Effect of dietary coenzyme Q and fatty acids on the antioxidant status of rat tissues

Summary.  Wistar rats were fed with different diets with or without supplement coenzyme Q₁₀ (CoQ₁₀) and with oil of different sources (sunflower or virgin olive oil) for six or twelve months. Ubiquinone contents (CoQ₉ and CoQ₁₀) were quantified in homogenates of livers and brains from rats fed with the four diets. In the brain, younger rats showed a 3-fold higher amount of ubiquinone than older ones for all diets. In the liver, however, CoQ₁₀ supplementation increased the amount of CoQ₉ and CoQ₁₀ in both total homogenates and plasma membranes. Rats fed with sunflower oil as fat source showed higher amounts of ubiquinone content than those fed with olive oil, in total liver homogenates, but the total ubiquinone content in plasma membranes was similar with both fat sources. Older rats showed a higher amount of ubiquinone after diets supplemented with CoQ₁₀. Two ubiquinone-dependent antioxidant enzyme activities were measured. NADH-ferricyanide reductase activity in hepatocyte plasma membranes was unaltered by ubiquinone accumulation, but this activity increased slightly with age. Both cytosolic and membrane-bound dicumarol-sensitive NAD(P)H:(quinone acceptor) oxidoreductase (DT-diaphorase, EC 1.6.99.2) activities were decreased by diets supplemented with CoQ₁₀. Animals fed with olive oil presented lower DT-diaphorase activity than those fed with sunflower oil, suggesting that the CoQ₁₀ antioxidant protection is strengthened by olive oil as fat source. Received May 22, 2002; accepted September 20, 2002; published online May 21, 2003 RID="*" ID="*" Correspondence and reprints: Departamento de Biología Celular, Fisiología e Inmunología, Universidad de Córdoba Edificio Severo Ochoa, Campus de Rabanales, 14071 Córdoba, Spain.     

Regeneration of lipophilic antioxidants by NAD(P)H:quinone oxidoreductase 1

Summary.  The aim of this work was to study the activity of NAD(P)H:(quinone acceptor) oxidoreductase 1 (EC 1.6.99.2) in the regeneration of lipophilic antioxidants, alpha-tocopherol, and reduced-coenzyme Q analogs. First, we tested whether or not two isoforms of the NAD(P)H:(quinone acceptor) oxidoreductase 1 designated as “hydrophilic” and “hydrophobic” (H. J. Prochaska and P. Talalay, Journal of Biological Chemistry 261: 1372–1378, 1986) show differential enzyme activities towards hydrophilic or hydrophobic ubiquinone homologs. By chromatography on phenyl Sepharose, we purified the two isoforms from pig liver cytosol and measured their reduction of several ubiquinone homologs of different side chain length. We also studied by electron paramagnetic resonance the effect of NAD(P)H:(quinone acceptor) oxidoreductase 1 on steady-state levels of chromanoxyl radicals generated by linoleic acid and lipooxygenase and confirmed the enzyme's ability to protect alpha-tocopherol against oxidation induced with H₂O₂-Fe²⁺. Our results demonstrated that the different hydrophobicities of the isoforms do not reflect different reactivities towards ubiquinones of different side chain length. In addition, electron paramagnetic resonance studies showed that in systems containing the reductase plus NADH, levels of chromanoxyl radicals were dramatically reduced. Morever, in the presence of oxidants, alpha-tocopherol was preserved by NAD(P)H:(quinone acceptor) oxidoreductase 1, supporting our hypothesis that regeneration of alpha-tocopherol may be one of the physiologic functions of this enzyme. Received May 20, 2002; accepted September 20, 2002; published online May 21, 2003 RID="*" ID="*" Correspondence and reprints: Departamento de Biología Celular, Fisiología e Inmunología, Facultad de Ciencias, Edificio Severo Ochoa, Campus de Rabanales, Universidad de Córdoba, 14014 Córdoba, Spain.     

