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Biomedical subjects

M Shahid

Publications and source records attributed to M Shahid.

At least 19 recordsLinked to original sources

Occurrence, prevalence and genetic environment of CTX-M beta-lactamases in Enterobacteriaceae from Indian hospitals.

OBJECTIVES: To determine occurrence, prevalence and CTX-M genotypes produced by Enterobacteriaceae from clinical samples from three geographically distant Indian hospitals and to detect linkage of IS26 with bla(CTX-M) and map its precise insertion position. METHODS: A total of 130, non-duplicate Escherichia coli and Klebsiella pneumoniae resistant to a third-generation cephalosporin (3GC) from three Indian centres were screened for extended-spectrum beta-lactamase (ESBL) production using phenotypic detection methods. All isolates were screened for bla(CTX-M) using multiplex PCR. Precise CTX-M genotype was identified using reverse-line hybridization. All CTX-M-producing isolates were screened for linkage of IS26 with bla(CTX-M). DNA sequencing was used to map the exact insertion position of this mobile element. RESULTS: Ninety-five of 130 3GC-resistant (73%) (73% of total E. coli, 72% of total K. pneumoniae) isolates were found to carry bla(CTX-M-15). No other CTX-M genotype was detected. IS26 linkage with bla(CTX-M-15) was detected in 31% of isolates carrying bla(CTX-M-15). DNA sequencing revealed variable insertion of this mobile element within tnpA of ISEcp1. RAPD-PCR typing demonstrated great diversity in isolates carrying bla(CTX-M-15); no predominant clone was identified. CONCLUSIONS: In contrast with other studies where greater diversity exists, CTX-M-15 was the only CTX-M ESBL produced in this Indian collection of unrelated E. coli and K. pneumoniae. This is the first systematic survey report from India detecting CTX-M-type beta-lactamases This is also the first report indicating such high mobility/diversity of insertion of IS26 in close association with bla(CTX-M) in a single bacterial collection.

Anti-Bacterial Agents↗

Intravitreal steroids may facilitate treatment of Eales' disease (idiopathic retinal vasculitis): an interventional case series.

PURPOSE: Eales' disease is an idiopathic peripheral perivasculitis leading to proliferative vascular retinopathy, recurrent vitreous haemorrhages, and tractional retinal detachment. It is an elusive cause of blindness in young, otherwise healthy individuals. We studied the effects of intravitreal triamcinolone acetonide (IVTA) in patients of Eales' disease, which may eventually reduce the side effects and cost of management, with results equivalent to or better than oral steroids. METHODS: Ethics approval and prior patient consent were obtained. Fluorescein fundus angiograms (FFAs) of 12 eyes of 12 Eales' disease patients were taken before enrolment. These patients received 0.1 ml of 40 mg/ml (4 mg) intravitreal triamcinolone through pars plana under topical anaesthesia. Regular weekly follow-ups were initiated to ascertain Snellen visual acuity, intraocular pressure (IOP) with Goldman tonometer, and triple mirror examination. Fluorescein fundus angiography was again performed in the 8th week to monitor response to treatment. Decrease in areas of late perivascular dye extravasation on fluorescein angiography was used as marker for improvement. RESULTS: Ten out of a total of 12 (83.33%) eyes treated with IVTA showed significant reduction of late leakage from retinal vessels on fluorescein fundus angiography. Two out of 12 eyes (16.67%) did not show considerable decrease in late perivascular fluorescein dye leakage after 8 weeks of intravitreal triamcinolone injection. Two patients (16.67%) had a significant rise in IOP after IVTA. CONCLUSION: Intravitreal steroids may be advocated for management of idiopathic retinal vasculitis without complications of systemic steroids, and minimize need for more invasive procedures.

Adult↗

Resistance due to aminoglycoside modifying enzymes in Pseudomonas aeruginosa isolates from burns patients.

