Absence of selective brain cooling in unrestrained baboons exposed to heat

To test whether baboons are capable of implementing selective brain cooling, we measured, every 5 min, the temperature in their hypothalamus, carotid arterial bloodstream, and abdominal cavity. The baboons were unrestrained and exposed to 22 degrees C for 7 days and then to a cyclic environment with 15 degrees C at night and 35 degrees C during the day for a further 7 days. During the latter 7 days some of the baboons also were exposed to radiant heat during the day. For three days, during heat exposure, water was withheld. At no time was the hypothalamus cooler than carotid arterial blood, despite brain temperatures above 40 degrees C. With little variation, the hypothalamus was consistently 0.5 degrees C warmer than arterial blood. At high body temperatures, the hypothalamus was sometimes cooler than the abdomen. Abdominal temperature was more variable than arterial blood and tended to exceed arterial blood temperature at higher body temperatures. Hypothalamic temperature cooler than a warm abdomen is not evidence for selective brain cooling. In species that can implement selective brain cooling, the brain is most likely to be cooler than carotid arterial blood when an animal is hyperthermic, during heat exposure, and also dehydrated and undisturbed by human presence. When we exposed baboons to high ambient temperatures while they were water deprived and undisturbed, they never implemented selective brain cooling. We conclude that baboons cannot implement selective brain cooling and can find no convincing evidence that any primate species can do so.     

A technique to study splanchnic metabolism in the unrestrained conscious pig

In recent years similarities recognized between porcine and human anatomy and physiology have made the pig an experimental animal of considerable value in biomedical research. We developed a pig model for unrestrained repeated sampling of portal and arterial blood, and selectively of small and large bowel veins. Catheters were inserted, under halothane anesthesia, in ten female crossbred (Yorkshire x Dutch Landrace) piglets (8 weeks; 20 +/- 2 kg). After recovery for at least 3 days the catheters were used for sampling of blood. Aortic and portal catheters patency rates were 60% at day 24. For the small bowel catheters, a patency rate of 30% was seen at day 24. The large bowel patency rate was 30% at day 10. These results are promising because they allow long-term metabolic splanchnic research in unrestrained piglets.     

Hemodynamics of conscious unrestrained baboons, including cardiac output

A method is described for comprehensive hemodynamic study of undisturbed baboons (Papio hamadryas) that incorporates cardiac output measurement by thermodilution. Instrumentation includes arterial, aortic, and central venous catheterization by a surgical technique that does not require entry to peritoneal or thoracic cavities. It provides a means for right atrial indicator delivery with aortic temperature recording of thermodilution curves. Accuracy was confirmed by comparison to measurement by Swan-Ganz catheters. Diurnal variations of systemic arterial pressure in long-term study of conscious baboons were shown to result from significant increases in cardiac output by day (P less than 0.001), despite concomitant falls in systemic vascular resistance. The cardiac output values obtained were 0.13 l.min-1.kg-1 at night and 0.16 l.min-1.kg-1 by day. Comparison of these results to previous reports of cardiac output in baboons highlights the inadequacies of methods that require physical restraint or anesthesia. This technique also leaves the baboons intact for subsequent breeding or experimental use after catheter removal without the need for further surgery.     

A modified technique for tail cuff pressure measurement in unrestrained conscious rats

In physiological experiments, it is essential to measure arterial pressure (AP) and heart rate (HR) in animals. Tail cuff pressure (TCP) measurement using photoelectric volume oscillometry has been commonly used. We designed a new technique for continuous measurement of AP and HR in conscious, unrestrained rats. This is based on the observation that fixation of the rat's tail with tape keeps the animal in position without struggling. The animal is free to move its body. To test the accuracy of this new technique, Sprague-Dawley rats underwent four AP and HR measurement techniques. These included a new unrestrained method (UR), which was compared to the following three methods: traditional restrained TCP method with restrainer, direct monitoring of AP and HR with femoral artery catheterization and a combination of photoelectric volume oscillometry (with body heating to 37 degrees C) and femoral arterial recording. The results show that the modified UR measurement provides accurate data on AP and HR. This method obtains a lower value of HR and similar mean AP when compared to direct monitoring from femoral arterial catheterization. Accordingly, the modified unrestrained TCP measurement can be used in conscious rats as a noninvasive method.     

