PURPOSE: To evaluate the ability of human airway epithelial cell layers and a simple rat isolated perfused lung (IPL) model to predict pulmonary drug absorption in rats in vivo. METHOD: The permeability of seven compounds selected to possess a range of lipophilicity was measured in two airway cell lines (Calu-3 and 16HBE14o-), in normal human bronchial epithelial (NHBE) cells and using a simple isolated perfused lungs (IPL) technique. Data from the cell layers and ex vivo lungs were compared to published absorption rates from rat lungs measured in vivo. RESULTS: A strong relationship was observed between the logarithm of the in vivo absorption half-life and the absorption half-life in the IPL (r = 0.97; excluding formoterol). Good log-linear relationships were also found between the apparent first-order absorption rate in vivo and cell layer permeability with correlation coefficients of 0.92, 0.93, 0.91 in Calu-3, 16HBE14o- and NHBE cells, respectively. CONCLUSION: The simple IPL technique provided a good prediction of drug absorption from the lungs, making it a useful method for empirical screening of drug absorption in the lungs. Permeability measurements were similar in all the respiratory epithelial cell models evaluated, with Calu-3 having the advantage for routine permeability screening purposes of being readily availability, robust and easy to culture.

• Keywords
• 16HBE14o-, NHBE, biopharmaceutics, calu-3, inhalation, isolated perfused lungs (IPL), permeability, pulmonary,
• MeSH
• Respiratory Tract Absorption * MeSH
• Models, Biological MeSH
• Cell Line MeSH
• Cells, Cultured MeSH
• Humans MeSH
• Lung metabolism   MeSH
• Alveolar Epithelial Cells metabolism   MeSH
• Rats, Wistar MeSH
• Primary Cell Culture MeSH
• Animals MeSH
• Check Tag
• Humans MeSH
• Male MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Comparative Study MeSH

Together with the development of peritoneal dialysis (PD), appropriate animal models play an important role in the investigation of physiological, pathophysiological and clinical aspects of PD. However, there is still not an ideal experimental PD animal model. In this study, 45 Sprague-Dawley rats were divided into three groups. Group 1 (n=15) was receiving daily peritoneal injection through the catheter connected to the abdominal cavity, using PD solution containing 3.86 % D-glucose. Group 2 (n=15) was receiving daily peritoneal injection of 0.9 % physiological saline through a catheter. Group 3 (n=15), which was subjected to sham operation, served as controls. Our results showed that WBC counts in peritoneal effluent of Group 1 were slightly higher than those of Group 2 and control group, respectively (p<0.05). However, there was no episode of infection in any group. In addition, there was no significant difference in neutrophils fractions among these three groups. Hematoxylin-eosin and Masson's trichrome staining demonstrated a dramatic increase in thickness of the mesothelium-to-muscle layer of peritoneum exposed to high glucose (Group 1) compared to Group 2 and controls (p<0.01). These data indicated that we established a novel rat model of PD with a modified catheter insertion method. This model is more practical, easy to operate, not too expensive and it will facilitate the investigate of long-term effects of PD.

• MeSH
• Dialysis Solutions pharmacology   MeSH
• Catheters MeSH
• Rats * MeSH
• Models, Animal * MeSH
• Peritoneal Dialysis methods   MeSH
• Peritoneum metabolism   pathology   MeSH
• Rats, Sprague-Dawley MeSH
• Uremia etiology   MeSH
• Animals MeSH
• Check Tag
• Rats * MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Names of Substances
• Dialysis Solutions MeSH

Gliflozins (sodium-glucose transporter-2 inhibitors) exhibited renoprotective effects not only in diabetic but also in non-diabetic patients with chronic kidney disease (CKD). Controversial results were reported in experimental non-diabetic models of CKD. Therefore, we examined empagliflozin effects in three CKD models, namely, in fawn-hooded hypertensive (FHH) rats, uninephrectomized salt-loaded (UNX + HS) rats, and in rats with Goldblatt hypertension (two-kidney, one-clip 2K1C) that were either untreated or treated with empagliflozin (10 mg/kg/day) for eight weeks. Plethysmography blood pressure (BP) was recorded weekly, and renal parameters (proteinuria, plasma urea, creatinine clearance, and sodium excretion) were analyzed three times during the experiment. At the end of the study, blood pressure was also measured directly. Markers of oxidative stress (TBARS) and inflammation (MCP-1) were analyzed in kidney and plasma, respectively. Body weight and visceral adiposity were reduced by empagliflozin in FHH rats, without a significant effect on BP. Experimentally induced CKD (UNX + HS and 2K1C) was associated with a substantial increase in BP and relative heart and kidney weights. Empagliflozin influenced neither visceral adiposity nor BP in these two models. Although empagliflozin increased sodium excretion, suggesting effective SGLT-2 inhibition, it did not affect diuresis in any experimental model. Unexpectedly, empagliflozin did not provide renoprotection because proteinuria, plasma urea, and plasma creatinine were not lowered by empagliflozin treatment in all three CKD models. In line with these results, empagliflozin treatment did not decrease TBARS or MCP-1 levels in either model. In conclusion, empagliflozin did not provide the expected beneficial effects on kidney function in experimental models of CKD.

