Adaptive genetic changes related to haemoglobin concentration in native high-altitude Tibetans.

NEW FINDINGS: What is the topic of this review? Tibetans have genetic adaptations that are hypothesized to underlie the distinct set of traits they exhibit at altitude. What advances does it highlight? Several adaptive signatures in the same genomic regions have been identified among Tibetan populations resident throughout the Qinghai-Tibetan Plateau. Many highland Tibetans exhibit a haemoglobin concentration within the range expected at sea level, and this trait is associated with putatively adaptive regions harbouring the hypoxia-inducible factor pathway genes EGLN1, EPAS1 and PPARA. Precise functional variants at adaptive loci and relationships to physiological traits, beyond haemoglobin concentration, are currently being examined in this population. Some native Tibetan, Andean and Ethiopian populations have lived at altitudes ranging from 3000 to >4000 m above sea level for hundreds of generations and exhibit distinct combinations of traits at altitude. It was long hypothesized that genetic factors contribute to adaptive differences in these populations, and recent advances in genomics provide evidence that some of the strongest signatures of positive selection in humans are those identified in Tibetans. Many of the top adaptive genomic regions highlighted thus far harbour genes related to hypoxia sensing and response. Putatively adaptive copies of three hypoxia-inducible factor pathway genes, EPAS1, EGLN1 and PPARA, are associated with sea-level range, rather than elevated, haemoglobin concentration observed in many Tibetans at high altitude, and recent studies provide insight into some of the precise adaptive variants, timing of adaptive events and functional roles. While several studies in highland Tibetans have converged on a few hypoxia-inducible factor pathway genes, additional candidates have been reported in independent studies of Tibetans located throughout the Qinghai-Tibetan Plateau. Various aspects of adaptive significance have yet to be identified, integrated, and fully explored. Given the rapid technological advances and interdisciplinary efforts in genomics, physiology and molecular biology, careful examination of Tibetans and comparisons with other distinctively adapted highland populations will provide valuable insight into evolutionary processes and models for both basic and clinical research.

Genetic variation in South Indian castes: evidence from Y-chromosome, mitochondrial, and autosomal polymorphisms.

BACKGROUND: Major population movements, social structure, and caste endogamy have influenced the genetic structure of Indian populations. An understanding of these influences is increasingly important as gene mapping and case-control studies are initiated in South Indian populations. RESULTS: We report new data on 155 individuals from four Tamil caste populations of South India and perform comparative analyses with caste populations from the neighboring state of Andhra Pradesh. Genetic differentiation among Tamil castes is low (RST = 0.96% for 45 autosomal short tandem repeat (STR) markers), reflecting a largely common origin. Nonetheless, caste- and continent-specific patterns are evident. For 32 lineage-defining Y-chromosome SNPs, Tamil castes show higher affinity to Europeans than to eastern Asians, and genetic distance estimates to the Europeans are ordered by caste rank. For 32 lineage-defining mitochondrial SNPs and hypervariable sequence (HVS) 1, Tamil castes have higher affinity to eastern Asians than to Europeans. For 45 autosomal STRs, upper and middle rank castes show higher affinity to Europeans than do lower rank castes from either Tamil Nadu or Andhra Pradesh. Local between-caste variation (Tamil Nadu RST = 0.96%, Andhra Pradesh RST = 0.77%) exceeds the estimate of variation between these geographically separated groups (RST = 0.12%). Low, but statistically significant, correlations between caste rank distance and genetic distance are demonstrated for Tamil castes using Y-chromosome, mtDNA, and autosomal data. CONCLUSION: Genetic data from Y-chromosome, mtDNA, and autosomal STRs are in accord with historical accounts of northwest to southeast population movements in India. The influence of ancient and historical population movements and caste social structure can be detected and replicated in South Indian caste populations from two different geographic regions.

Genetic similarities within and between human populations.

