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Functional characterization and visualization of a GABAA receptor-GFP chimera expressed in Xenopus oocytes.

The GABAA receptor is a ligand-gated chloride channel belonging to the superfamily of ligand-gated ion channels of which the nicotinic acetylcholine (nACh) receptor is prototypic. In the central nervous system the GABAA receptor mediates fast neuronal inhibition. To facilitate the study of this receptor, a GABAA receptor-green fluorescent protein (GABAAR-GFP) chimera was constructed by fusing green fluorescent protein (GFP) to the C-terminus region of the GABAA receptor alpha1 subunit. When expressed in Xenopus oocytes, this chimera responded in a manner indistinguishable from the wild-type GA

Integrated Network Pharmacology Approach for Drug Combination Discovery: A Multi-Cancer Case Study.

Despite remarkable efforts of computational and predictive pharmacology to improve therapeutic strategies for complex diseases, only in a few cases have the predictions been eventually employed in the clinics. One of the reasons behind this drawback is that current predictive approaches are based only on the integration of molecular perturbation of a certain disease with drug sensitivity signatures, neglecting intrinsic properties of the drugs. Here we integrate mechanistic and chemocentric approaches to drug repositioning by developing an innovative network pharmacology strategy. We developed

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… In colon adenocarcinoma, we found pentobarbital in combination with navitoclax as one of the top combinations. The anticancer efficacy of pentobarbital on animal models of colon adenocarcinoma was demonstrated in 2005 by Thaker and colleagues [ 46 ]. … They demonstrated that the treatment with pentobarbital of mice affected by artificially induced colon adenocarcinoma (through HT29 cell line injection) reduced the tumor weight (primary tumor and liver metastases) in respect of the control group, while showing similar tumor incidence. …

Prevention of medication-related osteonecrosis of the jaw after tooth extraction by local administration of antibiotics and atelocollagen sponge: A preliminary study.

BACKGROUND/PURPOSE: Medication-related osteonecrosis of the jaw (MRONJ) often develops after extraction of a tooth with a local infection. Therefore, it is necessary to develop extraction methods that can prevent MRONJ. Before examining whether antibiotics and atelocollagen administered in the extraction socket can prevent the development of MRONJ after tooth extraction, this study was conducted to determine the appropriate antibiotics concentration and to conduct a clinical study with a small number of cases as a preliminary study. MATERIALS AND METHODS: First, a mixture of minocycline and a

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… Japanese white rabbits of 11-week-old (Kitayama LaBes Co., Ltd., Ina-city, Japan) weighing 2–3 kg were anesthetized with pentobarbital, and four cavities of 5 mm diameter and 3 mm depth were formed in the frontal bone of five rabbits ( Fig. 1 ) One ml of minocycline at each concentration was injected … Two rabbits were euthanized after 24 h and three after 48 h were euthanized with a lethal dose of pentobarbital, sutured wounds were opened, each specimen was removed and frozen at −81 °C, and local antimicrobial concentrations were measured at a later date. …

Apoptosis in primary cultures of E14 rat ventral mesencephala: time course of dopaminergic cell death and implications for neural transplantation.

Transplantation using fetal nigral grafts has been performed by various groups worldwide in over 200 Parkinson's disease (PD) patients in an attempt to restore dopaminergic (DA) input to the striatum. However, the proportion of the implanted DA neurons that survives, whether using suspension, partially dissociated, or solid grafts, is small, often as low as 5 to 10%, which is insufficient to allow a full functional recovery. A significant proportion of the transplanted neurons in animal models of PD has been shown to die via apoptosis, but the reason for this is unclear. Since the methods used

Protein kinase C-delta mediates neuronal apoptosis in the retinas of diabetic rats via the Akt signaling pathway.

OBJECTIVE: Protein kinase C (PKC)-delta, an upstream regulator of the Akt survival pathway, contributes to cellular dysfunction in the pathogenesis of diabetes. Herein, we examined the role of PKC-delta in neuronal apoptosis through Akt in the retinas of diabetic rats. RESEARCH DESIGN AND METHODS: We used retinas from 24- and 35-week-old male Otsuka Long-Evans Tokushima fatty (OLETF) diabetic and Long-Evans Tokushima Otsuka (LETO) nondiabetic rats. To assess whether PKC-delta affects Akt signaling and cell death in OLETF rat retinas, we examined 1) PKC-delta activity and apoptosis; 2) protein

Anesthetized Long Evans rats show similar protein expression and long-term potentiation as Fischer 344 rats but reduced short-term potentiation in motor cortex.

