[Preliminary evaluation of immunogenicity and protective effect of multicomponent recombinant protein vaccine EPRHP014 against tuberculosis].

Objective: To evaluate the immunogenicity and protective effect of a multicomponent recombinant protein vaccine EPRHP014 constructed independently and provide a scientific basis for developing new tuberculosis (TB) vaccine and effective prevention and control of TB. Methods: Three full-length Mycobacterium (M.) tuberculosis protein antigens (EsxH, Rv2628, and HspX) and two epitope-predicted and optimized epitope-dominant protein antigens (nPPE18 and nPstS1) were selected, from which five protein antigens were used to construct a protein antigen composition EPRHP014, including a fusion expression multi-component protein antigen (EPRHP014f) and a multi-component mixed protein antigen (EPRHP014m) formed with the five single protein using clone, purification, and purification respectively. Multicomponent protein vaccines EPRHP014f and EPRHP014m were prepared with aluminum adjuvant, and the BCG vaccine was used as a control. ELISA detected the titer of serum-specific antibodies, the secretion of various cytokines was detected by ELISpot and Luminex, and immune protection was observed by the M. tuberculosis growth inhibition test in vitro. The results were statistically analyzed by t-test or rank sum test, and P<0.05 was considered a statistically significant difference. Results: Mice Immunized with EPRHP014m and EPRHP014f could produce highly effective IgG antibodies and their subtypes IgG1 and IgG2a, and the antibody titers were similar to those of mice immunized with BCG, with no statistical significance (P>0.05). The number of spot-forming cells (SFC) secreting IFN-γ and IL-4 induced by EPRHP014f group was significantly higher than those by EPRHP014m group and BCG group (P<0.05), but there was no significant difference in the number of SFC for IFN-γ and IL-4 induced between EPRHP014m group and BCG group (P>0.05). The secretion levels of GM-CSF and IL-12p70 induced by the EPRHP014m group were higher than those of the BCG group (P<0.05), but there was no significant difference in the levels of IL-6 and IL-10 induced between EPRHP014m group and BCG group (P>0.05). There was no significant difference in the secretions of IL-6, IL-10, IL-12, and GM-CSF between the EPRHP014f and BCG groups (P>0.05). EPRHP014m group, EPRHP014f group, and BCG group had obvious antibacterial effects in vitro, and the difference was insignificant (P>0.05). Conclusion: Both EPRHP014f and EPRHP014m can induce strong humoral and cellular immune responses in mice after immunization, and have a strong ability to inhibit the growth of M. tuberculosis in vitro, indicating that the antigen composition EPRHP014 has good potential in the development and application of TB vaccine.

[Analysis on drug sensitivity spectrum of 167 multidrug-resistant Mycobacterium tuberculosis in China].

Objective: To investigate the drugs-sensitivity spectrum of multidrug-resistant tuberculosis (MDR-TB) in China and provide a scientific evidence for the drug selection in clinical therapy and the control of MDR-TB. Methods: A total of 167 strains of MDR-TB were included in this study. Every strain was genotyped by lysX gene sequencing and their sensitivity to 13 different anti-TB drugs was tested by using MicroDST(TM) and BACTEC(TM) MGIT 960(TM) liquid-culturing method. The association between drug resistance and genotypes as well as cross drug resistance was also analyzed. The results were analyzed by means of the comparison of enumeration data between two groups with χ(2) test. Results: The overall resistance rate of 167 MDR-TB strains to 11 anti-TB drugs, except isoniazide and rifampicin, was 95.81%, the rates of pre-extensive drug-resistance (pre-XDR) and extensive drug-resistance were 31.14%(52/167) and 6.59% (11/167), respectively. The streptomycin resistance rate of Beijing genotypes was significantly higher than that of the non-Beijing genotypes ( χ(2)=30.682, P<0.05), while the pre-XDR proportion in Beijing genotypes was lower than that in non-Beijing genotypes (χ(2)=5.332, P<0.05). The resistance rates of Ofloxacin and Pyrazinamide in the modern Beijing genotype were significantly higher than those in classical ones (χ(2)=4.105 and χ(2)=3.912, P<0.05). In addition, the cross-resistance rate to rifampicin and rifabutin was 86.23%. A significant difference in drug-resistance rate to rifabutin was seen among groups with different levels of rifampicin resistance (χ(2)=45.912, P<0.05). There was positive correlation not only between ofloxac resistance and moxifloxac resistance, but also between amikacin resistance and kanamycin resistance, with the coefficient of 0.87 and 0.91, respectively. Conclusions: In this study, we observed that there were high incidences of the resistance to 11 anti-TB drugs in 167 clinical MDR-TB strains and the cross resistance phenomena between drugs of the same type were quite serious. The majority of MDR-TB strains belonged to Beijing genotype, which was highly associated with streptomycin resistance.

