Sham-operated Control Group Research Articles

Is castration leading to biological aging in dogs? Assessment of lipid peroxidation, inflammation, telomere length, mitochondrial DNA copy number, and expression of telomerase and age-related genes

BackgroundBiological aging is a complex process influenced by various factors, including reproductive status and castration. This study aimed to evaluate the impact of castration on biological aging in dogs.MethodFifteen male crossbred dogs were randomly divided into a sham-operation control group (n = 5) and a castrated group (n = 10). Blood samples were collected at weeks 0, 4, 8, 12, 16, and 18 post-surgery. Malondialdehyde (MDA as indicator of Lipid peroxidation), C-reactive protein (as an indicator of inflammation), telomere length, mitochondrial DNA (mtDNA) copy number, and the expression of age-related (P16, P21, TBX2) and telomerase-related (TERT) genes were assessed in blood samples.ResultsPlasma MDA levels were higher in the control group at weeks 16 and 18, while CRP levels were higher only at week 18. Telomere length and mtDNA copy number were lower in the control group at week 18. Gene expression analysis showed that P16 was lower in the control group at weeks 8 and 12, P21 and TERT were lower at weeks 16 and 18, and TBX2 was lower at weeks 16 and 18. The TBX2/P16 ratio was lower in the control group at weeks 16 and 18 but higher at week 12, while the TBX2/P21 ratio did not differ between groups.ConclusionCastration appears to have a protective effect against biological aging in dogs, as evidenced by lower lipid peroxidation, inflammation, and age-related changes in telomere length, mtDNA copy number, and gene expression.

Investigating the Impact of Estrogen Levels on Voiding Characteristics, Bladder Structure, and Related Proteins in a Mouse Model of Menopause-Induced Lower Urinary Tract Symptoms.

Lower urinary tract symptoms (LUTS) are common in postmenopausal women. These symptoms are often linked to decreased estrogen levels following menopause. This study investigated the relationship between estrogen levels, alterations in bladder tissue structure, bladder function, and the incidence of urinary frequency. An age-appropriate bilateral ovariectomized mouse model (OVX) was developed to simulate conditions of estrogen deficiency. Mice were divided into three groups: a sham-operated control group, OVX, and an estradiol-treated group. The assessments included estrogen level measurement, urination frequency, cystometry, histological analysis, immunofluorescence staining, and real-time quantitative PCR. Additionally, we quantified the expression of the mechanosensitive channel proteins Piezo1 and TRPV4 in mouse bladder tissues. Lower estrogen levels were linked to increased voiding episodes and structural changes in mouse bladder tissues, notably a significant increase in Collagen III fiber deposition. There was a detectable negative relationship between estrogen levels and the expression of Piezo1 and TRPV4, mechanosensitive proteins in mouse bladder tissues, which may influence voiding frequency and nocturia. Estrogen treatment could improve bladder function, decrease urination frequency, and reduce collagen deposition in the bladder tissues. This study explored the connection between estrogen levels and urinary frequency, potentially setting the stage for novel methods to address frequent urination symptoms in postmenopausal women.

Effects of gastric bypass on the digestibility and postprandial metabolic fate of 15N dietary protein in rats.

Roux-en-Y Gastric Bypass may be associated with an alteration of protein bioavailability in relation to intestinal remodeling. Our study aimed to test this hypothesis by Roux-en-Y Gastric Bypass. Diet-induced obese rats underwent Roux-en-Y Gastric Bypass surgery (RYGB rats) while a Sham-operated control group was used. All rats received a 15N-labeled protein meal 1 or 3 months after surgery and were euthanized 6h later. Protein digestibility, 15N recovered in organs and urea pool, fractional protein synthesis rate, and intestinal morphometry were assessed. Protein digestibility was similar in all groups (94.2±0.3%). The small intestine was hypertrophied in RYGB rats 1 month after surgery, weighing 9.1±0.2g vs. 7.0±0.3g in Sham rats (P = 0.003). Villus height and crypt depth were increased in the alimentary limb and ileum of RYGB rats. However, Roux-en-Y Gastric Bypass had no impact on the fractional synthesis rate. In the gastrointestinal tract, 15N retention only differed in the ileal mucosa and was higher in RYGB rats at 1 month (0.48±0.2% vs. 0.3±0.09%, P = 0.03). 15N recovery from the liver, muscle, and skin was lower in RYGB rats at 1 month. 15N recovery from urinary and plasma urea was higher in RYGB rats at both times, resulting in increased total deamination (13.2±0.9% vs. 10.1±0.5%, P<0.01). This study showed that Roux-en-Y Gastric Bypass did not affect protein digestibility. Dietary nitrogen sequestration was transitorily and moderately diminished in several organs. This was associated with a sustained elevation of postprandial deamination after Roux-en-Y Gastric Bypass, whose mechanisms merit further studies.