BACKGROUND & OBJECTIVE: Enzymatic modifications of aminoglycosides result in high-level resistance in numerous bacterial species. However, the data on this aspect are elementary in our country. The present study was therefore designed to determine resistance rates and patterns, and to find out the prevalent aminoglycoside modifying enzymes (AMEs) in clinical isolates of Pseudomonas aeruginosa from hospitalized burn patients. METHODS: Forty two, non-repeat, clinical isolates of P. aeruginosa obtained during a period from February to July 2003, were analysed for the presence of antibiotics resistance. On the basis of aminoglycoside susceptibility patterns, resistance phenotypes and possible AMEs were inferred according to interpretative reading. Seven isolates collected during the same period and previously characterized to harbour a 48.5 kb plasmid, encoding multiple drug resistance, were also analysed for aminoglycoside susceptibility patterns, and AMEs encoded by the plasmid were inferred. RESULTS: Ninety six per cent of the isolates were multi drug-resistant and majority (71.4%) were resistant to 5 or more antibiotics. Markedly high resistance to tobramycin (83.6%) and amikacin (55.1%) was noted, whereas gentamicin resistance was present in 32.6 per cent isolates. The enzyme N-acetyl transferases (AAC) viz. AAC(6')-I was the most common isolated AME followed by AAC(3)-II in 42.8 and 20.4 per cent of isolates respectively. The plasmid harbouring isolates belonged to AAC(6') phenotype and the enzyme encoded was inferred to be AAC(6')-I. INTERPRETATION & CONCLUSION: Markedly high resistance to tobramycin and amikacin was noted in the present study. AAC(6')-I was the most common AME and was inferred to be plasmid encoded in R-plasmid harbouring isolates. This is among the premier reports regarding the aminoglycoside resistance due to AMEs especially plasmid encoded, in P. aeruginosa from India. Further studies are required from different parts of the country to findout the prevalence of aminoglycoside resistance due to AMEs in P. aeroginosa isolates.

Aminoglycosides↗

Phenotypic detection of extended-spectrum and AmpC beta-lactamases by a new spot-inoculation method and modified three-dimensional extract test: comparison with the conventional three-dimensional extract test.

OBJECTIVES: To develop an easy, rapid and reproducible spot-inoculation method for phenotypic detection of extended-spectrum beta-lactamases (ESBLs) and AmpC beta-lactamases and to make the existing three-dimensional extract test more convenient for use in routine diagnostic laboratories. METHODS: ESBL and AmpC producing and non-producing isolates of Escherichia coli, Klebsiella pneumoniae and Pseudomonas aeruginosa, as identified by the conventional three-dimensional extract test, were used to evaluate the modified procedures. Whole bacterial cells and freeze-thaw preparations, as beta-lactamase sources, were strategically applied to culture plates near ceftazidime and cefoxitin discs on a lawn inoculum of E. coli ATCC 25922. Technical variations of the test included placing the beta-lactamase-containing inoculum into slits, wells and trenches, or onto the surface as spots at varying distances from the discs, and adding clavulanate or cloxacillin to the three-dimensional inoculum to confirm the presence of ESBLs and AmpC beta-lactamases, respectively. RESULTS: All the methods adopted for ESBL and AmpC detection by using the whole bacterial cells gave positive results. However, the best results were given by the spot-inoculation method. In modifications using the enzymic extracts, the enhanced growth of surface organisms was better appreciated in the designed modifications compared with the conventional methods. CONCLUSIONS: The method described here is simple and cost-effective. Furthermore, up to 16 isolates may be tested on a single culture plate, thus it is a less labour-intensive and more economic technique than other reported phenotypic methods.

Bacterial Proteins↗

Novel germline mutations in breast cancer susceptibility genes BRCA1, BRCA2 and p53 gene in breast cancer patients from India.