The Role of AMPARs in the Maturation and Integration of Caudal Ganglionic Eminence-Derived Interneurons into Developing Hippocampal Microcircuits.

In the hippocampal CA1, caudal ganglionic eminence (CGE)-derived interneurons are recruited by activation of glutamatergic synapses comprising GluA2-containing calcium-impermeable AMPARs and exert inhibitory regulation of the local microcircuit. However, the role played by AMPARs in maturation of the developing circuit is unknown. We demonstrate that elimination of the GluA2 subunit (GluA2 KO) of AMPARs in CGE-derived interneurons, reduces spontaneous EPSC frequency coupled to a reduction in dendritic glutamatergic synapse density. Removal of GluA1&2&3 subunits (GluA1-3 KO) in CGE-derived interneurons, almost completely eliminated sEPSCs without further reducing synapse density, but increased dendritic branching. Moreover, in GluA1-3 KOs, the number of interneurons invading the hippocampus increased in the early postnatal period but converged with WT numbers later due to increased apoptosis. However, the CCK-containing subgroup increased in number, whereas the VIP-containing subgroup decreased. Both feedforward and feedback inhibitory input onto pyramidal neurons was decreased in GluA1-3 KO. These combined anatomical, synaptic and circuit alterations, were accompanied with a wide range of behavioural abnormalities in GluA1-3 KO mice compared to GluA2 KO and WT. Thus, AMPAR subunits differentially contribute to numerous aspects of the development and maturation of CGE-derived interneurons and hippocampal circuitry that are essential for normal behaviour.

Afferent specific role of NMDA receptors for the circuit integration of hippocampal neurogliaform cells.

Appropriate integration of GABAergic interneurons into nascent cortical circuits is critical for ensuring normal information processing within the brain. Network and cognitive deficits associated with neurological disorders, such as schizophrenia, that result from NMDA receptor-hypofunction have been mainly attributed to dysfunction of parvalbumin-expressing interneurons that paradoxically express low levels of synaptic NMDA receptors. Here, we reveal that throughout postnatal development, thalamic, and entorhinal cortical inputs onto hippocampal neurogliaform cells are characterized by a large NMDA receptor-mediated component. This NMDA receptor-signaling is prerequisite for developmental programs ultimately responsible for the appropriate long-range AMPAR-mediated recruitment of neurogliaform cells. In contrast, AMPAR-mediated input at local Schaffer-collateral synapses on neurogliaform cells remains normal following NMDA receptor-ablation. These afferent specific deficits potentially impact neurogliaform cell mediated inhibition within the hippocampus and our findings reveal circuit loci implicating this relatively understudied interneuron subtype in the etiology of neurodevelopmental disorders characterized by NMDA receptor-hypofunction.Proper brain function depends on the correct assembly of excitatory and inhibitory neurons into neural circuits. Here the authors show that during early postnatal development in mice, NMDAR signaling via activity of long-range synaptic inputs onto neurogliaform cells is required for their appropriate integration into the hippocampal circuitry.

The Association between Distal Findings and Proximal Colorectal Neoplasia: A Systematic Review and Meta-Analysis.

