Stomach Tissue Samples Research Articles

Alpha-1-syntrophin protein is differentially expressed in human cancers

We studied the expression of α1-syntrophin (SNTA1) protein in histologically confirmed esophageal, stomach, lung, colon, rectal and breast cancerous tissue samples. Our results suggest a significant decrease in the expression level of SNTA1 protein in both esophageal squamous cell carcinoma (ESCC) and esophageal adenocarcinoma (EAC) compared with their respective controls while a significant increase in expression of SNTA1 protein compared with the normal tissue was observed in breast carcinoma samples. No significant difference in expression of SNTA1 protein was observed in stomach, lung, colon and rectal cancers. Our results suggest that SNTA1 has a role in carcinogenesis and could possibly be used as a novel diagnostic or prognostic marker in esophageal and breast cancers.

Combined expression of miR‐122a, miR‐1, and miR‐200b can differentiate degraded RNA samples from liver, pancreas, and stomach

The effect of RNA degradation on the diagnostic utility of microRNA has not been systematically evaluated in clinical samples. We asked if the microRNA profile is preserved in degraded RNA samples derived from mouse and human tissue. We selected tissue-specific microRNA candidates from published human microarray data, and validated them using quantitative reverse transcription polymerase chain reaction (QRTPCR) analyses on flash-frozen, normal mouse liver, pancreas, and stomach tissue samples. MiR-122a, miR-1, and miR-200b were identified as tissue-specific, and the 3-microRNA-based QRTPCR could predict the tissue origin for mouse tissue samples that were left at room temperature for 2 h with an accuracy of 91.7%. When we applied this 3-microRNA predictor to clinical specimens with various degree of RNA degradation, the predictor differentiated degraded RNA samples from liver, pancreas, and stomach with an accuracy of 90% (26/29). Expression levels of miR-122a, miR-1, and miR-200b were modestly changed after the extended (2-4 h) storage at room temperature, but the magnitudes of expression changes were small compared to the expression differences between various tissues of origin. This proof-of-principle study demonstrates that RNA degradation due to extended storage at room temperature does not affect the predictive power of tissue-specific microRNA QRTPCR predictor.

Expression of Cystatin C in human stomach neoplasms

Cystatin C is a member of the cysteine protease inhibitor family and functions to decrease protease production. A recent study showed that it is aberrantly expressed in many malignant tumors in association with tumor invasion and metastasis. Our study aimed to detect Cystatin C expression in stomach neoplasm tissues and adjacent reparative normal tissues. Samples of cancerous and non-cancerous stomach tissue were obtained via surgery as matched pairs from 12 patients with stomach neoplasms and preserved in paraffin. The expression of Cystatin C in the samples was investigated by immunohistochemistry. Fisher's exact test was used to analyze the relationship between stomach neoplasms and adjacent normal tissues. Additionally, mRNA was extracted and analysed by reverse transcriptase-polymerase chain reaction. The intensity of Cystatin C immunostaining was increased in the tumor tissues compared to the adjacent normal tissues. Cystatin C mRNA expression was increased in stomach neoplasms compared to the normal tissues (p<0.05). The results indicate that the expression of Cystatin C may serve as a marker for stomach neoplasms.

Study on trace elements behaviour in cancerous and healthy tissues of colon, breast and stomach: Total reflection X-ray fluorescence applications

In this work Total-reflection X-ray fluorescence (TXRF) was used to analyse healthy and cancerous tissues of the same individual along several contiguous thin sections of each tissue. Thirty two samples (16 pairs) of breast tissue, 30 samples (15 pairs) of intestine tissue and 10 samples (5 pairs) of stomach tissue were analysed. The samples were obtained in Civil Hospitals of Germany (Dortmund) and Portugal (Lisbon). The elemental distribution of P, S, Cl, K, Ca, Cr, Mn, Fe, Ni, Cu, Zn, Se, Br, Rb, Sr and Pb in these samples was studied. Descriptive statistics based on bar graphics and hypotheses tests and also an automatic classification based on hierarchical grouping analysis was used for the several analysed tissues. It was shown that the behaviour of the elements is tissue dependent. Some elements, like P and K exhibit the same behaviour in all the analysed tissue types. They have increased concentrations in all cancerous tissues. Unlike, other elements like Br show completely different behaviour depending on the tissue: similar concentration in healthy and cancerous stomach, decreased levels in colon cancerous tissues and enhanced concentrations in breast was observed. Moreover cancer tissues present decreased Se concentrations on colon and increased on breast.