Mutations in breast cancer susceptibility genes, BRCA1 and BRCA2 account for more than 80% of hereditary breast and ovarian cancers. p53 tumor suppressor gene that controls cellular growth and differentiation is also known to be mutated in more than 50% of human cancers including breast cancer. We have carried out a study on BRCA1 and BRCA2 along with p53 gene mutations in both sporadic as well as familial breast cancer patients from India where breast cancer is fast emerging as a major cancer among premenopausal urban women. We examined 124 untreated primary breast cancer patients comprising 100 sporadic and 24 familial cases including 56 age-matched healthy controls for the presence of BRCA1, BRCA2 and the p53 gene mutations using PCR-SSCP and direct nucleotide sequencing. Certain frequently mutated exons such as 2, 5, 11, 13 and 20 of BRCA1, exons 2, 9, 11 (for 6174delT), 18 and 20 of BRCA2 and 4-9 exons of p53 gene were analyzed in sporadic breast cancer while all 22 coding exons of BRCA1 including its flanking intronic regions along with above mentioned exons of BRCA2 and p53 gene were analyzed in familial breast cancer patients. We identified six patients (25%) with BRCA1 mutation of which three were found to be of novel type one in exon 16 (4956insG) and two in exon 7 (Lys110Thr) (Ser114Pro) out of 24 familial breast cancer patients studied from two different geographic regions/populations of India. Two sisters from a single family (12.5%) out of eight families from Goa with Portuguese colonial origin showed presence of founder Ashkenazi Jewish BRCA1 mutation (185delAG) along with (IVS7 561-34T>C; IVS18 5271 + 66G > A). While from New Delhi, four (25%) of 16 breast cancer families showed BRCA1 mutations; a frame shift protein truncating (4956insG), a transition nonsense (Gln1395Stop) and two amino acid substitutions (Lys110Thr) and (Ser114Pro). Only one (4%) p53 mutation (Val97Ile) in its exon 4 along with BRCA1 mutation (4956insG) could be detected. No major sequence variation in BRCA2 gene was observed except for G203A at 5' UTR of exon 2, a common population polymorphism in two Goan patients who also showed silent nucleotide change for amino acid serine at codon 1436 of BRCA1 gene. None of the 100 sporadic breast cancer patients revealed any protein truncating or deleterious BRCA1 or BRCA2 gene mutation. Interestingly, three (3%) p53 mutations in its exon 5 were detected in sporadic breast cancer patients. Although three novel BRCA1 mutations including a founder Ashkenazi Jewish BRCA1 mutation were recorded in Indian women with familial breast cancer, the overall prevalence of BRCA gene mutations in Indian women with a family history of breast cancer appears to be low.

Adult↗

Multidrug-resistant Pseudomonas aeruginosa strains harbouring R-plasmids and AmpC beta-lactamases isolated from hospitalised burn patients in a tertiary care hospital of North India.

The present study was designed to determine resistance rates and patterns in Pseudomonas aeruginosa isolates obtained from hospitalised burn patients in an Indian tertiary care hospital. To that end, we isolated plasmid(s) from the multidrug-resistant isolates, demonstrated the plasmid-mediated resistance by curing and transformation experiments, and screened all the isolates for the occurrence of AmpC beta-lactamases. Thirty isolates of P. aeruginosa were analysed for the presence of antibiotic resistance. Plasmid-curing experiments and AmpC beta-lactamase detection were performed on all the isolates and seven isolates showing the most common antibiotic resistance pattern were selected for plasmid isolation and transformation experiments. All 30 isolates were multidrug-resistant and the majority (83.3%) of isolates were resistant to seven or more antibiotics, out of 11 antibiotics tested including anti-pseudomonal and non-anti-pseudomonal antimicrobial drugs. The most striking feature was the presence of resistance to amikacin. A 48.5-kb plasmid was isolated from the isolates. Curing and transformation experiments showed that resistance to amikacin was plasmid-mediated. Phenotypic screening for the occurrence of AmpC beta-lactamases showed that 20% of isolates were AmpC producers whereas 10% of isolates were characterised as 'indeterminate' for AmpC enzyme. In conclusion, a markedly high (56.7%) resistance to amikacin was noted in the present study. Amikacin resistance was determined to be plasmid-encoded and the presence of an AmpC beta-lactamase was inferred in 20% of isolates. This is among the first reports regarding the emergence of plasmid-mediated resistance to amikacin and the occurrence of AmpC beta-lactamases in P. aeruginosa strains from India.

Amikacin↗

Testing the validity of c-fos expression profiling to aid the therapeutic classification of psychoactive drugs.