OBJECTIVES: Whether screening participants with distal hyperplastic polyps (HPs) detected by flexible sigmoidoscopy (FS) should be followed by subsequent colonoscopy is controversial. We evaluated the association between distal HPs and proximal neoplasia (PN)/advanced proximal neoplasia (APN) in asymptomatic, average-risk patients. METHODS: We searched Ovid Medline, EMBASE, and the Cochrane Library from inception to 30 June 2016 and included all screening studies that examined the relationship between different distal findings and PN/APN. Data were independently extracted by two reviewers with disagreements resolved by a third reviewer. We pooled absolute risks and odds ratios (ORs) with a random effects meta-analysis. Seven subgroup analyses were performed according to study characteristics. Heterogeneity was characterized with the I2 statistics. RESULTS: We analyzed 28 studies (104,961 subjects). When compared with normal distal findings, distal HP was not associated with PN (OR=1.16, 95% confidence interval (CI)=0.89-1.51, P=0.14, I2=40%) or APN (OR=1.09, 95% CI=0.87-1.36, P=0.39, I2=5%), while subjects with distal non-advanced or advanced adenoma had higher odds of PN/APN. Higher odds of PN/APN were observed for more severe distal lesions. Weaker association between distal and proximal findings was noticed in studies with higher quality, larger sample size, population-based design, and more stringent endoscopy quality-control measures. The Egger's regression tests showed all P>0.05. CONCLUSIONS: Distal HP is not associated with PN/APN in asymptomatic screening population when compared with normal distal findings. Hence, the presence of distal HP alone detected by FS does not automatically indicate colonoscopy referral for all screening participants, as other risk factors of PN/APN should be considered.

Subluminous phase velocity of a focused laser beam and vacuum laser acceleration.

It has been found that for a focused laser beam propagating in free space, there exists, surrounding the laser beam axis, a subluminous wave phase velocity region. Relativistic electrons injected into this region can be trapped in the acceleration phase and remain in phase with the laser field for sufficiently long times, thereby receiving considerable energy from the field. Optics placed near the laser focus are not necessary, thus allowing high intensities and large energy gains. Important features of this process are examined via test particle simulations. The resulting energy gains are in agreement with theoretical estimates based on acceleration by the axial laser field.

Lactated Ringer's solution alleviates brain trauma-precipitated lactic acidosis in hemorrhagic shock.

To determine the influence of brain trauma on blood acid-base and lactate-pyruvate responses to hemorrhage, and the effect of lactated Ringer's solution on these responses, 30 anesthetized rats were assigned to four groups: hemorrhage (n = 7), hemorrhage following fluid percussion brain trauma (trauma-hemorrhage group) (n = 7), hemorrhage treated with lactated Ringer's solution (hemorrhage-resuscitation group) (n = 8), and hemorrhage following brain trauma treated with lactated Ringer's solution (trauma-hemorrhage-resuscitation group) (n = 8). The hemorrhage group showed no significant changes in pH, HCO3, and base excess after hemorrhage. Base excess and pH were significantly reduced after the hemorrhage in the trauma-hemorrhage group but were raised after resuscitation in the hemorrhage-resuscitation group. Acid-base values showed no difference between the trauma-hemorrhage-resuscitation and hemorrhage groups. The trauma-hemorrhage-resuscitation group also had a significantly higher base excess than the trauma-hemorrhage group. Lactate rose significantly after hemorrhage in the hemorrhage group and was even higher in the trauma-hemorrhage group, but there were no differences between the hemorrhage versus hemorrhage-resuscitation or trauma-hemorrhage-resuscitation groups. Both brain trauma and lactated Ringer's solution increased pyruvate with marked reduction in the ratio of lactate to pyruvate. These data indicate that brain trauma precipitates blood lactate accumulation and metabolic acidosis after hemorrhage, and infusion of lactated Ringer's solution can relieve these disturbances.

Traumatic brain injury attenuates the effectiveness of lactated Ringer's solution resuscitation of hemorrhagic shock in rats.

