Applying artificial intelligence to determination of legal age of majority from radiographic data.

Forensic odontologists use biological patterns to estimate chronological age for the judicial system. The age of majority is a legally significant period with a limited set of reliable oral landmarks. Currently, experts rely on the questionable development of third molars to assess whether litigants can be prosecuted as legal adults. Identification of new and novel patterns may illuminate features more dependably indicative of chronological age, which have, until now, remained unseen. Unfortunately, biased perceptions and limited cognitive capacity compromise the ability of researchers to notice new patterns. The present study demonstrates how artificial intelligence can break through identification barriers and generate new estimation modalities. A convolutional neural network was trained with 4003 panoramic-radiographs to sort subjects into 'under-18' and 'over-18' age categories. The resultant architecture identified legal adults with a high predictive accuracy equally balanced between precision, specificity and recall. Moving forward, AI-based methods could improve courtroom efficiency, stand as automated assessment methods and contribute to our understanding of biological ageing.

Radiographic morphology of canines tested for sexual dimorphism via convolutional-neural-network-based artificial intelligence.

The permanent left mandibular canines have been used for sexual dimorphism when human identification is necessary. Controversy remains whether the morphology of these teeth is actually useful to distinguish males and females. This study aimed to assess the sexual dimorphism of canines by means of a pioneering artificial intelligence approach to this end. A sample of 13,046 teeth radiographically registered from 5838 males and 7208 females between the ages of 6 and 22.99 years was collected. The images were annotated using Darwin V7 software. DenseNet121 was used and tested based on binary answers regarding the sex (male or female) of the individuals for 17 age categories of one year each (i.e. 6-6.99, 7.7.99… 22.22.99). Accuracy rates, receiver operating characteristic (ROC) curves and confusion matrices were used to quantify and express the artificial intelligence's classification performance. The accuracy rates across age categories were between 57-76% (mean: 68%±5%). The area under the curve (AUC) of the ROC analysis was between 0.58 and 0.77. The best performances were observed around the age of 12 years, while the worst were around the age of 7 years. The morphological analysis of canines for sex estimation should be restricted and allowed in practice only when other sources of dimorphic anatomic features are not available.

SNPs, adipokynes and adiposity in children with asthma.

OBJECTIVES: Asthma and obesity are complex disorders influenced by environmental and genetic factors. We performed an integrative review of genetic polymorphisms and adipokines effects in children and adolescents with asthma and obesity. DATA SOURCES: Articles focused on these issues were collected from SciELO, PubMed, LILACS, Embase and ScienceDirect electronic databases, in 2009-2020 period. STUDY SELECTIONS: 22 articles were selected, including clinical trials, analyses approaches, case-control studies, meta-analysis and Mendelian randomization studies. RESULTS: Leptin concentrations were higher in obesity and asthma. The high value of BMI and Leptin indicated severe asthma. Adiponectin may be reduced in obese children. The high value of BMI and low level of Adiponectin may indicate severe asthma. Some linkage of PRKCA gene, asthma and BMI was observed. FTO T allele rs62048379 was positively associated with overweight/obesity, related to protein and PUFA:SFA ratio intake and influences the choice of more energy-dense foods. FTO rs9939609 effects are more pronounced among children with insufficient vitamin D levels. CONCLUSION: Leptin may be a potential predictor for asthma control in children. BMI and Adiponectin could have certain predictive value for asthma. FTO gene was related to a higher mean BMI Z-score and accelerated developmental age per allele. Strong genetic heterogeneity influencing on asthma and obesity susceptibilities is evident and related to distinct genetic features. GWAS with childhood obesity in asthma contributed to greater insights, mainly on later childhood. Standardized definitions for asthma and overweight/obesity in studies approaching adipokines and SNPs would provide stronger evidence in deciding the best management.

Pattern ERGs suggest a possible retinal contribution to the visual acuity loss in acute optic neuritis.