RATIONALE: Different stimuli, including pharmacological stimuli, induce different neuroanatomical profiles of c-fos expression. Can these profiles be used in classifying psychoactive drugs and predicting therapeutic utility? OBJECTIVE: To test the validity of c-fos expression profiling to aid therapeutic classification. METHODS: Anxiolytics, antidepressants, antipsychotics and psychostimulants were compared. (i) A meta-analysis was performed and profiles compiled from literature reports of changes in c-fos expression in rat brain regions, measured by in situ hybridisation histochemistry or immunohistochemistry, after acute injection of psychoactive drugs. (ii) Male rat brains were profiled for changes in c-fos mRNA expression induced by acute injection of psychoactive drugs. RESULTS: (i) The meta-analysis showed that anxiolytics activate few (mostly stress-related) brain regions; antidepressants activate more regions, including the central amygdaloid nucleus; antipsychotics activate more regions still, including the nucleus accumbens and striatal areas; and psychostimulants activate the greatest number of all, including the most cortical regions (especially the piriform cortex). Profiles also varied within drug classes. (ii) Our experimental profiles confirmed and extended meta-analysis profiles, showing more downregulation. (iii) Sites activated by mirtazapine (an antidepressant not previously profiled) matched those of the antidepressant imipramine. CONCLUSIONS: (i) Differences between drug classes support their classification by means of c-fos profiling. Differences within classes may reflect mechanistic variations. (ii) Greater downregulation in our experiments might be because of inclusion of low, clinically relevant, drug doses and fuller coverage of brain regions. (iii) The agreement between mirtazapine and imipramine increases our confidence in the validity of c-fos expression profiling to aid drug classification and predict therapeutic utility.

Animals↗

Prevalence of aspergillosis in bronchogenic carcinoma.

Bronchoalveolar lavage of 42 patients of bronchogenic carcinoma was studied to find out the prevalence of aspergillosis. Sera of the patients were also analysed for presence of anti-Aspergillus antibodies by Immunodiffusion (ID), Enzyme linked immunosorbent assay (ELISA) and dot blot assay (DBA). Aspergillus was isolated in culture from 6 (14.2%) patients of bronchogenic carcinoma. Aspergillus fumigatus was the predominant species isolated. All the strains of Aspergillus were sensitive to itraconazole, ketoconazole and amphotericin B while resistance (33.3%) was found with fluconazole. Anti-aspergillus antibodies were detected equally by ID, ELISA and DBA in 9 (21.4%) cases. The present study revealed prevalence and seroprevalance of Aspergillus in bronchogenic carcinoma to be 14.2% and 21.4% respectively. Consistent reactivity against 18 kDa Aspergillus fumigatus antigen was noted in serologically positive cases. Antibodies against 18 kDa protein antigen in western blotting may be used as a reference marker for diagnosis of aspergillosis in bronchogenic carcinoma. It is also suggested that the simplest serological technique like ID may be performed along with culture for diagnosing Aspergillosis in patients of bronchogenic carcinoma since ID, ELISA and DBA showed similar sensitivity.

Adolescent↗

Role of dexamethasone in acute bacterial meningitis in adults.

OBJECTIVE: To evaluate the role of dexamethasone as adjunctive therapy in adult acute bacterial meningitis (ABM) in two groups of patients treated with antibiotics alone or a combination of antibiotics and dexamethasone. DESIGN: Systematic sampling interventional open cohort study. SETTING: Department of Medicine (Medical Unit II), Jinnah Postgraduate Medical Centre, Karachi. PATIENTS: Sixty eight patients aged 12-85 years admitted in Medical Unit II and diagnosed to have ABM. Patients were divided into two groups. Group A received anti-microbial therapy for 14 days (a combination of benzyl penicillin 6 million units I/V 6 hourly and chloramphenicol 1 gm I/V 6 hourly) and group B received the same antimicrobial therapy with dexamethasone 0.6 mg/kg/day in 3 divided doses for 4 days. MAIN OUTCOME MEASURES: Differences in mortality and morbidity in the two groups and differences in the CSF inflammatory parameters between the two groups of patients. RESULTS: There was early resolution of fever, headache and altered consciousness in group B as compared to group A. Cranial nerves involvement was lower in group B. There was no difference in the occurrence of other focal neurological deficits between the two groups. CSF inflammatory parameters (glucose, protein and WBC count) were significantly better in group B by day 5. No complications attributable to dexamethasone were seen in group B. CONCLUSION: There was early resolution of symptoms and CSF inflammatory parameters in the group that received dexamethasone as adjuvant therapy. Mortality was lower in the group treated with dexamethasone but the difference was not statistically significant. Dexamethasone should be administered to all adults patients with ABM.

Adolescent↗

Amoebic versus pyogenic liver abscess.

