The analysis of variation of mtDNA hypervariable region 1 suggests that Eastern and Western Pygmies diverged before the Bantu expansion

The Eastern Pygmies from Zaire and Western Pygmies from Cameroon, Congo, and the Central African Republic represent the two principal groups of African Pygmies. In the "recent divergence" hypothesis in which Western Pygmies are thought to be the result of hybridization between the ancestors of Eastern Pygmies and Bantu farmers who penetrated the equatorial belt and came into contact with Pygmies around 2-3 kiloyears ago. On the basis of recent archaeological research in the tropical rain forest, we propose a "pre-Bantu divergence" hypothesis, which posits the separation between the ancestors of Eastern and Western Pygmies earlier than 18 kiloyears ago. In order to test the two hypotheses, we analyzed the variation of the hypervariable region 1 of the mitochondrial DNA in the Mbenzele, Western Pygmies of the Central African Republic, and compared our results with those of previous mtDNA and Y chromosome studies. Distribution, sequence variation, and age of haplogroups along with genetic distances among populations, estimates of divergence times, and simulations based on the coalescent approach were found to be congruent with the pre-Bantu divergence but failed to support the recent divergence hypothesis.     

Microsatellite variation in Central Africa: an analysis of intrapopulational and interpopulational genetic diversity

As a part of a research project on molecular variation in Central Africa, we have analyzed 10 microsatellites (CD4, CSFO, D3S1358, D18S51, D21S11, F13A1, FES, TH01, TPOX, and VWA) in the Bamileke and Ewondo from Cameroon and the Sanga and Mbenzele Pygmies from the Central African Republic (a total of 390 chromosomes). A statistically significant trend towards heterozygote deficiency was detected in the Mbenzele Pygmies. This was established through the use of powerful exact tests for the Hardy-Weinberg equilibrium. A certain degree of isolation and a small effective size may explain this finding. However, the lack of any substantial reduction in allelic diversity in the Mbenzele does not support the possibility that this group has a smaller effective size in evolutionary terms. A possible explanation based on ethnographic studies suggests that the gene flow from non-Pygmies to Pygmies could have been interrupted only in relatively recent times. The analysis of association between genotypes at pairs of independent loci indicates that the level of subheterogeneity is markedly lower in the Bamileke than in other sampled populations. This may be explained by the combined effect of larger population size, more rigid respect of clanic exogamy, and higher matrimonial mobility of the Bamileke. Finally, we have analyzed interpopulational relationships among our sampled populations and other Central African populations. The results are consistent with a previous study of protein loci (Spedini et al. 1999), which suggests the recent history of the Bamileke and Ewondo has led them to aquire a substantial genetic similarity. Furthermore, the Mbenzele Pygmies diverge from Biaka Pygmies, despite their common origin and geographical proximity. This is probably due to the differentiating effect of genetic drift, which is enhanced by the small effective size of Pygmy populations.     

Pygmy locomotion

The hypothesis that Pygmies may differ from Caucasians in some aspects of the mechanics of locomotion was tested. A total of 13 Pygmies and 7 Caucasians were asked to walk and run on a treadmill at 4–12 km · h^–1. Simultaneous metabolic measurements and three-dimensional motion analysis were performed allowing the energy expenditure and the mechanical external and internal work to be calculated. In Pygmies the metabolic energy cost was higher during walking at all speeds (P < 0.05), but tended to be lower during running (NS). The stride frequency and the internal mechanical work were higher for Pygmies at all walking (P < 0.05) and running (NS) speeds although the external mechanical work was similar. The total mechanical work for Pygmies was higher during walking (P < 0.05), but not during running and the efficiency of locomotion was similar in all subjects and speeds. The higher cost of walking in Pygmies is consistent with the allometric prediction for smaller subjects. The major determinants of the higher cost of walking was the difference in stride frequency (+9.45, SD 0.44% for Pygmies), which affected the mechanical internal work. This explains the observed higher total mechanical work of walking in Pygmies, even when the external component was the same. Most of the differences between Pygmies and Caucasians, observed during walking, tended to disappear when the speed was normalized as the Fronde number. However, this was not the case for running. Thus, whereas the tested hypothesis must be rejected for walking, the data from running, do indeed suggest that Pygmies may differ in some aspects of the mechanics of locomotion.     