Differential distribution of ascorbic acid,peroxidase activity,and hydrogen peroxide along the root axis in Allium cepa L. and its possible relationship with cell growth and differentiation

Summary.  In this paper we show an asymmetrical distribution of apoplastic and symplastic ascorbic acid content, peroxidase activities and hydrogen peroxide along the root axis in Allium cepa L. For most of these metabolites, a marked gradient from the root apex to the onion base was observed and was different for apoplastic and symplastic compartments. In total homogenates, ascorbic acid content was higher in the zones closer to the apex and decreased towards the root base. However, an opposite pattern was observed in the apoplastic fraction. Peroxidase activities with guaiacol, ferulic acid, ascorbic acid, and coniferyl alcohol were also different depending on the evaluated zone and the fraction used (apoplastic or symplastic). In general, each activity had a specific and unique pattern. Immunodetection of peroxidase proteins in Western blots using anti-horseradish peroxidase and anti-ascorbate peroxidase antibodies revealed different bands at the different zones of the root. Hydrogen peroxide was detected by electron microscopy and was mainly found in cell walls of epidermis (or rhizodermis), meristem, and elongating cells. The number of cell walls showing hydrogen peroxide decreased dramatically towards the root base. The results suggest that the different zones of the root show specific requirements for ascorbic acid and hydrogen peroxide. Also, each fragment of the root seems to express specific peroxidase proteins. Different processes that take place at every part of the root, as cell proliferation and elongation near the root apex and gradual lignification and differentiation towards the root base are the key to explain the results. Received May 10, 2002; accepted September 20, 2002; published online May 21, 2003 RID="*" ID="*" Correspondence and reprints: Departamento de Biología Celular, Fisiología e Inmunología, Edificio C-6, Campus de Rabanales, Universidad de Córdoba, 14014 Córdoba, Spain.     

NAD+/NADH and/or CoQ/CoQH2 ratios from plasma membrane electron transport may determine ceramide and sphingosine-1-phosphate levels accompanying G1 arrest and apoptosis

To elucidate possible biochemical links between growth arrest from antiproliferative chemotherapeutic agents and apoptosis, our work has focused on agents (EGCg, capsaicin, cis platinum, adriamycin, anti-tumor sulfonylureas, phenoxodiol) that target tNOX. tNOX is a cancer-specific cell surface NADH oxidase (ECTO-NOX protein), that functions in cancer cells as the terminal oxidase for plasma membrane electron transport. When tNOX is active, coenzyme Q(10) (ubiquinone) of the plasma membrane is oxidized and NADH is oxidized at the cytosolic surface of the plasma membrane. However, when tNOX is inhibited and plasma membrane electron transport is diminished, both reduced coenzyme Q(10) (ubiquinol) and NADH would be expected to accumulate. To relate inhibition of plasma membrane redox to increased ceramide levels and arrest of cell proliferation in G(1) and apoptosis, we show that neutral sphingomyelinase, a major contributor to plasma membrane ceramide, is inhibited by reduced glutathione and ubiquinone. Ubiquinol is without effect or stimulates. In contrast, sphingosine kinase, which generates anti-apoptotic sphingosine-1-phosphate, is stimulated by ubiquinone but inhibited by ubiquinol and NADH. Thus, the quinone and pyridine nucleotide products of plasma membrane redox, ubiquinone and ubiquinol, as well as NAD(+) and NADH, may directly modulate in a reciprocal manner two key plasma membrane enzymes, sphingomyelinase and sphingosine kinase, potentially leading to G(1) arrest (increase in ceramide) and apoptosis (loss of sphingosine-1-phosphate). As such, the findings provide potential links between coenzyme Q(10)-mediated plasma membrane electron transport and the anticancer action of several clinically-relevant anticancer agents.     

Asymmetric synthesis of all stereoisomers of 6-methylpipecolic acids

Summary.  Asymmetric synthesis of all four stereoisomers of 6-methylpipecolic acids with high enantiomeric purity via iterative AD reaction, starting from 1,6-heptadiene, has been described. Received March 25, 2002 Accepted June 15, 2002 Published online January 30, 2003 Authors' address: Hiroki Takahata, Faculty of Pharmaceutical Sciences, Tohoku Pharmaceutical University, Sendai 981-8558, Japan, E-mail: takahata@tohoku-pharm.ac.jp RID="*" ID="*"  The stereo-configurations of 8 and 9 in Fig. 3 and 12 in Fig. 4 show only major isomers. RID="**" ID="**"  The absolute configurations of 11 and 14     

Ubiquinol inhibition of neutral sphingomyelinase in liver plasma membrane: specific inhibition of the Mg(2+)-dependent enzyme and role of isoprenoid chain