• Keywords
• SGLT-2 inhibition, fawn-hooded hypertensive rat, one-clip hypertension, proteinuria, two-kidney, uninephrectomized salt-loaded,
• Publication type
• Journal Article MeSH

OBJECTIVE: Experimental lesions in the inferior alveolar nerve (IAN) are used for the study of peripheral and central alterations. The objective of our study was to contribute to a more precise description of the approach to the IAN and creating a lesion. MATERIAL AND METHODS: Twenty-six males of Wistar laboratory rats were used for the study. The animals were divided into three groups: control group (6 rats), experimental group (12 rats - a part of the bone above the mandibular canal was removed under general anaesthesia using extraoral approach, after exposing a part of the IAN, the nerve was excised in a length of 3 mm), and a sham group (8 rats - the nerve was only dissected but not transected). Persisting denervation was verified using surgical revision and histological and immunohistochemical analysis after the observation period (4 weeks). RESULTS: No evidence of re-innervation after 4 weeks. We found no statistically significant differences in mean weight gains between individual groups during the observation period. CONCLUSION: The described technique used in the study is one of the possible ways to create a nerve lesion at the site of the main trunk of the nerve. At the same time, the study provides a more precise description of the anatomical situation and approach to the IAN in the mandibular canal.

• Keywords
• Nervus alveolaris inferior, laboratory rat, nerve transection technique, neuroanatomical study,
• MeSH
• Denervation methods   MeSH
• Rats MeSH
• Models, Animal MeSH
• Mandibular Nerve physiopathology   surgery   MeSH
• Neuroanatomy MeSH
• Rats, Wistar MeSH
• Nerve Regeneration physiology   MeSH
• Animals MeSH
• Check Tag
• Rats MeSH
• Male MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Evaluation Study MeSH

BACKGROUND: There is increasing evidence that peroxisome proliferator-activated receptors (PPARs) may be involved in the regulation of angiogenesis. In this study, we examined whether rosiglitazone, a PPARγ agonist, can restore angiogenesis in a rat hindlimb ischemia model of diabetes. METHODS: Male wistar rats were divided into four groups (n=6 each): control, diabetic and control and diabetic rats who received rosiglitazone (8 mg/kg/day). Diabetes was induced by streptozotocin (55 mg/kg; ip). After 21 days, serum concentrations of nitric oxide (NO), vascular endothelial growth factor (VEGF) and soluble VEGF receptor-2 (VEGFR-2) were measured and neovascularization in ischemic legs was evaluated by immunohistochemistry. RESULTS: Capillary density and capillary/fiber ratio in hindlimb ischemia of diabetic animals were significantly lower than the control group (P<0.05). Rosiglitazone significantly restored neovascularization in diabetic animals (P<0.05). CONCLUSIONS: rosiglitazone enhances neovascularization in diabetic ischemic skeletal muscle and could be considered for treatment of peripheral artery disease in diabetic subjects.

• MeSH
• Models, Biological MeSH
• Diabetes Mellitus, Experimental physiopathology   MeSH
• Neovascularization, Physiologic drug effects   MeSH
• Rats MeSH
• Rats, Wistar MeSH
• Rosiglitazone MeSH
• Thiazolidinediones pharmacology   MeSH
• Hindlimb blood supply   MeSH
• Animals MeSH
• Check Tag
• Rats MeSH
• Male MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Names of Substances
• Rosiglitazone MeSH
• Thiazolidinediones MeSH

