Clinical Correlation of Altered Molecular Signatures in Epileptic Human Hippocampus and Amygdala.

Widespread alterations in the expression of various genes could contribute to the pathogenesis of epilepsy. The expression levels of various genes, including major inhibitory and excitatory receptors, ion channels, cell type-specific markers, and excitatory amino acid transporters, were assessed and compared between the human epileptic hippocampus and amygdala, and findings from autopsy controls. Moreover, the potential correlation between molecular alterations in epileptic brain tissues and the clinical characteristics of patients undergoing epilepsy surgery was evaluated. Our findings revealed significant and complex changes in the expression of several key regulatory genes in both the hippocampus and amygdala of patients with intractable epilepsy. The expression changes in various genes differed considerably between the epileptic hippocampus and amygdala. Different correlation patterns were observed between changes in gene expression and clinical characteristics, depending on whether the patients were considered as a whole or were subdivided. Altered molecular signatures in different groups of epileptic patients, defined within a given category, could be viewed as diagnostic biomarkers. Distinct patterns of molecular changes that distinguish these groups from each other appear to be associated with epilepsy-specific functional consequences.

Efficiency of mitochondrial genes and nuclear Alu elements in detecting human DNA in blood meals of Anopheles stephensi mosquitoes: a time-course study.

BACKGROUND: The time required for PCR detection of DNA in human blood meals in vector mosquitoes may vary, depending on the molecular markers used, based on the size and copy number of the amplicons. Detailed knowledge of the blood-feeding behavior of mosquito populations in nature is an essential component for evaluating their vectorial capacity and for assessing the roles of individual vertebrates as potential hosts involved in the transmission of vector-borne diseases. METHODS: Laboratory experiments were conducted to compare the time course of PCR detection of DNA in human blood meals from individual blood-fed Anopheles stephensi mosquitoes, using loci with different characteristics, including two mitochondrial DNA (mtDNA) genes, cytB (228 bp) and 16S ribosomal RNA (rRNA) (157 bp) and nuclear Alu-repeat elements (226 bp) at different time points after the blood meal. RESULTS: Human DNA was detectable up to 84-120 h post-blood-feeding, depending on the length and copy number of the loci. Our results suggest that 16S rRNA and Alu-repeat markers can be successfully recovered from human DNA up to 5 days post-blood-meal. The 16S rDNA and Alu-repeat loci have a significantly (P = 0.008) slower decline rate than the cytB locus. Median detection periods (T50) for the amplicons were 117, 113 and 86.4 h for Alu-repeat, 16S rDNA and cytB, respectively, suggesting an inverse linear relationship between amplicon size/copy number and digestion time. CONCLUSION: This comparative study shows that the Alu-repeat locus is the most efficient marker for time-course identification of human DNA from blood meals in female mosquitoes. It is also a promising tool for determining the anthropophilic index (AI) or human blood index (HBI), i.e. the proportion of blood meals from humans, which is often reported as a relative measure of anthropophagy of different mosquito vectors, and hence a measure of the vector competence of mosquito species collected in the field.

MGMT Gene rs1625649 Polymorphism in Iranian Patients with Brain Glioblastoma: A Case Control Study.

Glioblastoma (GBM) is the most common and aggressive primary malignant brain tumor with poor prognosis and high potential of dispersion to other brain tissues in adult. Effective and modern choices of treatment including chemotherapy with alkylating agents marginally extend survival of GBM. However, alkylating agents can lead to highly harmful mismatch during DNA replication causing apoptosis and cell death. Accordingly, O6-Methylguanine-DNA methyltransferase (MGMT) removes alkyl adducts, thereby causing resistance to alkylating drugs. Single-Nucleotide Polymorphisms (SNPs) in MGMT promoter region may play a role in the regulation of MGMT expression and prediction of glioma development risk. In order to evaluate the clinical significance of rs1625649 SNP in the MGMT promoter region of glioblastoma, genomic DNA from a series of 54 patients with GBM and 50 healthy individuals in Iranian population were collected for tetra ARMS PCR amplification. None of the "A" or "C" alleles were associated with tumor occurrence, the "AA" genotype was more frequent in healthy subjects, and the "AC" genotype was 4.6 times more common in patients with GBM. The longest survival time was observed in the "CC" genotype; however, this difference was not statistically significant. On the other hand, homozygous rs1625649 (AA genotype) was significantly associated with a better survival than the cases with heterozygous rs1625649 (CA genotype) or wild type rs1625649 (CC genotype), predicting better response to temozolomide-based chemotherapy.