The proportion of human genetic variation due to differences between populations is modest, and individuals from different populations can be genetically more similar than individuals from the same population. Yet sufficient genetic data can permit accurate classification of individuals into populations. Both findings can be obtained from the same data set, using the same number of polymorphic loci. This article explains why. Our analysis focuses on the frequency, omega, with which a pair of random individuals from two different populations is genetically more similar than a pair of individuals randomly selected from any single population. We compare omega to the error rates of several classification methods, using data sets that vary in number of loci, average allele frequency, populations sampled, and polymorphism ascertainment strategy. We demonstrate that classification methods achieve higher discriminatory power than omega because of their use of aggregate properties of populations. The number of loci analyzed is the most critical variable: with 100 polymorphisms, accurate classification is possible, but omega remains sizable, even when using populations as distinct as sub-Saharan Africans and Europeans. Phenotypes controlled by a dozen or fewer loci can therefore be expected to show substantial overlap between human populations. This provides empirical justification for caution when using population labels in biomedical settings, with broad implications for personalized medicine, pharmacogenetics, and the meaning of race.

Human population genetic structure and diversity inferred from polymorphic L1(LINE-1) and Alu insertions.

BACKGROUND/AIMS: The L1 retrotransposable element family is the most successful self-replicating genomic parasite of the human genome. L1 elements drive replication of Alu elements, and both have had far-reaching impacts on the human genome. We use L1 and Alu insertion polymorphisms to analyze human population structure. METHODS: We genotyped 75 recent, polymorphic L1 insertions in 317 individuals from 21 populations in sub-Saharan Africa, East Asia, Europe and the Indian subcontinent. This is the first sample of L1 loci large enough to support detailed population genetic inference. We analyzed these data in parallel with a set of 100 polymorphic Alu insertion loci previously genotyped in the same individuals. RESULTS AND CONCLUSION: The data sets yield congruent results that support the recent African origin model of human ancestry. A genetic clustering algorithm detects clusters of individuals corresponding to continental regions. The number of loci sampled is critical: with fewer than 50 typical loci, structure cannot be reliably discerned in these populations. The inclusion of geographically intermediate populations (from India) reduces the distinctness of clustering. Our results indicate that human genetic variation is neither perfectly correlated with geographic distance (purely clinal) nor independent of distance (purely clustered), but a combination of both: stepped clinal.

Local adaptation and population differentiation at the interleukin 13 and interleukin 4 loci.

A 25.6 kb region at chromosome 5q31, covering the entire human interleukin 13 (IL-13) and interleukin 4 (IL-4) genes, has been reported to be associated with bronchial asthma. We have examined nucleotide variations at this locus in African, European American, and Japanese populations, using 120 diallelic variants. A block of strong linkage disequilibrium (LD) (mid R:D'mid R:>0.7) spans a 10 kb region containing IL-4 in European American and Japanese populations, and is present but less clear in African samples. Two major haplotypes at IL-4 account for >80% of haplotypes in European Americans and Japanese. These haplotypes are common and quite diverged from each other and the ancestral haplotype, resulting in highly significant deviations from neutrality. F(ST) statistics show that European American and Japanese populations are unusually distinct at the IL-4 locus. The most common haplotype in the European American population is much less common in the Japanese population, and vice versa. This implies that natural selection has acted on IL-4 haplotypes differently in different populations. This selected variation at IL-4 may account for some genetic variance underlying susceptibility to asthma and other allergic diseases. The strong LD observed in the IL-4 region may allow more efficient disease-association studies using this locus.

Apoptosis: an introduction for the endodontist.

Apoptosis plays an important role in many aspects of endodontics, yet there is a paucity of information in this regard in the endodontic literature. Apoptosis is a single deletion of scattered cells by fragmentation into membrane-bound particles that are phagocytosed by other cells. It is a key process in the embryological development of the tooth, periodontal ligament and supporting oral tissue in the progression of oral disease, bone resorption, immunological response and inflammation, and in wound healing and certain pharmacological effects. The understanding of the ability of clinical materials to induce or inhibit apoptosis and the investigation of apoptosis as it relates to the pathogenesis of pulpal and periradicular pathology may eventually lead to new treatment approaches for the endodontist. The purpose of this review is to familiarize the clinical endodontist with current knowledge on apoptosis as it relates to the pulp and periradicular tissues.