Minor micro-rheological alterations in the presence of an artificial saphenous arteriovenous shunt, as an arteriovenous malformation model in the rat.

Arteriovenous malformations (AVMs) are vascular anomalies characterized by abnormal shunting between arteries and veins. The progression of the AVMs and their hemodynamic and rheological relations are poorly studied, and there is a lack of a feasible experimental model. To establish a model that cause only minimal micro-rheological alterations, compared to other AV models. Sixteen female Sprague Dawley rats were randomly divided into control and AVM groups. End-to-end anastomoses were created between the saphenous veins and arteries to mimic AVM nidus. Hematological and hemorheological parameters were analyzed before surgery and on the 1st, 3rd, 5th, 7th, 9th, and 12th postoperative weeks. Compared to sham-operated Control group the AVM group did not show important alterations in hematological parameters nor in erythrocyte aggregation and deformability. However, slightly increased aggregation and moderately decreased deformability values were found, without significant differences. The changes normalized by the 12th postoperative week. The presented rat model of a small-caliber AVM created on saphenous vessels does not cause significant micro-rheological changes. The alterations found were most likely related to the acute phase reactions and not to the presence of a small-caliber shunt. The model seems to be suitable for further studies of AVM progression.

Evaluation of individual visual implant elastomer tag marking of Japanese eel (Anguilla japonica) larvae

Visual implant elastomer (VIE) marking was evaluated as a tool for individual tagging of larval Japanese eels, Anguilla japonica, during rearing conditions. Combinations of eight colors of VIE tag were used to individually mark leptocephalus larvae (104–110 days post hatched) by injecting them with either a mixture of a silicone color material and a curing agent, or only color material. Injections were at three locations on the dorso-lateral body surface for individual identification, and a sham-operated control group was also prepared. Tag retention was 100% with or without curing agent, and individuals could be tracked until larvae completely metamorphosed into glass eels. After some initial mortality, the survival and growth rates were not different between each treatment group. These results suggested that VIE tagging could provide good individual larva identification method for research during rearing conditions.

Moxibustion Promoted Axonal Regeneration and Improved Learning and Memory of Post-stroke Cognitive Impairment by Regulating PI3K/AKt and TACC3

BackgroundThis study aimed to investigate whether moxibustion could affect PI3K/Akt pathway to regulate Transforming acidic coiled-coil containing protein 3 (TACC3) and promote axonal regeneration to improve learning and memory function in middle cerebral artery occlusion (MCAO) rats. MethodsSixty SD rats were randomly divided into 4 groups: sham-operated control group (SC), model control group (MC), model + moxibustion group (MM), and model + inhibitor + moxibustion group (MIM). The rats in MC, MM, and MIM groups were made into MCAO models, and PI3K inhibitor LY294002 was injected into the rats in MIM group before modeling; while the rats in SC group were only treated with artery separation without monofilament inserting. After that, the rats in MM and MIM groups were intervented with moxibustion. We used the Zea-Longa scale, micro-Magnetic Resonance Imaging (micro-MRI), Morris water maze (MWM), TUNEL, western blot (WB), immunofluorescence and immunohistochemistry to evaluate the neurological deficits, cerebral infarct volume, learning and memory, apoptotic cell percentage in the hippocampal, the expression level of axonal regeneration and PI3K/AKt related proteins, the expression level of TACC3. The detection of 2 h after surgery showed the result before moxibustion and 7 days after the intervention showed the results after moxibustion. ResultsAfter 7 d of intervention, the scores of Zea-Longa and the cerebral infarct volume, the escape latency, the percentage of apoptosis cells of MM group were lower than that of MC and MIM groups; the frequency of rats crossed the previous platform location, PI3K, p-Akt/t-Akt and TACC3, the level of GAP-43 in MM group was more than MC and MIM groups (P < 0.05). While no statistical difference existed between MIM group and MC group (P > 0.05). ConclusionMoxibustion can promote axonal regeneration and improve learning and memory of Post-stroke cognitive impairment via activating the PI3K/AKT signaling pathway and TACC3.

Regulatory role of BNP in the spinal center of rats with nonbacterial prostatitis.