PURPOSE: Macular involvement in optic neuritis (ON) is well-recognised but poorly understood and may be of clinical relevance. This study explores macular structure-function correlates in acute ON. METHODS: This cross-sectional cohort study recruited ON patients within 14 days of symptom onset. Subjects underwent pattern electroretinography (PERG), pattern visual evoked potentials (PVEP) and optical coherence tomography (OCT) imaging. PERG P50 and N95 components were correlated with OCT data. RESULTS: Twenty-six individuals with ON were recruited, comprising eleven multiple sclerosis (MS-ON), six myelin oligodendrocyte glycoprotein associated (MOG-ON) and nine with isolated ON. These were compared with 28 healthy controls. PVEPs were undetectable in 11 (42%) of individuals with ON. When detectable, PVEP P100 was delayed (median 136 ms range 110-173 ms) and amplitude reduced (median 6 µV, range 3-14 µV) in ON compared with controls (both p < 0.001). PERG P50 component amplitudes, largely reflecting macular function, were reduced in affected eyes (median 2.3 µV; range 0.8-5.0 µV) compared with controls (3.3 µV; range 2.8-5.7 µV) and compared with fellow eyes (p < 0.001). The N95:P50 ratio was below the reference range in the affected eyes of five patients. Eight cases (32%) had subnormal P50 amplitudes (< 2.0 µV), and these patients had poorer visual acuity (p = 0.020). P50 amplitudes were positively correlated with an increase in inner nuclear layer thickness (rs = 0.36; p = 0.009) and macular ganglion cell and inner plexiform layer (mGCIPL) thickness (rs = 0.44, p = 0.022). CONCLUSION: PERG P50 component reduction reveals dysfunction of inner macular layers in acute ON and correlates with structural alterations on OCT. These early macular pathologic processes are likely to contribute to the visual loss.

Severe cutaneous adverse drug reactions: diagnostic approach and genetic study in a Brazilian case series.

Summary: Background. Severe cutaneous adverse reactions (SCAR) are potentially fatal reactions. Genetic predisposition is involved in their pathogenesis related to drugs and ethnicities, however in a mixed population these relationships are still unknown. The aim of this study was to describe phenotypes, suspect drugs and HLA-alleles related to SCAR, identified by a systematized approach in a Brazilian case series. Methods. Patients who were diagnosed with SCAR between March 2011 and July 2019 at our university hospital were included. European Network for Drug Allergy (ENDA) questionnaire was used to collect clinical and laboratory data and algorithms for assessment of drug causality were applied. Socio-demographic variables included age, gender and skin color/ethnicity. Drug patch tests (DPT) and HLA-A, -B, -DRB1 typing were carried out. Results. A total of 74 patients were included: 36 (48.64%) with SJS/TEN, 32 (43.24%) DRESS/DIHS, 3 (4.05%) AGEP, 2 (2.70%) overlap(DRESS/SJS and DRESS/AGEP) and 1 (1.35%) GBFDE. The median age was31.5 years (IQR = 14-52.25), most were female (n = 44/59.46%) and brown (n = 38/51.35%). Anticonvulsants (n = 32/43.24%) were the largest group involved and antibiotics (n = 26/35.13%) were the second most common. Two patients with DRESS died during the acute phase. Positive DPT were shown only in anticonvulsant associated DRESS. HLA related to abacavir, allopurinol and carbamazepine were identified. Conclusions. A systematized approach allowed the phenotypic characterization of SCAR. The HLA-A*31:01, B*57:01 and B*58:01 alleles were identified, reinforcing the causality in SCAR by CBZ, ABC and ALLO in the Brazilian population.

Genomic ancestry and metabolic syndrome in individuals with type 1 diabetes from an admixed population: a multicentre, cross-sectional study in Brazil.