VEGF gene polymorphisms in Iranian patients with intracranial glioblastoma.

BACKGROUND: Glioblastoma is one of the most common malignant brain tumors in adults with poor prognosis. Neovascularization is one of the characteristics of these tumors, which is associated with overexpression of vascular endothelial growth factor (VEGF). Accordingly, single nucleotide polymorphisms of this gene could play an important role in structural and functional alterations leading to overexpression of this gene in GBM. METHODS: A total number of 49 patients with GBM and 50 healthy controls were included in the current study. The Genomic DNA was extracted from brain tumor/tissue samples, and after purification assessment, the alleles, and genotypes of rs3025039 and rs2010963 polymorphisms of the VEGF gene were investigated using T-ARMS-PCR. RESULTS: The "T" allele of rs3025039 was 2.79 times more frequent in GBM patients compared to controls (P=0.01). Moreover, the "CT" genotype was 2.83 times more common among patients (P=0.015), while the "CC" was more frequent in controls (P=0.009). The mean overall survival was significantly different between three genotypes of rs3025039, with the longest survival time in "CT" genotype (15.10±5.21, P=0.041). Besides, rs2010963, was significantly associated with GBM occurrence, with the "G" allele being 1.96 times more frequent in patients (P=0.01), as well as the "GG" genotype, which was 7.87 times more common in patients (P<0.001). CONCLUSIONS: Polymorphisms of VEGF could potentially play a role in pathogenesis of GBM, as the allele and genotype distributions of rs3025039 and rs2010963 SNPs were significantly associated with GBM occurrence.

Lateral femoral circumflex artery contribution to the articular and periarticular hip circulation: relevance to the anterior hip approach-a cadaveric study.

BACKGROUND: The purpose of this cadaveric study was to anatomically demonstrate the contribution of the lateral femoral circumflex artery (LFCA) to the articular and peri-articular hip circulation with an emphasis on the vascularization of hip abductors specially tensor fasciae latae muscle (TFLM). METHODS AND MATERIALS: Thirty hips from 26 fresh cadavers were dissected after injection of the aorta or common iliac artery by colored silicon, to study the contribution of the LFCA to the articular and periarticular hip circulation. Furthermore, the aorta was injected in another 18 fresh cadavers after unilaterally ligating the ascending branch of the LFCA (a-LFCA) as the vascular pedicle of the TFLM, to evaluate the collateral circulation to the TFLM, if available. RESULTS: In all specimens but one, the a-LFCA was found as the single major vascular pedicle of the TFLM. When ligated, only 4 out of 18 hips demonstrated colored cast vessels in the cut surface of the muscle. The ascending branch had also consistent but variable contribution to the gluteus medius and gluteus minimus muscles in 80% of the hips. Furthermore, the a-LFCA consistently supplied the anterior and anterolateral aspects of the hip capsule. In 35% of hips, the a-LFCA contributed to the femoral head and neck circulation via one or two small anterior retinacular arteries. CONCLUSION: LFCA contribution to the articular and periarticular hip circulation is only delivered by the ascending branch. The TFLM is specially perfused by the a-LFCA; its ligation significantly decreases the muscle perfusion. It is poorly supplied by the collateral circulation.

Investigating the Relationship between Fingerprint Pattern and Development of Oral Squamous Cell Carcinoma.

Statement of the Problem: Oral squamous cell carcinoma (OSCC) constitutes more than 90% of oral malignancies. The main risk factors of OSCC include cigarette smoking and alcohol. However, since not all smokers or alcohol drinkers develop this disease, other factors have also been suggested including genetic characteristics of every person to be implicated in the probability of developing OSCC. Purpose: Our aim in this study is to investigate the possible relationship between fingerprint patterns and the probability of developing OSCC. Materials and Method: In a cross sectional study, we had 140 patients in 2 groups as OSCC and cancer free. Fingerprints were recorded by fingerprint scanner device. The fingerprint patterns were categorized into three major groups and four subgroups. Groups were tested by chi-square. Results: The relationship between the main fingerprint patterns and incidence of OSCC became significant (p= 0.037). The frequency of the main pattern of Arch was significantly higher in the experimental group than in the control group (p< 0.05). Considering the main patterns of Loop and Whorl, no significant difference existed between the two groups. Furthermore, the frequency of subtype patterns of Double Whorl and Central Pocket Whorl was significantly higher in the control group than in the experimental group (p< 0.05). Conclusion: Since dermatoglyphics is contingent upon genetic variations, fingerprint can be used for investigating the susceptibility of people in developing different diseases, though further studies are required in this regard. This method is in no way a substitute for gold standard methods for diagnosis.