A growing body of evidence has shown that activating spinal cord glial cells (typically astrocytes and microglial cells) is closely related to hyperpathia and persistent pain. To investigate the expression of GFAP and CR3/CD11b in cornu dorsale medullae spinalis of rats with nonbacterial prostatitis, to explore the therapeutic efficacy and action mechanism of intrathecal injection of BNP alleviating chronic neuropathic pain. Eighteen male SPF SD rats were randomly divided into sham operation control group, nonbacterial prostatitis group (NBP) and intrathecal injection BNP group, the NBP model was established by intraprostatic injection of CFA, and the spinal cord of L6-S1 segment was extracted seven days after intrathecal injection of BNP; The expression of GFAP and CR3/CD11b in dorsal horn of spinal cord were detected by immunofluorescence and Western blot. The cumulative optical density values of GFAP and CR3/CD11b immunofluorescence assay in the NBP group were higher than those in the sham operation group, with statistical significance (p⁢ï⁢¼⁢ 0.01); The expression of GFAP and CR3/CD11b in intrathecal injection BNP group were lower than those in NBP group, the differences were statistically significant (p⁢ï⁢¼⁢ 0.01). Western blot results showed that the expression of GFAP and CR3/CD11B in NBP group were higher than those in sham operation group, with statistical significance (p⁢ï⁢¼⁢ 0.05). The expression of GFAP and CR3/CD11B in intrathecal injection BNP group were lower than those in NBP group, the differences were statistically significant (p⁢ï⁢¼⁢ 0.05). Intrathecal injection of BNP can down-regulate the expressions of GFAP and CR3/CD11b in L6-S1 spinal cord of NBP rat model and to further inhibit chronic pain caused by NBP.

#1116 Hyperuricemia worsens bacterial sepsis by suppressing neutrophil functions in CKD

Abstract Background and Aims Patients with kidney disease, especially those on dialysis, are more susceptible to infections such as peritonitis, sepsis, pneumonia, and are at a higher risk for infection-related mortality. Pathogenic mechanisms underlying the secondary immunodeficiency in kidney disease include gut dysbiosis and barrier dysfunction, persistent inflammation, immune paralysis due to increased levels of immunoregulatory metabolites (e.g. uric acid) and proteins (e.g. FGF23) [1]. Currently, it is unknown whether CKD-related hyperuricemia may contribute to the immune paralysis in this context. We hypothesized that hyperuricemia (HU) may aggravate the innate immune response to infection in CKD in mice. Method Mono- and polybacterial sepsis was induced by LPS injection or cecal ligation and puncture in Alb-creERT2;Glut9lox/lox mice with HU and/or CKD, and in Glut9lox/lox (healthy) control mice. After 24 hours, systemic inflammation was evaluated by measuring serum cytokine levels, immune cell activation and recruitment, and neutrophil extracellular trap release. Tissue injury in liver, heart, and kidney was assessed by performing RT-qPCR and immunohistochemistry. In addition, kidney function was determined by measuring the glomerular filtration rate (GFR). Additionally, mice were treated with urate-lowering therapy to reduce serum uric acid levels and the immune response and functional outcomes after sepsis is quantified. Results We found that both mono- and polybacterial sepsis caused systemic inflammation as indicated by an increased number of blood neutrophils (see Figure), neutrophil activation and pro-inflammatory cytokine levels as compared to the PBS-treated or sham-operated control groups in Glut9lox/lox healthy mice, respectively. Mice with sepsis also displayed a slightly worsened kidney function. Interestingly, HU significantly increased neutrophil numbers but decreased their activation status and the ability of neutrophils to form neutrophil extracellular traps in Alb-creERT2;Glut9lox/lox mice with sepsis as compared to the control groups, which was further aggravated in mice with CKD-related HU (CKD+HU). This impaired inflammatory response was partially reversible by lowering serum uric acid with febuxostat. Conclusion Our results identify hyperuricemia related or unrelated to kidney disease as immune regulator in bacterial infection by suppressing neutrophil functions in mice. Specifically targeting uric acid may help to overcome the secondary immunodeficiency related to kidney disease during infection but enhances sterile inflammation [2].

#3023 Dietary protein intake modulates the tubular handling of the uremic retention solute indoxyl sulfate