AIMS: To evaluate the relationship between self-reported colour-race, genomic ancestry, and metabolic syndrome in an admixed Brazilian population with type 1 diabetes. METHODS: We included 1640 participants with type 1 diabetes. The proportions of European, African and Amerindian genomic ancestries were determined by 46 ancestry informative markers of insertion deletion. Two different sets of analyses were performed to determine whether self-reported colour-race and genomic ancestry were predictors of metabolic syndrome. RESULTS: Metabolic syndrome was identified in 29.8% of participants. In the first model, the factors associated with metabolic syndrome were: female gender (odds ratio 1.95, P < 0.001); diabetes duration (odds ratio 1.04, P < 0.001); family history of type 2 diabetes (odds ratio 1.36, P = 0.019); and acanthosis nigricans (odds ratio 5.93, P < 0.001). Colour-race was not a predictive factor for metabolic syndrome. In the second model, colour-race was replaced by European genomic ancestry. The associated factors were: female gender (odds ratio 1.95, P < 0.001); diabetes duration (odds ratio 1.04, P < 0.001); family history of type 2 diabetes (odds ratio 1.39, P = 0.011); and acanthosis nigricans (odds ratio 6.12, P < 0.001). Physical exercise (≥3 times a week) was a protective factor (odds ratio 0.77, P = 0.041), and European genomic ancestry was not associated with metabolic syndrome but showed an odds ratio of 1.77 (P = 0.05). CONCLUSIONS: Although a higher level of European genomic ancestry was observed among participants with metabolic syndrome in the univariate analysis, this association did not persist after multivariable adjustments. Further prospective studies in other highly admixed populations remain necessary to better evaluate whether the European ancestral component modulates the development of metabolic syndrome in type 1 diabetes.

Recruit fitness and police academy performance: a prospective validation study.

BACKGROUND: Police academies need fit recruits to successfully engage in training activities. In a previous retrospective study, we documented that recruits with poor fitness at entry to the academy had significantly lower graduation rates, and we also suggested evidence-based entry-level fitness recommendations. AIMS: To validate our findings in a prospective cohort of police recruits. METHODS: Recruits entering Massachusetts municipal police academies during 2015-16 were followed prospectively until they dropped out, failed or successfully graduated their academy classes. Entry-level fitness was quantified at the start of each training class using: body composition, push-ups, sit-ups, sit-and-reach and 1.5-mile run time. The primary outcome of interest was the odds of failure (not successfully graduating from an academy). We used logistic regression to assess the probability of not graduating, based on entry-level fitness. RESULTS: On average, successful graduates were leaner and possessed better overall entry-level fitness. After adjusting for age, gender and body mass index, several fitness measures were strongly associated with academy failure: fewer sit-ups completed (OR 9.6 (95% CI 3.5-26.3) (≤15 versus 41-60)); fewer push-ups completed (OR 6.7 (95% CI 2.5-17.5) (≤20 versus 41-60)); and slower run times (OR 18.4 (95% CI 6.8-50.2) (1.5 miles in > 15 min 20 s versus 10 min 37 s to 12 min 33 s)). The prospective study results supported previously suggested minimum entry-level fitness (95% graduation rate) and target (98% graduation rate) recommendations. CONCLUSIONS: Push-ups completed and 1.5-mile run time at police academy entry were successfully validated as predictors of successful academy graduation, while sit-ups were also a strong independent predictor in the prospective study.

Lower levels of dehydroepiandrosterone sulfate are associated with more advanced liver fibrosis in chronic hepatitis C.