Surgical Anatomy of the Rectus-Sparing Approach for Periacetabular Osteotomy: A Cadaveric Study.

BACKGROUND: The Bernese periacetabular osteotomy (PAO) is a widely used technique for the management of acetabular dysplasia and other hip deformities in adolescents and young adults. Originally, the approach was described with a release of both origins of the rectus femoris muscle1. In the more recently described rectus-sparing approach, both heads remain attached2,3. It has been proposed that this modification may decrease pain, ease postoperative rehabilitation, and avoid heterotopic ossifications, without limitations of the surgical overview. DESCRIPTION: Both the original and the rectus-sparing approach are modifications of the Smith-Petersen approach. The skin incision and further dissection remain identical in both approaches for the protection of the lateral femoral cutaneous nerve, the osteotomy of the anterior superior iliac spine (or takedown of the inguinal ligament), the exposure of the iliac fossa, and the medial retraction of the abdominal and iliopsoas muscles. In both variants, the further dissection traverses the iliopectineal bursa. In contrast to the original approach, in which the rectus muscle becomes part of the medial flap after releasing both heads, the rectus-sparing approach involves the undetached rectus muscle becoming part of the lateral flap while the medial flap includes the sartorius and iliacus-iliocapsularis muscles. The anterior capsule and deep structures can be accessed through the interval between the rectus femoris and iliopsoas muscles or lateral to the rectus muscle. The remaining surgical steps are again similar in both techniques. According to preference, the surgeon starts with the pubic osteotomy or with the ischial cut first, the latter avoiding additional bleeding from the pubic osteotomy. For the ischial osteotomy, the bone is accessed by making an anteroposterior tunnel between the medial capsule and the iliopsoas tendon anteriorly and between the medial capsule and the obturator externus muscle posteriorly. While the ischial osteotomy is an incomplete separation, the pubic osteotomy is a complete separation. It sections the superior pubic ramus medial to the iliopectineal eminence, in a somewhat oblique fashion. The third and fourth cuts are made in the iliac bone in such a way as to keep the posterior column intact. By connecting the posterior iliac and ischial cuts as the last osteotomy step, the acetabulum is freed and repositioned as needed. The aim of our cadaver dissection is primarily to describe part of the rectus-sparing approach and to test this modification for eventual disadvantages over the classic approach. The remaining steps of the procedure correspond to the approach as described earlier1,4, appreciating that several modifications of the procedure are in use. ALTERNATIVES: Nonsurgical treatment may be an alternative in borderline dysplasia; however, it needs to be reconsidered and eventually changed to surgical treatment when symptoms persist or come back. Other current techniques for surgical treatment of adolescent and adult hip dysplasia include triple and rotational or spherical osteotomies5-7. RATIONALE: The Bernese PAO is performed through a single incision. All cuts are performed from the inner side of the pelvis, avoiding interference with the vascularity of the acetabular and periacetabular bone, which mainly comes from the outside of the pelvis8. The procedure needs minimal hardware for fixation, and partial weight-bearing can be allowed. The PAO provides a wide range of acetabular reorientation options. Wide capsulotomy and intra-articular procedures are possible, as well as additional femoral corrections. Childbirth via natural delivery is possible even after bilateral PAO9. Nerve injuries can be guarded against with careful surgical execution of the osteotomies4.

The Critical Role of Prenatal Genetic Study in Prevention of Primary Immunodeficiency in High-risk Families: The Largest Report of 107 Cases.