Early calcification patterns of the iliac arteries and their relation to the arterial structure

Summary Gross calcifications of the common iliac and internal iliac arteries represent a common finding in newborn children and infants. In both arteries, the calcific deposits regularly appear in certain areas of the arterial luminal surface only, whereas the other parts of the arterial wall remain free of gross lesions even in cases with a pronounced calcification. In the common iliac artery, the lateral wall of the vessel and the adjacent sectors of the anterior and posterior wall represent the predilection site of calcific deposits. In the internal iliac artery, the gross calcifications have been regularly demonstrated in the dorso-medial wall. The predominant localisation of the calcification in these parts of the vessels and its absence in the others depend on the definite structural features of the arterial tube and different affinity for calcium of the individual structural elements. In both iliac arteries, only the primary internal elastic membrane undergoes early calcification. However, unlike the most muscular arteries, this membrane is not developed in the whole arterial circumference of the common iliac and internal iliac arteries, but is absent in large areas of their arterial luminal layer. In these areas, the subendothelial or subintimal elastic layers are formed by the networks of longitudinally arranged elastic fibers or membraneous elastic structures which arise from the elastic networks with the further growth. These elastic elements always stay free of calcific deposits. The structural features found in both iliac arteries may be important for the development of the later pathological changes.     

Pasteurella multocida infections in baboons (Papio cynocephalus)

The respiratory pathogenPasteurella multocida was isolated from infections of the laryngeal air sacs of two baboons and from abscesses in the neck or femoral area of two other baboons. The infections were associated with surgical procedures involving the cervical area, chronic catheterization, and chair restraint. The organism was also detected among the commensal pharyngeal flora in 2 of 15 clinically healthy, wildborn adult baboons. These findings suggest that the organism is harbored naturally in baboons and that exudative infections can occur secondary to specific procedures.     

Functional small-diameter neovessels created using endothelial progenitor cells expanded ex vivo

Arterial conduits are increasingly preferred for surgical bypass because of inherent functional properties conferred by arterial endothelial cells, especially nitric oxide production in response to physiologic stimuli. Here we tested whether endothelial progenitor cells (EPCs) can replace arterial endothelial cells and promote patency in tissue-engineered small-diameter blood vessels (4 mm). We isolated EPCs from peripheral blood of sheep, expanded them ex vivo and then seeded them on decellularized porcine iliac vessels. EPC-seeded grafts remained patent for 130 days as a carotid interposition graft in sheep, whereas non-seeded grafts occluded within 15 days. The EPC-explanted grafts exhibited contractile activity and nitric-oxide-mediated vascular relaxation that were similar to native carotid arteries. These results indicate that EPCs can function similarly to arterial endothelial cells and thereby confer longer vascular-graft survival. Due to their unique properties, EPCs might have other general applications for tissue-engineered structures and in treating vascular diseases.     

Nitric oxide from endothelium and smooth muscle modulates responses to sympathetic nerve stimulation: implications for endotoxin shock.

The influence of nitric oxide (NO) on vascular responses to transmural stimulation (TNS) of noradrenergic nerves was studied in isolated rings of rat iliac arteries. TNS produced frequency-dependent contractions in all vessels. The NO synthase inhibitor NG-monomethyl-L-arginine (L-NMMA) significantly enhanced TNS responses in intact vessels, but not in those in which the endothelium had been removed. However, in endothelium-denuded rings incubated for 8 hours, L-NMMA increased the contractions induced by nerve stimulation, an effect which was prevented by treatment with dexamethasone or cycloheximide, and enhanced by incubation with lipopolysaccharide and gamma-interferon. Addition of L-arginine reversed the effect of L-NMMA in intact rings; however, it significantly decreased below control values TNS-induced contractions in vessels without endothelium. The results indicate that a) the arterial response to noradrenergic nerve stimulation is modulated by NO originating either in endothelial cells or in smooth muscle cells after induction of NO synthase activity, and b) once NO synthase is induced, the limiting step in NO production is the availability of the substrate L-arginine. An overproduction of vascular NO in the presence of endotoxin or other inflammatory stimuli may prevent the vascular response to sympathetic stimuli and contribute to the vasodilation observed in inflammation or endotoxic shock.     

Effect of alterations in femoral artery flow on abdominal vessel hemodynamics in swine

In support of an in vivo investigation in swine of the influence of changes in fluid dynamic wall shear on arterial macromolecular permeability, a procedure has been developed to alter the flows in the porcine posterior arterial vasculature by opening and closing a reversible arteriovenous shunt placed on one of the femoral arteries. Laparoscopic techniques were used to place appropriately modified Transonic Systems ultrasonic flow probes on both external and circumflex iliac arteries, and on the terminal aorta. Flow measurements were made prior to shunt placement, and with the shunt open and closed, to measure the influence of altered external iliac artery flow on the distribution to the infrarenal abdominal vessels. Similar experiments were carried out to relate the flow rates in the external iliac arteries to those in the femoral arteries, which are more accessible. Based on the relationships among the measured flow rates, rules have been developed to estimate the major infrarenal flows in the pig, at baseline and with the shunt opened and closed, from only the flow rates measured at the two femoral arteries.     