A number of studies describe strain-related differences in the motor behavior of rats. Inbred albino F344 rats are found to be impaired in procedural spatial learning, skilled reaching, and over ground locomotion in relation to pigmented out bred Long Evans (LE) rats. These deficits could be related to the functional differences in the motor cortex of the two strains, and the objective of the present study was to examine this hypothesis. Synaptic transmission was examined in the two rat strains, using long-term potentiation (LTP) and short-term potentiation (STP), two electrophysiological meas

GABAC receptors are localized with microtubule-associated protein 1B in mammalian cone photoreceptors.

Protein MAP1B was recently reported to link GABA(C) receptors to the cytoskeleton at neuronal synapses. This interaction was demonstrated in the mammalian retina, where GABA(C) receptors were thought to be exclusively expressed in bipolar cells. Our previous studies on cultured photoreceptors suggested however the presence of GABA(C) receptors in cones. To further investigate GABA(C) receptor expression in cones, we measured GABA responses in mammalian photoreceptors in situ, and we examined the distribution of the receptor and that of protein MAP1B in the mammalian outer retina. Photoreceptor

Selective neuronal death in the contralateral hippocampus following unilateral kainate injections into the CA3 subfield.

Intracerebral or intraperitoneal injections of kainic acid, an agonist at a class of glutamate receptors, have been extensively used to model temporal lobe epilepsy. In the present study we compared the types and distributions of selectively vulnerable neurons in the ipsi- and contralateral hippocampi following unilateral kainate injections into the CA3 subfield in order to examine whether "proximal" or "distant" neuronal damage resembled the pathology, and possibly also the mechanism, of human temporal lobe epilepsy. The degeneration of principal cells in the different hippocampal subfields w

[Exploring the effect of acupuncture at the "Qiaomai" points on the immune function of central and peripheral macrophages in insomnia rats based on the "Defense Qi Theory"].

OBJECTIVES: To observe the effect of acupuncture at the "Qiaomai" points on the immune function activity of central and peripheral macrophag … ht was recorded before modeling, before intervention, and after intervention. The threshold dose of pentobarbital sodium in sleep experiment before and after treatment were also recorded. The thymus index and spleen index were calculated. Flow cytometry was used to determine the CD68+, CD86+, and CD163+ positive cells in peripheral blood. Immunohistochemistry w …

[Therapeutic effect of Ziziphi Spinosae Semen extracts on chronic unpredictable mild stress-induced depression and insomnia-like behavior in mice].

This paper aims to study the effect of Ziziphi Spinosae Semen extracts on chronic unpredictable mild stress(CUMS)-induced depression-like an … 15th day of modeling, the drug was administered intragastrically once a day for one week. Then, the pentobarbital sodium cooperative righting experiment, open field experiment, and elevated plus maze experiment were carried out, respectively. The contents of neurotransmitters 5-hydroxytryptamine(5-HT) and 5-hydroxyindoleacetic acid(5-HIAA) in serum and thalamus …

Efficacy of needling Baihui (GV20), Neiguan (PC6), Shenmen (HT7) and Taichong (LR3) on cerebral cortical blood oxygen level in rats with insomnia.

OBJECTIVE: To evaluate the effect of needling Baihui (GV20), Neiguan (PC6), Shenmen (HT7) and Taichong (LR3) on cerebral cortical blood oxyg … oup were needled with four non-acupoint sites. After 7 d of treatment of rats in each group, sodium pentobarbital sleep experiment was used to evaluate the sleep latency (SL) and sleep time (ST). The percentage of rats entering the open arm (OE%) and the percentage of time entering the open arm (OT%) in each group were monitored by elevated cross maze, and the …

[Serotonin receptor 1A-modulated dephosphorylation of glycine receptor α3: a new molecular mechanism of breathing control for compensation of opioid-induced respiratory depression without loss of analgesia].

To control the breathing rhythm the medullary respiratory network generates periodic salvo activities for inspiration, post-inspiration and expiration. These are under permanent modulatory control by serotonergic neurons of the raphe which governs the degree of phosphorylation of the inhibitory glycine receptor α3. The specific activation of serotonin receptor type 1A (5-HTR(1A)), which is strongly expressed in the respiratory neurons, functions via inhibition of adenylate cyclase and the resulting reduction of the intracellular cAMP level and a gradual dephosphorylation of the glycine recepto

Vascular actions of MDMA involve alpha1 and alpha2-adrenoceptors in the anaesthetized rat.