Abstract Background and Aims Nutritional intervention constitutes a key part of the clinical management of individuals with chronic kidney disease (CKD). The gut emerged as an important source of uremic retention solutes, including indoxyl sulfate, p-cresyl sulfate and TMAO among the others. Indoxyl sulfate (IS) is produced through the gut microbial fermentation of tryptophan into indole and its conversion into indoxyl sulfate in the liver. Previous authors hypothesised that higher amounts of gut microbial metabolites can be sensed by kidney's proximal tubule cells through the use of receptors and signaling pathways. Particular evidence points to the involvement of kidney Organic Anion Transporters’(OATs') in controlling the gut-kidney communication. According to the so-called remote sensing hypothesis, kidney tubular cells respond to uremic toxins plasma fluctuations altering the activity of OAT1. Few studies have investigated whether in CKD the protein content in the diet affects gut generation, plasma retention and excretion mechanisms of indolic uremic toxins including kidney cells OAT1 expression. Method Eighteen male Sprague-Dawley rats (Janvier, France) aged 7-8 weeks and weighing between 270 and 388 g were randomly assigned to a low protein (LP) diet (n = 10) or a high protein (HP) diet (n = 8) after undergoing a 5/6 nephrectomy to develop CKD. For each diet, a sham-operated control group (n = 7 and n = 8 correspondingly) was used. After 7 weeks, urine was collected, and 8 weeks after the disease was induced, euthanasia was performed. Samples of kidney and blood were taken. mRNA and protein expression of OAT1 were sought via qPCR and Western blot. Forty patients with CKD were recruited in a cross-sectional study investigating eating habits and uremic toxins levels. Eating habits were searched via a food diary. Protein and energy intake were therefore calculated. Blood and 48 h-urine were collected. Blood creatinine and urea were determined with standard laboratory techniques. Total plasmatic and urinary IS concentrations were measured using LC-MS/MS. The fractional excretion (FE) i.e. the percentage of IS excreted relatively to the kidney filtered load, was calculated to assess IS remote sensing. Results Rats with chronic kidney disease (CKD) had considerably higher plasmatic levels of IS (p &amp;lt; .001) than sham rats. Rats with CKD on an HP diet, however, did not differ in plasma IS from rats fed an LP diet (p = .63). On the other hand, CKD rats fed an HP diet showed a substantial rise in 24-hour urinary IS (p &amp;lt; .001). The FE of IS was significantly higher (p = .005) in CKD rats on a HP diet and correlated with 24 h-urinary IS (Spearman r = .51, p value=.03) and with protein intake (Spearman r = .52, p value=.03) in CKD rats. OAT1 mRNA and protein expression was significantly higher in CKD rats given a HP diet (p = .025 and p = .009, respectively). By dividing the CKD patients into two groups by the median of protein intake, the FE of IS was significantly higher (p = .018) in the patients consuming a larger portion of proteins. Conclusion A diet higher in protein content in 5/6 nephrectomized CKD rats as well as in CKD patients likely leads to an increased production of indole in the colon. This results in increased IS fractional excretion but unchanged IS plasma retention. These findings offer more evidence for the mechanisms behind the remote sensing and signaling of uremic toxins derived from indole. In summary, a low-protein diet is still advised for people with chronic kidney disease.

Protective effects and mechanisms of dapagliflozin on renal ischemia/reperfusion injury

BackgroundRenal diseases have a significant negative impact on human health and the quality of life. Renal ischemia/reperfusion (I/R) injury is considered as one of the leading causes of renal dysfunction and tissue damage. Oxidative stress and inflammation are responsible for cellular apoptosis playing critical roles in renal I/R injury. Recent studies suggested that dapagliflozin—a medication used to treat Type 2 Diabetes—may exert protective effects on I/R injury in kidneys by alleviating oxidative stress and inflammation. Our study evaluated the protective effects of dapagliflozinon in renal I/R injury. MethodsA group of 32 male Sprague-Dawley rats were divided into four groups: 1) control group without any manipulation; 2) sham-operated control group with surgery but without I/R injury; 3) experimental group with 30-min I/R injury; and 4) therapeutic group with 30-min IR injury and dapagliflozin therapy. The fourth therapeutic group received 1 mg/kg dapagliflozin delivered once daily by oral gavage. All rats were evaluated by measurements of neutrophil gelatinase-associated lipocalin (NGAL), creatinine kinase (CR), blood urea nitrogen (BUN), kidney injury molecule-1 (KIM-1), myoglobin (MYO), creatinine kinase (CK), lactate dehydrogenase (LDH) LD, GSH, superoxide dismutase (SOD), MDA, interleukin-6 (IL-6), and tumor necrosis factor-a (TNF-a and glutathione peroxidase (GSH-Px) levels. TUNEL and flow cytometry assays evaluated apoptosis. ResultsOverall, the 30-min exposure to I/R injury significantly elevated levels of NGAL, CR, BUN, CK, LDH, KIM-1, and MYO (all p < 0.05). Inflammatory cytokine levels (IL-6 and TNF-a) were also increased after I/R injury (p > 0.05). At the same time, I/R injury decreased levels of SOD and GSH-Px (p > 0.05). In contrast, administration of dapagliflozin following I/R injury reduced renal damage, enhanced antioxidant capacity, and suppressed inflammatory responses (all p > 0.05), thus improving renal function, while reducing oxidative stress status and inflammatory responses. Further investigations revealed that dapagliflozin exerted its protective effects on renal tissues by activating the phosphatidylinositol 3-kinase-protein kinase B (PI3K-AKT) signaling pathway, inhibiting cellular apoptosis, and promoting proliferation and autophagy through bone morphogenetic protein 4 (BMP4). ConclusionThese findings documented that dapagliflozin protected kidneys from I/R injury suggesting its therapeutic potential.

Passive exercise is an effective alternative to HRT for restoring OVX induced mitochondrial dysfunction in skeletal muscle.