OBJECTIVES: To study the differences between the clinical presentation and complications of amoebic and pyogenic liver abscess. To correlate the diagnostic significance of Entamoeba Indirect Haemagglutination test (E.IHA) in establishing the diagnosis of amoebic liver abscess. DESIGN: Open cohort observational study. SETTING: Department of Medicine (Medical Unit II) Jinnah Postgraduate Medical Centre, Karachi. PARTICIPANTS: Fifty two patients aged 13-70 years admitted in Medical Unit II and diagnosed to have liver abscess. INTERVENTIONS: Group A comprised of patients clinically diagnosed to have amoebic liver abscess and received Metronidazole 500mg iv x 8 hourly for atleast 10 days or more if patient developed complications. Group B comprised of patients diagnosed to have pyogenic abscess and received Cefotaxime 1g iv x 8 hourly for the same duration or more if complicated (antibiotic reviewed in accordance with culture and sensitivity report). MAIN OUTCOME MEASURES: Differences in the clinical presentation, complications and diagnostic parameters between the two groups of patients. RESULTS: It was not possible to differentiate between amoebic and pyogenic liver abscess on clinical grounds, routine investigations and imaging techniques. Aspiration of pus, especially if the abscess was multiple, was most helpful in differentiating the two types of abscesses. Serological test of E.IHA was highly specific and sensitive for amoebic liver abscess. CONCLUSION: Majority of liver abscesses in Karachi are due to Entamoeba Histolytica. Pyogenic abscess though less frequent, must be excluded by pus aspiration and culture and sensitivity. E.IHA is a good rapid method of discriminating between the two types of abscesses.

Adolescent↗

Status of chromosome breaks and gaps in breast cancer. a follow-up study.

Genetic susceptibility and environmental factors are believed to be responsible for chromosomal instabilities and higher incidence of breast cancer. We conducted a follow-up study to find the levels of chromosome breaks and gaps in 20 premenopausal women with breast cancer before surgery, 1 month after surgery, and 3 years after surgery with respect to 20 age- and gender-matched controls. The mean level of chromosome breaks and gaps was found to be significantly higher (P<0.001) in breast cancer patients (before surgery) as compared with the controls. The chromosome breaks and gaps after 1 month of surgery were observed significantly decreased (P<0.005) when compared with that of patients before the surgery. Further significant increase in chromosome breaks and gaps was found after 3 years of surgery as compared with both the patients after 1 month of surgery (P<0.05) and controls (P<0.005). The significant increase in chromosome breaks and gaps in breast cancer patients (before surgery) may be due to the effects of genetic susceptibility to environmental carcinogens and endogenous factors. However, the decrease in this level after 1 month of surgery may be due to the removal of cancerous tissues, which in turn removes the effect of mutagens and clastogenic factors. Further increase in chromosome breaks and gaps after 3 years of surgery may be due to the long-term effects of therapeutic agents and genetic susceptibility to environmental carcinogens in the patients. The study furthermore suggests that the high level of chromosome breaks and gaps after 3 years of surgery may be a risk factor for the development of secondary tumor in patients.

Adult↗

Mechanism and impact of allosteric AMPA receptor modulation by the ampakine CX546.

Glutamate release at central synapses is transduced into a characteristic fast postsynaptic response by AMPA receptor gating and agonist affinity. The effect of two classes of modulators of AMPA receptor desensitization, the benzothiadiazides (cyclothiazide and IDRA 21) and the benzoylpiperidines (CX516 and CX546), were studied on gating kinetics of recombinant, native AMPA receptors and on synaptic currents. CX546 reduced the degree of desensitization more potently than CX516 or IDRA 21, but not as efficiently as cyclothiazide. In presence of CX516/CX546, desensitization of GluR2(flip) receptors was inhibited more than of GluR1(flip), whereas they had no effect upon response shape or conductance. CX546 increased agonist affinity threefold on nondesensitizing AMPA receptors by slowing agonist unbinding. Analysis of modulatory action suggests that, in contrast to cyclothiazide or IDRA 21, the Ampakine CX546 binds specifically to the agonist bound nondesensitized receptor, most likely acting by destabilizing the desensitized receptor conformation. All modulators tested showed higher efficiency on native receptors as compared to homomeric receptors. At the glutamatergic synapse, evoked synaptic amplitudes were weakly potentiated, while EPSC decay was slowed by nearly a factor of three in the presence of CX546 or cyclothiazide. In the presence of CX546, the current induced by short pulses of glutamate from recombinant GluR2 receptors decayed with a time course that was approximately twentyfold faster than EPSCs. The unique properties of CX546 may be beneficial for therapeutical use.