Chronic infection with the hepatitis C virus induces liver fibrosis, but it is unknown why some patients progress to advanced fibrosis while others remain with mild disease. Recently, an inverse association between serum levels of dehydroepiandrosterone sulphate (DHEA-S) and liver fibrosis in patients with nonalcoholic fatty liver disease was described, and it was postulated that dehydroepiandrosterone (DHEA) has antifibrotic effects. Our aim was to compare serum DHEA-S levels with liver fibrosis in hepatitis C patients. We collected serum samples from hepatitis C patients at the same day they underwent a liver biopsy. S-DHEA was compared to different stages of fibrosis. Binary logistic regression models were applied to evaluate independent variables associated to fibrosis. We included 287 patients (43.9% male). According to fibrosis stages 0, 1, 2, 3 and 4, median serum DHEA-S levels were 103 (26-462), 73 (5-391), 46 (4-425), 35 (6-292) and 28 (2-115) µg/dL, respectively (P < .001). Median serum DHEA-S levels were 74 (5-462) vs 36 (2-425) µg/dL for mild (F0-1) vs significant (F2-4) fibrosis, respectively (P < .001). Median serum DHEA-S levels were 64 (4-462) vs 31 (2-292) µg/dL for non advanced (F0-2) vs advanced fibrosis (F3-4), respectively (P < .001). The same association was found when the subgroup of HCV patients with and without steatosis or steatohepatitis was analysed. The association between lower DHEA-S levels and advanced fibrosis was independent of age, gender, diabetes mellitus, obesity and steatosis. Lower circulating DHEA-S levels are associated with more advanced stages of liver fibrosis in hepatitis C patients.

Health-related physical fitness and quality of life in men with congenital hypogonadotropic hypogonadism.

Congenital hypogonadotropic hypogonadism is a rare disorder characterised by impaired testosterone secretion since birth, and represents a valuable model for studying the effects of testosterone replacement therapy (TRT) in humans. This cross-sectional study aimed to investigate all health-related physical fitness (HRPF) components and quality of life in a series of eight men with hypogonadotropic hypogonadism under regular TRT. The study group was compared to a control group of 16 healthy subjects paired for age, body mass index and physical activity. Body composition, aerobic capacity, muscular strength and endurance, and joint flexibility were evaluated in two different 7-day interval time points, based on the pharmacokinetics of testosterone in the hypogonadal group. Quality of life was assessed by the WHOQOL-brief questionnaire. Both groups had similar performances in all HRPF components evaluated, independently of plasma testosterone levels (p > .05). Quality of life was also similar in the four domains analysed (p > .05). The results of this pilot study suggest that regular testosterone replacement was efficient in providing HRPF and quality of life in a series of congenitally hypogonadal men to levels like those observed in healthy men. In addition, acute fluctuations in plasma testosterone did not correlate with changes in muscle strength and endurance.

Killer cell immunoglobulin-like receptor (KIR) gene diversity in a population naturally exposed to malaria in Porto Velho, Northern Brazil.

Killer cell immunoglobulin-like receptors (KIR) are expressed mainly in natural killer cells and specifically recognize human leukocyte antigen (HLA) class I molecules. The repertoire of KIR genes and KIR-HLA pairs is known to play a key role in the susceptibilities to and the outcomes of several diseases, including malaria. The aim of this study was to investigate the distribution of KIR genes, KIR genotypes and KIR-HLA pair combinations in a population naturally exposed to malaria from Brazilian Amazon. All 16 KIR genes investigated were present in the studied population. Overall, 46 KIR genotypes were defined. The two most common genotypes in the Porto Velho communities, genotypes 1 and 2, were present at similar frequencies as in the Americas. Principal component analysis based on the frequencies of the KIR genes placed the Porto Velho population closer to the Venezuela Mestizos, USA California hispanic and Brazil Paraná Mixed in terms of KIR gene frequencies. This analysis highlights the multi-ethnic profile of the Porto Velho population. Most of the individuals were found to have at least one inhibitory KIR-HLA pair. Seventy-five KIR-HLA pair combinations were identified. The KIR-2DL2/3_HLA-C1, KIR3DL1_HLA-Bw4 and KIR2DL1_HLA-C2 pairs were the most common. There was no association between KIR genes, KIR genotypes or KIR-HLA pair combinations and malaria susceptibility in the studied population. This is the first report on the distribution of KIR and known HLA ligands in the Porto Velho population. Taken together, these results should provide baseline information that will be relevant to population evolutionary history, malaria and other diseases studies in populations of the Brazilian Amazon.