Comparison of Diaket and MTA when used as root-end filling materials to support regeneration of the periradicular tissues.

AIM: The objective of this study was to evaluate on a comparative basis the potential for mineral trioxide aggregate (MTA) and Diaket to promote periradicular tissue regeneration when used as surgical root-end filling materials. METHODOLOGY: Seven dogs weighing between 15 and 25 kg were anesthetized prior to having the root canals of their mandibular premolars accessed, cleaned, shaped and obturated. Coronal access cavities were restored with IRM. Surgical access to the root ends was established and the root ends were resected and prepared with ultrasonic tips. Root-end fillings of either MTA or thickly mixed Diaket were randomly assigned to the preparations. Reflected tissues were repositioned and sutured with 4-0 vicryl sutures. Sixty days postsurgery, the animals were killed, perfused with 10% neutral buffered formalin and the third and fourth premolars removed in block sections. The specimens were demineralized and sectioned at 6-microm intervals for histological assessment. Sections were stained with either haematoxylin and eosin or Gomori's one step trichrome stain and examined under the light microscope. All evaluations were made by two calibrated examiners and gradings were scored based on established criteria. The raw data was evaluated statistically using anova after adjusting for the animal block effect. RESULTS: Statistical evaluation indicated that there were no statistical differences between the observed regenerative responses of the tissues to the two root-end filling materials. CONCLUSIONS: Both Diaket and MTA can support almost complete regeneration of the periradicular periodontium when used as root-end filling materials in periradicular surgery on noninfected teeth.

One-visit apexification: technique for inducing root-end barrier formation in apical closures.

Numerous procedures and materials have been utilized to induce root-end barrier formation. Mineral trioxide aggregate (MTA) was introduced to dentistry as a root-end filling material. It has been advocated for filling root canals, repairing perforations, pulp capping, and root-end induction. Mineral trioxide aggregate reacts with tissue fluids to form a hard tissue apical barrier. As a result, MTA shows promise as a valuable material for use in one-visit apexification treatment, primarily for treating immature teeth with necrotic pulps.

Interleukin-1beta production in periradicular lesions in a human immunodeficiency virus/acquired immune deficiency syndrome model compared with a noninfected host.

This study elucidates the role of interleukin-1 (IL-1) in developing periradicular lesions in immunocompetent and immunocompromised (human immunodeficiency virus/acquired immune deficiency syndrome) hosts. Eight cats were immunosuppressed with steroids before infection with feline immunodeficiency virus (FIV). Eight uninoculated cats served as controls. Periradicular lesions were induced around the canine teeth. At 1 and 4 wk periradicular exudate was sampled via the root canals. IL-1beta levels were measured with ELISA. Data were analyzed using the Mann-Whitney U test and the Wilcoxon signed rank test. Statistically significant differences existed in cytokine levels between the FIV and non-FIV groups (p < 0.001). Cytokines were below detectable levels in the FIV group. A significant decrease in IL-1beta levels at 4 wk compared with 1 wk occurred in the non-FIV group (p < 0.05). In conclusion decreased IL-1beta production was obtained in the FIV group. In the non-FIV group decreases in IL-1beta levels were encountered at the chronic stage of the periradicular lesion compared with the acute stage.

Feline immunodeficiency virus model to study human immunodeficiency virus/acquired immune deficiency syndrome conditions.

This study was designed to induce rapid progression of the feline immunodeficiency virus (FIV) infection in cats. Predictably inducing the FIV disease state in the cat would yield an excellent tool to study endodontic disease processes under immunosuppressed conditions. Eight cats were immunosuppressed with steroids before infection with FIV. Another eight cats, age- and sex-matched littermates, served as uninoculated seronegative controls. Complete blood counts were taken for 10 mo in the FIV group, and 10 wk in the control group, including lymphocyte subsets. ELISAs were used to detect FIV infection. Statistical analysis was performed with generalized estimating equation models. All cats were positive at one point in time. The FIV group had significantly lower peripheral blood CD4+ counts compared with the control group. Therefore the FIV model presented gives the desired outcome and simulates what occurs in human immunodeficiency virus infection.