Postmenopausal women are more prone to develop muscle weakness, which is strongly associated with impairment of mitochondrial function in skeletal muscle. This study aimed to examine the impact of a passive exercise modality, whole-body vibration training (WBVT), on muscle mitochondrial function in ovariectomized (OVX) mice, in comparison with 17β-estradiol (E2) replacement. Female C57BL/6J mice were assigned to four groups: sham operation control group (Sham), ovariectomized group (OVX), OVX with E2 supplement group (OVX+E), and OVX with WBVT group (OVX+W). The estrous cycle, body weight, body composition, and muscle strength of the mice were monitored after the operation. Serum E2 level was assessed by enzyme-linked immunosorbent assay (ELISA). The ATP levels were determined using a luciferase-catalyzed bioluminescence assay. The activity of mitochondrial respiration chain complexes was evaluated using high-resolution respirometry (O2K). Expression levels of oxidative phosphorylation (OXPHOS), peroxisome proliferator-activated receptor gamma coactivator 1 alpha (PGC-1α), and mitochondrial transcription factor A (TFAM) were detected using western blotting. We observed decreased muscle strength and impaired mitochondrial function in the skeletal muscle of OVX mice. The vibration training alleviated these impairments as much as the E2 supplement. In addition, the vibration training was superior to the ovariectomy and the estradiol replacement regarding the protein expression of PGC-1α and TFAM. WBVT improves the OVX-induced decline in muscle strength and impairment of mitochondrial function in the skeletal muscle. This passive exercise strategy may be useful as an alternative to E2 replacement for preventing menopausal muscular weakness. Further studies are needed to understand the effects of WBVT on various physiological systems, and precautions should be taken when implementing it in patient treatment.

Subcellular distribution of prohibitin 1 in rat liver during liver regeneration and its cellular implication.

The function of prohibitin 1 (Phb1) during liver regeneration (LR) remains relatively unexplored. Our previous research identified downregulation of Phb1 in rat liver mitochondria 24 h after 70% partial hepatectomy (PHx), as determined by subcellular proteomic analysis. To investigate the potential role of Phb1 during LR. We examined changes in Phb1 mRNA and protein levels, subcellular distribution, and abundance in rat liver during LR following 70% PHx. We also evaluated mitochondrial changes and apoptosis using electron microscopy and flow cytometry. RNA-interference-mediated knockdown of Phb1 (PHBi) was performed in BRL-3A cells. Compared with sham-operation control groups, Phb1 mRNA and protein levels in 70% PHx test groups were downregulated at 24 h, then upregulated at 72 and 168 h. Phb1 was mainly located in mitochondria, showed a reduced abundance at 24 h, significantly increased at 72 h, and almost recovered to normal at 168 h. Phb1 was also present in nuclei, with continuous increase in abundance observed 72 and 168 h after 70% PHx. The altered ultrastructure and reduced mass of mitochondria during LR had almost completely recovered to normal at 168 h. PHBi in BRL-3A cells resulted in increased S-phase entry, a higher number of apoptotic cells, and disruption of mitochondrial membrane potential. Phb1 may contribute to maintaining mitochondrial stability and could play a role in regulating cell proliferation and apoptosis of rat liver cells during LR.

Betulinic acid reduces inflammation in rats with sepsis‐induced myocardial dysfunction by inhibiting TLR4/MyD88/NF‐κB signaling pathway

AbstractIn this study, we aimed to investigate the effect and underlying mechanism of betulinic acid (BA) on myocardial dysfunction in septic rats. Twenty‐four male Sprague–Dawley rats were randomly allocated into four groups (n = 6 rats/group): the sham operation (Control) group, the cecal ligation and puncture (CLP) group, the BA 10 mg/kg group, and the BA 50 mg/kg group. The septic rat model was induced through cecal ligation and puncture in the CLP and BA groups, except for the Control group. Then, cardiac function parameters were assessed using echocardiography, myocardial injury markers were quantified via biochemical assays, and myocardial histopathological injuries were observed through H&amp;E staining. Inflammatory factors in the serum were measured using ELISA assays, immunohistochemistry and qRT‐PCR were performed to determine macrophage numbers and the expression of iNOS, CD86, Arg‐1, and Mrc1 in myocardial tissue. The protein expression levels of TLR4, Myd88, and NF‐κB in myocardial tissue were assessed through western blot analysis. The results showed that BA significantly improved cardiac function, reduced myocardial injury, and attenuated inflammation in CLP rats. Specifically, BA decreased LVEDD and LVESD while increasing LVEF and LVFS. Furthermore, BA upregulated the levels of BNP, cTnT, CK‐MB, LDH, IL‐6, IL‐1β, and TNF‐α in CLP rat serum. Additionally, BA reduced macrophage infiltration, inhibited M1/M2 gene expression, and downregulated TLR4, Myd88 and NF‐κB protein expression in CLP rats myocardial tissues. In conclusion, BA can inhibit myocardial inflammation and prevent sepsis‐induced myocardial dysfunction by inhibiting TLR4/MyD88/NF‐κB signaling, thereby promoting M2 macrophage polarization in myocardial tissues.