In this work, the specificity of ubiquinol as inhibitor of the neutral sphingomyelinases present at the plasma membrane (Mg(2+)-dependent and -independent) and structural requirements for such inhibition have been studied. Our results have shown that ubiquinol specifically inhibits Mg(2+)-dependent neutral sphingomyelinase activity in isolated liver plasma membranes, but no significant participation of the Mg(2+)-independent enzyme was observed. Both the reduction state of the (hydro)quinone ring and the length of the hydrophobic side chain were important determinants in neutral sphingomyelinase inhibition. Ubiquinols inhibited the nSMase more efficiently than ubiquinones, and hydrophobic homologs with six or nine isoprene units were the most effective inhibitors. Inhibition of nSMase by ubiquinols displayed similarities with inhibition by manumycin and the hydroquinones F11334's, suggesting that these compounds could act as structural analogs of ubiquinol. Beyond its participation in mitochondrial energy metabolism, and as antioxidant, this novel role for ubiquinol as a neutral sphingomyelinase inhibitor should be considered an important factor to regulate lipid signaling at the plasma membrane that could be related to its beneficial effects on cells, tissues, and organisms.     

Neutral Magnesium-Dependent Sphingomyelinase from Liver Plasma Membrane: Purification and Inhibition by Ubiquinol

Plasma membranes isolated from pig liver contained almost no acid sphingomyelinase but significant neutral magnesium-dependent sphingomyelinase that was activated by phosphatidylserine. We report here the purification to apparent homogeneity of neutral sphingomyelinase of about 87 kDa from liver plasma membranes. The purified enzyme strictly required magnesium and had a neutral optimal pH. In contrast with neutral sphingomyelinase purified from other sources (such as brain), the enzyme purified from from liver plasma membrane was not inhibited by GSH and, strikingly, it was not activated by phosphatidylserine. Liver sphingomyelinase was inhibited by several lipophilic antioxidants in a dose-dependent way. Ubiquinol-10 was more effective than alpha-tocopherol, alpha-tocopherylquinone, alpha-tocopherylquinone, and ubiquinone-10, and inhibition was noncompetitive. Differential inhibition of neutral sphingomyelinase by antioxidants did not correlate with different levels of protection against lipid peroxidation. The purified sphingomyelinase was not inhibited significantly by ubiquinone-10 and ubiquinol- 10, but ubiquinol-0 and ubiquinone-0 inhibited by 30 and 60% respectively. Our results demonstrate a direct inhibitory effect of ubiquinol on the plasma membrane n-SMase and support the participation of this molecule in the regulation of ceramide-mediated signaling.     

Behavior of storage lipids during development and germination of olive ( Olea europaea L.) pollen

Summary.  The presence of abundant oil bodies in the mature olive pollen grain has led us to focus on the behavior of these lipid bodies during pollen development and in vitro pollen germination. The appearance, increase, and accumulation of lipid bodies have been determined by following the sequential development of the pollen grain. Semithin slices of anthers and pollen grains were stained with Sudan Black B in order to identify neutral lipids. Ultrastructural studies were also carried out. Our results show a notable increase in lipid bodies between the young-pollen-grain stage and the mature-pollen-grain stage. Substantial polarization of lipid bodies was observed after 1 or 2 h of pollen incubation in germination medium. During pollen tube growth, the lipid bodies are located near the germinative aperture after 3 h of incubation, as well as inside the pollen tube, thus suggesting that the lipid bodies move from the pollen grain to the pollen tube. After 7 h of germination the presence of lipid bodies inside the pollen tube is no longer substantial. Our results support the idea that lipid bodies are involved in pollen germination, stigma penetration, and pollen tube growth. These results are discussed in connection with their implications for the pollen germination process. Received June 4, 2002; accepted October 29, 2002; published online April 8, 2003 RID="*" ID="*" Correspondence and reprints: Departamento de Bioquímica, Biología Celular y Molecular de Plantas, Estación Experimental del Zaidín, Consejo Superior de Investigaciones Científicas, Profesor Albareda 1, 18008 Granada, Spain.     