DNA variation in a 5-Mb region of the X chromosome and estimates of sex-specific/type-specific mutation rates.

We describe a new approach for the study of human genome variation, based on our solid-phase fluorescence chemical mismatch-cleavage method. Multiplex screening rates >/=80 kb/36-lane gels are achieved, and accuracy of mismatch location is within +/-2 bp. The density of differences between DNA from any two humans is sufficiently low, and the estimate of their position is accurate enough, to avoid sequencing of most polymorphic sites when defining their allelic state. Furthermore, highly variable sequences, such as microsatellites, are distinguished easily, so that separate consideration can be given to loci that do and do not fit the definition of infinite mutation sites. We examined a 5-Mb region of Xq22 to define the haplotypes of 23 men (9 Europeans, 9 Ashkenazim, and 5 Pygmies) by reference to DNA from one Italian man. Fifty-eight 1.5-kb segments revealed 102 segregating sites. Seven of these are shared by all three groups, two by Pygmies and Europeans, two by Pygmies and Ashkenazim, and 19 by Ashkenazim and Europeans. Europeans are the least polymorphic, and Pygmies are the most polymorphic. Conserved allelic associations were recognizable within 40-kb DNA segments, and so was recombination in the longer intervals separating such segments. The men showed only three segregating sites in a 16.5-kb unique region of the Y chromosome. Divergence between X- and Y-chromosome sequences of humans and chimpanzees indicated higher male mutation rates for different types of mutations. These rates for the X chromosomes were very similar to those estimated for the X-linked factor IX gene in the U.K. population.     

Increased oxidative damage of RNA in liver injury caused by hepatitis B virus (HBV) infection

To evaluate the urinary levels of 8-oxo-7,8-dihydro-2′deoxyguanosine (8-oxo-dGsn) and 8-oxo-7,8-dihydroguanosine (8-oxo-Gsn) in liver injury patients with hepatitis B virus (HBV) infection and to explore the relationship between urinary 8-oxo-dGsn or 8-oxo-Gsn and degree of liver damage. We enrolled 138 liver injury patients with HBV infection and 169 age- and sex-matched healthy controls in this study. A sensitive and accurate isotope-diluted liquid chromatograph mass spectrometer/mass spectrometer (LC-MS/MS) method was used to measure the urinary levels of 8-oxo-Gsn and 8-oxo-dGsn. Simultaneously, pathological analysis of liver biopsy tissues was carried out, and immunohistochemistry was carried out for 8-oxo-Guo, 8-oxo-dGuo and MTH1 protein in some liver injury tissues. We analysed the correlation between the degrees of inflammation and fibrosis and levels of 8-oxo-Gsn and 8-oxo-dGsn. We also analysed the levels of urinary 8-oxo-Gsn and 8-oxo-dGsn with clinical data of HBeAg, HBsAg, and HBV genotype and detected the levels of plasma aspartate aminotransferase, alanine aminotransferase (AST), platelet, alkaline phosphatase, prothrombin time (PT) and HBV DNA, and calculated the aspartate amino transferase-to–platelet ratio index (APRI) score. Nonparametric correlations were used to evaluate the correlation between 8-oxo-Gsn, 8-oxo-dGsn or APRI and various laboratory biochemical indicators. Results showed that the levels of urinary 8-oxo-Gsn and 8-oxo-dGsn in patients with liver injury were significantly higher than those of healthy controls (both p?p?=?.013, p?=?.026 and p?=?.049). The receiver operating characteristic curves of 8-oxo-Gsn were 0.696 (0.632–0.759) and 0.731 (0.672–0.790) for inflammatory activity and fibrosis, respectively. Patients with higher levels of urinary 8-oxo-Gsn are more likely to have a high degree of fibrosis and urinary 8-oxo-Gsn may have a great potential in assessing liver fibrosis.     

