedure restored a cerebrospinal fluid (CSF) circulation otherwise impaired by aqueduct obstruction. Later, a new feeder arose and a second embolization was ultimately needed in order to achieve VGAM regression. Throughout four years of follow up, the child attained all developmental marks. Discussion/conclusionVGAMs are prone to hydrocephalus development as there is both an underlying venous congestion and a mechanical, obstructive component. Although there is a rationale for addressing both components, the underlying AV shunts and subsequent venous pressure elevations usually determine failure of traditional CSF shunting strategies. It is therefore challenging to manage HCP in patients who failed to improve following endovascular embolizations. For such cases, ETV stands as an elegant minimal invasive alternative with potential to provide a more physiologic drainage route and thus better allow for neurological development.

Postoperative functional evaluation of gastrectomy for gastric cancer.

To improve the quality of surgical procedures for gastric cancer, it is essential to consider many components comprehensively, including gastric motility, small intestinal absorption, hormones that affect gastric motility and appetite, presence or absence of vagus nerve preservation, esophageal regurgitation on endoscopic findings, in addition to whether or not there is a physiological route for food passage through the duodenum. Furthermore, proper functional evaluation cannot be performed without considering the form and amount of energy in the nutritional supplement to be loaded, and the posture at the time of investigation. The results of functional evaluation vary according to the method selected from many available options, but we believe that use of the most physiologically accurate, appropriate and selectable option will enable us to arrive at the best resection/reconstruction technique. We have reported that it is important to consider the preservation of three elements when performing gastrectomy: 1) reduction of the extent of gastrectomy, 2) preservation of the pylorus, and 3) preservation of the vagus nerve; among which preservation of the remnant stomach is the most important. Furthermore, the selection of a reconstruction method that maintains secretion of hormones beneficial to gastric motility preserves the energy balance inherent in the human body, and also provides better quality of life.

Genetics and Genomic basis of Stay-green Trait

Staygreen is one such trait in which genotypes possessing this trait maintain more photo synthetically active leaves (&amp; less senescent) than genotypes not possessing this trait. Delay of leaf senescence also known as stay-green character, has been identified as an important component in the genetic improvement of several crops to promote stress tolerance and yield gain. Although the stay-green phenotype is superficially similar in all species and genotypes, the genetic and physiological routes the traits are diverse. Photosynthetically active leaves for longer period depends on the concentration of chlorophyll pigment absorbing sunlight for photosynthesis. An multi dimensional approach for studying the senescence pathway rather than studying only the physiological role made a significant role in improvement. Hence new approaches like genomics, proteomics and metabolomics studies are necessary to understand the various transcription factors involved in regulating the leaf senescence process. Therefore, this review has aimed to bring light to major aspects of the stay-green character, showing its potential use in plant breeding.

Endoscopic pancreatic stenting using the combination technique of injection needle and needle knife for complete pancreaticojejunal anastomotic obstruction.

Watch a video of this article. A 69-year-old woman who underwent pancreaticoduodenectomy presented with chronic diarrhea and exacerbation of diabetes mellitus. Magnetic resonance cholangiopancreatography revealed dilatation of the main pancreatic duct (MPD) (Fig. 1a). Exocrine and endocrine pancreatic insufficiency due to pancreaticojejunal anastomotic stenosis (PJS) were considered as possible causes of the symptoms. Therefore, endoscopic retrograde pancreatography (ERP) was planned. On endoscope (PCF-H290ZI; Olympus Medical Systems, Tokyo, Japan) insertion, the anastomotic site (AS), which was characterized by membranous obstruction, was identified (Fig. 2a). ERP using a conventional cannulation method was considered difficult because of complete anastomotic obstruction (CAO). As we confirmed that there were no major blood vessels around the AS on computed tomography (Fig. 1b), ERP using an injection needle (23-G TOP endoscopic injection needle; TOP, Tokyo, Japan) was performed. Four punctures were required for ERP and dilatation of the MPD was observed (Fig. 2b). The AS was incised using a needle knife (KD-1L-1; Olympus Medical Systems) (Fig. 2c). After incision of the AS and guidewire placement, it was dilated using a dilator (ES dilator; Zeon Medical, Tokyo, Japan) and a 5-Fr × 5-cm pancreatic stent (Advanix Pancreatic Stent; Boston Scientific Japan, Tokyo, Japan) was placed without adverse events (Fig. 2d) (Video S1). For CAO in PJS, endoscopic ultrasound-guided pancreatic duct drainage is preferred because endoscopic pancreatic stenting under ERP (EPS) is technically challenging.1-3 However, EPS has the advantage that it can be safely performed using a physiological route.4 Similar to our technique, for CAO in hepaticojejunostomy, cholangiography and guidewire placement were simultaneously performed through a 20-G injection needle, and the access route to the bile duct was secured using a diathermic dilator via a guidewire.5 This technique should also be considered because it does not involve a blind incision with the risk of perforation. EPS using these techniques may be a treatment option for CAO. This study was conducted in accordance with the ethical standards described in the latest revision of the Declaration of Helsinki. Informed consent for patient participation and publication was received. All data relevant to the study are included in the article. Authors declare no conflict of interest for this article. Please note: The publisher is not responsible for the content or functionality of any supporting information supplied by the authors. Any queries (other than missing content) should be directed to the corresponding author for the article.

Coupling Complete Blood Count and Steroidomics to Track Low Doses Administration of Recombinant Growth Hormone: An Anti-Doping Perspective.

