Carlos Gilberto Carlotti Junior

Avaliação da isquemia cerebral pela respiração mitocondrial: modelo experimental

Brain ischemia occurs in several diseases. One of the critical factors for recovery of patients is the duration of the ischemic process. Brain activity depends on the energetic supply, it suggests that the study of mitochondrial function can be useful for evaluation of neuronal damage. The purpose of the present research was to study the mitochondrial respiration by occlusion of the left middle cerebral artery by intraluminal suture technique. Adults Wistar rats were subdivided in 4 groups: control, 15, 30 and 60 minutes of occlusion. Results showed that there was no significant difference between the group of 15 minutes and the control group. The group of 30 minutes had significant decrease of state III of mitochondrial respiration compared with control group. The group of 60 minutes had significant decrease in state III and IV of mitochondrial respiration compared with control group. Mitochondrial respiration allowed an early and effective evaluation of focal ischemic process of the rat brain.

Amygdala gene expression of NMDA and GABAA receptors in patients with mesial temporal lobe epilepsy

Temporal lobe epilepsy (TLE) is the most common form of partial epilepsy and affects 40% of the patients. Seizures arising from the mesial temporal lobe structures (i.e., amygdala and hippocampus) are common, whereas neocortical seizures are rare. In recent years, many studies aimed to identify the pattern of gene expression of neurotransmitters involved in molecular mechanisms of epilepsy. We used real‐time PCR to quantify the expression of GABAA (subunits α1, β1, β2) and NMDA (subunits NR1, NR2A, and NR2B) receptor genes in amygdalae of 27 patients with TLE and 14 amygdalae from autopsy controls. The NR1 subunit was increased in patients with epilepsy when compared with controls. No differences were found in expression of NMDA subunits NR2A and NR2B or in α1, β1, and β2 subunits of GABAA receptors. Our results suggest that the NR1 subunit of NMDA receptors is involved in the amygdala hyperexcitability in some of the patients with TLE.

Caspase-3 and Bcl-2 expression in glioblastoma: an immunohistochemical study

The unfavorable prognosis of malignant gliomas can also be explained by the incomplete knowledge of their molecular pathways. Studies regarding the regulatory process of apoptosis in glioblastoma (GBM), the most common malignant glioma, are few, and better knowledge of the expression of pro and anti-apoptotic proteins could collaborate with the development of new treatments founded on molecular basis. The objective of this study was to evaluate by immunohistochemistry the expression of caspase-3 and Bcl-2 in 30 samples of GBMs. The expression of caspase-3 (mean 17.67%) was lower than Bcl-2 (mean 30.92%), a statistically significant result (p<0.0001), suggesting low apoptotic activity in these tumors. Other studies of proteins related to the intrinsic and extrinsic pathway of apoptosis are required to provide additional information of this mechanism in GBMs.

Olfactory groove meningiomas: surgical technique and follow-up review

BACKGROUND Olfactory groove meningiomas comprise 4-10% of the intracranial meningiomas. Generally they give signs of brain compression due to great size they reach before diagnosis. In this study, the clinical outcome of patients with olfactory groove meningiomas surgically treated was analyzed. METHOD 17 patients operated on from 1988-2006. Female: 16, Male: 1. Age: 19-76 years-old (mean=53.12 +/- 13.11). FOLLOW-UP 1-209 months (mean=51.07 +/- 12.73. Bifrontal/bifrontal-bi-orbital approaches were used. Outcome was analyzed using survival/recurrence-free Kaplan-Mayer curves. RESULTS 16 had WHO grade 1; one grade 2 meningiomas. Resection Simpsons grade 1 was in achieved in 64.7%, grade 2 in 29.4% and grade 3 in 5.9%. There was no recurrence during the follow-up. Global and operative mortality were 11.8%. Main postoperative complications were osteomyelitis (11.8%) and pneumonia (5.9%). CONCLUSION Extensive approaches allowed total resection of most olfactory groove meningiomas with no recurrence during the follow-up, but operative mortality and local complications were high.

Toxoplasmose do sistema nervoso central em paciente sem evidência de imunossupressão: relato de caso

O quadro clinico da toxoplasmose adquirida em pacientes imunocompetentes habitualmente nao inclui manifestacoes neurologicas focais, o que e frequente em pacientes imunodeprimidos, como aqueles com sindrome da imunodeficiencia adquirida. Este trabalho tem como objetivo relatar o caso de uma paciente adulta que apresentou abscessos cerebrais por Toxoplasma gondii, sem evidencia de qualquer fator causador de imunossupressao.The clinical manifestations of acquired toxoplasmosis in the immunocompetent patient rarely include localized neurological signs, which are frequent in the immunosuppressed patient (Aids). The objective of this paper is to report the case of a woman with Toxoplasma gondii brain abscess, without an identified cause of immunosuppression.