Traumatic brain injury suppresses spontaneous cardiovascular compensation for hemorrhage, prompting us to examine the possibility that trauma to the brain modifies hemodynamic response to therapy in hemorrhage. Thirty rats that were anesthetized were randomly assigned to four groups--hemorrhagic shock (H), hemorrhagic shock after brain trauma (TH), hemorrhagic shock treated with lactated Ringer's (LR) solution (HR), and hemorrhagic shock after brain trauma treated with LR (THR). After hemorrhage, group TH had significantly lower mean arterial pressure (MAP), cardiac index (CI) and stroke volume index (SVI) than group H. Throughout the postresuscitative period, group HR had significantly higher MAP, CI, SVI and central venous pressure than group H. At 50 and 70 minutes after the start of hemorrhage, group THR showed significantly lower MAP, CI and SVI than group HR. This difference in hemodynamics is not because of transcapillary refill effect, because brain trauma did not cause changes in hematocrit and plasma protein levels. As heart rate, preload and afterload were not significantly different between groups THR and HR, the attenuation of fluid resuscitation can be attributed mainly to a depressed cardiac function. Furthermore, neither brain trauma nor fluid replacement altered the content of water in the brain in hemorrhaged rats. These data indicate that brain trauma not only suppresses spontaneous hemodynamic recovery from hemorrhage, but also impedes the efficacy of LR resuscitation. The results of the current study suggested that a more aggressive fluid replacement may be needed to treat hemorrhagic shock in individuals with brain injury.

Influences of traumatic brain injury on the outcomes of delayed and repeated hemorrhages.

Effects of traumatic brain injury (fluid percussion) on outcomes of hemorrhage, either delayed (70.5 min after the injury) or repeated (0.5 min after injury, then followed by a delayed hemorrhage), were examined in 4 groups of 10 anesthetized rats. Comparisons were made for delayed hemorrhage following sham injury (A1) vs. injury (A2) and for repeated hemorrhage following sham injury (B1) vs. injury (B2). No significant differences were observed in MAP, Cl, HR, SVI, SVRI, and CVP between groups A1 and A2. Hemodynamic recovery was significantly better in B1 than B2 at 70 min after the initial hemorrhage. The respective values of MAP and Cl for the groups B1 and B2 at 70 min were 65 +/- 3 vs. 56 +/- 4 mmHg and 220 +/- 15 vs. 182 +/- 15 ml/min/kg. Brain trauma did not affect survival rate (90 vs. 100%) following delayed hemorrhage, but significantly worsened the outcome of repeated hemorrhage. The 130 min survival rates for groups B1 and B2 were 50% and 10% (P = 0.08), respectively; their survival curves were significantly different (P = 0.02). Our data indicate that brain trauma has greater impact on responses to immediate hemorrhage than delayed hemorrhage, suggesting that traumatic brain injury may have a time-dependent effect on the response to hemorrhage.

Suppression by traumatic brain injury of spontaneous hemodynamic recovery from hemorrhagic shock in rats.

The effects of brain trauma on cardiovascular and endocrine responses to hemorrhage were investigated. Forty anesthetized rats were randomly assigned to one of four groups of 10 rats each: a control group (Group C): a group with induction of hemorrhage at 16.2 ml/kg/10 min (Group H); a group with fluid-percussion brain injury at a peak pressure of 1.7 atm and an impulse duration of 25 msec (Group T); and a group receiving hemorrhagic shock following brain trauma (Group TH). Group C and T rats showed no significant alterations in cardiovascular function. At the end of hemorrhage there were no significant differences between Groups TH and H in the nadirs of mean arterial blood pressure (MABP) (mean values +/- standard error of the mean: 42 +/- 2 vs. 40 +/- 4 mm Hg) and stroke volume index (SVI) (0.61 +/- 0.11 vs. 0.66 +/- 0.10 ml/bt/kg); however, 1 hour post-hemorrhage recovery was blunted in Group TH compared to Group H (MABP 56 +/- 4 vs. 65 +/- 3 mm Hg; cardiac index 182 +/- 15 vs. 220 +/- 15 ml/min/kg; and SVI 0.71 +/- 0.06 vs. 0.81 +/- 0.06 ml/bt/kg). Since the two groups showed no significant differences in heart rate, preload (central venous pressure), and afterload (systemic vascular resistance), the reduced cardiac index recovery in Group TH is believed due to the attenuation of cardiac contractile performance. The Group TH preparation potentiated hormonal responses to hemorrhage with significantly higher epinephrine and aldosterone levels than in Group H. Brain trauma enhanced the norepinephrine response to hemorrhage, even at an injury level that by itself did not result in an increase in this hormone. Group TH rats also had significantly lower blood pH and HCO3 levels. The data suggest that brain trauma suppresses MABP and cardiac index recovery after hemorrhage mainly by inhibiting cardiac contractile performance, probably due to high catecholamine levels and severe metabolic acidosis.