Insights into the demographic history of African Pygmies from complete mitochondrial genomes

Pygmy populations are among the few hunter-gatherers currently living in sub-Saharan Africa and are mainly represented by two groups, Eastern and Western, according to their current geographical distribution. They are scattered across the Central African belt and surrounded by Bantu-speaking farmers, with whom they have complex social and economic interactions. To investigate the demographic history of Pygmy groups, a population approach was applied to the analysis of 205 complete mitochondrial DNA (mtDNA) sequences from ten central African populations. No sharing of maternal lineages was observed between the two Pygmy groups, with haplogroup L1c being characteristic of the Western group but most of Eastern Pygmy lineages falling into subclades of L0a, L2a, and L5. Demographic inferences based on Bayesian coalescent simulations point to an early split among the maternal ancestors of Pygmies and those of Bantu-speaking farmers (～ 70,000 years ago [ya]). Evidence for population growth in the ancestors of Bantu-speaking farmers has been observed, starting ～ 65,000 ya, well before the diffusion of Bantu languages. Subsequently, the effective population size of the ancestors of Pygmies remained constant over time and ～ 27,000 ya, coincident with the Last Glacial Maximum, Eastern and Western Pygmies diverged, with evidence of subsequent migration only among the Western group and the Bantu-speaking farmers. Western Pygmies show signs of a recent bottleneck 4,000-650 ya, coincident with the diffusion of Bantu languages, whereas Eastern Pygmies seem to have experienced a more ancient decrease in population size (20,000-4,000 ya). In conclusion, the results of this first attempt at analyzing complete mtDNA sequences at the population level in sub-Saharan Africa not only support previous findings but also offer new insights into the demographic history of Pygmy populations, shedding new light on the ancient peopling of the African continent.     

Polymorphism and mapping of the IGF1 gene,and absence of association with stature among African Pygmies

Summary Probes detecting restriction fragment length polymorphisms (RFLPs) in the insulinlike growth factor (IGF1) gene were isolated and allele frequencies in different human populations determined. No difference was detected between the distribution of IGF1 alleles in Pygmies versus non-Pygmy black Africans, despite the proposal that a defect in the IGF1 gene might be responsible for Pygmy short stature. This was supported by the absence of a correlation of IGF1 genotype with height in the C.A.R. Pygmies. Polymerase chain reaction (PCR) and direct sequencing failed to demonstrate an alteration in the region upstream the IGF1 start site in Pygmies. Linkage analysis demonstrated that IGF1 is tightly linked to the phenylalanine hydroxylase gene on chromosome 12q22–24.1.     

Limited dispersal in mobile hunter–gatherer Baka Pygmies

Hunter–gatherer Pygmies from Central Africa are described as being extremely mobile. Using neutral genetic markers and population genetics theory, we explored the dispersal behaviour of the Baka Pygmies from Cameroon, one of the largest Pygmy populations in Central Africa. We found a strong correlation between genetic and geographical distances: a pattern of isolation by distance arising from limited parent–offspring dispersal. Our study suggests that mobile hunter–gatherers do not necessarily disperse over wide geographical areas.     

Population genetics of red cell enzymes in Pygmies: a conclusive account

In the course of a long-term research project, three groups of Pygmies and some non-Pygmy Central Africans have been examined for the following red cell enzyme markers: ACP, PGM1, PGM2, PEPA, PEPB, and PEPC, AK, ADA, and PHI. Several other red cell enzymes (ESD, CA1 and CA2, GPT, GLO, and DIA1) have been studied in only some of these groups. This paper reports all the information we obtained, including what we have already published. The following conclusions can be drawn from the whole body of data: (1) Gene patterns of Pygmies are those typical of other Africans (e.g.: lack of ADA2 and AK2 genes, low GPT2 gene frequency, polymorphism of the CA2 locus, and presence at polymorphic frequencies of PEPA2 allele. (2) Superimposed on this African genetic makeup, a number of Pygmy characters were identified, namely, a private polymorphism for the PGM26 Pygmy allele and possibly one for the PEPC2 allele, and particularly high ACPR and low PGM12 gene frequencies. (3) Some markers, especially PGM1 and ACP, turned out also to discriminate efficiently among different groups of Pygmies.     

