Correlation of γ-radioactivity and Scanning Electron Microscopy in measurement of retention of 111Indium-labeled canine endothelial cells on synthetic vascular grafts

Endothelial cell (EC) seeding is postulated as a mechanism of improving patency in small caliber vascular grafts. However the majority of seeded EC are lost within 24 hours of restoration of blood flow in previous canine studies . We postulate that the cells have insufficient time to fully develop their attachment to the graft surface prior to exposure to hemodynamic stress. We allowed EC to incubate on fibronectin-coated ePTFE grafts for four different time periods after seeding and measured EC retention after perfusion in a canine ex vivo shunt circuit.Autologous canine EC, were enzymatically harvested, grown to confluence, and labeled with 30 μCi 111 Indium-oxine/80 cm 2 flask. Four groups of 5 cm x 4 mm ID ePTFE vascular prostheses were coated with 1.5 μg/cm.2 human fibronectin, and seeded with 1.5 x 105 EC/ cm.2. After seeding grafts in Group 1 were incubated in complete growth medium for 90 minutes, Group 2 were incubated for 24 hours, Group 3 for 72 hours and Group 4 for 6 days. Grafts were then placed in the canine ex vivo circuit, constructed between femoral artery and vein, and subjected to blood flow of 75 ml per minute for 6 hours. Continuous counting of γ-activity was made possible by placing the seeded graft inside the γ-counter detection crystal for the duration of perfusion. EC retention data after 30 minutes, 2 hours and 6 hours of flow are shown in the table.

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Influence of adenosine on cerebral blood flow during hypoxic hypoxia in the newborn piglet

Journal of Applied Physiology ◽

10.1152/jappl.1990.68.4.1534 ◽

1990 ◽

Vol 68 (4) ◽

pp. 1534-1541 ◽

Cited By ~ 33

Author(s):

N. Laudignon ◽

E. Farri ◽

K. Beharry ◽

J. Rex ◽

J. V. Aranda

Keyword(s):

Blood Flow ◽

Cerebral Blood Flow ◽

High Performance ◽

Severe Hypoxia ◽

Group 4 ◽

Csf Levels ◽

Group 3 ◽

Group 2 ◽

Arterial Po2 ◽

Group 1

This study investigated the role of adenosine in the regulation of neonatal cerebral blood flow (CBF) during moderate (arterial PO2 = 47 +/- 9 Torr) and severe (arterial PO2 = 25 +/- 4 Torr) hypoxia. Twenty-eight anesthetized and ventilated newborn piglets were assigned to four groups: 8 were injected intravenously with the vehicle (controls, group 1); 13 received an intravenous injection of 8-phenyltheophylline (8-PT), a potent adenosine receptor blocker, either 4 mg/kg (group 2, n = 6, mean cerebrospinal fluid (CSF) levels less than 1 mg/l) or 8 mg/kg (group 3, n = 7, mean CSF levels less than 3.5 mg/l); and 7 received an intracerebroventricular injection of 10 micrograms 8-PT (group 4). During normoxia, CBF was not altered by vehicle or 8-PT injections. In group 1, 10 min of moderate and severe hypoxia increased total CBF by 112 +/- 36 and 176 +/- 28% (SE), respectively. Compared with controls, the cerebral hyperemia during moderate hypoxia was not altered in group 2, attenuated in group 3 (to 53 +/- 13%, P = NS), and completely blocked in group 4 (P less than 0.01). CBF increase secondary to severe hypoxia was attenuated only in group 4 (74 +/- 29%, P less than 0.05). CSF concentrations of adenosine and adenosine metabolites measured by high-performance liquid chromatography increased during hypoxia. Arterial O2 content was inversely correlated (P less than 0.005) to maximal CSF levels of adenosine (r = 0.73), inosine (r = 0.87), and hypoxanthine (r = 0.80).(ABSTRACT TRUNCATED AT 250 WORDS)

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Brain Interstitial Adenosine and Sagittal Sinus Blood Flow during Systemic Hypotension in Piglet

Journal of Cerebral Blood Flow & Metabolism ◽

10.1038/jcbfm.1988.138 ◽

1988 ◽

Vol 8 (6) ◽

pp. 822-828 ◽

Cited By ~ 24

Author(s):

T. S. Park ◽

David G. L. Van Wylen ◽

Rafael Rubio ◽

Robert M. Berne

Keyword(s):