Immediate hypertensive response to fluid percussion brain injury may be related to intracerebral hemorrhage and hypothalamic damage.

Fluid percussion brain injury is associated with an immediate rise in mean arterial pressure (MAP). However, the cerebral morphologic basis for this response is still not clear. Thirty-four anesthetized rats were injured using a lateral craniotomy preparation. In 19 rats, impact level was set at 1.73 +/- 0.04 atm, and impact duration was kept at 25 msec to examine the relationship between postinjury hypertensive response and cerebral lesions. MAP was monitored for 1 hour after impact. Fluid percussion produced an increase in MAP from 99 +/- 3 to 134 +/- 4 mm Hg (p less than 0.001), with an increment range of -2 to 87 mm Hg (36 +/- 5 mm Hg) or 0 to 96% increase. The MAP peak occurred at 15 +/- 2 seconds and then rapidly returned to the preimpact level. Histopathological findings, principally hemorrhage, were graded and ranked from 1 to 19 according to relative severity and hypothalamic involvement. There was a significant correlation between MAP rise and the injury ranking (r = 0.52, p = 0.02). No appreciable damage was observed in the brainstem caudal to the diencephalon. Fifteen rats were subjected to higher injury levels. The overall impact magnitude ranged from 1.3 to 3.5 atm. A linear relationship was found between impact magnitude (X, atm) and increment in MAP (Y, mm Hg) (Y = 28.1*X - 14.0, r = 0.62, p less than 0.001). Our study indicates that the immediate postinjury hypertensive response is closely correlated with the impact magnitude and may be related to intracerebral hemorrhage and hypothalamic damage but not necessarily to caudal brainstem damage.

Neuroendocrine abnormalities in patients with traumatic brain injury.

This article provides an overview of hypothalamic and pituitary alterations in brain trauma, including the incidence of hypothalamic-pituitary damage, injury mechanisms, features of the hypothalamic-pituitary defects, and major hypothalamic-pituitary disturbances in brain trauma. While hypothalamic-pituitary lesions have been commonly described at postmortem examination, only a limited number of clinical cases of traumatic hypothalamic-pituitary dysfunction have been reported, probably because head injury of sufficient severity to cause hypothalamic and pituitary damage usually leads to early death. With the improvement in rescue measures, an increasing number of severely head-injured patients with hypothalamic-pituitary dysfunction will survive to be seen by clinicians. Patterns of endocrine abnormalities following brain trauma vary depending on whether the injury site is in the hypothalamus, the anterior or posterior pituitary, or the upper or lower portion of the pituitary stalk. Injury predominantly to the hypothalamus can produce dissociated ACTH-cortisol levels with no response to insulin-induced hypoglycemia and a limited or failed metopirone test, hypothyroxinemia with a preserved thyroid-stimulating hormone response to thyrotropin-releasing hormone, low gonadotropin levels with a normal response to gonadotropin-releasing hormone, a variable growth hormone (GH) level with a paradoxical rise in GH after glucose loading, hyperprolactinemia, the syndrome of inappropriate ADH secretion (SIADH), temporary or permanent diabetes insipidus (DI), disturbed glucose metabolism, and loss of body temperature control. Severe damage to the lower pituitary stalk or anterior lobe can cause low basal levels of all anterior pituitary hormones and eliminate responses to their releasing factors. Only a few cases showed typical features of hypothalamic or pituitary dysfunction. Most severe injuries are sufficient to damage both structures and produce a mixed endocrine picture. Increased intracranial pressure, which releases vasopressin by altering normal hypothalamic anatomy, may represent a unique type of stress to neuroendocrine systems and may contribute to adrenal secretion by a mechanism that requires intact brainstem function. Endocrine function should be monitored in brain-injured patients with basilar skull fractures and protracted posttraumatic amnesia, and patients with SIADH or DI should be closely monitored for other endocrine abnormalities.