Histologic assessment of healing after the use of a bioresorbable membrane in the management of buccal bone loss concomitant with periradicular surgery.

Histological and morphometric assessment of periradicular wound healing was made after the use of a bioresorbable membrane over a buccal dehiscence. The third and fourth premolar teeth of nine dogs were resected and buccal defects created. Teeth were assigned randomly to the membrane or control group. One tooth in each quadrant received a membrane, covering both roots. The other tooth received no further treatment and served as a control. The animals were killed and specimens were assessed at two time periods: 9 wk and 27 wk. The 27-wk membrane group exhibited significantly more (p = 0.004) connective tissue height than the control group or either of the 9-wk groups. The amount of regenerated alveolar bone was significantly greater for the 27-wk membrane group than for the control (p = 0.001) and 9-wk groups. Mean junctional epithelium measurements were significantly greater (p = 0.012) for the control. The use of a bioresorbable membrane enhances bone regeneration when a buccal defect exists at the time of periradicular surgery.

Coronal leakage following three obturation techniques.

AIM: To compare coronal bacterial and India ink leakage in three different obturation techniques with the smear layer having been removed. METHODOLOGY: Seventy extracted single-rooted teeth were instrumented to an apical preparation size 7 Profile Series 29 (Tulsa Dental Products, Tulsa, OK, USA). The smear layer was removed and 20 teeth were randomly obturated with lateral compaction, 20 teeth with vertical compaction, and 20 teeth with Thermafil (Tulsa Dental Products, Tulsa OK, USA). Ten teeth were used for positive (five teeth) and negative (five teeth) controls. Teeth were stored for 90 days in 100% humidity, then subjected coronally to Proteus vulgaris for 21 days to assess bacterial leakage. Following bacterial challenge, India ink was placed coronally for a further 21 days, then scored according to depth of dye leakage. RESULTS: Vertical compaction leaked significantly less than lateral compaction during bacterial challenge. However, when dye was used there were no significant differences. CONCLUSIONS: Bacterial leakage and dye leakage demonstrated considerable variability. The use of a dye following bacterial testing may highlight the failure of experimental devices and vertical root fractures, thus avoiding false positive results found with bacterial testing alone.

Calcific metamorphosis: a challenge in endodontic diagnosis and treatment.

Calcific metamorphosis (CM) is seen commonly in the dental pulp after traumatic tooth injuries and is recognized clinically as early as 3 months after injury. Calcific metamorphosis is characterized by deposition of hard tissue within the root canal space and yellow discoloration of the clinical crown. Opinion differs among practitioners as to whether to treat these cases upon early detection of CM or to observe them until symptoms or radiographic signs of pulpal necrosis are detected. In this review, the clinical, radiographic, and histopathologic appearance of CM is described; a review of the literature is presented to address these issues in an attempt to establish a sound rationale for treatment. Approximately 3.8% to 24% of traumatized teeth develop varying degrees of CM. Studies indicate that of these, approximately 1% to 16% will develop pulpal necrosis. Most of the literature does not support endodontic intervention unless periradicular pathosis is detected or the involved tooth becomes symptomatic. It may be advisable to manage cases demonstrating CM through observation and periodic examination.

The effect of FIV infection on CD4+ and CD8+ counts in periradicular lesions.