A possible relationship of human leucocyte antigens with psoriasis vulgaris and geographic tongue.

BACKGROUND: Geographic tongue (GT) is the most frequent oral lesion in psoriatic patients (PP), and genetic involvement in these conditions has been described. The association of psoriasis with GT is still not clear, and the study of human leucocyte antigen (HLA) may help clarify this relation. OBJECTIVE: The aim of this study was to investigate the association of HLA alleles with psoriasis vulgaris and GT. METHODS: Fifty-eight Brazilian PP, 29 GT patients and 125 healthy controls individuals were selected. Information on demographic and clinical characteristics was collected. All patients underwent an oral examination and blood collection for HLA typing. RESULTS: HLA-A did not show significant differences in frequencies among the groups. HLA-B*57 allele was more frequently found in PP and was not found in GT. HLA-B*58 allele was more frequently found in GT. HLA-C*06 and -C*18 alleles were associated with psoriasis. No significant differences in HLA-DRB1 and HLA-DQB1 were observed. CONCLUSION: HLA-B*58 was associated with GT and HLA-B*57 was possibly associated with psoriasis. This suggested that some GT cases may represent true oral psoriasis and some may represent only GT. Therefore, it is necessary to make this distinction and increase our sample size to improve the correct diagnosis and treatment of these conditions.

Health-related physical fitness in middle-aged men with and without metabolic syndrome.

AIM: Objective of the study was to compare health-related physical fitness (HRPF) between men with and without metabolic syndrome (MS) and to evaluate the risk of being unfit associated with MS. METHODS: The study included 79 middle-aged civil servant men (46.2 ± 8.4 years) who underwent a physical annual evaluation to access HRPF as follows: BMI; cardiorespiratory fitness by Ebbeling test (VO(2max)), flexibility by sit-and-reach test (SRT), muscular strength by handgrip test (HDT) and vertical jump test (VJT) and muscular endurance by push-up test (PUT). MS was defined by the ATP III (2009) criteria. Comparisons were performed with the Mann-Whitney test and univariate General Linear Model was used for age-adjusted analysis. Odds ratio (OR-95% CI) was calculated to evaluate the odds of the MS group to be unfit and the odds of having MS according to the HRPF levels. RESULTS: Nineteen volunteers (24.1%) with MS were identified. After age adjustment, VO(2max) and BMI were significantly different in the MS group than in the non-MS group: 39.7 vs. 44.8 mL.kg-1.min⁻¹ and 29.4 vs. 25.7 kg/m² (P<0.05) and PUT tended to be lower in men with MS (16 vs. 21 repetition; P=0.06). Blood pressure ≥ 130/85 mmHg was the most prevalent MS criterion, associated with lower VO(max) (40.3 vs. 45.6 mL.kg⁻¹.min⁻¹) and SRT (22.2 vs. 28 cm), and higher BMI (28.9 vs. 25.3 kg/m²) (P<0.05). The OR of being unfit for VO2max and BMI in the MS group were 6.5 (1.9-22.6) and 5.7 (1.2-26.8). The odds of having MS increased by 23% (3-45%) for each BMI unit increase, irrespectively to age. CONCLUSION: MS group showed lower VO(2max), PUT, higher BMI and a greater risk of being unfit compared to the non-MS one. The proportion of MS was 3.4-fold higher within those with lower VO(2max). Small reductions on BMI may produce significant decrease on MS prevalence.

A 7-month multi-disciplinary healthy lifestyle intervention effectively improved cardiometabolic risk profile of firefighters.