OC-065 Stem cell analysis by label retention within Barrett's oesophagus

Introduction Stem cells are thought to be the only long lived cells in the gastrointestinal epithelium. It is thought they are the target for mutations that may lead to the premalignant Barrett9s Metaplasia (BM). These cells are thought to divide slowly allowing DNA dyes to be retained within the stem cells over a long period of time. Here, we have used this characteristic to identify, for the first time, the location of stem cells within the glands of Barrett9s oesophagus, an important premalignant condition. Methods We utilised iododeoxyuridine (IUdR) a thymidine analogue which is incorporated into the DNA of replicating cells, as a marker to identify the label retaining cells (LRC) as putative stem cells. Four patients with BM diagnosed with oesophageal adenocarcinoma undergoing oesophagectomy were infused with IudR prior to their surgery (7–69 days). After resection, tissue samples of normal oesophagus, BM, normal stomach and tumour were extracted for immunocytochemistry and FACS. Results Infusion 3 days or less in vitro in transformed cells and explants revealed appropriate abundant staining of the proliferative compartments. Results from FACS analysis showed the percentage of BRdU labelled cells was much greater in BM tissue (7.85%) compared with fundus (0.69%) or squamous (0.38%). However labelling at 7 days in both patients ex vivo showed positive discrete staining of LRC9s within various gastrointestinal tissue types including BM. LRCs were seen in the basal layer of the squamous epithelium, likewise LRCs were seen at the neck/foveolar regions of the gastric tissue conforming to the suggested gastric SC zone. LRC9s were seen at several locations in metaplastic tissue both at the base of the gland and in the neck region at 7, 11, 29 and 67 days, this ties in with preliminary data showing LGR5 (a stem cell marker) is also expressed at the base of the Barrett9s gland. Conclusion This is the first report of LRC9s within the human oesophagus and our findings correlate with the hypothesised location of SCs in the squamous epithelium and gastric mucosa. Furthermore LRCs were located in the base of the BM gland implying that this is the location of BM SC9s. Analysis and molecular characterisation of LRCs and BM SC9s and their niche are underway, involving the use of other GI SC markers.

Protective immunization with homologous and heterologous antigens against Helicobacter suis challenge in a mouse model

Helicobacter (H.) suis colonizes the stomach of more than 60% of slaughter pigs and is also of zoonotic importance. Recently, this bacterium was isolated in vitro, enabling the use of pure cultures for research purposes. In this study, mice were immunized intranasally or subcutaneously with whole bacterial cell lysate of H. suis or the closely related species H. bizzozeronii and H. cynogastricus, and subsequently challenged with H. suis. Control groups consisted of non-immunized and non-challenged mice (negative control group), as well as of sham-immunized mice that were inoculated with H. suis (positive control group). Urease tests on stomach tissue samples at 7 weeks after challenge infection were negative in all negative control mice, all intranasally immunized mice except one, and in all and 3 out of 5 animals of the H. cynogastricus and H. suis subcutaneously immunized groups, respectively. H. suis DNA was detected by PCR in the stomach of all positive control animals and all subcutaneously immunized/challenged animals. All negative control animals and some intranasally immunized/challenged mice were PCR-negative. In conclusion, immunization using antigens derived from the same or closely related bacterial species suppressed gastric colonization with H. suis, but complete protection was only achieved in a minority of animals following intranasal immunization.

Ghrelin improves burn-induced multiple organ injury by depressing neutrophil infiltration and the release of pro-inflammatory cytokines

Mechanisms of burn-induced skin and remote organ injury involve oxidant generation and the release of pro-inflammatory cytokines. In this study the possible antioxidant and anti-inflammatory effects of ghrelin were evaluated in a rat model of thermal trauma. Wistar albino rats were exposed to 90 degrees C bath for 10 s to induce thermal trauma. Ghrelin, was administered subcutaneously (10 ng/kg/day) after the burn injury and repeated twice daily. Rats were decapitated at 6 h and 48 h after burn injury and blood was collected for the analysis of pro-inflammatory cytokines (TNF-alpha and IL-1beta), lactate dehydrogenase (LDH) activity and antioxidant capacity (AOC). In skin, lung and stomach tissue samples malondialdehyde (MDA) and glutathione (GSH) levels, myeloperoxidase (MPO) and Na(+)-K(+)-ATPase activity were measured in addition to the histological analysis. DNA fragmentation ratio in the gastric mucosa was also evaluated. Burn injury caused significant increase in both cytokine levels, and LDH activity, while plasma AOC was found to be depleted after thermal trauma. On the other hand, in tissue samples the raised MDA levels, MPO activity and reduced GSH levels, Na(+)-K(+)-ATPase activity due to burn injury were found at control levels in ghrelin-treated groups, while DNA fragmentation in the gastric tissue was also reduced. According to the findings of the present study, ghrelin possesses a neutrophil-dependent anti-inflammatory effect that prevents burn-induced damage in skin and remote organs and protects against oxidative organ damage.