Blood Flow ◽

Interstitial Fluid ◽

Cerebrovascular Resistance ◽

Sagittal Sinus ◽

Group 4 ◽

Brain Dialysis ◽

Group 3 ◽

Group 2 ◽

The Brain ◽

Dialysis Technique

We sampled, using the brain dialysis technique, interstitial fluid adenosine from the frontal cortex of newborn piglets subjected to hemorrhagic hypotension while measuring sagittal sinus blood flow, cerebrovascular resistance (CVR), and cerebral O2 delivery. In group 1 (n = 8), MABP was reduced in successive steps from 76 to 30 mm Hg with decrements of ∼ 10 mm Hg. At 60 mm Hg, CVR decreased by 19% (p < 0.001), but sagittal sinus blood flow and interstitial fluid adenosine remained unchanged. At 50 mm Hg, both sagittal sinus blood flow and CVR decreased by 19% (p < 0.001) and interstitial fluid adenosine rose 4.7-fold (p < 0.05). At 40 and 30 mm Hg, sagittal sinus blood flow decreased further but CVR remained steady, whereas interstitial fluid adenosine rose 10- and 16-fold, respectively. In group 2 (n = 7), an abrupt reduction of MABP from 80 to 47 mm Hg produced no change in sagittal sinus blood flow and a 29% decrease in CVR (p < 0.01). Interstitial fluid adenosine increased twofold (p < 0.01). In group 3 (n = 7), an abrupt reduction of MABP from 79 to 40 mm Hg decreased sagittal sinus blood flow and CVR by 24 and 30%, respectively (p < 0.01). Interstitial fluid adenosine rose threefold (p < 0.01). In groups 1, 2, and 3, the increases in interstitial fluid adenosine accompanied decreases in cerebral O2 delivery. In group 4 (n = 7), artificial CSF with a Po2 of 152 mm Hg was perfused through the brain dialysis cannula during graded hypotension. In this group, interstitial fluid adenosine rose only at an MABP of 20 mm Hg. These data support the concept that adenosine participates in the regulation of CBF during hypotension in piglets.

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Circulatory Rejuvenated EPCs Derived from PAOD Patients Treated by CD34+ Cells and Hyperbaric Oxygen Therapy Salvaged the Nude Mouse Limb against Critical Ischemia

International Journal of Molecular Sciences ◽

10.3390/ijms21217887 ◽

2020 ◽

Vol 21 (21) ◽

pp. 7887

Author(s):

Yin-Chia Chen ◽

Jiunn-Jye Sheu ◽

John Y. Chiang ◽

Pei-Lin Shao ◽

Shun-Cheng Wu ◽

...

Keyword(s):

Blood Flow ◽

Cytochrome C ◽

Nude Mouse ◽

Hyperbaric Oxygen ◽

Nude Mice ◽

Group 4 ◽

Protein Expressions ◽

Group 3 ◽

Group 2 ◽

Group 1

This study tested whether circulatory endothelial progenitor cells (EPCs) derived from peripheral arterial occlusive disease (PAOD) patients after receiving combined autologous CD34+ cell and hyperbaric oxygen (HBO) therapy (defined as rejuvenated EPCs) would salvage nude mouse limbs against critical limb ischemia (CLI). Adult-male nude mice (n = 40) were equally categorized into group 1 (sham-operated control), group 2 (CLI), group 3 (CLI-EPCs (6 × 105) derived from PAOD patient’s circulatory blood prior to CD34+ cell and HBO treatment (EPCPr-T) by intramuscular injection at 3 h after CLI induction) and group 4 (CLI-EPCs (6 × 105) derived from PAOD patient’s circulatory blood after CD34+ cell and HBO treatment (EPCAf-T) by the identical injection method). By 2, 7 and 14 days after the CLI procedure, the ischemic to normal blood flow (INBF) ratio was highest in group 1, lowest in group 2 and significantly lower in group 4 than in group 3 (p < 0.0001). The protein levels of endothelial functional integrity (CD31/von Willebrand factor (vWF)/endothelial nitric-oxide synthase (eNOS)) expressed a similar pattern to that of INBF. In contrast, apoptotic/mitochondrial-damaged (mitochondrial-Bax/caspase-3/PARP/cytosolic-cytochrome-C) biomarkers and fibrosis (Smad3/TGF-ß) exhibited an opposite pattern, whereas the protein expressions of anti-fibrosis (Smad1/5 and BMP-2) and mitochondrial integrity (mitochondrial-cytochrome-C) showed an identical pattern of INBF (all p < 0.0001). The protein expressions of angiogenesis biomarkers (VEGF/SDF-1α/HIF-1α) were progressively increased from groups 1 to 3 (all p < 0.0010). The number of small vessels and endothelial cell surface markers (CD31+/vWF+) in the CLI area displayed an identical pattern of INBF (all p < 0.0001). CLI automatic amputation was higher in group 2 than in other groups (all p < 0.001). In conclusion, EPCs from HBO-C34+ cell therapy significantly restored the blood flow and salvaged the CLI in nude mice.