OBJECTIVE: We investigated the effect of a 7-month healthy lifestyle intervention on cardiometabolic risk factors (CMRF) among male career military firefighters (FFs). METHODS: 49 FFs participated in a 7-month workplace multi-disciplinary healthy lifestyle intervention designed to reduce CMRF through exercise, diet, and improved sleep. Medical assessments, accelerometry, and surveys at the beginning and end determined program effectiveness. RESULTS: At the end of the intervention period, there was a significant improvement in measures of body composition and blood glucose. The prevalence of hypertension also decreased significantly (p < 0.01). The 57% of participants who fully adhered to the program had significantly greater improvements across multiple CMRF. Participants increased their physical activity and improved their diet following the intervention. CONCLUSION: This healthy lifestyle intervention was effective in changing behavior and lowering cardiometabolic risk among FFs.

Ethnicity and route of HCV infection can influence the associations of HLA with viral clearance in an ethnically heterogeneous population.

Approximately 20% of hepatitis C virus (HCV) infected individuals clear the virus. Host factors that influence the course of HCV infection are still under investigation, and the data on the association of human leukocyte antigen (HLA) alleles and HCV clearance are scarce and controversial. The aims of this study were to investigate whether HLA alleles are associated with clearance of HCV infection in a highly admixed Brazilian population and whether these associations could be influenced by ethnicity and route of infection. HLA-A, -B, -C, -DRB1 and -DQB1 genotyping were performed in 135 HCV-infected Brazilian patients among which 45 cleared HCV infection (cases) and 90 had persistent viral infection (controls). Controls were matched by sex, ethnicity (withes and non-whites) and route of infection (high infectious dose or low infectious dose). No significant association was identified between HLA alleles and the outcome of HCV infection when analyzing the sample as a single group. However, a new protective association of HLA-DQB1*04 (P = 0.006; P(c) = 0.030) and a rarely described association of HLA-DRB1*08 (P = 0.004; P(c) = 0.048) were found only among white patients. The DRB1*11 allele, previously reported in homogeneous population, was associated with HCV clearance (P = 0.020) only among patients with expected high-dose exposure. These findings confirm the influence of ethnicity on the associations of HLA with spontaneous viral clearance of HCV infection and emphasize the possible influence of route of infection in this process.

Morphometry of the pituitary gland and hypothalamus in long-term survivors of childhood trauma.

PURPOSE: Chronic pituitary dysfunction is increasingly recognized as a sequela of traumatic brain injury (TBI). Our aim was to rule out any late morphometric changes of the pituitary gland and hypothalamus in survivors of TBI during childhood requiring intensive care. METHODS: We assessed morphometric abnormalities of the sella region and hypothalamus in patients who sustained TBI during childhood. The patients showed no clinical hormonal dysfunction at the acute phase and pituitary hormone levels at the time of our study were within normal limits. From the 18 enrolled patients in the magnetic resonance study, five were removed due to morphological changes or anatomical variations. We studied the MRI of 13 male survivors (mean age 27 years, mean time after trauma 20 years) and compared them to 13 male control subjects who were matched in terms of age (mean age, 26 years), education and ethnicity. Analyses of the pituitary gland and sella on a midsagittal T2- and T1-weighted image were performed. We used voxel-based morphometry (VBM), an unbiased MRI morphometric method to investigate hypothalamic region in this group of patients. RESULTS: There was only a trend towards a reduced pituitary gland width in the patient group compared to controls. However, no significant morphological and morphometric abnormality was seen and VBM showed no hypothalamic grey matter loss. CONCLUSION: In the absence of hormonal dysfunction, no persisting morphometric changes of the pituitary gland and hypothalamus were seen in survivors of childhood TBI requiring intensive care.

Report from the killer immunoglobulin-like receptor (KIR) anthropology component of the 15th International Histocompatibility Workshop: worldwide variation in the KIR loci and further evidence for the co-evolution of KIR and HLA.