Argonaute 2 restored erectile function and corpus cavernosum mitochondrial function by reducing apoptosis in a mouse model of cavernous nerve injury.

To determine whether the overexpression of the Argonaute RNA-induced silencing complex catalytic component 2 (Ago2) improves erectile function in mice after cavernous nerve injury (CNI). Lentiviruses containing Ago2 open reading frame (ORF) mouse clone (Ago2 O/E) were used to overexpress Ago2, and lentiviruses ORF negative control particles (NC) were used as a negative control. Three days before preparing the CNI model, we injected lentiviruses into the penises of 8-week-old male C57BL/6 mice. Animals were then divided into four groups: the sham operation control group and the CNI+phosphate-buffered saline, CNI+NC, and CNI+Ago2 O/E groups. One week later, erectile function was assessed by electrically stimulating cavernous nerves bilaterally and obtaining intracavernous pressure parameters. Penile tissue was also collected for molecular mechanism studies. Ago2 overexpression improved erectile function in mice after CNI-induced erectile dysfunction (ED). Immunofluorescence staining and Western blot analysis showed that under Ago2 overexpressing conditions, the contents of endothelial cells, pericytes, and neuronal cells increased in the penile tissues of CNI mice, and this was attributed to reduced apoptosis and ROS production. In addition, we also found that Ago2 overexpression could restore penile mitochondrial function, thereby improving erectile function in CNI-induced ED mice. Our findings demonstrate that Ago2 overexpression can reduce penile cell apoptosis, restore penile mitochondrial function, and improve erectile function in CNI-induced ED mice.

Olive oil Ameliorates renal Ischemia-reperfusion-mediated hepatic and renal changes in stressed rats

Background: Renal ischemia-reperfusion injury (IRI) does not only affect kidneys, but also affects remote organs especially the liver. Stress may lead to further progression of renal and hepatic insults. This study was planned to explore the efficacy of olive oil (OO) on the assumed renal and hepatic changes of immobilization stress following renal IRI. Methods: Seventy-seven adult male Wistar albino rats were divided into the following five groups; Sham-Operated Control group, Renal Ischemia-Reperfusion group, Stressed Renal Ischemia-Reperfusion group, OO-Supplemented Renal Ischemia-Reperfusion group) and OO-Supplemented Stressed Renal Ischemia-Reperfusion Group (OO+Stressed+IR). All animals were subjected to determination of renal and hepatic biochemical functions and histopathological examination. Results: Renal IR significantly increased plasma creatinine and urea levels. This was associated with significant rise in serum levels of both ALT and AST, total bilirubin together with plasma level of TNF-[Formula: see text], whereas the plasma level of albumin (ALB) was significantly reduced. In addition, histological disruptions of kidneys and livers were observed. Chronic immobilization stress aggravated the effects of renal IR. Also, renal and hepatic morphological changes were more worsened. Whilst, OO supplementation resulted in significant amelioration of renal and hepatic functions. Also, the kidney and liver morphologic lesions were attenuated. Conclusion: Renal IR not only affected the renal functional and structural integrity but also the remote organ, the liver. Chronic immobilization stress rendered the kidney and liver more prone to injury. OO improved renal and hepatic dysfunction and morphological damage mediated by renal IRI, in control rats and in those exposed to chronic stress which can be exerted partially via its antioxidant, anti- inflammatory and nitric oxide (NO) reducing activities.

Preventive and therapeutic effects of the peel powder of P. granatum in a rat sepsis model

The aim of the present study was to evaluate the treatment potential of Punica granatum L. peel powder in an experimentally induced sepsis model in rats. Sepsis was induced in 10-week-old, male, Wistar Albino (n=24) rats. The animals were divided into four groups: Sham-operated (S) Group, Control (C) Group, Treatment-1 (T1) Group, and Treatment-2 (T2) Group. To induce the sepsis model, the cecal ligation and puncture procedure was performed. The P. granatum peel powder (200 mg/kg; per os) was applied one hour before (T1) and 10 hours after (T2) surgery in a volume of 2 mL. At the end of the experimental procedure, microbial and histopathological analyses were performed. The histopathological scores on the liver, lungs, heart, kidney, spleen, and pancreas were evaluated. Escherichia coli, Staphylococcus aureus and E. coli + S. aureus were isolated from blood cultures. Severe bacteria were detected in the blood of the group C animals. It was found that there were fewer bacteria in groups T1 (n=2) and T2 (n=4) compared with group C. There were no lesions in the pancreas tissues of any groups. Vascular changes, degeneration, and necrosis were common in the organs in all cases of group C compared to group S. The findings in group T1 were similar to those in group C, however, it was seen in fewer animals. It was determined that there was a general improvement in group T2, and in addition the existing lesions were moderate in severity. In conclusion, P. granatum L. peel powder prevented CLP-induced lung injury in experimental rats. Thus, P. granatum L. peel powder may be an alternative therapeutic agent against lung tissue injury induced by sepsis. The recovery from inflammation was better in group T2 than in the other groups. According to the results of the current study, P. granatum peel powder was found to be effective in the treatment of sepsis with antimicrobial and anti-inflammatory functions.