A three generation study with genetically modified Bt corn in rats: Biochemical and histopathological investigation

For the last ten years, in accordance with the increased use of genetically modified (GM) foods for human and livestocks, a large number of feeding studies have been carried out. However, the evidence is still far from proving whether the long-term consumption of GM foods posses a possible danger for human or animal health. Therefore, this study was designed to evaluate the effects of transgenic corn on the rats that were fed through three generations with either GM corn or its conventional counterpart. Tissue samples of stomach, duodenum, liver and kidney were obtained for histopathological examinations. The average diameter of glomeruli, thickness of renal cortex and glomerular volume were calculated and number of affected animals/number of examined animals for liver and kidney histopathology were determined. Amounts of urea, urea nitrogen, creatinine, uric acid, total protein, albumin and globulin were determined; enzyme activities of aspartate aminotransferase, alanine aminotransferase, alkaline phosphatase, gamma glutamyltransferase, creatine kinase and amylase were measured in serum samples. No statistically significant differences were found in relative organ weights of rats within groups but there were some minimal histopathological changes in liver and kidney. Changes in creatinine, total protein and globulin levels were also determined in biochemical analysis.

Synthesis and evaluation of radioiodine-labelled CP-118,954 for the in-vivo imaging of acetylcholinesterase

Alzheimer's disease (AD) is characterized by reduced acetylcholinesterase (AChE) activity in the post-mortem tissues of AD patients. Therefore, AChE has been an attractive target for the diagnosis of AD. In the present study, 5,7-dihydro-3-[2-(1-(phenylmethyl)-4-piperidinyl)ethyl]-6H-pyrrolo[3,2-f]-1,2-benzisoxazol-6-one (CP-118,954), a potent AChE inhibitor, was labelled with radioiodine and evaluated as an AChE imaging agent for SPECT. Radioiodine-labelled CP-118,954 was prepared from CP-144,885 and [(125)I]iodobenzyl bromide, and anti-AChE activities of iodine-substituted CP-118,954 were measured. Metabolism studies were carried out in samples of blood and whole brain of mice injected with 2-[(123)I]iodo-CP-118,954 ((123)I-1). Tissue distribution studies were also performed in mice injected with I-1, and samples of blood, thyroid, stomach, and brain tissue (cerebellum, striatum and cortex) were removed, weighed and counted. Of the ligands, 2-iodo-CP-118,954 exhibited higher binding affinity for AChE (IC50=24 nM) than the other positional isomers. 2-[(125)I]Iodo-CP-118,954 was found to have a lipophilicity (log P=2.1) favouring brain permeability and metabolic stability in mouse brain, but a marginal target (striatum) to non-target (cerebellum) uptake ratio (1.1) in mouse brain. This result demonstrates that 2-[(125)I]iodo-CP-118,954 may be unsuitable for AChE imaging. These findings suggest that radioligands suitable for AChE imaging should have not only a specific structure but also a sub-nanomolar to low nanomolar IC50.

Distribution of trace metal concentrations in paired cancerous and non-cancerous human stomach tissues.

To assess whether trace metal concentrations (which influence metabolism as both essential and non-essential elements) are increased or decreased in cancerous tissues and to understand the precise role of these metals in carcinogenesis. Concentrations of trace metals including Cd, Ni, Cu, Zn, Fe, Mg and Ca in both cancerous and non-cancerous stomach tissue samples were determined by atomic absorption spectrometry (AAS). Tissue samples were digested using microwave energy. Slotted tube atom trap was used to improve the sensitivity of copper and cadmium in flame AAS determinations. From the obtained data in this study, the concentrations of nickel, copper and iron in the cancerous human stomach were found to be significantly higher than those in the non-cancerous tissues, by using t-test for the paired samples. Furthermore, the average calcium concentrations in the cancerous stomach tissue samples were found to be significantly lower than those in the non-cancerous stomach tissue samples by using t-test. Exceedingly high Zn concentrations (207-826 mg/kg) were found in two paired stomach tissue samples from both cancerous and non-cancerous parts. In contrast to the literature data for Cu and Fe, the concentrations of copper, iron and nickel in cancerous tissue samples are higher than those in the non-cancerous samples. Furthermore, the Ca levels are lower in cancerous tissue samples than in non-cancerous tissue samples.