We have investigated the effects of methylenedioxymethamphetamine (MDMA, 'ecstasy'), i.v., on diastolic blood pressure (DBP) in pithed and pentobarbitone anaesthetized rats. In pithed rats, the non-selective 5-HT receptor antagonist methiothepin (0.1 mg kg(-1)) and the alpha2-adrenoceptor antagonists methoxyidazoxan and yohimbine (1 mg kg(-1)) showed significant alpha1-adrenoceptor antagonist potency, but methiothepin did not show alpha2-adrenoceptor antagonist potency. MDMA (1 and 5 mg kg(-1)) produced pressor responses which were significantly reduced by the alpha(1)-adrenoceptor antagonist

The Herba Patriniae (Caprifoliaceae): A review on traditional uses, phytochemistry, pharmacology and quality control.

ETHNOPHARMACOLOGICAL RELEVANCE: Herba Patriniae has been used for thousands of years in China as a traditional Chinese medicine with heat-clearing and detoxicating effects. It is applied widly for the treatment of rheumatoid arthritis, diarrhea, acute hepatitis, pelvic inflammatory disease and ulcerative colitis in clinic. Two species, namely Patrinia scabiosaefolia Fisch. (PS) and Patrinia villosa Juss. (PV) from the Caprifoliaceae family, are considered as Herba Patriniae in the pharmaceutical industry. AIM OF THE REVIEW: This paper aims to comprehensively outline the traditional uses, bota

Management of Severe Acute Pancreatitis.

Sepsis‑induced abnormalities in brain function or sepsis‑associated encephalopathy (SAE) can manifest as cognitive dysfunction and other neuropsychiatric symptoms; however, the underlying mechanisms remain unclear. The aim of the present study was to elucidate the possible effects and mechanism of capsaicin, a transient receptor potential vanilloid 1 (TRPV1) agonist, on the pathological features of SAE. A model of SAE in C57BL/6 mice was generated using cecal ligation and puncture (CLP). Capsaicin (1 mg/kg) was injected subcutaneously before surgery. Cognitive function in mice was evaluated us

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… Mice were euthanized by deep anesthesia with sodium pentobarbital [120 mg/kg, intraperitoneal (i.p.)] followed by cervical dislocation. The researchers made every effort to minimize the number and suffering of the animals for all experiments. … Hematoxylin and eosin (H&E) staining A total of six mice per group were anesthetized with sodium pentobarbital (65 mg/kg; i.p.), and perfused transcardially with 4% paraformaldehyde in PBS resulting in mortality. …

Fingolimod улучшает амилоидное осаждение и нейродегенерацию в модели APP/PS1 мыши болезни Альцгеймера.

ВВЕДЕНИЕ: Иммунная система играет важнейшую роль в регулировании метаболизма амилоид-бета (Aβ) в болезни Альцгеймера (AD). Как T, так и B лимфоциты участвуют в патогенезе AD. Спингосин-1-фосфат (S1P) рецепторный модулятор финголимод, используемый для лечения множественного склероза, может способствовать обживанию лимфоцитов, что может уменьшить инфильтрацию периферийных лимфоцитов в мозг. МЕТОДЫ: В рамках данного исследования 8-месячные мыши АПП/PS1 в течение 2 месяцев подвергались пероральному воздействию фингимода при дозе 1 мг/кг в сутки или соленого раствора в качестве средства контроля. П

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… At 10 months of age, the animals were euthanized with an overdose of pentobarbital (0.08 g/kg) and perfused with filtered phosphate-buffered saline (PBS) through the heart. … After anesthesia with an intraperitoneal injection of 0.7 % pentobarbital sodium, each mouse underwent intracardial perfusion using 0.1 % NaNO 2 in 0.9 % saline. …

Тирапазамин, загруженный CalliSfheres микросферы: подготовка и характеристика как химомболизационный агент для рака печени.