Synoptic and meteorological characterisation of olive pollen transport in Córdoba province (south-western Spain)

The main goal of the present study was to provide a detailed analysis of olive pollen transport dynamics in the province of Córdoba (south-western Spain) by applying back-trajectory analysis. Pollen data from 2006 and 2007 were analysed at four monitoring sites: Córdoba city in the centre of the province, Baena and Priego de Córdoba located in the south, and El Cabril reserve (Hornachuelos Natural Park) in the north. Particular attention was paid to nine episodes of high pollen counts. Synoptic surface maps were used, and kinematic back-trajectories (3D) were computed using the hybrid single particle Lagrangian integrated trajectory model (HYSPLIT) at 500 m above ground level, run with a time-step of 6 h over a period of 36 h. Findings were analysed in conjunction with daily and bi-hourly airborne pollen data, field phenological data and hourly surface meteorological data recorded at nearby stations: temperature, relative humidity, rainfall, wind direction and wind speed. The results identified two pollen source areas over the Córdoba province, the largest one located in the south, affecting Baena, Priego de Córdoba and Córdoba city, and one smaller located in the west, which determines mainly the pollen cycle over the north of the province, El Cabril. In addition, two air mass circulations were found, one coming from the south and crossing the main olive pollen sources very close to the surface and being frequently associated with higher pollen counts, and the other coming from the west and, in the episodes investigated, influencing mainly the north of the province.     

Cytoskeletal 10 nm filaments in cells of the algal phyla Chlorophyta,Charophyta, and Chrysophyta and their developmentally regulated and species-specific association with prosomes

Summary.  10 nm diameter filaments were observed in whole-mount preparations of algae of diverse phyla: Acetabularia acetabulum and A. major (Chlorophyta), Chara australis and Nitella flexilis (Charophyta), and Poterioochromonas malhamensis (Chrysophyta). A polyclonal antibody raised against a basic, 50 kDa DNA-binding protein of A. acetabulum stains the filaments of A. acetabulum and A. major as well as of C. australis and N. flexilis. While in the perinuclear region of A. acetabulum and A. major and throughout the cytoplasm of P. malhamensis the 10 nm filaments have a smooth appearance, in the stalk of A. acetabulum and A. major they are densely covered by globular structures; in C. australis and N. flexilis they are less frequently associated with such material. The morphology of a part of the globular particles is quite reminiscent of prosomes. A monoclonal antibody elicited against prosomes isolated from A. acetabulum indeed decorates the globular particles on the A. acetabulum and A. major filaments. The possible role of these filament-particle associations is discussed. Received August 10, 2001; accepted October 30, 2002; published online April 8, 2003 RID="*" ID="*" Correspondence and reprints: Max-Planck-Institut für Zellbiologie, 68526 Ladenburg, Federal Republic of Germany. E-mail: sberger@zellbio.mpg.de RID="**" ID="**" Present address: Long Island University, Southampton, New York, U.S.A. RID="+" ID="+" Present address: Leica Microsystems Wetzlar GmbH, Wetzlar, Federal Republic of Germany     

A multi-level approach using Gambusia affinis as a bioindicator of environmental pollution in the middle-lower basin of Suquía River

The Suquía River middle-lower basin (Córdoba, Argentina) is subjected to a strong anthropogenic impact because it receives pollutants from different sources. The main goal of this study was to evaluate the introduced fish species Gambusia affinis as a bioindicator of environmental pollution in the middle-lower basin of the Suquía River. The assessment was performed by measuring biomarkers at different levels of biological organization, at two sampling sites (before and after Córdoba city), and at dry and wet seasons. Water quality evaluation was made through a water-quality index (based on physico-chemical parameters), heavy-metals and pesticides concentrations in water. The water quality varied between sampling sites, showing the most degraded conditions downstream Córdoba city. The same pattern of variation was detected in the biomarkers studied, mainly in: gill and liver histopathological indexes, copulatory organ (gonopodium) morphology and vitellogenin expression in males and females. The present study characterized the environmental conditions in the middle-lower basin of the Suquía River and revealed the low freshwater quality at the most polluted site. Although G. affinis is an introduced species, it could be considered a good sentinel of water resource quality of invaded Neotropical basins. Our results demonstrated the importance of addressing the environmental quality monitoring through an integrated analysis of water quality parameters together with histological, morphological and molecular parameters. Thus, our study provides a good model for application in other basins of South America.     

