Whole mitochondrial DNA sequencing in Alpine populations and the genetic history of the Neolithic Tyrolean Iceman.

The Tyrolean Iceman is an extraordinarily well-preserved natural mummy that lived south of the Alpine ridge ~5,200 years before present (ybp), during the Copper Age. Despite studies that have investigated his genetic profile, the relation of the Iceman´s maternal lineage with present-day mitochondrial variation remains elusive. Studies of the Iceman have shown that his mitochondrial DNA (mtDNA) belongs to a novel lineage of haplogroup K1 (K1f) not found in extant populations. We analyzed the complete mtDNA sequences of 42 haplogroup K bearing individuals from populations of the Eastern Italian Alps - putatively in genetic continuity with the Tyrolean Iceman-and compared his mitogenome with a large dataset of worldwide K1 sequences. Our results allow a re-definition of the K1 phylogeny, and indicate that the K1f haplogroup is absent or rare in present-day populations. We suggest that mtDNA Iceman´s lineage could have disappeared during demographic events starting in Europe from ~5,000 ybp. Based on the comparison of our results with published data, we propose a scenario that could explain the apparent contrast between the phylogeographic features of maternal and paternal lineages of the Tyrolean Iceman within the context of the demographic dynamics happening in Europe from 8,000 ybp.

Detecting Sex-Biased Gene Flow in African-Americans through the Analysis of Intra- and Inter-Population Variation at Mitochondrial DNA and Y- Chromosome Microsatellites.

This study reports on variations at the mitochondrial DNA (mtDNA) hypervariable region 1 (HVR-1) and at seven Y-chromosome microsatellites in an African-American population sample from Chicago, IL, USA. Our results support the hypothesis that the population studied had undergone a European male-biased gene flow. We show that comparisons of intra-and inter-population diversity parameters between African-Americans, Europeans and Africans may help detect sex-biased gene flow, providing a complement to quantitative methods to estimate genetic admixture.

A multi-perspective view of genetic variation in Cameroon.

In this study, we report the genetic variation of autosomal and Y-chromosomal microsatellites in a large Cameroon population dataset (a total of 11 populations) and jointly analyze novel and previous genetic data (mitochondrial DNA and protein coding loci) taking geographic and cultural factors into consideration. The complex pattern of genetic variation of Cameroon can in part be described by contrasting two geographic areas (corresponding to the northern and southern part of the country), which differ substantially in environmental, biological, and cultural aspects. Northern Cameroon populations show a greater within- and among-group diversity, a finding that reflects the complex migratory patterns and the linguistic heterogeneity of this area. A striking reduction of Y-chromosomal genetic diversity was observed in some populations of the northern part of the country (Podokwo and Uldeme), a result that seems to be related to their demographic history rather than to sampling issues. By exploring patterns of genetic, geographic, and linguistic variation, we detect a preferential correlation between genetics and geography for mtDNA. This finding could reflect a female matrimonial mobility that is less constrained by linguistic factors than in males. Finally, we apply the island model to mitochondrial and Y-chromosomal data and obtain a female-to-male migration Nnu ratio that was more than double in the northern part of the country. The combined effect of the propensity to inter-populational admixture of females, favored by cultural contacts, and of genetic drift acting on Y-chromosomal diversity could account for the peculiar genetic pattern observed in northern Cameroon.

HLA class I variation in the West African Pygmies and their genetic relationship with other African populations.

We have studied the polymorphism of HLA class I in two West African Pygmy populations, namely, the Bakola from Cameroon and the Mbenzele from the Central African Republic. A unique number of HLA alleles and haplotypes showed specific patterns of these populations. In this study, we identify two alleles (B*37, B*41) and three haplotypes (A*30-B*37, A*66-B*41 and A*68-B*58) that appear to be 'private' or typical of Western Pygmies. These data reflect similarities with the AKA Pygmies from the Central African Republic. On the other hand, we failed to identify alleles that are found at high frequencies among other sub-Saharan populations (B*42, B*51). Allelic and haplotypic frequency distributions show differences between the two Pygmy groups, e.g. B*35 was very common in the Mbenzele but has been found to be absent in the Bakola. In contrast, B*53, which is found in the Bakola, has been found to be rare in the Mbenzele Pygmies. In order to analyse the genetic relationships of the Bakola and Mbenzele Pygmies with other sub-Saharan populations, HLA gene frequencies were subjected to the Neighbour-Joining tree analysis. The Mbenzele, Bakola and AKA were found to be relatively close to each other and isolated from other sub-African populations. However, both the genetic distances and the within-group variation suggests that the Bakola are more admixed with Bantu farmers than Mbenzele.

A fast amplification-reverse hybridization assay kit to detect the most frequent deficient variants in the alpha-1-antitrypsin gene.