Influence of Piper sarmentosum Aqueous Extract on the Expression of Osteocalcin in Glucocorticoid-induced Osteoporotic Rats

Secondary osteoporosis is mainly caused by prolong used of glucocorticoid treatment. The Piper sarmentosum leaf aqueous extract was found to exhibit bone formatting osteocalcin activity against Dexamethasone induced osteoporotic rats. Thirty-two Sprague-Dawley rats were divided equally into four groups - G1: Sham-operated control group given intramuscular (IM) olive oil as vehicle and normal saline orally as vehicle; G2: Adrenalectomized (Adrx) control group given IM dexamethasone (DEX) (120 μg/kg/day) and normal saline orally as vehicle; G3: Adrx group given IM DEX (120 μg/kg/day) and aqueous extract of Piper sarmentosum leaves (125 mg/kg/day) orally; and G4: Adrx group given IM DEX (120 μg/kg/day) and glycyrrhizic acid (GCA) (120 mg/kg/day) orally. Immunohistochemical method with gold labelling was used to label the osteocalcin protein. Silver brightener was used, sprinkled on gold with a size of 5 nm so that the resulting image can be seen more clearly using a light microscope. The osteocalcin protein was measured quantitatively based on nomenclature report of the ASBMR Histomorphometry Committee (American Society for Bone Mineral Research). The activity shown by immunohisto-gold expression and localization of osteocalcin was comparable with the reference, glycyrrhizic acid, a potent inhibitor of 11β-hydroxysteroid dehydrogenase enzyme in RANKL-OPG pathway. As a conclusion, Piper sarmentosum may one day be utilized as an alternate treatment for individuals receiving long-term glucocorticoid therapy to prevent osteoporosis, therefore osteoporotic fractures.

P25/CDK5-mediated Tau Hyperphosphorylation in Both Ipsilateral and Contralateral Cerebra Contributes to Cognitive Deficits in Post-stroke Mice.

Post-stroke cognitive impairment (PSCI) develops in approximately one-third of stroke survivors and is associated with ingravescence. Nonetheless, the biochemical mechanisms underlying PSCI remain unclear. The study aimed to establish an ischemic mouse model by means of transient unilateral middle cerebral artery occlusions (MCAOs) and to explore the biochemical mechanisms of p25/cyclin-dependent kinase 5 (CDK5)-mediated tau hyperphosphorylation on the PSCI behavior. Cognitive behavior was investigated, followed by the detection of tau hyperphosphorylation, mobilization, activation of kinases and/or inhibition of phosphatases in the lateral and contralateral cerebrum of mice following ischemia in MACO mice. Finally, we treated HEK293/tau cells with oxygen-glucose deprivation (OGD) and a CDK5 inhibitor (Roscovitine) or a GSK3β inhibitor (LiCl) to the roles of CDK5 and GSK3β in mediating ischemia-reperfusion-induced tau phosphorylation. Ischemia induced cognitive impairments within 2 months, as well as causing tau hyperphosphorylation and its localization to neuronal somata in both ipsilateral and contralateral cerebra. Furthermore, p25 that promotes CDK5 hyperactivation had significantly higher expression in the mice with MCAO than in the shamoperation (control) group, while the expression levels of protein phosphatase 2 (PP2A) and the phosphorylation level at Tyr307 were comparable between the two groups. In addition, the CDK5 inhibitor rescued tau from hyperphosphorylation induced by OGD. These findings demonstrate that upregulation of CDK5 mediates tau hyperphosphorylation and localization in both ipsilateral and contralateral cerebra, contributing to the pathogenesis of PSCI.

SAT236 Pharmacokinetics (PK) And Pharmacodynamics (PD) Of The Parathyroid Hormone Analog PTH (1-34) (Teriparatide) Delivered Via An Orally Administered Robotic Pill (RT-102)