AIM: The purpose of this study was to elucidate whether a decrease/increase in T-cell populations is present in the development of periradicular disease in the immunocompromised feline model. METHODOLOGY: Eight cats were immunosuppressed with steroids prior to infection with feline immunodeficiency virus (FIV). Another eight cats, age and sex matched littermates, were monitored and tested at equivalent periods of time and served as uninoculated, seronegative controls. Periradicular lesions were induced using local bacterial inoculations into the pulp of the canine teeth and assessed after one- and four-week periods, corresponding to the acute and chronic stages of the periradicular disease. Block sections were obtained and specimens were prepared for H & E and immunohistochemical staining for CD4+ and CD8+ receptors. Cells were quantified using a computer imaging system and data analysed using generalized estimating equation (GEE) regression models. RESULTS: Significantly lower CD4+ counts and CD4+/ CD8+ ratios were observed at all time periods in the periradicular region of the FIV group (P = 0.0006). No significant difference in CD8+ counts was observed between the two groups. In both groups there was a significant difference in the CD4+ counts between one week and baseline, and 1 week and 4 weeks. There was no significant difference between baseline and 4 weeks for either group. CONCLUSION: FIV infection reflected decreased CD4+ counts at the periradicular level, however, inflammation and progression of the lesion, appeared to be comparable to the non-immunocompromised controls.

Serious fireworks-related eye injuries.

OBJECTIVE: To analyze and compare epidemiological and clinical information on serious fireworks-related eye injuries from two affiliates of the United States Eye Injury Registry. METHOD: Retrospective review. RESULTS: In the Eye Injury Registry of Alabama (EIRA) database, 185 of the 4150 injuries (4.4%) were caused by fireworks. In the Hungarian Eye Injury Registry database, only two of the 1245 cases (0.1%, p=0.000001) were fireworks-related. In the EIRA, 79% of patients were males and 87% were under 31 years. A bystander was injured in 67% of the cases, being an average of 23 feet away; 39% of bystanders had a final vision < or =19/200. No injured person wore eye protection. Bottle rockets caused 80% of the 185 injuries. Overall, 20% of eyes had <5/200 final visual acuity. Twenty-five percent of bottle rocket-injured eyes, compared to 64% of those injured by other devices, had > or =20/40 final vision (p=0. 000004). CONCLUSIONS. The rate of fireworks-related serious eye injuries has not decreased in Alabama in the last 16 years; most patients are young males. Since bystanders are at a measurable risk even at a distance of 100 feet, wearing eye protection is recommended to both bystanders and operators. Bottle rockets cause most of the injuries and the more severe ones, and should be the prime target for prevention. The benefit of a strict and enforced legislative ban on private fireworks displays is demonstrated by the much lower incidence figure in Hungary. Such a ban should be considered in other countries where fireworks-related eye injuries are common.

Restoring teeth that are endodontically treated through existing crowns. Part II: Survey of restorative materials commonly used.

OBJECTIVE: A survey was undertaken to categorize the materials used for the restoration of endodontic access openings through complete-coverage crowns after completion of nonsurgical root canal treatment. METHOD AND MATERIALS: The survey package consisted of a cover letter stating instructions, rationale, and purpose for the questionnaire, a questionnaire of 8 short-answer questions, and a stamped, self-addressed envelope. A randomized sample of active dentists (300 general practitioners, 300 prosthodontists, and 300 endodontists), was selected. Collected data were analyzed with the chi-square analysis. RESULTS: Most general practitioners (93%), endodontists (61%), and prosthodontists (75%) reported that they frequently or always permanently restore teeth after nonsurgical root canal treatment. Empress was the all-ceramic system used most commonly by prosthodontists (42%) and general practitioners (38%). A statistically significant difference in restorative material preference was found (P < 0.0001), depending on the type of crown used. CONCLUSION: Amalgam alone and in combination with bonding agents are materials of choice for restoration of access openings through all-metal complete crowns, while resin composite is the choice for all types of complete crowns involving porcelain. Endodontists preferred "other" materials.

Restoring teeth that are endodontically treated through existing crowns. Part I: Survey of pulpal status on access.