Hypercholesterolemia and the baboon cerebral circulation

We investigated cerebrovascular vasodilator responses to increased arterial CO2 and the cerebrovascular response to infused 5-hydroxytryptamine (5-HT) in normal and hypercholesterolemic baboons. After 6-8 weeks of feeding an atherogenic diet the plasma cholesterol levels were increased without change in the triglycerides. The hypercholesterolemic animals showed a higher basal systemic arterial blood pressure than the normal controls without significant decrease in cerebrovascular prostacyclin production, altered basal cerebral blood flow or altered cerebrovascular response to infused 5-HT. However, the vasodilator response to hypercapnia was significantly decreased from the control value of 2.78 ml/min per mmHg increase in PCO2, to 1.62 ml/min per mmHg. Thus functional impairment of cerebral hemodynamics occurred before atherosclerotic alteration in the cerebral vessels could have been present.     

Sequential vascularized iliac bone graft and a superficial circumflex iliac artery perforator flap with a single source vessel for established mandibular defects

The major problems in dealing with established mandibular loss are severe soft-tissue contracture and a limited number of recipient vessels. The skin portion of the iliac osteocutaneous flap often necrotizes in cases without perforators of the deep circumflex iliac vessel. To overcome these problems, eight patients with established mandibular loss and no skin perforators of the deep circumflex iliac vessel were treated with a sequential vascularized iliac bone graft and a superficial circumflex iliac perforator flap with a single recipient vessel. Regarding the recipient vessels, the ipsilateral cervical vessels were used for four patients, and the contralateral facial and ipsilateral superficial temporal vessels were used for two cases each. The superficial circumflex iliac perforator flaps were 7 to 28 cm in length and 3 to 15 cm in width. The iliac bone grafts ranged from 7 to 13 cm in length, and three cases were repaired with the inner cortex of the iliac bone. There were no serious complications, such as flap necrosis or bone infection and resulting absorption. The advantages of this method are that both pedicles are very close to each other and of suitable diameter for anastomosis. Simultaneous flap elevation and preparation for the recipient site is possible. The skin flap and vascularized bone graft can be obtained from the same donor site. A single source vessel can nourish both the large skin area and bone sequentially. Longer dissection of the superficial circumflex iliac system to the proximal femoral division is unnecessary. A large flap can survive with a short segment of the superficial circumflex iliac system. Only the vascularized inner cortex of the iliac bone needs to be used, and the outer iliac cortex can be preserved, which results in less morbidity at the donor site.     

Responses of baboons to traditionally pyrogenic agents

It is not clear whether baboons develop fever in response to endotoxin or other pyrogens. We injected various pyrogens intravenously in 12 unrestrained baboons (Papio ursinus) and measured their body temperature using intra-abdominal radiotelemeters. Serum iron concentration was also measured. The baboons developed fever after injection of killed Staphylococcus aureus (5 X 10(7) organisms/kg). No significant fever was measured after injection of lipopolysaccharide (Salmonella typhosa) (0.1, 8, 40, and 100 micrograms/kg), bovine serum albumin (4 mg/kg), killed Salmonella minnesota (5 X 10(7) organisms/kg), and killed Salmonella typhi (5 X 10(7) organisms/kg). A significant decrease in serum iron concentration was found only after injection of S. aureus and lipopolysaccharide, 100 micrograms/kg. The phagocytic synthesis of interleukin-1 following pyrogen stimulation in baboons and some other primates appears to differ from that in man and in nonprimates.     

Numerical simulation of aortic bifurcation flows: The effect of flow divider curvature

Two dimensional steady flow calculations in computational regions obtained from radiographs of human aortic bifurcations correlate well with unsteady measurements of wall shear in flow-through casts of the same vessels. The results suggest that wall slope may be an important factor affecting the variability of shear along the medial walls of this arterial segment. If extremes of shear stress promote atherogenesis, then variations in the curvature of the proximal iliac arteries may affect the susceptibility of these vessels to vascular disease on their medial aspect.     