Características dos pacientes com traumatismo craniencefálico atendidos no Hospital das Clínicas da Faculdade de Medicina de Ribeirão Preto

The chart of 3468 patients with head injury assisted in the Hospital das Clinicas - Ribeirao Preto Medical School, from 1990 through 1992 were analyzed aiming to determine their main characteristics. Regarding sex, there was predominance of male. Accidental fall among children and traffic accidents among adults were the main causes of trauma. Daily distribution of assistance revelead an increase between 8 and 12 PM and during the week there was a constant flow from Tuesday to Friday and progressively increased on Saturday to Sunday. Approximately 75% of the patients presented mild head injury (score equal or superior to 13 in the Glasgow Coma Scale). Headache among children and vomiting, headache and alcoholic abuse among adults were the most frequent signs and symptoms at admission. At discharge 87.2% of patients had no symptoms and mortality was 5.7%. Peculiarities of head injury in Ribeirao Preto are discussed.The chart of 3468 patients with head injury assisted in the Hospital das Clínicas-Ribeirão Preto Medical School, from 1990 through 1992 were analyzed aiming to determine their main characteristics. Regarding sex, there was predominance of male. Accidental fall among children and traffic accidents among adults were the main causes of trauma. Daily distribution of assistance revealed an increase between 8 and 12 PM and during the week there was a constant flow from Tuesday to Friday and progressively increased on Saturday to Sunday. Approximately 75% of the patients presented mild head injury (score equal or superior to 13 in the Glasgow Coma Scale). Headache among children and vomiting, headache and alcoholic abuse among adults were the most frequent signs and symptoms at admission. At discharge 87.2% of patients had no symptoms and mortality was 5.7%. Peculiarities of head injury in Ribeirão Preto are discussed.Foram analisados os prontuarios de 3468 pacientes com traumatismo craniencefalico (TCE) atendidos no Hospital das Clinicas da Faculdade de Medicina de Ribeirao Preto (HC-FMRP) no periodo de 1990 a 1992, objetivando estabelecer seu perfil. Os pacientes do sexo masculino predominaram. As causas mais frequentes de TCE foram quedas acidentais em criancas e acidentes de trânsito em adultos. A distribuicao circadiana do atendimento mostrou um pico entre 20 e 24 horas e a distribuicao semanal mostrou um afluxo constante de segunda a quinta-feira e um aumento progressivo no sabado e no domingo. A maioria dos pacientes (74,5%) apresentou TCE leve (escore igual ou superior a 13 na Escala de Coma de Glasgow). Os sinais e sintomas mais comuns foram cefaleia nas criancas e vomitos, cefaleia e alcoolismo agudo nos adultos. Na alta hospitalar 87,2% dos pacientes apresentavam-se assintomaticos. A mortalidade foi de 5,7%. As peculiaridades do TCE na regiao sao discutidas em relacao a literatura.

Expression of HSP70 in cerebral ischemia and neuroprotetive action of hypothermia and ketoprofen

Heat shock proteins (HSPs) are molecular chaperones that bind to other proteins to shepherd them across membranes and direct them to specific locations within a cell. Several injurious stimuli can induce Hsp70 expression, including ischemia. This study aimed to investigate the pattern of expression of protein (immunohistochemistry) and gene (real-time PCR) Hsp70 in experimental focal cerebral ischemia in rats by occlusion of the middle cerebral artery for 1 hour and the role of neuroprotection with hypothermia (H) and ketoprofen (K). The infarct volume was measured using morphometric analysis defined by triphenyl tetrazolium chloride. It was observed increases in the protein (p=0.0001) and gene (p=0.0001) Hsp70 receptor in the ischemic areas that were reduced by H (protein and gene: p<0.05), K (protein: p<0.001), and H+K (protein: p<0.01 and gene: p<0.05). The Hsp70 increases in the ischemic area suggests that the Hsp70-mediated neuroexcitotoxicity plays an important role in cell death and that the neuroprotective effect of both, H and K are directly involved with the Hsp70.