Identification of Protein Expression Signatures Associated with Helicobacter pylori Infection and Gastric Adenocarcinoma Using Recombinant Antibody Microarrays

Antibody microarray based technology is a powerful emerging tool in proteomics, target discovery, and differential analysis. Here, we report the first study where recombinant antibody fragments have been used to construct large scale antibody microarrays, composed of 127 different antibodies against mostly immunoregulatory antigens. The arrays were based on single framework recombinant antibody fragments (SinFabs) designed for high on-chip stability and functionality and were used for the analysis of malignant and normal stomach tissue samples from Helicobacter pylori-positive and -negative patients. Our results demonstrate that distinct tumor- as well as infection-associated protein expression signatures could be identified from these complex tissue proteomes, as well as biomarkers such as IL-9, IL-11, and MCP-4, previously not found in these diseases. In a longer perspective, this study may improve the understanding of H. pylori-induced stomach cancer and lead to development of improved diagnostics.

Calcimimetic R-568 Decreases Extraosseous Calcifications in Uremic Rats Treated with Calcitriol

Calcimimetics decrease parathyroid hormone (PTH) levels in uremic patients with secondary hyperparathyroidism without increasing serum calcium (Ca). The aim of this study was to evaluate the effect of calcimimetic R-568 alone or in combination with calcitriol on vascular and other soft tissue calcifications in uremic rats with secondary hyperparathyroidism. Sham-operated and 5/6 nephrectomized Wistar rats were studied. 5/6 Nephrectomized rats were treated with vehicle, calcitriol (80 ng/kg every other day), R-568 (1.5 and 3 mg/kg per d), and both calcitriol and R-568 1.5 mg/kg, as above. Rats were killed after 14 or 56 d of treatment. Blood was drawn for biochemical measurements. Aortic, heart, kidney, lung, and stomach tissue samples were processed for histopathology and measurement of tissue Ca and phosphorus content. PTH concentrations were significantly reduced by all treatments. Treatment with calcitriol induced significant vascular calcification (aortic Ca increased to 4.2+/-1.2 mg/g at day 14 and to 11.4+/-0.7 mg/g at day 56; P<0.05 versus vehicle). Treatment with R-568 did not induce vascular calcification. Concurrent administration of R-568 with calcitriol reduced the aortic Ca (1.9+/-0.2 mg/g at day 14 and 7.5+/-1.4 mg/g at day 56) in relation to calcitriol alone. Soft tissue calcifications mirrored aortic mineralizations. Survival was significantly (P<0.001) reduced in calcitriol-treated rats, and mortality was attenuated (P=0.01) by concurrent treatment with R-568. In uremic rats, R-568 reduces elevated PTH levels without inducing vascular calcification, prevents calcitriol-induced vascular calcification, and decreases mortality.

Protective immunization against “ Candidatus Helicobacter suis” with heterologous antigens of H. pylori and H. felis

“ Helicobacter ( H.) heilmannii” type 1 colonizes the human stomach. It has been shown to be identical to “ Candidatus H. suis”, a Helicobacter species colonizing the stomach of more than 60% of slaughter pigs. This bacterium is, until now, not isolated in vitro. The effect of vaccination on “ Candidatus H. suis” infection was studied in a mouse model. Mice were vaccinated intranasally or subcutaneously with whole bacterial cell lysate of Helicobacter pylori or Helicobacter felis and subsequently challenge infected with “ Candidatus H. suis”. Intranasal and subcutaneous immunisation caused a decrease in faecal excretion of “ Candidatus H. suis” DNA. Urease tests on stomach tissue samples at 16 weeks after challenge infection were negative in all H. felis intranasally immunized animals and in the majority of the animals of the other immunisation groups. Since PCR on stomach tissue samples at 16 weeks after challenge infection could still detect “ Candidatus H. suis DNA” in all immunisation-challenge groups, complete clearance of challenge bacteria was not achieved.

Enhancing capillary liquid chromatography/tandem mass spectrometry of biogenic amines by pre-column derivatization with 7-fluoro-4-nitrobenzoxadiazole.