Long-term effects of nimodipine on pial microvasculature and systemic circulation in conscious rats.

The chronic cranial window preparation allows repeated measurements of the same pial vessels in unanesthetized rats for several weeks and correlation with 24-h monitoring of hemodynamic variables. Nimodipine (20 mg) or placebo was given via two subcutaneous pellets. Large arterioles dilated 26 and 16%, at hour 1 and days 6-13, respectively (P less than 0.02). There was an increase in number of small arterioles throughout the whole observation period with the maximal increment of 47% (P less than 0.05) at days 6-13. Maximal vasodilation with 10% CO2 indicated that the increase in number of small arterioles after administering nimodipine was not caused by the opening of previously closed vessels. The total length of small arterioles and venules increased 47 and 23% at days 6-13, respectively (P less than 0.001). These increases seem to be caused by the increases in the numbers of vessels, because the average length of the small vessels did not appear to change. This suggests that nimodipine reduces cerebral vascular resistance by causing cerebral microvessel neovascularization. Our data demonstrate that the administration of nimodipine (20 mg) is potent in dilating pial arterioles in the short-term without affecting systemic arterial pressure, and that its long-term effect results in new vessel growth.

Traumatic brain injury creates biphasic systemic hemodynamic and organ blood flow responses in rats.

Traumatic brain injury affects systemic circulation as well as directly damages the brain. The present study examined the effects of fluid percussion brain injury on systemic hemodynamics and organ arterial blood flow in rats. Rats were prepared for fluid percussion injury under anesthesia. Twenty-four hours later, rats were anesthetized (1.0% halothane in N2O:O2) and prepared for radioactive microsphere measurement of cardiac output and organ blood flow. After baseline blood flow and physiological measurements were established, the rats were injured (2.47 +/- 0.02 atm, n = 17) or not injured (n = 20). Additional blood flow determinations were made at two of the following four time (T) points: 5, 15, 30, and 60 min after the injury or sham injury. Fluid percussion brain injury produced an immediate systemic hypertension followed by a hypotension and low cardiac output. Organ blood flows remained constant or increased for 30 min and then declined. Decreased blood flow was most pronounced in the kidneys and the spleen and was less severe in the liver. The reduced cardiac output was redistributed to favor blood flow through the heart and pancreas. These data suggest that traumatic brain injury creates a hyperdynamic period followed by a hypodynamic state with a heterogeneous hypoperfusion among organs.

Triiodothyronine (T3) antagonizes adverse effects of high circulating reverse-T3 (rT3) during hemorrhagic shock.

To examine whether triiodothyronine (T3) could counteract the lethal effect of exogenous reverse T3 (rT3) in hemorrhagic shock, 21 anesthetized, heparinized mongrel dogs were given 15 micrograms/kg of rT3 IV. Thirty minutes later, the dogs were bled rapidly into a reservoir to achieve and maintain a mean arterial pressure of 40 mm Hg. After 60 minutes at 40 mm Hg (compensated shock), the reservoir line was clamped for 30 minutes (uncompensated shock). The shed blood was then reinfused over 30 minutes, and the dogs were monitored for an additional 60 minutes. At the start of uncompensated shock, 11 dogs were given at least 15 micrograms/kg of T3 IV, and 10 animals received saline. Before T3 treatment, there were no significant intergroup differences in the measured hemodynamic and blood gas variables. In the untreated group, 8 of 10 dogs (80%) died during uncompensated shock, in comparison to 3 of 11 dogs (27%) that received T3 (P less than 0.01). Long-term survival in the T3 group was 5/11 (45%), significantly higher than that (1/10, 10%) in the untreated group (P less than 0.05). These results, interpreted in relationship to previous studies, suggest that the therapeutic efficacy of T3 in canine hemorrhagic shock may be related to antagonism of adverse effects of endogenous rT3.