The killer immunoglobulin-like receptor (KIR) anthropology component of the 15th International Histocompatibility Workshop (IHIWS) sought to explore worldwide population variation in the KIR loci, and to examine the relationship between KIR genes and their human leukocyte antigen (HLA) ligands. Fifteen laboratories submitted KIR genotype and HLA ligand data in 27 populations from six broad ethnic groups. Data were analyzed for correlations between the frequencies of KIR and their known HLA ligands. In addition, allelic typing was performed for KIR2DL2 and 3DL1 in a subset of populations. Strong and significant correlations were observed between KIR2DL2, 2DL3 genotype frequencies and the frequency of their ligand, HLA-C1. In contrast, only weak associations were seen for 3DL1, 3DS1 and the HLA-Bw4 ligand. Although some aspects of the correlations observed here differ from those reported in other populations, these data provide additional evidence of linked evolutionary histories for some KIR and HLA loci. Investigation of allele-level variation for the B haplotype locus KIR 2DL2 showed that two alleles, *001 and *003, predominate in all populations in this study. Much more allelic variation was observed for the A haplotype locus 3DL1, with several alleles observed at moderate frequencies and extensive variation observed between populations.

Decrease in oxidative stress and histological changes induced by physical exercise calibrated in rats with osteoarthritis induced by monosodium iodoacetate.

OBJECTIVE: The purpose of this study was to evaluate the effects of impact exercise on the joint cartilage of rats with osteoarthritis (OA) induced by monosodium iodoacetate (MIA). METHODS: Eighteen male rats were divided into three groups of six animals each: control, OA, and OA plus exercise (OAE). The OAE group trained on a treadmill for 8 weeks. Afterward, the right joints of the animals were washed with saline solution and joint lavage was used for biochemical analyses of myeloperoxidase (MPO) and enzyme superoxide dismutase (SOD) activities and total thiol content. The same limb provided samples of the articular capsule for analyses of MPO activity and total thiol content. The left joint was used for histological analysis. RESULTS: Our results indicate that MPO activity was increased in both OA groups in the lavage as well as the articular capsule, regardless of exercise status. SOD activity was increased in animals with OA, especially in the animals that had run on the treadmill. On the other hand, thiol content in the articular capsule and joint lavage decreased in the OA group, while the OAE group had values similar to those of the control group. The histological data indicate that animals that were submitted to running exercise showed a higher preservation rate of proteoglycan content in the superficial and intermediate areas of the joint cartilage. CONCLUSION: Our results show that physical training contributes to the preservation of joint cartilage in animals with OA and to increase the defense mechanism against oxidative stress.

Evaluations of bioantioxidants in cryopreservation of umbilical cord blood using natural cryoprotectants and low concentrations of dimethylsulfoxide.

Transplantation using hematopoietic stem cells from umbilical cord blood (UCB) is a life-saving treatment option for patients with select oncologic diseases, immunologic diseases, bone marrow failure, and others. Often this transplant modality requires cryopreservation and storage of hematopoietic stem cells (HSC), which need to remain cryopreserved in UCB banks for possible future use. The most widely used cryoprotectant is dimethylsulfoxide (Me(2)SO), but at 37 degrees C, it is toxic to cells and for patients, infusion of cryopreserved HSC with Me(2)SO has been associated with side effects. Freezing of cells leads to chemical change of cellular components, which results in physical disruption. Reactive oxygen species (ROS) generation also has been implicated as cause of damage to cells during freezing. We assessed the ability of two bioantioxidants and two disaccharides, to enhance the cryopreservation of UCB. UCB was processed and subjected to cryopreservation in solutions containing different concentrations of Me(2)SO, bioantioxidants and disaccharides. Samples were thawed, and then analysed by: flow cytometry analysis, CFU assay and MTT viability assay. In this study, our analyses showed that antioxidants, principally catalase, performed greater preservation of: CD34+ cells, CD123+ cells, colony-forming units and cell viability, all post-thawed, compared with the standard solution of cryopreservation. Our present studies show that the addition of catalase improved the cryopreservation outcome. Catalase may act on reducing levels of ROS, further indicating that accumulation of free radicals indeed leads to death in cryopreserved hematopoietic cells.