Assessing temperature and humidity conditions for dairy cattle in Córdoba,Argentina

Temperature and humidity conditions affect livestock production in Central Argentina. This study evaluates the risk of thermal stress affecting dairy production. The temperature-humidity index (THI) was used to analyze the regional and seasonal effects of temperature and humidity. Statistically, the THI was found to be normally distributed. The probability of occurrence of a daily THI higher than 72 was 40% for Río Cuarto during January. Regional variability of THI indicates a low risk of harmful extreme thermal stress conditions. The probability of THI being 78 or above ranges between 4% and 10% for the main dairy region of Córdoba during January. Also, in January and February, dairy production losses between 3 and 4 l cow(-1) day(-1) could be expected with a frequency of 5% in Río Cuarto and 15% in Villa de María de Río Seco.     

Ubiquinol regeneration by plasma membrane ubiquinone reductase

Summary Several enzyme systems have been proposed to play a role in the maintenance of ubiquinol in membranes other than the inner mitochondrial membrane. The aim of this study was to investigate the mechanisms involved in NADH-driven regeneration of antioxidant ubiquinol at the plasma membrane. Regeneration was measured by quantifying the oxidized and reduced forms of ubiquinone by electrochemical detection after separation by high-performance liquid chromatography. Plasma membrane incubation with NADH resulted in the consumption of endogenous ubiquinone, and a parallel increase in ubiquinol levels. The activity showed saturation kinetics with respect to the pyridine nucleotides and was moderately inhibited byp-hydroxymercuribenzoate. Only a slight inhibition was achieved with dicumarol at concentrations reported to fully inhibit DT-diaphorase. Salt-extracted membranes displayed full activity of endogenous ubiquinol regeneration, supporting the participation of an integral membrane protein. In liposomes-reconstituted systems, the purified cytochromeb ₅ reductase catalyzed the reduction of the natural ubiquinone homologue coenzyme Q₁₀ at rates accounting for the activities observed in whole plasma membranes, and decreased the levels of lipid peroxidation. Our data demonstrate the role of the cytochromeb ₅ reductase in the regeneration of endogenous ubiquinol.Abbreviations AAPH 2,2-azobis-(2-amidinopropane) hydrochloride - CoQ coenzyme Q, ubiquinone - CoQH₂ reduced coenzyme Q, ubiquinol - pHMB p-hydroxymercuribenzoate     

Evidence that neutral spingomyelinase of cultured murine neuroblastoma cells is oriented externally on the plasma membrane

The activity of the neutral, Mg²⁺-stimulated sphingomyelinase of cultured neuroblastoma cells (N1E-115) is enriched in the plasma membrane fraction and is reduced following treatment of intact or broken cells with trypsin, α-chymotrypsin, papain, and protease. Two protease-sensitive enzymes of the cell interior (lactate dehydrogenase and NADPH-cytochrome c reductase) are not affected by protease treatment of intact cells. These results indicate that the neutral, Mg²⁺-stimulated sphingomyelinase is oriented externally on the plasma membrane of the cultured neuroblastoma cell.     

Comparison of airborne herb pollen types in Córdoba (Southwestern Spain) and Poznan (Western Poland)

This study sought to compare airborne pollen counts for a number of common herbaceous species (Plantago, Chenopodiaceae–Amaranthaceae, Rumex, and Urticaceae) in two cities with differing weather conditions, Córdoba (Southwestern Spain) and Poznan (Western Poland). Pollen seasons for these species were studied from 1995 to 2005. Aerobiological sampling was performed using a Hirst type 7-day spore trap, in accordance with the procedure developed by the European Aerobiology Network. A Spearman correlation test was used to test for correlations between meteorological parameters and daily airborne pollen counts. The Spearman correlation test and the Wilcoxon signed ranks test were also used to compare mean daily pollen counts for the two study sites. In Córdoba, the pollen season generally started around two months earlier than in Poznan, and also lasted longer. These findings were attributed to the presence of a larger number of species in Córdoba, with overlapping pollen seasons, and also to more favorable weather conditions. Trends in pollen season start dates were fairly stable over the study period, with a slight tendency to delayed onset in Córdoba and a modest advance in start date in Poznan. The pollen season end date also remained reasonably stable over the study, with only a slight tendency for the season to end earlier in Córdoba and later in Poznan. A clear trend towards declining annual pollen counts was recorded over the study period for all pollen types in both cities.     