Тирапазамин (TPZ), гипоксикоксический цитотоксичный агент, доказал, что он осуществляет синергическую деятельность по уничтожению опухоли с транскатерской эмболизацией (TAE) против рака печени. Это позволяет TPZ перейти к транскатетерной терапии рака печени, особенно в сочетании с микросферами, поглощающими наркотики. Мы описываем методы подготовки и характеристики загруженных TPZ микросфер (CSMTPZ) с точки зрения их свойств в качестве химемболизирующего агента, включая 1) подготовка КШМТПЗ и определение уровня загрузки наркотиков, 2) определение витро высвобождения ТЗТ, 3) оценка размера и вн

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… . • The animals were anesthetized with an intravenous injection of sodium pentobarbital (30 mg/kg body weight). • After the preparation and sterilization of skin, an incision was made below the xiphoid process to expose the liver. …

CD28 Deficiency Ameliorates Blast Exposure-Induced Lung Inflammation, Oxidative Stress, Apoptosis, and T Cell Accumulation in the Lungs via the PI3K/Akt/FoxO1 Signaling Pathway.

Although CD28 is associated with the expression of inflammatory mediators, apoptosis-related protein, immunosuppression, and tumorigenesis, the effects of CD28 deficiency on blast exposure-induced lung injury have not been investigated. In this study, we have explored the effects of CD28 on blast exposure-induced lung injury and studied its potential molecular mechanisms. A mouse model of blast exposure-induced acute lung injury was established. Sixty C57BL/6 wild-type (WT) and CD28 knockout (CD28-/-) mice were randomly divided into control or model groups. Lung tissue samples were collected 2

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… Briefly, mice were anesthetized by the abdominal injection of 2% pentobarbital sodium (1.5 ml/kg). The anesthetized mice were placed in a protective cover with their chest exposed, and aluminum foil was placed in the middle layer. … Briefly, after 12 hours of fasting and 4 hours of water deprivation preoperatively, mice were intraperitoneally anesthetized with 2% sodium pentobarbital (1.5 ml/kg) and fixed in a prone position on the operating table. …

Caspase-Dependent HMGB1 Release from Macrophages Participates in Peripheral Neuropathy Caused by Bortezomib, a Proteasome-Inhibiting Chemotherapeutic Agent, in Mice.

Given the role of macrophage-derived high mobility group box 1 (HMGB1) in chemotherapy-induced peripheral neuropathy (CIPN) caused by paclitaxel, we analyzed the role of HMGB1 and macrophages in the CIPN caused by bortezomib, a proteasome-inhibiting chemotherapeutic agent used for the treatment of multiple myeloma. Repeated administration of bortezomib caused CIPN accompanied by early-stage macrophage accumulation in the dorsal root ganglion. This CIPN was prevented by an anti-HMGB1-neutralizing antibody, thrombomodulin alfa capable of accelerating thrombin-dependent degradation of HMGB1, anta

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… Determination of Protein Levels of HMGB1 and Its Targeted Receptors On day 14 after the onset of bortezomib treatment, mice were anesthetized with i.p. pentobarbital at 10 mg/kg following i.p. administration of a mixture of midazolam at 4 mg/kg and medetomidine at 0.3 mg/kg. … Analysis of Macrophage Accumulation in the Sciatic Nerve and DRG by Immunofluorometry Under anesthesia with i.p. pentobarbital at 10 mg/kg following i.p. administration of a mixture of midazolam at 4 mg/kg and medetomidine at 0.3 mg/kg, mice were transcardially perfused with ice-cold saline in a volume …

Bortezomib alters sour taste sensitivity in mice.

Chemotherapy-induced taste disorder is one of the critical issues in cancer therapy. Bortezomib, a proteasome inhibitor, is a key agent in multiple myeloma therapy, but it induces a taste disorder. In this study, we investigated the characteristics of bortezomib-induced taste disorder and the underlying mechanism in mice. Among the five basic tastes, the sour taste sensitivity of mice was significantly increased by bortezomib administration. In bortezomib-administered mice, protein expression of PKD2L1 was increased. The increased sour taste sensitivity induced by bortezomib returned to the co

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… Exfoliation of epithelial tissue of circumvallate papillae (CP) Mice were perfused transcardially with saline under deep anesthesia (pentobarbital sodium, 25 mg/kg, i.p. ). … 3′ | β-actin | F | 5′- AGGTCATCACTATTGGCAACGA-3′ | 171 bp | NM_007393 R | 5′- CACTTCATGATGGAATTGAATGTAGTT-3′ | Tissue preparation for frozen sections Mice were perfused transcardially with 4% paraformaldehyde in 0.1 M phosphate buffer (pH 7.4) containing 0.2% picric acid under deep anesthesia (pentobarbital …