Iliac biopsy for histomorphometric analysis of trabecular bone in cynomolgus monkeys and baboons

A humane, repeatable surgical technique was developed to harvest trabecular bone for histomorphometry from nonhuman primates. Using a direct surgical approach to the iliac crest, bone specimens with a mean cross sectional area of trabecular bone plus marrow (AT) of 12.02 and 18.07 mm2 were harvested from cynomolgus monkeys (Macaca fascicularis) and baboons (Papio anubis) respectively. The method provides bone samples with little artifact or bone dust contamination, and easy orientation and reproducibility of the plane of section. None of the cynomolgus monkeys were affected adversely by the surgery, but the baboons were quieter than normal for 12-24 hours afterward. The technique offers excellent biopsy specimens, repeatability and a minimum of stress to the subjects.     

New methodology for measuring blood pressure in awake baboons with use of behavioral training techniques

Adult male baboons were behaviorally conditioned to extend an arm outside of the living cage and to accept repeated cuff inflations for manual auscultatory blood pressure measurements. Frequency distributions of systolic and diastolic blood pressure for both normotensive and renovascular hypertensive baboons generally were normally distributed. The procedure accurately tracked rapid changes in blood pressure after oral administration of antihypertensive drugs. Advantages over direct arterial cannulation for blood pressure measurement during extended, chronic experiments are discussed.     

儿童重症监护病房医院感染暴发白色念珠菌血流感染4例分析

摘要 目的：探讨儿童重症监护病房白色念珠菌血流感染暴发的临床表现、危险因素、控制措施等,为预防和控制院内白色念珠菌血流感染暴发提供科学依据。方法：以2018年7月我院儿童重症监护病房发生的4例白色念珠菌血流感染暴发患儿为研究对象,分析患儿临床情况、临床特征、危险因素、暴发原因以及采取的预防控制措施。结果：4例医院感染暴发白色念珠菌血流感染患儿均存在基础疾病、有机械通气史、存在中心静脉或动脉置管、静脉或动脉置管前后均使用碘伏消毒、曾使用广谱抗生素、输血制品,白色念珠菌血流感染后最突出的临床表现均是发热。药敏方面,医院感染暴发的4例白色念珠菌感染患儿对唑类及5-氟胞嘧啶均耐药,但对两性霉素B均敏感。经拔除血管置管、减少或者避免广谱抗菌药的应用,根据药敏使用卡泊芬净及两性霉素B抗真菌等积极治疗,1例患儿放弃治疗后死亡,3例患儿顺利出院。通过Fisher确切概率法分析可知,留置中心静脉或动脉置管是儿童重症监护病房发生医院感染暴发白色念珠菌血流感染的危险因素（P<0.05）。结论：留置中心静脉或动脉置管是儿童重症监护病房发生医院感染暴发白色念珠菌血流感染的危险因素,医院感染暴发白色念珠菌血流感染患儿最突出的临床表现是发热,唑类及5-氟胞嘧啶耐药的患儿使用卡泊芬净及两性霉素B可能获得较好的治疗效果。     

The role of the pericytes of the adventitial microcirculation in the arterial intimal thickening

Segments of rat femoral arteries, with one collateral each, occluded between ligatures and dissected from surrounding tissue, developed intimal thickening, with or without ligation of their collaterals. Numerous newly-formed capillaries from the surrounding arterial microcirculation growing into the adventitia, tunica media and intimal thickening were demonstrated by means of serial longitudinal sections, predominantly in the ostium of the collateral. When the ligatures were applied without damaging the microcirculation surrounding the artery and the normal continuity of the adventitial vessels was unchanged, earlier presence of intimal thickening was observed. When the fibrous layers of the adventitia were removed at the moment of the arterial ligation, the continuity between newly-formed vessels of the neoadventitia and those growing into the media and neointima was much more evident. It was then noted that the pericytes constituted a major component of the intimal thickening. The introduction of contrast material in microcirculation confirmed the connections between newly-formed adventitial and intimal vessels. At the beginning of the experiment, autoradiographic studies showed an increased DNA synthesis in the cells of preformed postcapillary venules and capillaries of surrounding arterial microcirculation and later in those of the newly-formed vessels growing into the arterial wall. These results indicate that newly-formed capillaries derived from surrounding arterial microcirculation penetrate the wall of the occluded arterial segments and contribute to the intimal thickening formation. It is likely that the pericytes and endothelial cells (EC) of these ingrowing vessels are sources of myointimal cells at the intimal thickening and of endothelium at the luminal surface, respectively.     