This study aims to investigate the role of prenatal diagnosis (PND) in Iranian couples with a previous history of primary immunodeficiency disorders (PIDD) in their family. All referred couples with a family history of PIDD and a tendency for PND were included in this project. Based on gestational age, chorionic villus sampling (CVS) was performed to analyze the molecular defect of the fetus according to the previous gene defect of the affected case in the family. Postnatal confirmation was performed by immunological screening tests. In a total of 100 cases, CVS was not evaluated in 19 patients due to unwillingness (n=5), late prenatal referral (n=7), miscarriage before CVS (n=3), and female fetus with x-linked diseases in previous children (n=4). In the remaining 81 patients, heterozygous and homozygous mutations were found in 33 and 23 cases, respectively. The hemizygous mutation was obtained in 6 and no pathogenic mutations were found in 19 individuals. Postnatal evaluations revealed that a total of 65 babies were healthy, 32 fetuses were aborted (3 cases before CVS, 2 spontaneous abortions of a healthy and as affected fetus in the CVS subgroup, and 27 cases were aborted due to therapeutic causes). One fetus from the heterozygous subgroup was spontaneously aborted with severe combined immunodeficiency (SCID) and one fetus from the homozygous subgroup that was supposed to be healthy was affected by the autosomal dominant-chronic granulomatous disease (AR-CGD). The diagnostic error was 1.2%. PND is highly recommended in families with a history of PID in their previous child to prevent an affected baby being born and to reduce the government, family, and personal burden of these diseases.

Morphology of asterion and its proximity to deep vein sinuses in Iranian adult skull.

Background: The asterion is located at the posterior lateral side of the skull at the junction of the parietal, temporal and occipital bones.Methods: We examined the morphology of the asterion, its association with deep vein elements, the mastoid apex and inion in 105 adult cadavas (210 hemicraniums) including 146 males and 64 females at the anatomy lab of the Legal Medicine Organization.Results: Two types of asterion were observed. Type I was found in 14.7%, and type II in 85.3% of cases. In 70% of cases, the asterion was at or above the venous sinus. The distance between the asterion and the mastoid appendage on the right side was 47.03 mm and on the left side was 46.5 mm. The distance between the asterion and the inion at the right side was 70.55 mm and on the left side was 70.2 mm.Conclusion: The asterion in 70% of cases was at or above the level of the transverse sinus. For this reason, in posterior fossa surgical approaches, the first burr hole is preferred to start in the lower parts of the asterion.

Cell injury and receptor expression in the epileptic human amygdala.

Neuropathological findings in the amygdala obtained from patients with mesial temporal lobe epilepsy (MTLE) indicate varying degrees of histopathological alterations, such as neuronal loss and gliosis. The mechanisms underlying cellular damage in the amygdala of patients with MTLE have not been fully elucidated. In the present study, we assess cellular damage, determine the receptor expression of major inhibitory and excitatory neurotransmitters, and evaluate the correlation between the expression of various receptors and cell damage in the basolateral complex and the centromedial areas in the amygdala specimens resected during brain surgery on 30 patients with medically intractable MTLE. Our data reveal an increased rate of cell damage and apoptosis as well as decreased expression levels of several GABAergic receptor subunits (GABAARα1, GABAARß3, and GABABR1) and GAD65 in the amygdalae obtained during epilepsy surgery compared to autopsy specimens. Analyses of the expression of glutamate excitatory receptor subunits (NR1, NR2B, mGluR1α, GluR1, and GluR2) reveal no significant differences between the epileptic amygdalae and autopsy control tissues. Furthermore, the increased occurrence of apoptotic cells in the amygdala is negatively correlated with the reduced expression of the studied GABAergic receptor subunits and GAD65 but is not correlated with the expression of excitatory receptors. The present data point to the importance of GABAergic neurotransmission in seizure-induced cell injury in the amygdala of patients with MTLE and suggest several GABA receptor subunits as potential druggable target structures to control epilepsy and its comorbid disorders, such as anxiety.

Anthropometric measurements in Iranian men.

There is inevitable need for data regarding anthropometric measurements of each community's population. These anthropometric data have various applications, including health assessment, industrial designing, plastic & orthopedic surgery, nutritional studies, anatomical studies and forensic medicine investigations. Anthropometric parameters vary from race to race throughout the world, hence providing an anthropometric profile model of residents of different geographic regions seems to be necessary. To our knowledge, there is no report of bone parameters of the Iranian population. The present study was carried out to provide data on anthropomorphic bone parameters of the Iranian population, as a basis for future relevant studies. We calculated most of the known anthropometric parameters including skull, mandible, clavicle, scapula, humerus, radius, ulna, sacrum, hip, femur, tibia and fibula of 225 male corpses during a period of 2 years (2014-2016). Data expression was done as mean ± standard deviation. The results consist the first documented report on anthropometric bone measurement profile of Iranian male population, that can be considered a valuable source of data for future research on Iranian population in this regard.