OBJECTIVE: The purpose of this study was to identify the pulpal findings encountered by practitioners when accessing complete-coverage crowns that require nonsurgical root canal treatment and the relevance of coronal leakage to the success of the RTC. METHOD AND MATERIALS: The survey package consisted of a cover letter stating the instructions, rationale, and purpose for the questionnaire, a questionnaire with 8 short-answer questions, and a stamped, self-addressed envelope. A randomized sample of active dentists (300 general practitioners, 300 prosthodontists, and 300 endodontists) was selected. Collected data were analyzed with the chi-square test. RESULTS: A 60% response rate was obtained. Statistically significant differences were found among the practitioner groups, depending on the question. General practitioners and endodontists obtain access through crowns and maintain these crowns as final restoration significantly more often than do prosthodontists. Practitioners responded that teeth with complete crowns require nonsurgical root canal treatment after 5 to 10 years. CONCLUSION: Respondents believe that leakage must be addressed when endodontic access cavities in artificial crowns are restored after nonsurgical root canal treatment. General practitioners perform nonsurgical root canal treatment more frequently than do prosthodontists. Practitioners indicated that when teeth with complete crowns require nonsurgical root canal treatment, treatment is most often performed 5 to 10 years after placement of the crown.

Effects of calcium hydroxide and transforming [correction of tumor] growth factor-beta on collagen synthesis in subcultures I and V of osteoblasts.

Collagen protein synthesis by osteoblasts is influenced by transforming growth factor-beta (TGF-beta 1) and is essential to bone formation. The effectiveness of TGF-beta 1 depends on efficient delivery of the growth factor to target cells, adequate binding to cell surface receptors, and an optimum environment for promotion of collagen synthesis. The effects of calcium hydroxide (Ca(OH)2), TGF-beta 1, and Ca(OH)2/TGF-beta 1 co-administration on total protein, collagen protein, and noncollagen protein synthesis by early (subculture I) and late (subculture V) osteoblast cultures were tested. TGF-beta 1 significantly increased all protein synthesis in subculture I osteoblasts (p = 0.001; p < 0.001; p = 0.019). Ca(OH)2/TGF-beta 1 co-administration significantly increased total protein and collagen protein levels in subculture I osteoblasts as well (p = 0.048; p = 0.012). TGF-beta 1 increased total protein and collagen protein synthesis significantly in subculture V cells (p = 0.025; p = 0.01). These data indicate that co-administration of Ca(OH)2 and TGF-beta 1 enhances collagen synthesis by osteoblasts and may have implications for the clinical setting.

TGF-beta 1 alone and in combination with calcium hydroxide is synergistic to TGF-beta 1 production by osteoblasts in vitro.

AIM: To examine the effects of calcium hydroxide (Ca(OH)2), transforming growth factor-beta (TGF-beta 1), and Ca(OH)2/TGF-beta 1 coadministration on TGF-beta 1 and interleukin-6 (IL-6) synthesis by early (subculture 1) and late (subculture 5) osteoblast cultures. METHODOLOGY: Early and late cultures were established using bone cells harvested from 21-day-old fetal rat calvaria. Cell cultures of both early and late osteoblasts were divided into four groups: group 1, control; group 2, cells challenged with Ca(OH)2; group 3, cells challenged with TGF-beta 1; and group 4, cells challenged with Ca(OH)2 and TGF-beta 1 in combination. TGF-beta 1 and IL-6 levels for all groups were determined using ELISA methodology. RESULTS: ANOVA and Tukey HS analyses revealed that osteoblasts of groups 3 and 4 significantly increased (P < 0.001) TGF-beta 1 synthesis in both early and late cultures of osteoblasts. IL-6 was not detected in any of the groups considered in this study. CONCLUSIONS: Exogenous TGF-beta 1 has an autocrine effect on cell cultures of osteoblasts. Administration of TGF-beta 1 alone or in combination with Ca(OH)2 increases the synthesis of TGF-beta 1 in osteoblast cultures. Ca(OH)2 and TGF-beta 1 are compatible when placed in a culture of osteoblasts. Ca(OH)2 provides a favourable environment for the anabolic effects of TGF-beta 1.