BACKGROUND: There is worldwide growing awareness of alpha-1-antitrypsin deficiency (AATD), a major hereditary disorder in Caucasians. The gold standard for its laboratory diagnosis is thin-layer isoelectric focusing, which should be performed in reference laboratories. OBJECTIVES: The aim of this study was to check the characteristics of a commercially available amplification-reverse hybridization assay kit in detecting at a molecular level the alpha-1-antitrypsin (AAT) Z and S variants, i.e. the most frequent variants associated with AATD, by comparing its performance with DNA restriction fragment length polymorphism. METHODS: We studied samples from 36 subjects enrolled in the Italian National Registry for Severe Alpha-1-antitrypsin Deficiency. Based on previous plasma isoelectric focusing typing, we selected samples with the following phenotypes: MM (9 samples), MS (9 samples), SZ (3 samples), MZ (11 samples), ZZ (3 samples), and a rare variant (1 sample). DNA was extracted by the standard method. The presence of the AAT Z and S gene variants was determined by the amplification-reverse hybridization test kit, following the manufacturer's instructions, and by the restriction fragment length polymorphism technique, according to established procedures. RESULTS: We found that the identification of the AAT Z and S gene variants obtained by the amplification-reverse hybridization test kit was completely in agreement with that obtained by the restriction fragment length polymorphism technique. CONCLUSIONS: We conclude that the test kit provides a fast, easy and unambiguous identification of Z and S alleles. Because of its transferability to routine laboratories, the test kit may be useful in identifying cases of severe AATD, thus resulting in increasing awareness of this rare disorder.

The carpal tunnel syndrome. Anatomo-clinical correlations.

We review a series of 1,280 operated cases of carpal tunnel syndrome (CTS), with an unexpected high frequency of transverse muscular fibers within the carpal canal (11%), particularly in a group of young male patients under 30 years of age. This abnormal presence of muscle fibers could explain the early development of CTS in young male workers.

Testing a biochemical model of human genetic resistance to falciparum malaria by the analysis of variation at protein and microsatellite loci.

We recently proposed a biochemical model of genetic resistance to falciparum malaria based on the role of oxidant stress (of parasitic origin) in inducing the irreversible oxidation of hemoglobin and its binding to the erythrocyte membrane (Destro-Bisol et al. 1996). To test the model, we analyzed the relationships between the polymorphisms at the hemoglobin beta chain (HBB) and red cell glutathione peroxidase (GPX1) loci in 18 populations that had been subjected to endemic malaria (Cameroon and Central African Republic). The erythrocytes of GPX1*2 heterozygotes should be more efficient in sheltering the cell membrane from irreversible oxidation and binding of hemoglobin caused by the oxidant stress exerted by Plasmodium falciparum. According to our model, the GPX1*2 allele has an epistatic effect on the HBB*A/*S genotype by lowering its protection against falciparum malaria. In turn, this should decrease the fitness of the HBB*A/*S-GPX1*2/*1 genotype. Our predictions were confirmed. In fact, we observed a clear trend toward a dissociation between the HBB*A/*S and GPX1*2/*1 genotypes in the overall data. To test alternative hypotheses, we also analyzed the genetic variation at 9 protein and 10 autosomal microsatellite loci at both the single- and the 2-locus level. We also discuss the possible relevance of an alternative biochemical pathway. The results further support the conclusions of our study because the dissociation between the GPX1*2/*1 and HBB*A/*S genotypes does not appear to be related either to a general decrease in heterozygosity or to an increased risk of sudden death in HBB*A/*S individuals.

In vivo and in vitro neurotoxic action of plasma ultrafiltrate from uraemic patients.

BACKGROUND: In order to investigate the aetiology of uraemic neuropathy, we evaluated the neurotoxic activity of plasma from uraemic patients. To this end we prepared a concentrate (1:1000) of 2-60 kDa MW compounds from paired filtration dialysis ultrafiltrate and evaluated its activity on peripheral nerve conduction in vivo and in vitro. METHODS: The in vivo neurotoxicity was tested on rat sciatic nerve by intraneural injection of the uraemic concentrate, followed, 1 to 6 days later, by electrophysiological assessment of motor response and maximum conduction velocity. In vitro experiments were performed on isolated frog sciatic nerve in the presence of uraemic concentrate, and the neurotoxicity was evaluated from the rate of the decrease in the amplitude of the evoked maximal action potential. RESULTS: In the in vivo experiments, the sciatic nerves injected with the uraemic concentrate showed a decrease in maximum conduction velocity and a progressive impairment in evoked motor response. In the in vitro experiments uraemic concentrate induced a dose-dependent neurotoxic effect. CONCLUSIONS: Our study demonstrates the presence in plasma of uraemic patients of a compound of 2-60 kDa MW with neurotoxic activity.

A seven year anthropobiological survey on the distribution of abnormal hemoglobins in Cameroon.