Suspected Methadone Toxicity: from Hospital to Autopsy Bed.

High mortality rates have been reported for methadone in both adults and children. We aimed to determine the pattern of toxicity, possible underlying diseases and treatment challenges in patients referred to our centre with early diagnosis of methadone toxicity and who later died. Medical files of all methadone-poisoned patients who had been admitted to a referral centre of toxicology between March 2011 and March 2016, died during the hospital stay and sent for autopsy to Legal Medicine Organization were retrospectively evaluated. In a total of 94 patients, autopsy findings and laboratory evaluations showed that cause of death was pure methadone toxicity in 57 (60.6%). Other causes of death were ischaemic heart disease in ten, co-ingestions (toxicities including methadone) in eight, brain haemorrhage, multi-organ failure and pneumosepsis (each in four), meningitis/encephalitis in three and head trauma and other toxicities (other than methadone but including an opioid, each in two) patients. Time of cardiopulmonary arrest was significantly different between those with pure methadone toxicity and those who died due to other causes (p = 0.01). Patients who had died due to co-ingestions and other toxicities were younger (p = 0.029) and took more bolus doses of naloxone (p = 0.042). In methadone users, especially in older ages and those with trivial response to naloxone administration, loss of consciousness should not be strictly attributed to methadone toxicity. In such patients, thorough evaluation for other possible causes of loss of consciousness is mandatory.

Three years evaluation of retained foreign bodies after surgery in Iran.

BACKGROUND: Medical errors such as retained foreign bodies (RFB) during surgery are not well studied. To define risk factors associated with this type of error, we performed retrospective study. METHODS: We reviewed medical records for unintentional foreign object remaining in the body during surgery such as gender, age, surgery ward, and incident reports referred to several forensic medicine administrations as well as adverse effects of retained foreign bodies and methods for detection of them over a 3-years period from January 2008 through May 2011. RESULTS: Thirty eight patients were involved in the study to have retained foreign bodies (73% sponges and 27% other instruments including 7 (18.42%) cases of other bandages, 2 cases (5.26%) of scissor and 1 case (2.63%) of forceps). The general surgery ward was most commonly involved (47%) followed by the gynecology surgery ward (34%). Men (58%) were more involved with RFB than women (42%). CONCLUSION: Considering the most frequent type of retained foreign bodies and also more frequent involved surgery wards besides detection methods for RFB, a mixed of preventing protocols such as regular counting of devices, post-operative X-ray with radiopaque markers and exact evaluation of surgery site should be employed to reduce the occurrence of retained foreign bodies and its complications.

Prevalence of HCV, HBV, and HIV Seropositivity among Cadavers Referred to Autopsy Hall of Legal Medicine Bureau of Tehran, Iran.

A large number of dead bodies are referred to forensic autopsy halls for medicolegal examination. They can be a source of transmission of infectious diseases through direct contact or autopsy tools. The main aim of this study was to estimate the virus infection rates in the dead bodies. One thousand consecutive dead bodies that had been referred to autopsy hall of Legal Medicine Bureau of Tehran, Iran, during 2016, were included. The blood samples were analyzed in the laboratory for detection of HBs Ag, HBs Ab, HIV Ab, and HCV Ab, after providing informed consent from legal next of kin of the dead bodies. The general characteristics of the dead bodies were also collected by a checklist. Forty-seven cases of HIV seropositivity, 80 cases of HBs Ag seropositivity, and 97 cases for HCV Ab seropositivity were found. Among them, 27 cases of HIV, 40 cases of anti-HBC positive, and 94 cases of RIBA testing positive for HCV were proved through confirmatory tests. In other words, 2.6% of the dead bodies were infected with HIV, 3.8% with HBV, and 9% with HCV. The total infection rate was 15.5%. This is a worrying risk for pathologist and autopsy technicians.

Virginity Testing Beyond a Medical Examination.