Angelman syndrome (AS) is a rare genetic disorder characterized by severe intellectual disability, seizures, lack of speech, and ataxia. The gene responsible for AS was identified as Ube3a and it encodes for E6AP, an E3 ubiquitin ligase. Currently, there is very little known about E6AP's mechanism of action in vivo or how the lack of this protein in neurons may contribute to the AS phenotype. Elucidating the mechanistic action of E6AP would enhance our understanding of AS and drive current research into new avenues that could lead to novel therapeutic approaches that target E6AP's various functions. To facilitate the study of AS, we have generated a novel rat model in which we deleted the rat Ube3a gene using CRISPR. The AS rat phenotypically mirrors human AS with loss of Ube3a expression in the brain and deficits in motor coordination as well as learning and memory. This model offers a new avenue for the study of AS. Autism Res 2020, 13: 397-409. © 2020 International Society for Autism Research,Wiley Periodicals, Inc. LAY SUMMARY: Angelman syndrome (AS) is a rare genetic disorder characterized by severe intellectual disability, seizures, difficulty speaking, and ataxia. The gene responsible for AS was identified as UBE3A, yet very little is known about its function in vivo or how the lack of this protein in neurons may contribute to the AS phenotype. To facilitate the study of AS, we have generated a novel rat model in which we deleted the rat Ube3a gene using CRISPR. The AS rat mirrors human AS with loss of Ube3a expression in the brain and deficits in motor coordination as well as learning and memory. This model offers a new avenue for the study of AS.

• Keywords
• Angelman syndrome, E6AP, Ube3a, cognitive deficits, rat model,
• MeSH
• Angelman Syndrome genetics   physiopathology   MeSH
• Gene Deletion * MeSH
• Phenotype MeSH
• Rats MeSH
• Humans MeSH
• Disease Models, Animal MeSH
• Brain physiopathology   MeSH
• Memory MeSH
• Rats, Sprague-Dawley MeSH
• Ubiquitin-Protein Ligases genetics   MeSH
• Animals MeSH
• Check Tag
• Rats MeSH
• Humans MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Names of Substances
• Ube3a protein, rat MeSH Browser
• Ubiquitin-Protein Ligases MeSH

Hyperhomocysteinemia is a common occurrence in many neurodegenerative diseases, including tauopathies. We developed and validated a simple and sensitive liquid chromatography-tandem mass spectrometry method for the analysis of homocysteine (Hcy) in rat plasma. Hcy was analyzed using ultra-performance liquid chromatography on a C8 column with detection by positive ESI tandem mass spectrometry. For optimal retention and separation, we used ion-pair reagent-heptafluorobutyric acid. The method utilizes heavy labeled internal standard and does not require any derivatization or extraction step. The procedure was validated in compliance with the European Medicines Agency guideline. The limit of detection was 0.15 µmol/L and the limit of quantification was 0.5 µmol/L. The method showed excellent linearity with regression coefficients higher than 0.99. The accuracy was in the range of 93-98%. The inter-day precision (n = 5 days), expressed as % relative standard deviation, was in the range 3-8%. Using this method, we analyzed plasma samples from two transgenic lines of the rat model for tauopathies.

• MeSH
• Chromatography, Liquid methods   MeSH
• Homocysteine blood   MeSH
• Rats MeSH
• Limit of Detection MeSH
• Linear Models MeSH
• Rats, Inbred SHR MeSH
• Rats, Transgenic MeSH
• Rats, Wistar MeSH
• Reproducibility of Results MeSH
• Drug Stability MeSH
• Case-Control Studies MeSH
• Tandem Mass Spectrometry methods   MeSH
• Tauopathies blood   MeSH
• Animals MeSH
• Check Tag
• Rats MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Names of Substances
• Homocysteine MeSH

Several animal models of genetic hypertension have been developed but not all of them possess a closely related control strain. Therefore, a new model based on Wistar rats is described in which both hypertensive and normotensive lines were bred from a single parental pair. Several basic data on the two lines (called the Prague Hypertensive Rat, PHR, and the Prague Normotensive Rat, PNR) are given. PNR had a longer survival compared with PHR. At the age of 7 weeks, systolic blood pressure was 161 +/- 14 mmHg in PHR males and 109 +/- 9 mmHg in PNR males. Its further increase with age was very slow in PNR but very steep in PHR. Typical left ventricular cardiac hypertrophy developed in PHR in which cardiac output was not significantly different from that of PNR but total peripheral resistance was higher. Kidney weight was also greater in PHR than in PNR. There was no difference in basic renal functions except of proteinuria which was higher in PHR than in PNR. No differences were observed in extracellular and interstitial fluid volumes whereas plasma and blood volumes were slightly but significantly greater in PHR than in PNR suggesting a shift of extracellular fluid towards the intravascular compartment. This hypertensive model the parameters of which resemble to those of human essential hypertension should be especially suitable for cross-transplantation studies.