This paper describes a capillary liquid chromatography/tandem mass spectrometry (LC/MS/MS) determination of biogenic amines enhanced by pre-column derivatization with 7-fluoro-4-nitrobenzoxadiazole (NBD-F). Biogenic amines including tryptamine, N-methylsalsolinol, histamine, and agmatine were studied. The biogenic NBD-amine derivatives could be quantitatively enriched in-line on 20 x 0.25 mm capillary columns packed in-house with 5 microm C(8) silica particles. In an electrospray ionization (ESI) source these derivatives were ionized effectively, and collision-induced dissociation (CID) produced predominant characteristic ions allowing sensitive MS/MS detection. Agmatine, a potential neurotransmitter/modulator, was taken as a reference compound to study the analytical figures of merit of the procedure. The detection limit of agmatine was estimated to be 0.6 ng/mL (signal-to-noise (S/N) = 3). A linear calibration curve in the range 15-1000 ng/mL agmatine with an r value of 0.9997 was obtained. Tissue samples of rat brain, stomach, and intestine were analyzed. Minimum sample pre-treatment was needed. Each analysis was accomplished within ca. 12 min. The concentration of agmatine was found to be 0.246, 3.31, and 0.058 microg/g wet tissue in the brain, stomach, and intestine, respectively.

Investigation of Normal and Malignant Tissue Samples from the Human Stomach Using Fourier Transform Raman Spectroscopy

Fourier transform (FT) Raman studies of 40 tissue samples from the human stomach, including 22 normal and 18 malignant tissue samples, were performed. These stomach tissue samples had been classified as normal or malignant on the basis of pathological studies and biopsy detection. The results indicate that a series of major bands in the FT-Raman spectrum can be used to distinguish the malignant tissue from the normal tissue. Subtraction spectra support the result of the spectroscopic identification. Statistical analysis is also in agreement with the FT-Raman measurements.

O6-Alkylguanines, Dietary N-Nitroso Compounds, and Their Precursors in Gastric Cancer

Several N-nitroso compounds, present in foods and beverages or formed in the stomach from their precursors, act as alkylating agents. By using a highly reliable technique (high-resolution gas chromatography-mass spectrometry with negative-ion chemical ionization and selected ion recording), we measured a series of specific O6-alkylguanines in snap-frozen paired stomach tissue samples (tumor and noninvolved mucosa) obtained at surgery from 24 gastric cancer patients identified in Florence, Italy. Samples of noninvolved mucosa had higher levels of total O6-alkylguanines and more frequently detectable levels (54%) than tumor samples (29.2%). O6-propylguanine and O6-methylguanine were the single adducts most frequently detected in noninvolved mucosa and tumor tissue, respectively. Tumor samples showed higher levels of total O6-alkylguanines in female patients (p = 0.03) and among those with a diffuse histological type (p = 0.06) or seronegative for Helicobacter pylori CagA antibodies (p = 0.06). Mean dietary nitrate intake was significantly higher in patients with detectable levels of adducts in tumor samples (p = 0.03). Estimated intakes of dimethylamine and N-nitrosodimethylamine correlated with total levels of O6-alkylguanines in noninvolved gastric mucosa. These findings, although based on a small series of cases, support a role for N-nitroso compounds from dietary sources in the etiology of gastric cancer.

Improved hepatic and pancreatic localisation of the equine alpha-1-proteinase inhibitor family of serpins using an antigen enhancement technique and a monoclonal antibody

Equine alpha- l-proteinase inhibitor (API) consists of three, occasionally four, serum glycoproteins. This study investigated the immunohistochemical localisation of equine API in paraformaldehyde fixed, paraffin embedded equine tissue samples of liver, lung, stomach, pancreas, jejunum and colon in five horses using affinity purified sheep polyclonal and protein A purified mouse monoclonal antibodies, whose specificities were verified by Western blotting. Exposing tissue sections to boiling citrate buffer greatly enhanced antigen recovery and improved immunostaining with both antibodies, resulting in discovery of novel tissue distribution patterns for the horse. In the horses studied, all hepatocytes showed some degree of cytoplasmic staining, many having perinuclear intense granular inclusions. This finding is contrary to findings in human studies where hepatocytes of Pi MM phenotype have proven difficult to stain for human API, despite evidence at the molecular level suggesting hepatocytes as the major source of serum API. This discrepancy may be due to the use of different tissue fixation and antigen recovery techniques. In all other tissues examined, the distribution of equine API was similar to human studies.