Endogenous scavenger levels (vitamin E and A) of patients with chronic alcoholic liver disease under different environmental conditions.

Serum level of endogenous scavengers (E and A vitamin) was studied in groups of patients with various chronic alcoholic liver diseases and in a healthy control group on polluted and non-polluted areas. Vitamin levels in patients with chronic liver disease are diminished in comparison to the healthy in general, but mainly in the cirrhotic group. Diminution of vitamin E levels was observed in earlier phase of liver disease than that of vitamin A levels. Patients and healthy control on polluted area showed more expressed diminution of vitamin levels than the same groups on non-polluted area. Free radical parameter (RBC diene conjugate content) and characteristic alcoholic parameters (serum GOT, gamma-GT, cholesterol level and liver GOT, gamma-GT content in biopsy specimen) were used to explain the differences between the same investigated groups on polluted and non-polluted areas. As conclusion can be supposed that industrial pollution of environment has a worsening effect in diseases with free radical mechanism.     

Metabolic Responses to Oral Glucose in the Kalahari Bushmen

The plasma glucose, immunoreactive insulin, and growth hormone levels after a 50-g oral glucose load have been measured in 15 adult Bushmen subjects living in the Kalahari region of Southern Africa. Compared with 10 non-obese white controls, they showed relative glucose intolerance and significantly impaired insulin secretion. Growth hormone responses showed no significant differences between the two groups. Factors such as inadequate or unusual nutrition and stress do not appear to account completely for the abnormalities in carbohydrate metabolism observed in the Bushmen. Of interest are the clinical and hormonal similarities that seem to exist between the Bushmen and the Central African Pygmies.     

Y chromosome DNA polymorphisms in two African populations.

Y chromosome-specific DNA polymorphisms were detected using probe p49f after restriction with TaqI enzyme on samples coming from two African populations: Bantus and Pygmies. All the main TaqI alleles at five Y loci already found in Caucasians are also found in these two populations; 12 of the 16 Caucasian haplotypes were found in these two African populations, and two new haplotypes are Pygmy specific. A proposed phylogeny of the various haplotypes that was derived by using the parsimony criterion established that haplotypes XIII and XVIII, respectively the most frequent one and only one present in Pygmies, are probably ancestral.     

Biological characterization of Campylobacter fetus lipopolysaccharides

Abstract Ten patients with chronic liver disease, seven healthy seropositive individuals with a remote history of rubella, and three patients with acute rubella were examined for serum levels of IgG subclasses and subclass antibodies against rubella virus structural proteins. One patient with AICAH had no detectable total or rubella specific IgG3 or IgG4. The liver disease patients were hypergammaglobulinemic and had greatly raised IgG1 levels. Patients with acute rubella lacked antibodies to the rubella virus E2 protein and showed no IgG4 antibody response. The liver disease patients showed a somewhat weaker IgG4 antibody response against the core (C) protein than healthy controls. However, differences are suggested within the subclasses in antibody reactivity against the individual rubella virus antigens. It is concluded that test systems that discriminate reactivities against individual antigens have to be used for characterization of viral antibody subclass profiles.     

Studies on four restriction fragment length polymorphisms of the type I collagen genes in two Italian populations

Type I collagen, the most abundant of the collagen protein family, is encoded by two genes, COL1A1 and COL1A2. Two random population samples, one from central Italy and one from southern Italy, were studied for 1 restriction fragment length polymorphism (RFLP) of COL1A1 (RsaI) and 3 RFLPs of COL1A2 (EcoRI, RsaI and MspI). A considerable heterogeneity for COL1A1/RsaI was found not only between Italians and English but even among Italians. The potential usefulness of these RFLPs and haplotypes as anthropogenetic markers, particularly in distinguishing Caucasoids from Negroids, has been discussed.     