Apart from religious values, virginity is important in different communities because of its prominent role in reducing sexually transmitted diseases and teen pregnancies. Even though virginity testing has been proclaimed an example of violence against women by the World Health Organization, it is still conducted in many countries, including Iran. 16 in-depth, semi-structured interviews were conducted with participants aged 32 to 60 years to elucidate the perceptions and experiences of Iranian examiners of virginity testing.The perception and experience of examiners were reflected in five main themes. The result of this study indicated that virginity testing is more than a medical examination, considering the cultural factors involved and its overt and covert consequences. In Iran, testing is performed for both formal and informal reasons, and examiners view such testing with ambiguity about the accuracy and certainty of the diagnosis and uncertainty about ethics and reproductive rights. Examiners are affected by the overt and covert consequences of virginity testing, beliefs and cultural values underlying virginity testing, and informal and formal reasons for virginity testing.

Comparison of physical examination and conventional radiography in diagnosis of nasal fracture.

Nasal bone fracture is the most common fracture which would result from facial trauma. So, the present study performed to select the most reliable way to diagnose new fractures considering CT scan results as a gold standard in this matter. All the people refer to a forensic medicine center were, at first, physically examined by general practitioners. Plain lateral radiography and ENT consult were requested afterwards. CT scan was requested to get trusty results in case of any imbalance between Radiographic finding and physical examination. The results finally were evaluated and compared. CT scan was tried for 61 (6%) patients with positive clinical findings for new fracture which were not supported by radiologic studies. New fracture was identified in 55 participants out of the above number. Trusting physical examination and its preference to the radiologic findings has special value in cases where fracture is not detectable by radiography and there is no access to CT scan.

Anatomic variations in femoral head circulation.

The purpose of this cadaveric study was to clearly define the blood supply to the femoral head to help further reduce the incidence of iatrogenic avascular necrosis (AVN). Thirty-five hips of twenty-eight fresh cadavers were injected with colored silicone. Anterior and posterior dissection was performed to assess the vessels contributing to femoral head vascularity. The medial femoral circumflex artery (MFCA) was found to be the main blood supply to the hip in twenty-nine specimens; the inferior gluteal artery (IGA) was found to be the main blood supply in six. The MFCA consistently provided at least one smaller-calibre inferomedial retinacular artery. The foveal artery provided no significant vascular contribution in any specimen. The quantity and calibre of superior retinacular vessels demonstrated their dominance in head vascularity, although inferior retinacular arteries were consistently present. To reduce the risk of iatrogenic AVN, branches of both the IGA and MFCA traversing the interval between the quadratus femoris and piriformis muscles must be protected during surgery. Since all intracapsular vessels penetrated the capsule near its distal attachment, distal capsulotomy carries a significantly higher risk of AVN than proximal capsulotomy, particularly posterolaterally and inferomedially.

Survey of sudden death from aneurysmal subarachnoid hemorrhage in cadavers referred to Legal Medicine Organization of Tehran, 2001-2005.

BACKGROUND AND AIM: Some cases with aneurysmal subarachnoid hemorrhage (SAH) never reach the medical centre alive and they are not included in most studies of SAH. To explain the clinical profile of sudden death from aneurysmal SAH, we examined the epidemiology and clinicopathologic characteristics of patients with aneurysmal SAH who never reached medical attention or died within first 24 hours of the onset of first symptoms. MATERIALS AND METHODS: Using the autopsy records in Legal Medicine Organization of Tehran, we identified all cadavers who were diagnosed with aneurysmal SAH between 2001 and 2005. RESULTS: There were 85 women and 60 men with a mean age of 50 years. Twenty patients (14%) died without reaching medical care and 58 (40%) died within first 24 hours of the onset of first symptoms. The remnant had died after 24 hours to 28 days of hospitalization. In comparing patients with sudden death versus remnant, the main variables were the frequency of posterior circulation aneurysms that was found in 59% compared with 19.4% in those who had died after 24 hours, intraventricular hemorrhage 53.8% versus 19.4%, and pulmonary edema 92.3% versus 34.3% (P = 0.01). CONCLUSION: In our population, the frequency of sudden death from aneurysmal SAH has not changed during the last 5 years. The typical clinical profile of sudden death in SAH includes intraventricular hemorrhage, pulmonary edema, and a ruptured posterior circulation aneurysm. Intracerebral hemorrhage is rarely connected to sudden death from aneurysmal SAH.