• MeSH
• Hypertension etiology   genetics   physiopathology   MeSH
• Rats, Inbred Strains MeSH
• Blood Pressure MeSH
• Rats MeSH
• Kidney physiopathology   MeSH
• Disease Models, Animal MeSH
• Rats, Inbred SHR genetics   physiology   MeSH
• Body Fluids physiology   MeSH
• Organ Size MeSH
• Animals MeSH
• Check Tag
• Rats MeSH
• Male MeSH
• Female MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH

The aim of this work is to present the efficacy of a previously introduced computational procedure, developed for evaluation of vascular responsiveness. On this reason, as an example a common study of noradrenaline (NA) effect on a rat renal artery under in vitro conditions was arbitrarily selected. The response of the arterial segment to NA doses (0.1-10 microg) was digitally recorded on a PC and employed to develop mathematical model of NA effect. Using the model, the following NA effect variables were determined: the vessel sensitivity parameter, mean effect time and rate constant, respectively, characterizing the effect intensity, duration, and regression and also classic response variables: the maximal effect and time of the maximal effect. The two-way analysis of variance followed by Bonferroni's test revealed a significant influence of the increasing NA dose on the vessel sensitivity parameter and mean effect time. These findings indicated nonlinearity of processes underlying NA effect on the rat renal artery over the given range of NA doses. The procedure exemplified has the potential for use as an effective adjunct to routine studies of vascular responsiveness as it enables the extraction of meaningful information which cannot by obtained by common manual evaluation procedures.

• MeSH
• Renal Artery drug effects   MeSH
• Time Factors MeSH
• Rats MeSH
• Models, Cardiovascular * MeSH
• Nonlinear Dynamics * MeSH
• Norepinephrine pharmacology   MeSH
• Computer Simulation MeSH
• Rats, Wistar MeSH
• Vasoconstriction drug effects   MeSH
• Vasoconstrictor Agents pharmacology   MeSH
• Dose-Response Relationship, Drug MeSH
• Animals MeSH
• Check Tag
• Rats MeSH
• Male MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Names of Substances
• Norepinephrine MeSH
• Vasoconstrictor Agents MeSH

Studies suggest age- and sex-dependent structural and functional patterns of human cerebral lateralization underlie hemisphere specialization and its alterations in schizophrenia. Recent works report sexual dimorphism of neurons in the hippocampal formation and specialization of hemispheres in rats. Our experiments indicate for the first time functional lateralization of the high-affinity choline uptake (HACU) system directly associated with a synthesis of acetylcholine in the hippocampus of Wistar rats. The markedly increased HACU activity was found in the left compared to the right hippocampus of adult male but not female animals. Lineweaver-Burk plot analysis revealed a statistically significant increase of Vmax in the left hippocampus of 14-day-old when compared to 7-day-old males. It appears that laterality of HACU occurs during late postnatal maturation, and its degree is markedly enhanced after puberty and attenuated during aging. Quinolinic acid (QUIN), an endogenous agonist of N-methyl-D-aspartate type glutamate receptors, was used in this study to evaluate the neurodevelopmental hypothesis of schizophrenia. It is known that elevated levels of QUIN accompany viral infections, increasing the risk of developing schizophrenia. Bilateral intracerebroventricular application of QUIN (250 nmoles/ventricle) to pups aged 12 days significantly impaired the cholinergic hippocampal system of adolescent male and female rats and reversed lateralization of male HACU. Morphological analysis indicated marked changes in brain lesion sizes (extensive 24 h and moderate 38 days after the operation). Asymmetry of lesions was observed in the majority of cases, but the left hemisphere was not generally more vulnerable to QUIN effects than the right side. Moreover, no lateral differences were found between lesioned hippocampi in the specific binding of [3H]hemicholinium-3 (10%-15% loss of binding sites when compared to sham-operated animals). In summary, our results indicate a symmetrical drop in the number of choline carriers of lesioned male rats but a asymmetrical decrease in the activity of remaing carriers, suggesting defects in processes of sexual brain differentiation, leading under normal conditions to the higher activity of carriers in the left hippocampus. The data demonstrate viral infection-mediated alterations in normal patterns of brain asymmetry and are discussed in relation to animal models of neurodevelopmental and neurodegenerative diseases.

• MeSH
• Functional Laterality * MeSH
• Hippocampus physiopathology   MeSH
• Rats MeSH
• Disease Models, Animal * MeSH
• Neurodegenerative Diseases physiopathology   MeSH
• Rats, Wistar MeSH
• Receptors, Cholinergic physiology   MeSH
• Sex Factors MeSH
• Animals MeSH
• Check Tag
• Rats MeSH
• Male MeSH
• Female MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Names of Substances
• Receptors, Cholinergic MeSH