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Prolonged ex-vivo normothermic kidney perfusion: The impact of perfusate composition

PLoS ONE ◽

10.1371/journal.pone.0251595 ◽

2021 ◽

Vol 16 (5) ◽

pp. e0251595

Author(s):

Merel B. F. Pool ◽

Tim L. Hamelink ◽

Harry van Goor ◽

Marius C. van den Heuvel ◽

Henri G. D. Leuvenink ◽

...

Keyword(s):

Ex Vivo ◽

Thiobarbituric Acid ◽

Renal Perfusion ◽

Group 4 ◽

Normothermic Machine Perfusion ◽

Group 3 ◽

Group 2 ◽

The Individual ◽

Scintigraphic study of the splenic circulation in rats

Revista do Colégio Brasileiro de Cirurgiões ◽

10.1590/0100-69912014005009 ◽

2014 ◽

Vol 41 (5) ◽

pp. 345-350

Author(s):

Fábio Gontijo Rodrigues ◽

Andy Petroianu ◽

Simone Odília Fernandes Diniz ◽

Valbert Nascimento Cardoso ◽

Eduardo Vilhena de Parreira

Keyword(s):

Blood Flow ◽

Venous Drainage ◽

Group 4 ◽

Abdominal Organs ◽

Splenic Parenchyma ◽

Flow Through ◽

Group 3 ◽

Group 5 ◽

Group 2 ◽

Iliac Bifurcation

Objective: To evaluate the splenic parenchymal blood distribution through scintigraphic study.Methods: Thirty Wistar rats were randomly divided into six groups (n = 5). Group 1 (spleen - 30 minutes) and Group 2 (spleen - 90 minutes) underwent laparotomy with direction of blood flow to the spleen by ligature of the aorta near the iliac bifurcation and splanchnic vessels, keeping blood flow only in the splenic artery; Group 3 (spleen and stomach - 30 minutes) and Group 4 (spleen and stomach - 90 minutes) underwent laparotomy with direction of blood flow to the spleen and stomach by ligature of the aorta near the iliac bifurcation and splanchnic vessels, maintaining the flow through the splenic, gastric and splenogastric vessels; Group 5 (control - 30 minutes) and Group 6 (control - 90 minutes) underwent laparotomy and ligation of the aorta near the iliac bifurcation, keeping the flow to the abdominal organs. After arterial ligation, the animals received an injection of 0.2 ml of sodium pertechnetate in the aorta. Scintigraphic images were taken and the animals had their spleens removed for radioactivity counting with an automatic counter device.Results: There was no difference in the amounts of radiation from the spleen between groups, indicating retention of the radioisotope by the spleen, even after the period of 90 minutes.Conclusion: The blood flow through the spleen is not continuous. The blood diffuses through the splenic parenchyma and its venous drainage is slow, not following a predictable sequence.

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Interstitial Fluid Adenosine and Sagittal Sinus Blood Flow during Bicuculline-Seizures in Newborn Piglets

Journal of Cerebral Blood Flow & Metabolism ◽

10.1038/jcbfm.1987.116 ◽

1987 ◽

Vol 7 (5) ◽

pp. 633-639 ◽

Cited By ~ 33

Author(s):

T. S. Park ◽

David G. L. Van Wylen ◽

Rafael Rubio ◽

Robert M. Berne

Keyword(s):