Within the frame of an anthropobiological survey on some populations of Cameroon (1985-1991), Hb beta data were collected from numerous ethnic groups including Bakaka, Bamileke, Daba, Fali, Guiziga, Kanuri, Mada, Mafa, Mundang, Uldeme, Podokwo, Tali, Tupuri, Fulbe, Mandara, Ewondo and Bassa. Hb beta *S allele frequencies ranged from 0.008 +/- 0.003 (among Fali) to 0.152 +/- 0.020 (among Mandara) and 0.152 +/- 0.044 (among Podokwo), whereas Hb beta *S was found to be absent among Tupuri. Hb beta *C was observed among Bamileke (0.001 +/- 0.001), Fali (0.003 +/- 0.002), Fulbe (0.002 +/- 0.002), Mafa (0.005 +/- 0.005), Mundang (0.005 +/- 0.005), Tupuri (0.010 +/- 0.007) and Podokwo (0.015 +/- 0.015). The possible reasons for these variations in allele frequencies are discussed.

Improving pictures in dermatopathology.

We have found that a remarkable improvement in the resolution of the details and in the richness of halftones can be obtained using 4" x 5" black-and-white professional films specifically prepared for scientific and documentary photography. Through modification of the developing time and developer dilution, a close control of the contrast and gray tones is also possible. Using this large-format film, a contact print can be made and top-quality pictures obtained even for a subject as difficult as skin pathology.

A bio-anthropological study on the Bakakas of Cameroon.

Bakakas are native Bantus belonging to the Mbo-Bakossi group, peopling the Cameroon's Littoral region. In the context of a wide bio-anthropological study project focused on the bio-historical processes involved in the areas, 278 adults of both sexes from the villages of Ebone and Bakwat (Bakaka Canton) were investigated for 14 erythrocyte and serum genetic polymorphisms (ACP1, ADA, EsD, GLO, Hb beta, GPX1, CAII, PGM1, SAHH, 6-PGD, Hp, Pi, Gc and Tf). With only a few exceptions (Hp and GLO systems), the genetic frequencies of the polymorphisms considered tend to fall within the range of variation known for the subsaharan populations. With reference to the malaria endemicity characterizing the Littoral environment, high frequencies for Hb beta*S allele and absence of the ACP1*R 'Negro allele' were recorded. The genetic distances among Bakakas and 14 other Central African populations were also calculated from six genetic loci.

Cerebral blood flow changes induced by electrical stimulation of the Gasserian ganglion after experimentally induced subarachnoid haemorrhage in pigs.

The effect of trigeminal electrical stimulation on cerebral blood flow has been studied in conditions of normal or reduced cerebral blood flow (CBF). Autologous blood was injected into the subarachnoid space of ten Pittmann-Moore pigs to induce subarachnoid haemorrhage (SAH) accompanied by cerebral blood flow (CBF) reduction. One week later, in six of ten animals, a considerable decrease of CBF was noted as evaluated by means of a recording-system monitoring over the right parieto-temporal calvarium the washout of 133Xenon injected into the internal carotid artery after the external carotid had been clamped. Continuous electrical stimulation of the Gasserian ganglion performed in the six animals with severely induced CBF reduction produced a remarkable cerebrovascular dilation and increase of CBF lasting over 3 h. Electrical stimulation of the Gasserian ganglion produced a similar pattern of vasodilation in six pigs in which no blood was injected and no reduction of CBF was evident. The mechanisms and the anatomical pathways which underlie these results are discussed.

Evaluation of the sensitivity of microfilter paper assays in an anthropological study: results of samples from Cameroon and Tanzania.

Microfilter absorbed whole blood samples from 223 Tanzanian babies and 189 adults from Cameroon have been examined. Blood specimens are difficult to obtain from African suburban and rural areas, and lack of storage and transportation facilities can prevent the collection of samples. We evaluated some microassays employing whole blood collected on filter paper. This method is a well established technique in neonatal screening for endocrinometabolic diseases. We also developed microassays for whole dried blood spots to type AB0 blood groups and HIV disease using commercial reagents. Phenotype and gene frequencies for AB0 and hemoglobin systems as well as our results concerning the typings of thyroxine (T4), thyroid stimulating hormone (TSH), human immunodeficiency virus (HIV) and hepatitis B virus (HBV) are reported.

A report of four different unusual 6-PGD electrotypes in Caucasian and Negro populations.

Four different unusual 6-phosphogluconate dehydrogenase (6-PGD) electrophoretic patterns found among the Italian (Rome), Bamileke (Cameroon), and North Bateke and Babenga Pygmy (Congo) populations are described.

The treatment of hemifacial spasm with percutaneous radiofrequency thermocoagulation of the facial nerve.

The authors present their first results with the treatment of hemifacial spasm by controlled percutaneous thermocoagulation of the facial nerve. Seven patients have been treated to date with good immediate results on the movements, although a slight paresis of the homolateral facial musculature, aesthetically acceptable, persists after treatment. A long-term follow-up at more than 10 months was achieved in only 3 patients, who showed a complete regression of the spasm with partial disappearance of the facial hemiparesis.