Postpartal ovarian activity of healthy cows and those affected by subclinical metabolic disorders

The postpartal ovarian activity was studied in healthy cows and in those affected by subclinical fatty liver disease and subclinical primary ketosis. Twenty-six dairy cows, 4–7 years old, with not less than 5000 l milk yield in the preceding lactation, were monitored for the following blood parameters: aspartate-amino-transferase (AST)/glutamicoxalacetic transaminase (GOT), albumin, total protein, total lipid, cholesterol, non-esterified fatty acid (NEFA) and triglyceride levels, glucose and ketones. The ratios albumin : total protein and NEFA : triglyceride were also calculated. Blood samples were collected once a week between days 255 and 260 of gestation and days 1–15 postpartum, and subsequently on the 30th, 45th and 60th days postpartum. Based on the biochemical parameters, the cows were assigned in retrospect to three groups: (1) healthy (n = 5), (2) subclinical fatty liver disease (n = 6), and (3) subclinical primary ketosis (n = 10). The postpartal ovarian activity was assessed by rectal palpation and by milk progesterone levels, determined by radioimmunoassay in samples collected 2–3 times weekly between days 5 and 70 postpartum.As early as 10–18 days postpartum, ovarian activity was evident in healthy cows. This was confirmed by the progesterone profiles indicating three regular ovarian cycles with normal hormone levels within 70 days postpartum. All five healthy cows were detected in estrus and only one failed to conceive after insemination.The six cows with subclinical fatty liver disease had no regular cycles and only a few small follicles were palpable on their ovaries. The milk progesterone fluctuated around the basal level (? 1.6 nmol/l). The first Graafian follicles appeared as late as days 30–35 postpartum and the luteal function commenced only between days 38 and 60, as indicated by palpation of corpora lutea and a concomitant rise in milk progesterone. The progesterone peaks were at the lower physiological limits characteristic of luteal function.The ovarian activity of the ten cows with subclinical primary ketosis was intermediate between that of healthy and fatty liver cows. The first follicles and corpora lutea were somewhat delayed, and the progesterone levels fluctuated considerably during the luteal phase. One cow in this group resumed ovarian inactivity during the 70-day postpartal observation period.     

Energetics of locomotion in African pygmies

The energy cost of walking (Cw) and running (Cr), and the maximal O2 consumption (VO2max) were determined in a field study on 17 Pygmies (age 24 years, SD 6; height 160 cm, SD 5; body mass 57.2 kg, SD 4.8) living in the region of Bipindi, Cameroon. The Cw varied from 112 ml.kg-1.km-1, SD 25 [velocity (v), 4 km.h-1] to 143 ml.kg-1.km-1, SD 16 (v, 7 km.h-1). Optimal walking v was 5 km.h-1. The Cr was 156 ml.kg-1.km-1, SD 14 (v, 10 km.h-1) and was constant in the 8-11 km.h-1 speed range. The VO2max was 33.7 ml.kg-1.min-1, i.e. lower than in other African populations of the same age. The Cr and Cw were lower than in taller Caucasian endurance runners. These findings, which challenge the theory of physical similarity as applied to animal locomotion, may depend either on the mechanics of locomotion which in Pygmies may be different from that observed in Caucasians, or on a greater mechanical efficiency in Pygmies than in Caucasians. The low Cr values observed enable Pygmies to reach higher running speeds than would be expected on the basis of their VO2max.     