Regulation and tissue distribution of the human sodium iodide symporter gene.

Iodide uptake by the thyroid gland is mediated by the sodium iodide symporter (NIS). In the present report, we have analysed the factors that modulate human NIS mRNA expression and iodide uptake in primary thyroid follicular cell (TFC) cultures. In addition, NIS mRNA tissue distribution was investigated. Primary thyroid follicular cell cultures were treated with human recombinant TSH with or without cytokines for 72 h. Subsequently, NIS gene expression and iodide uptake were analysed using reverse transcription-polymerase chain reaction (RT-PCR) and 125I uptake, respectively. Human tissue samples were investigated for NIS gene expression using both RT-PCR and Northern blotting. Human TSH increased both NIS gene expression and iodide uptake in TFC cultures in a dose-dependent manner. Using concentrations of 0.1 U/l of hTSH, a minor increase in NIS gene expression was detected without a detectable increase in iodide uptake. IL-1 alpha, TNF alpha and IFN gamma at concentrations of 10(5) U/l all inhibited TSH-induced NIS gene expression and iodide uptake. In these experiments, there was a good correlation between NIS mRNA expression and iodide uptake. Using RT-PCR higher levels of NIS mRNA were detected in Graves' disease (GD) compared to multi-nodular goitre tissue samples. Stomach and salivary gland tissue also expressed NIS mRNA, whereas low levels were found in the mammary gland and extraocular muscle tissue. No expression was detected in the ovary, oesophagus, colon, extraocular fat or skin. In contrast, Northern blot analysis failed to detect NIS in stomach, salivary gland, intestinal fat or non-toxic multi-nodular goitre tissue samples, although this was present in GD thyroid tissue. TSH upregulates sodium iodide symporter gene expression and iodide uptake in primary thyroid follicular cell cultures, and this induction is modulated by cytokines. Variable levels of sodium iodide symporter mRNA are present in different tissue samples, with high expression evident in Graves' disease thyroid tissue.

Is herpes simplex virus associated with peptic ulcer disease?

To test the hypothesis that herpes simplex virus type 1 (HSV-1) may be associated with peptic ulcer disease, we examined ulcerative lesions of the distal stomach and proximal duodenum for the presence of nucleic acids and antibodies specific for HSV-1. Utilizing in situ hybridization, immunocytochemistry, and polymerase chain reaction with sequencing, gastric or duodenal tissues from 4 of 22 patients (18%) with documented peptic ulcer disease demonstrated the presence of both specific HSV-1 nucleic acid sequences and proteins. HSV-1 was found restricted in clusters of cells near the margin of the ulcer but was absent at sites distal to the lesion. Several of such HSV-1-infected cells also contained cholecystokinin. These cholecystokinin-containing cells are of neuroendocrine origin and receive contact from the vagal nerve. Campylobacter pylori bacteria were not found in three of the four peptic ulcer tissues that harbored HSV-1. Further, none of the stomach or duodenal tissue samples from 33 patients undergoing clinical evaluation, but having no evidence of peptic ulcer disease, had HSV-1 materials. Thus, our data suggest that a subset of peptic ulcer disease may be associated with HSV-1 and raise the possibility that some peptic ulcers may be caused by this virus.

Age-related changes in regional blood flow in the rat.

The purpose of this investigation was to characterize the changes in regional blood flow and central hemodynamic measures that occur in the rat as a result of the aging process. The isotope-labeled microsphere technique was used to measure cardiac output and regional blood flows in conscious and anesthetized adult (12 mo) and senescent (24 mo) Fischer 344 virgin female rats. No significant changes were observed in central hemodynamic measurements or regional blood flows in conscious rats with the exception of a 25% reduction in splenic blood flow. Pentobarbital anesthesia significantly reduced cardiac index and heart rate but elevated total peripheral resistance and mean arterial blood pressure. There was a decrease in blood flow to skeletal muscle, spleen, duodenum, stomach, and brain tissue samples and increased hepatic arterial blood flow in both age groups. The use of anesthesia caused a greater reduction in the cardiac index and brain blood flow in the senescent anesthetized rats than in the adult rats. Heart and kidney blood flows were decreased by anesthesia in the senescent rats but not in the adult rats. Skeletal muscle blood flow, however, was significantly greater in the senescent anesthetized rats than in the younger anesthetized animals. Although body weight and organ weights of the liver, spleen, kidneys, stomach, heart, and brain were significantly greater for the senescent rats, no differences could be demonstrated in tibial length or lean body mass.