Blood Flow ◽

Interstitial Fluid ◽

Cerebrovascular Resistance ◽

Newborn Piglets ◽

Sagittal Sinus ◽

Group 4 ◽

Group 3 ◽

Group 2 ◽

The Brain ◽

Level Group

Changes of interstitial fluid adenosine concentrations and effects of O2 supply on interstitial fluid adenosine were studied by the brain dialysis technique in the frontal cortex of newborn piglets subjected to bicuculline-induced seizures. The O2 supply was changed globally by changing MABP and locally by varying PO2 in the artificial CSF perfusing the dialysis cannula. Sagittal sinus blood flow (SSBF), cerebrovascular resistance (CVR), and CMRO2 were also examined in the same animals. Seizures increased interstitial fluid adenosine 7.9-fold (p < 0.05) when ictal MABP was maintained at preictal level and perfusate PO2 was 24 mm Hg (group 1, n = 6). Interstitial fluid adenosine increased 11.8-fold (p < 0.05) during seizures associated with moderate systemic hypotension and the low perfusate PO2 (group 2, n = 6). By contrast, seizures increased interstitial fluid adenosine three-fold (p < 0. 05) when perfusate PO2 was increased to 182 mm Hg and ictal MABP was maintained at preictal level (group 3, n = 8). When ictal MABP was elevated from the preictal level and the perfusate was rich in oxygen, seizures failed to increase interstitial fluid adenosine (group 4, n = 7). In groups 1 and 3, the increase in interstitial fluid adenosine during seizures was associated with significant increases in SSBF and CMRO2, as well as significant decreases in CVR. These data suggest that the increase in O2 supply during seizures in piglets did not match completely the increase in O2 demand and resulted in enhanced release of adenosine into the interstitial space.

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Involvement of the coeliac ganglion in the luteotrophic effect of androstenedione in late pregnant rats

Reproduction ◽

10.1530/rep.1.00852 ◽

2006 ◽

Vol 131 (2) ◽

pp. 361-368 ◽

Cited By ~ 9

Author(s):

Marilina Casais ◽

Silvia Marcela Delgado ◽

Zulema Sosa ◽

Ana María Rastrilla

Keyword(s):

Ex Vivo ◽

Functional Regression ◽

Pregnant Rats ◽

Luteal Regression ◽

Group 4 ◽

System A ◽

Superior Ovarian Nerve ◽

Group 3 ◽

Group 2 ◽

Group 1

Using the ex vivo coeliac ganglion-superior ovarian nerve-ovary system at the end of pregnancy when luteal regression starts, we investigated whether, when administered systemically or when added directly to the ganglion compartment, androstenedione (A2) can reverse such regression, and whether the neural (noradrenaline (NA)) and endocrine (A2) joint action modifies the release of ovarian progesterone. The experimental groups were as follows: group 1 – A2 injected systemically 48 h before incubation of the system (A2)s; group 2 – A2 directly added to the ganglion compartment (A2)g; group 3 – A2 injected 48 h before incubation of the system with NA in the ganglion compartment (A2 + NA); group 4 – A2 plus NA added to the ganglion compartment (NA + A2)g. The controls were ex vivo systems without treatment (control), and with the addition of NA alone in the ganglion compartment (NA). The results were as follows. For (A2)s versus control, progesterone increased on days 19 and 21 of pregnancy at all the studied times and only at 180 min on day 20. For (A2 + NA) versus (A2)s, progesterone increased on days 19 and 21. For (A2 + NA) versus NA, progesterone increased at all the studied times on days 19 and 21 and at 180 min on day 20. For (A2)g versus control, progesterone significantly increased every pregnancy day. For (NA + A2)g versus (A2)g, progesterone decreased at 120 and 180 min on day 19. For (NA + A2)g versus NA, progesterone increased on days 20 and 21. We can conclude that A2 can reverse the functional regression of the corpus luteum either systemically or, what is more surprising, when directly added to the coeliac ganglion, whose action on the ovary is exerted via superior ovarian nerve.

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Morphological Changes in the Rat Granulosa Cell Surface During Differentiation Induced by Estradiol and Gonadotropins

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100079851 ◽

1977 ◽

Vol 35 ◽

pp. 484-485

Author(s):

P. Bagavandoss ◽

JoAnne S. Richards ◽

A. Rees Midgley

Keyword(s):

Cell Surface ◽

Granulosa Cell ◽

Morphological Changes ◽

Follicular Development ◽

Female Rats ◽

Functional Changes ◽

Group 4 ◽

Group 3 ◽

Group 5 ◽

Group 2

During follicular development in the mammalian ovary, several functional changes occur in the granulosa cells in response to steroid hormones and gonadotropins (1,2). In particular, marked changes in the content of membrane-associated receptors for the gonadotropins have been observed (1).We report here scanning electron microscope observations of morphological changes that occur on the granulosa cell surface in response to the administration of estradiol, human follicle stimulating hormone (hFSH), and human chorionic gonadotropin (hCG).Immature female rats that were hypophysectcmized on day 24 of age were treated in the following manner. Group 1: control groups were injected once a day with 0.1 ml phosphate buffered saline (PBS) for 3 days; group 2: estradiol (1.5 mg/0.2 ml propylene glycol) once a day for 3 days; group 3: estradiol for 3 days followed by 2 days of hFSH (1 μg/0.1 ml) twice daily, group 4: same as in group 3; group 5: same as in group 3 with a final injection of hCG (5 IU/0.1 ml) on the fifth day.