Simultaneous, quantitative measurement of local blood flow and glucose utilization in tissue samples in normal and injured feline brain.

Cerebral blood flow (CBF) and local cerebral glucose utilization (LCGU) were measured using radioactive microspheres and [14C]2-deoxyglucose, respectively, in 26 brain regions in control animals (n = 8) and in animals (n = 4) sustaining low-level experimental brain injury. Examination of the initial (resting) CBF measurement in the uninjured cats revealed two subgroups with significantly (p less than 0.01) different CBF levels. In uninjured cats with normal CBF levels (33.4 +/- 1.8 ml/100 g/min) there was a close linear relationship between CBF and LCGU (n = 0.71, p less than 0.01). In contrast, the remainder of the uninjured cats exhibited abnormally high levels of CBF (72.6 +/- 9.9 ml/100 g/min) and the absence of a close relationship between CBF and LCGU (r = 0.27). One hour following low-level (2.0 atm) fluid percussion brain injury, CBF was increased and LCGU was decreased, though not significantly. The relationship between CBF and LCGU remained intact (r = 0.66, p less than 0.01) in most brain regions. However, the relationship between CBF and LCGU in the hippocampus differed significantly from the relationship between the two parameters in the rest of the brain. Thus, the use of the radioactive microsphere method for CBF measurements allows multiple measurements of CBF and permits the assessment of the status of the cerebral vasculature prior to experimental manipulations such as traumatic brain injury. In view of our current findings of an abnormal relationship between CBF and LCGU in cats with high resting CBF levels, this is an important advantage. In addition, the combination of the microsphere and 2-DG techniques within the same tissue samples allows for the investigation of the effects of traumatic injury on the important relationship between CBF and LCGU.

The effects of traumatic brain injury on regional cerebral blood flow in rats.

Alterations in cerebral blood flow (CBF) are among the most important secondary pathophysiologic consequences of traumatic brain injury. The present study compared CBF in control rats (n = 20) and in rats that received a calibrated experimental traumatic brain injury (n = 17). The traumatized rats were anesthetized with ketamine (25 mg/kg) and xylazine (10 mg/kg), and prepared for fluid percussion injury (FPI). Twenty-four hours later, the rats were anesthetized with 1% halothane in nitrous oxide-oxygen (70:30) and the left atrium was catheterized via a thoroacotomy. The atrial cannula was used to inject 15 microns radiolabeled microspheres to measure CBF. Following surgery, the concentration of halothane was reduced to 0.5% and the rats were paralyzed with pancuronium bromide (0.1 mg/kg). Thirty minutes later, baseline microsphere determinations were made, and the rats were injured (2.47 +/- 0.08 atm). Each rat received additional injections of microspheres at two of the following four times (T): 5, 15, 30, and 60 min after the brain injury. The procedures for the control group rats were the same as described above except that the rats were not subjected to the craniotomy and the FPI. The traumatized group exhibited heterogeneous decreases in CBF following trauma. Global CBF in this group was 78% (p less than 0.01), 64% (p less than 0.05), 52% (p less than 0.001) of those in the control group at T5, 15, 30, and 60, respectively. In rats, the most prominent cerebral circulatory changes following fluid percussion injury were early reductions of CBF and an increasingly heterogeneous CBF pattern. Hemorrhage, edema, and elevated prostagandin levels are mechanisms that may contribute to these changes.