Allosteric Regulation↗

Effect of nitric oxide and malondialdehyde on sister-chromatid exchanges in breast cancer.

Nitric oxide (NO) and malondialdehyde (MDA) play a significant role in DNA damage, sister-chromatid exchanges (SCEs) and carcinogenesis. Here, we determine plasma NO and MDA to evaluate their role in carcinogenesis and their effect on the frequency of SCEs in 45 female breast cancer patients and in 35 age- and sex-matched controls. Plasma NO (P<0.01) and MDA (P<0.001) was significantly higher in the breast cancer group, and a direct correlation were found between plasma NO and MDA concentration and tumour grade. Patients with stage II disease showed the highest levels of both NO and MDA, compared with controls. Simultaneously, SCE frequency per lymphocyte in the breast cancer group was found to be significantly (P<0.001) higher; the greatest increase being found in patients with stage IV disease. Positive correlation was found between SCEs and both NO and MDA in the breast cancer group; however, both NO and MDA production decreased with increasing severity of the disease. Lower NO production in stage IV disease may be due to lower expression of nitric oxide synthase (NOS), further facilitating the production of superoxide anions (O2*-). The reaction between NO and O2*- results in peroxynitrite (OONO-) formation, which works efficiently at the molecular level and may induce higher SCE frequency. This work suggests that further cytogenetic and molecular study is required to provide definite answers for the therapeutic use of NO in breast cancer.

Adult↗

Gaucher disease with oculomotor apraxia and cardiovascular calcification (Gaucher type IIIC).

The authors describe four siblings from consanguineous parents who presented with oculomotor deficit in early childhood characterized by impaired volitional horizontal saccades, compensatory lateral head thrust, and preservation of vertical movement. When about 10 years of age, heavily calcified aortic and mitral valves required surgery. Fibroblast beta-glucocerebrosidase activity was markedly reduced. Genotype analysis indicated that the two patients who were tested were homozygous for the D409H (1342G-->C) mutation. Relating this rare phenotype of Gaucher disease to D409H mutation will facilitate management of the disease and counseling of families.

Adolescent↗

Molecular cloning and functional expression of the human glycine transporter GlyT2 and chromosomal localisation of the gene in the human genome.

Neurotransmitter transport systems are major targets for therapeutic alterations in synaptic function. We have cloned and sequenced a cDNA encoding the human type 2 glycine transporter GlyT2 from human brain and spinal cord. An open reading frame of 2391 nucleotides encodes a 797 amino acid protein that transports glycine in a Na+/Cl--dependent manner. When stably expressed in CHO cells, human GlyT2 displays a dose-dependent uptake of glycine with an apparent Km of 108 microM. This uptake is not affected by sarcosine at concentrations up to 1 mM. Radiation hybrid analysis mapped the GlyT2 gene to D11S1308 (LOD=8.988) on human chromosome 11p15.1-15.2.

Amino Acid Sequence↗

Treated bovine and autologous pericardium for aortic valve reconstruction.

BACKGROUND: To determine the differences in clinical behavior of bovine versus autologous pericardium, all consecutive patients undergoing aortic valve reconstruction were reviewed. METHODS: Between October 1988 and December 1995, 91 patients (mean age 30 years) underwent reconstruction with bovine (n = 27) or autologous (n = 64) pericardium. RESULTS: There were 2 hospital deaths, 5 late deaths, and no embolic events. Dysfunction of the aortic reconstruction required reoperation in 6 bovine (infection 1, fibrocalcific 5) and in 5 autologous (infection 3, annulus dilatation 1, commissural tear 1). Actuarial survival and freedom from structural deterioration at 8 years were 82.2%+/-9.6% and 76.2%+/-10.7% for bovine and 91.05%+/-3.96% and 96.8%+/-2.25% for autologous pericardium, respectively. The last Doppler echocardiographic study showed a mean regurgitation (1 to 4+) and gradient in the bovine pericardium of 1.25+/-and 20.7 mm Hg and in the autologous pericardium of 1+ and 7.7 mm Hg. CONCLUSIONS: Aortic valve reconstruction with pericardium can be safely performed with low thromboembolic rate. At 8 years follow up, there is a difference in favor of the autologous pericardium.

Actuarial Analysis↗