Becoming Italian,becoming American: ethnic affinity as a strategy of boundary making

Based on the case study of Albanian immigrant incorporation in a Little Italy in the Bronx, this paper develops the concept of ethnic affinity. It argues that boundary-work between ethnic Italians and Albanian immigrants resulted in intergroup relations that coupled Albanian occupational incorporation with the (re)construction of respective group identities as culturally similar – and distinct from Latino and African-American groups in the neighbourhood. Engaging recent literature on ethnic boundary making, I argue that ethnic affinity constitutes a new strategy of boundary reinforcing, in addition to established strategies of boundary crossing, blurring, and shifting (Zolberg and Woon 1999; Wimmer 2008). Developed in the context of shifting ethno-racial neighbourhood makeup, this affinity between ethnic Italians and Albanian immigrants relied on American constructions of shared European whiteness, overturning contemporary divides between Italians and Albanians in Europe. Ethnic affinity provides a conceptual framework that goes beyond notions of ethnic succession, passing, or assimilation.     

Oral health awareness in Croatian and Italian urban adolescents

Purpose of this study was to investigate and compare differences in oral health awareness between Croatian and Italian urban adolescents. The sample consisted of primary school last grade students aged between 13 and 15 years, 300 children from Zagreb (Croatia) and 298 children from Bari (Italy). Oral health awareness was evaluated using a self-administered standardized questionnaire. Self-perception of oral health proved to be different between the two groups (p < 0.001). The Croatians reported that their oral health was "excellent" or "very-good" more often than the Italians (68.6% vs. 50.2%). The reasons given for visiting a dentist were different (p < 0.001). The Italians cleaned their teeth more often than the Croatians ("two or more times a day", 83.1% vs. 72.2%, p < 0.003). Wooden toothpicks were preferred by the Croatians (p < 0.001), while floss was preferred by the Italians (p = 0.03). The awareness regarding the use of fluoridated toothpaste was higher in the Italian group (95.6% vs. 72.5%, p < 0.001). The Croatians were consuming sweetened foods more often than the Italians (p < 0.001). Croatian adolescents reported more indicators of a lower level of oral health awareness than the Italians, while on the contrary Croatians had higher esteem of their oral health. Defining national preventive strategies is essential for improving adolescents' attitudes toward oral health in both countries, particularly in Croatia.     

Subtyping of phosphoglucomutase locus 1 (PGM1) polymorphism in some populations of Rwanda: description of variant phenotypes, "haplotype" frequencies,and linkage disequilibrium data.

Some populations of Rwanda (South Twa Pygmies, Hutu, and Tutsi) have been analyzed by acid starch gel electrophoresis for the subtyping of PGM1 polymorphism. The new polymorphic third PGM11 allele, the PGM1(1Twa), which we recently detected in Twa Pygmies from North Rwanda, has not been found in this survey, whereas the rare PGM1(6) allele attains subpolymorphic frequencies in all groups. Comparison between the various populations of Rwanda shows that they differ significantly from each other with the exception of South Twa Pygmies and Tutsi. A relatively low frequency (9.6%) of the PGM1(2S) allele appears to be typical of North Twa Pygmies; a low frequency of PGM1(2F) (1.2%-3.6%) has been found in all these groups but not in the Hutu (6.4%); and a particularly high incidence of the PGM1(1F) allele (the highest so far reported) has been observed in the South Twa Pygmies (20%) and in the Tutsi (18%). The PGM1(1Twa) and PGM1(6) enzymes, which in acid starch gel are not distinguishable, can be clearly differentiated by isoelectric focusing. In addition, the same technique has shown that the rare PGM1(7) allele observed in one Hutu is different from that found at polymorphic frequency in the Japanese and from a rare PGM1(7) allele found in Germany. On the very likely hypothesis that the PGM1(1S), PGM1(1F), PGM1(2S), and PGM1(2F) result from variations at two different polymorphic sites, 1/2 and F/S, within the PGM1 structural gene, all the available population data have been analyzed to investigate whether preferential combinations (haplotypes) were identifiable. Whereas Caucasians show a prevalence of 2F and 1S combination with an 8.02% mean value of linkage disequilibrium expressed as % Dmax, from the very few and scattered African data, it is impossible to draw any inference at present.