Hypodermosis in the red deer Cervus elaphus in Cordoba, Spain

The prevalence of red deer hypodermosis and the life cycle of Hypodermia diana Brauer 1858 in three provinces of the south of Spain, Córdoba, Jaen and Ciudad Real, were studied by inspecting 254, thirty-eight and thirty-five deer from each province respectively. The prevalence of infestations was: Córdoba, 87.75%; Jaen, 92.10%, Ciudad Real, 91.42%. From this we deduced an overall prevalence of 88.67%, comprising 88.23% in male deer and 89.96% in female deer. The intensity of the parasitism in 213 of the 245 animals from Córdoba varied between one warble per animal to more than 100 in 28 of the inspected animals. The chronology of the life cycle during the hunting period (October to the end of February) was investigated.     

New Patagonian baurusuchids (Crocodylomorpha; Notosuchia) from the Bajo de la Carpa Formation (Upper Cretaceous; Neuquén,Argentina): New evidences of the early sebecosuchian diversification in Gondwana

The Bajo de la Carpa Formation (Santonian) of the Neuquén basin (southwestern Argentina) has one of the most complete crocodyliform records of South America, together with the Adamantina and Marília formations of Brazil. In this contribution we report two fragmentary individuals from two different localities of the Bajo de la Carpa Formation: a middle-sized individual (MLP 26-IV-30-2), represented by postcranial remains from the Boca del Sapo locality and a large-bodied individual (MACN Pv-RN 1150), consisting only of snout elements from Paso Córdoba locality. Despite the remains of both specimens are fragmentary, they display anatomical characters of a specialized notosuchian carnivore clade: Baurusuchidae. Our phylogenetic analysis recovers these new individuals as non-baurusuchine baurusuchids, although they take multiple phylogenetic positions on the different most parsimonious trees. These new remains highlight the relevance of the Bajo de la Carpa Formation for understanding the rise of notosuchians during the Late Cretaceous.     

Proton conduction through full-length gp91phox requires histidine 115

Summary.  The NADPH oxidase of neutrophils is a transmembrane electron transfer complex, containing a flavin adenine dinucleotide and two hemes, all of which are suggested to be contained within gp91 ^phox , one of four subunits of the enzyme. The transfer of electrons through the NADPH oxidase is associated with an efflux of protons. gp91 ^phox has previously been demonstrated to function as the proton conduction pathway. The mutation of histidines 111, 115, and 119 to leucines and of histidine 115 to leucine within the N-terminal 230-amino-acid fragment of gp91 ^phox has previously been demonstrated to result in the loss of proton conduction through this N-terminal fragment. In this paper we have investigated the role of these histidines in proton conduction by the full-length gp91 ^phox . Stable CHO cell lines were established which expressed full-length gp91 ^phox in which histidines 111, 115, and 119 had been mutated to leucines (CHO91H111/115/119) and in which histidine 115 had been mutated to leucine (CHO91H115L). The expression of gp91 ^phox and its cellular localisation in these cell lines were comparable between wild-type and the mutant gp91 ^phox . The mutation of histidines 111, 115, and 119 to leucines or just histidine 115 to leucine resulted in an almost total loss of both the arachidonate-activated influx and efflux of protons, in comparison with that observed for wild-type gp91 ^phox . Therefore, histidine 115 is required for proton conduction by both full-length gp91 ^phox and the N-terminal 230-amino-acid fragment of gp91 ^phox . Histidine 115 has recently been proposed to act as a coordinating ligand for the outer heme iron of the NADPH oxidase. On the basis of observations for cytochrome c oxidase, we propose a model for this dual role of histidine 115. Received May 2, 2002; accepted July 26, 2002; published online May 21, 2003 RID="**" ID="**" Present address: Bristol Institute for Transfusion Sciences, Bristol, United Kingdom. RID="*" ID="*" Correspondence and reprints: Department of Biochemistry, School of Medical Sciences, University of Bristol, University Walk, Bristol BS8 1TD, United Kingdom.     

Distribution of a 9.1-kb insertion-deletion polymorphism among native and admixed populations from Argentina

We studied five population samples from Argentina, four drawn from Native American groups of the northeast region (Wichí, Pilagá, Toba, and Mbyá-Guaraní) and one from two small villages of the Córdoba province. In this study we report genotypes and allele frequencies of the 9.1-kb insertion-deletion polymorphism located on chromosome 22. The frequency of the deletion allele ranges from 0.276 in the Mbyá-Guaraní to 0.470 in the Pilagá. The coefficient of population differentiation is fairly low (F(ST) = 0.013), does not reflect any geographic or linguistic pattern, and seems to be more related to stochastic processes than to directional forces.