Growth Hormone (GH) under its human recombinant homologue (rhGH), may be abused by athletes to take advantage of its well-known anabolic and lipolytic properties; hence it is prohibited in sports by the World Anti-Doping Agency. Due to the rapid turnover of rhGH, anti-doping screening tests have turned to monitor two endocrine biomarkers (IGF-I and P-III-NP), but unfortunately, they show population-wise variability, limiting the identification rate of rhGH users. Previous studies have evidenced the numerous effects of GH on human physiology, especially in hematopoiesis and steroidogenesis. In this work, aiming to discover novel physiological rhGH biomarkers, we analyzed the complete blood count and the steroidomics profile of healthy, physically active, young males treated either with EPO + rhGH or EPO + placebo. The time-trends of these two physiological routes have been analyzed through geometric trajectory analysis (GTA) and OPLS-DA. Individuals supplemented with micro-doses of rhGH exhibited different leukopoietic and steroidal profiles compared to the control population, suggesting a role of the rhGH in both pathways. In the article, hypotheses on the observed differences are discussed according to the most recent literature and compared to results in animal models. The use of leukopoietic and steroidal biomarkers together with endocrine biomarkers (IGF-1 and P-III-NP) allows to correctly classify over 98% of samples with no false positives, miss-classifying only one single sample (false negative) over a total of 56; a promising result, if compared to the current rhGH detection strategies.

A Feasibility Study of In Vivo Control and Tracking of Microrobot Using Taxicab Geometry for Direct Drug Targeting.

In vivo direct drug targeting aims at delivering drug molecules loaded on microrobots to the diseased site using the shortest possible physiological routes, which potentially improves targeting efficiency and reduces systemic toxicity. It is thus essential to consider realistic in-body limitations for direct drug targeting applications. Here, we present a novel controller for microrobot maneuver by considering four key in vivo constraints: non-Euclidean structure of capillaries, irreversibility of blood flow, invisibility of microvasculature, and inaccuracy of microrobot tracking. We use the taxicab geometry of capillaries as the a priori knowledge for steering and tracking a microrobot in lattice-like vessels. Furthermore, we introduce a minimax repulsive boundary function to prevent the microrobot from getting too close to the boundaries imposed by the direction of blood flow. We also propose a novel Kalman filtering algorithm to reduce tracking error, while avoiding possible obstacles such as vessel walls without knowing their actual locations. The proposed control method consists of four modules, namely a model predictive control module for tumor targeting, a Kalman filtering module for microrobot tracking, a blind obstacle detection module, and a vessel structure estimation module. The interplay of these four modules offers successful maneuver and tracking of the microrobot while avoiding obstacles in a blind manner by utilizing the taxicab geometry of blood vessels. We present a comprehensive in silico simulation study to verify our designed controller.

A Mouse Model of Sublethal Leptospirosis: Protocols for Infection with Leptospira Through Natural Transmission Routes, for Monitoring Clinical and Molecular Scores of Disease, and for Evaluation of the Host Immune Response

Leptospirosis is a zoonotic disease caused by pathogenic Leptospira species that are maintained in sylvatic and domestic environments by transmission among rodents and other carriers. Humans become infected after contact of breached skin or mucosa with contaminated water or soil. Understanding persistent or sublethal infection in a host is critical for controlling human risk of exposure to pathogenic Leptospira. Animal models that recapitulate disease progression after infection via natural transmission routes are more appropriate for validation of vaccines and therapeutics. Furthermore, the ability to measure shedding of live Leptospira in urine of reservoir and carrier hosts can be used to develop new diagnostic assays and sensors to evaluate human risk of exposure. We developed inbred mouse models of Leptospirosis, that bypass survival as a criterion, in which we can analyze both pathogen and host factors affecting sublethal infection (<1 month), including shedding of Leptospira in urine. Mice are infected with pathogenic Leptospira using a physiologic route, and the clinical, histological, and molecular scores of disease are measured. Furthermore, the host immune response to Leptospira is evaluated. This mouse model also provides a tool in which to test fundamental hypotheses related to host-pathogen interactions and the immune mechanisms engaged in protective and pathogenic immune responses. © 2020 Wiley Periodicals LLC Basic Protocol 1: Culture and maintenance of virulent Leptospira Basic Protocol 2: Infection of mice through a physiologic route and collection of clinical scores and biological samples Basic Protocol 3: Analysis of pathogenesis after Leptospira infection.

Oral Insulin Delivery Using Poly (Styrene Co-Maleic Acid) Micelles in a Diabetic Mouse Model.

The oral delivery of insulin is a convenient and safe physiological route of administration for management of diabetes mellitus. In this study, we developed a poly-(styrene-co-maleic acid) (SMA) micellar system for oral insulin delivery to overcome the rapid degradation of insulin in the stomach, improve its absorption in the intestine, and provide a physiologically-relevant method of insulin to reach portal circulation. The insulin was encapsulated into SMA micelles in a pH-dependent process. The charge and size of the nanoparticles were determined by dynamic light scattering. The insulin loading of the nanoparticles was measured by HPLC. The transport of the SMA-insulin through biological membranes was assessed in vitro using Caco-2 cells, ex vivo rat intestinal section, and in vivo in a streptozotocin-induced diabetes mouse model. SMA-insulin micelles were negatively charged and had a mean diameter of 179.7 nm. SMA-insulin efficiently stimulated glucose uptake in HepG-2 hepatic cells and was transported across the Caco-2 epithelial cells in vitro by 46% and ex vivo across intestinal epithelium by 22%. The animal studies demonstrated that orally-administered SMA-insulin can produce a hypoglycemic effect up to 3 h after administration of one dose. Overall, our results indicate that SMA micelles are capable of the oral delivery of bioactive compounds like insulin and can be effective tools in the management of diabetes.