Pre, intra and post-ischemic hypothermic neuroprotection in temporary focal cerebral ischemia in rats: morphometric analysis

OBJECTIVE To evaluate the neuroprotection of mild hypothermia, applied in different moments, in temporary focal cerebral ischemia in rats. METHODS Rats was divided into Control (C), Sham (S), Ischemic-control(IC), Pre-ischemic Hypothermia (IH1), Intra-ischemic Hypothermia (IH2), and Post-ischemic Hypothermia (IH3) groups. Morphometry was performed using the KS400 software (Carl Zeiss®) in coronal sections stained by Luxol Fast Blue. Ischemic areas and volumes were obtained. RESULTS Statistically, blue areas showed difference for C vs. IC, IC vs. IH1 and IC vs. IH2 (p=0.0001; p=0.01; p=0.03), and no difference between C vs. S, IC vs. IH3 and IH vs. IH2 (p=0.39; p=0.85; p=0.63). Red areas showed difference between C vs. IC, IC vs. IH1 and IC vs. IH2 (p=0.0001; p=0.009; p=0.03), and no difference between C vs. S, IC vs. IH3 and IH1 vs. IH2 (p=0.48; p=0.27; p=0.68). Average ischemic areas and ischemic volumes showed difference between IC vs. IH1 and IC vs. IH2 (p=0.0001 and p=0.0011), and no difference between IC vs. IH3 and IH1 vs. IH2 (p=0.57; p=0.79). CONCLUSION Pre-ischemic and intra-ischemic hypothermia were shown to be similarly neuroprotective, but this was not true for post-ischemic hypothermia.

Apoptosis in Glioma Cells Treated with PDT

BACKGROUND Despite significant advances in neurosurgical techniques, the median survival time of patients with glioblastoma has improved little over the past 50 years and remains less than one year. Photodynamic therapy (PDT) is presently established as a widely accepted modality for the treatment of a variety of solid tumors. OBJECTIVES This study evaluated the effect of PDT-Photogem(®) on five glioma cell lines (U87, U138, U251, U343, and T98G). METHODS The experiments were carried out in 25-cm(3) flasks with different groups of cells seeded at a density of 1 × 10(5) cells per flask. After 3 h, the medium was removed, and the cells were incubated for 4 h with Photogem (5 μg/mL). After the incubation time, the photosensitizer-containing medium was removed and the cells were irradiated with LED (630 nm, 25 mW/cm(2), 25 J/cm(2)) devices for 17 min. For the final steps of the PDT, the cells were returned to the incubator and kept at 37°C with 5% CO(2) for 24 h, the cell viability assay was assessed using the trypan blue method, and the expression of Caspase 3 mRNA levels was assessed by real-time quantitative PCR. RESULTS Upon PDT-Photogem(®) treatment, viable cells, as evaluated by the trypan blue dye-exclusion method, decreased in two cell lines (U87 and U138) but not in the other three. Apoptosis, as assessed by the expression of caspase-3 mRNA levels, was at least partly involved in the death mechanism of the cell lines. CONCLUSIONS Collectively, our results indicated that PDT-Photogem(®) can act in glioma cells, thus encouraging new experiments in this field.

Concurrent Chemoradiotherapy with Weekly Paclitaxel in Malignant Cerebral Glioma Treatment

Background: Anaplastic astrocytomas (AA) and glioblastomas (GB) are the most common malignant gliomas, and despite newly developed drugs and combined treatments, they still have an adverse prognosis. Paclitaxel is a cytotoxic agent with radiosensitizing properties and exerts objective growth inhibition in glioma tumor cells. Patients and Methods: From 1998 to 2002, 61 microneurosurgically treated patients were randomized to group I (18 GB, 14 AA) which received radiotherapy and weekly paclitaxel at dose of 100 mg/m2, and group II (21 GB, 8 AA) which received only radiotherapy as a complementary treatment. Results: Median overall survival was 27.96 months in group I and 23.06 months in group II with no statistical difference. The 12-month survival was 81% in group I and 76% in group II. Kaplan-Meier curves of both groups did not demonstrate any difference. Analysis of each histological subgroup (AA or GB) also showed no statistical difference in the survival curves. All 427 cycles were well tolerated with no treatment-associated deaths. Conclusions: Chemoradiotherapy with weekly paclitaxel is safe and tolerable although there was no increase in the overall survival and 12-month survival of malignant glioma patients. Further investigations modulating the paclitaxel entrance and delivery into the brain should be encouraged.