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In-vivo evaluation of the effect of cyanoacrylate on prosthetic vascular graft infection – does cyanoacrylate increase the severity of infection?

VASA ◽

10.1024/0301-1526/a000867 ◽

2020 ◽

Vol 49 (4) ◽

pp. 281-284

Author(s):

Atıf Yolgosteren ◽

Gencehan Kumtepe ◽

Melda Payaslioglu ◽

Cuneyt Ozakin

Keyword(s):

Vascular Graft ◽

Graft Infection ◽

Vascular Graft Infection ◽

Group 4 ◽

Significant Difference ◽

Group 3 ◽

Group 2 ◽

Prosthetic Vascular Graft Infection ◽

Group 1

Summary. Background: Prosthetic vascular graft infection (PVGI) is a complication with high mortality. Cyanoacrylate (CA) is an adhesive which has been used in a number of surgical procedures. In this in-vivo study, we aimed to evaluate the relationship between PVGI and CA. Materials and methods: Thirty-two rats were equally divided into four groups. Pouch was formed on back of rats until deep fascia. In group 1, vascular graft with polyethyleneterephthalate (PET) was placed into pouch. In group 2, MRSA strain with a density of 1 ml 0.5 MacFarland was injected into pouch. In group 3, 1 cm 2 vascular graft with PET piece was placed into pouch and MRSA strain with a density of 1 ml 0.5 MacFarland was injected. In group 4, 1 cm 2 vascular graft with PET piece impregnated with N-butyl cyanoacrylate-based adhesive was placed and MRSA strain with a density of 1 ml 0.5 MacFarland was injected. All rats were scarified in 96th hour, culture samples were taken where intervention was performed and were evaluated microbiologically. Bacteria reproducing in each group were numerically evaluated based on colony-forming unit (CFU/ml) and compared by taking their average. Results: MRSA reproduction of 0 CFU/ml in group 1, of 1410 CFU/ml in group 2, of 180 200 CFU/ml in group 3 and of 625 300 CFU/ml in group 4 was present. A statistically significant difference was present between group 1 and group 4 (p < 0.01), between group 2 and group 4 (p < 0.01), between group 3 and group 4 (p < 0.05). In terms of reproduction, no statistically significant difference was found in group 1, group 2, group 3 in themselves. Conclusions: We observed that the rate of infection increased in the cyanoacyrylate group where cyanoacrylate was used. We think that surgeon should be more careful in using CA in vascular surgery.

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Defensive activity of Hesperidin against Ciprofloxacin induced hepatic injury in rabbits.

International Journal of Pharmaceutical Quality Assurance ◽

10.25258/ijpqa.10.3.29 ◽

2019 ◽

Vol 10 (03) ◽

Author(s):

Hawraa M. Murad ◽

Tamadhur Hani Hussein ◽

Audai Sulaiman Khudhair ◽

Manal Muhi Murad ◽

Jawad Kadhim Faris

Keyword(s):

Body Weight ◽

Bacterial Infections ◽

Hepatoprotective Activity ◽

The Body ◽

Local Breed ◽

Group 4 ◽

Group 3 ◽

Group 2 ◽

Defensive Activity ◽

Antioxidant Effects

This study was conducted to find out hepatoprotective activity of hesperidin (HES) 100mg/kg body weight (b.w.) against ciprofloxacin (CPX) 100 mg/kg induced hepatotoxicity in local breed rabbits .CPX is a broad spectrum antibiotic used for treatment of many bacterial infections. Twenty four male rabbits were divided into four groups ,group1: control, (1 ml/kg Saline orally) group 2: CPX (100 mg/kg orally) for (14) consecutive days , group 3: HES (100 mg//kg) orally for (14) consecutive days group 4: CPX (100 mg/kg orally) plus HES (100 mg//kg orally ) for (14) consecutive days. All the rabbits were killed on the (15) day of the experiment, and then the blood, and livers samples were taken. CPX induced hepatotoxicity was proved by a significant (p less than 0.01) reduction in the body weight ,and a significant (p less than 0.01) increased serum aspartate transaminase (AST), alanine transaminase (ALT) , Malonaldehyde enzyme (MAD) and histopathological changes. Protective hepatic toxicity effect and oxidative damage caused by CPX significantly (p less than 0.01) increasing in body weight and significantly (p less than 0.01) decreasing AST , ALT, MAD and improving tissue morphology in HES (100 mg//kg) . These results assure that HES (100 mg//kg) antioxidant effects can protect CPX-induced hepatotoxicity in rabbits.

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