Differential constrictor responses of cephalic and caudal vasculature to α-adrenoceptor agonist after hind limb unloading

We examined the effects of hind limb unloading (HLU, 14 days) on constriction of carotid and iliac arterial beds in vivo in thiobutabarbital-anaesthetized rats and isolated carotid and iliac arteries in vitro. Both control and HLU rats had similar arterial pressure and carotid and iliac arterial flows. The HLU rats had increased carotid arterial but reduced iliac arterial constriction in response to methoxamine (α1-adrenoceptor agonist) in vivo. In contrast, constriction in response to methoxamine was reduced in the isolated carotid and unchanged in the iliac artery of HLU rats relative to control rats. Thus, HLU is associated with increased constriction of carotid arterial bed but reduced constriction of the isolated carotid artery, and reduced constriction of iliac arterial bed but unchanged constriction of the isolated iliac artery. These results show differential influence of HLU on constriction of cephalic and caudal arterial beds, and differential effect on constrictions of arterial beds relative to conduit arteries.     

Superiority of the deep circumflex iliac vessels as the supply for free groin flaps. Clinical work.

In this paper we present 16 cases of free transfer of compound flaps from the groin, 11 using the deep circumflex iliac vessels as a stem and 5 using the superficial circumflex iliac vessels as a stem. We found the deep vessels superior in many ways to the superficial vessels for this purpose; they are larger, permitting greater ease in anastomoses and providing more reliable blood flow. We believe that larger osteocutaneous or myocutaneous flaps can be transferred on the deep vessels than on the superficial ones-and that the deep circumflex iliac flap may supplant the conventional free going flap in most situations. The method evolved in response to patient need, not for surgical éclat.     

Restoring the endothelium of cryopreserved arterial grafts: co-culture of venous and arterial endothelial cells

The use of arterial homografts in clinical practice is becoming increasingly common, yet there is an urgent need to address one of the most well-established problems associated with their use: the loss of integrity of the endothelium following cryopreservation. The partial lack of endothelium causes contact between the extracellular matrix and blood flow, which, in turn, often gives rise to thrombosis and/or restenosis. Our objective was first to attempt to replace the arterial endothelial cells lost during the cryopreservation process by seeding autologous venous endothelial cells, and to evaluate the behaviour of venous and arterial endothelial cells in co-culture. The idea was to establish whether venous endothelial cells would be accepted by arterial endothelial cells and could therefore be used to restore the endothelial lining for the subsequent use of these vessels in in vivo grafting procedures. For the co-culture experiments, endothelial cells were obtained from the jugular vein and both iliac arteries of the minipig by treatment with 0.1% type I collagenase. The venous endothelial cells were fluorescently labelled with the membrane intercalating dye PKH26. Equal numbers of venous and arterial endothelial cells were mixed and co-cultured for 24h, 48h or 4 days. Cell viability, determined by 2% trypan blue staining and the TUNEL method, was established before and after fluorescence labelling. Cellular activity was determined by estimating PGI2 levels in the cultures. The proliferation index was established by [H(3)]thymidine (1muCi/ml) in the cell culture medium. For the in vivo tests, 5 cm length segments of minipig iliac artery were used to establish the groups: control (n = 6), fresh arterial segments; group I (n = 16), cryopreserved arterial segments and group II (n = 16), cryopreserved arterial segments seeded with autologous venous endothelial cells. The cryopreserved vessels in group II were seeded by flooding with a labelled venous endothelial cell suspension. Once seeded, the arterial segments were included in an in vitro flow circuit. All the specimens were processed for fluorescence and light microscopy, and scanning electron microscopy. The denuded endothelial surface was determined in each group. Cell death was evaluated by the TUNEL method. We confirmed the existence of intercellular PECAM1-type junctions between venous (PKH26+) and arterial cells in co-culture and the functional activity of the cells. The cryopreserved arterial segments showed a well-preserved wall structure. However, different size areas of marked endothelial denudation were detected. After seeding with labelled cells (PKH26+), these denuded areas of the cryopreserved artery were entirely covered by fluorescent cells. After seeding, a drop in the proportion of damaged endothelial cells was recorded. Despite some loss of seeded cells after inclusion in the in vitro flow circuit, the endothelial cell count was not significantly different to those recorded for control, non-cryopreserved specimens. In conclusion arterial and venous endothelial cells growing in co-culture modify their behaviour to form multilayers. The two cell populations form normal PECAM1 junctions and preserve their functional properties. Seeding autologous venous endothelial cells on the luminal surface of cryopreserved arterial segments serves to restore the integrity of the endothelial layer.