Abstract Disclosure: K. Horlen: Employee; Self; Rani Therapeutics. Stock Owner; Self; Rani Therapeutics. K. Kaminskaya: Employee; Self; Rani Therapeutics. Stock Owner; Self; Rani Therapeutics. A. Yamaguchi: Employee; Self; Rani Therapeutics. Stock Owner; Self; Rani Therapeutics. L.C. Fung: Employee; Self; Rani Therapeutics. Stock Owner; Self; Rani Therapeutics. H. Kennedy: Employee; Self; Rani Therapeutics. Stock Owner; Self; Rani Therapeutics. S. Nguyen: Employee; Self; Rani Therapeutics. Stock Owner; Self; Rani Therapeutics. A. Dasari: Employee; Self; Rani Therapeutics. Stock Owner; Self; Rani Therapeutics. A. Toledo Vo: Employee; Self; Rani Therapeutics. Stock Owner; Self; Rani Therapeutics. B. Archana: Employee; Self; Rani Therapeutics. Stock Owner; Self; Rani Therapeutics. N. Patel: Employee; Self; Rani Therapeutics. Stock Owner; Self; Rani Therapeutics. D. Mohan: Employee; Self; Rani Therapeutics. Stock Owner; Self; Rani Therapeutics. B. Syed: Employee; Self; Rani Therapeutics. Stock Owner; Self; Rani Therapeutics. A.K. Dhalla: Employee; Self; Rani Therapeutics. Stock Owner; Self; Rani Therapeutics. M. Imran: Employee; Self; Rani Therapeutics. Stock Owner; Self; Rani Therapeutics. M.A. Hashim: Employee; Self; Rani Therapeutics. Stock Owner; Self; Rani Therapeutics. Background: An orally administered robotic pill (RP) protects the drug from digestion during transit through the gastrointestinal tract before painlessly injecting the drug trans-enterically into the peritoneal space. Although parenteral, this unique delivery route can result in altered PK/PD profiles as drug absorption is primarily via mesenteric vasculature which drains into the portal vein. Here we report on two nonclinical studies which, 1) in awake canines, compared the PK profiles of orally delivered teriparatide via the RP (RT-102) versus subcutaneous (SC) injections; and 2) in an ovariectomized (OVX) rat model of osteoporosis, compared the PD (osteoanabolic) effect of intraperitoneal (IP) vs SC administered teriparatide on bone mineral density (BMD).Methods:PK Study: Beagle dogs (n = 43) received a single RT-102 capsule orally containing 20, 40, 80, or 160µg of teriparatide or a SC or intravenous (IV) injection of 20µg of the FDA-approved PTH(1-34) commercial product (Forteo®). Teriparatide concentrations in serial blood samples (collected for up to 300 minutes) were quantitated via a validated ELISA.PD Study: OVX Sprague-Dawley rats (n=50) underwent an 8-week bone depletion period followed by 6-weeks of daily treatment with teriparatide at a dose of 2, 5, or 15 µg/kg via IP injection or 5 µg/kg via SC injection (Forteo®) (n = 10 per treatment). A sham-operated control group received daily saline injections. BMD was assessed via dual-energy X-ray absorptiometry (DXA). Results:PK Study: RT-102 yielded steep increases in serum PTH concentrations followed by rapid declines to baseline levels, similar to the overall PK profile of Forteo® SC. RT-102 yielded more sustained exposures resulting in a prolonged Tmax compared to Forteo® SC (Tmax 37 to 48 vs 21 min). At 80 µg, PK curves of RT-102 tracked closely with SC, albeit with RT-102 yielding higher peak concentrations (Cmax = 3511 vs 1342 pg/mL). PD Study: DXA imaging of whole-body skeleton, femur, femur diaphysis, and lumbar vertebrae showed that intraperitoneally administered teriparatide yielded dose dependent increases in BMD in all areas examined. Animals receiving teriparatide via either IP or SC injections at equivalent doses showed no significant differences in the BMD increases between the two routes of administration. Conclusions: Orally administered RT-102 capsules to canines yielded steep rises in serum teriparatide concentrations followed by rapid elimination within three hours. When administered intraperitonially to mimic RT-102 delivery in a rodent model of osteoporosis, teriparatide was osteoanabolic similar to equivalent drug doses injected SC. These data provide the PK/PD proof-of-concept data for oral RT-102 capsules indicating their potential to replace current injection therapy for the treatment of osteoporosis. Presentation: Saturday, June 17, 2023

Surface material of acoustic transmitters influences the inflammatory response of rainbow trout (Oncorhynchus mykiss) during long-term implantation

Telemetry tags are a widely used technology for tracking animals that are difficult to observe in their natural environment. This technology has been increasingly used to monitor and study populations of high value salmonid species in Canadian waters. This study expands on a previous study of the impacts of tag implantation on the immune system of Rainbow Trout (Oncorhynchus mykiss). Pro-inflammatory cytokines and protein level markers were examined in fish that underwent peritoneal implantation of three tag types and compared to a sham surgery control group. The different materials on the surface of the tags showed differential immune induction extending over a two-month period. This included peritoneal total protein, IL-1β protein, the immunoglobulins IgT and IgM, as well as pro-inflammatory transcripts in the spleen. These results are suggestive of a prolonged, costly foreign body response which may be differentially induced by the different types of tag coating, with ceramic tags being least immunogenic. Examining tag impacts at the level of the immune system will facilitate the development of more biocompatible tags which will improve data fidelity. This will support more effective strategies for the management of fisheries resources.