Low genetic diversity in Wolbachia-Infected Culex quinquefasciatus (Diptera: Culicidae) from Brazil and Argentina

Culex quinquefasciatus is a vector of human pathogens, including filarial nematodes and several viruses. Although its epidemiological relevance is known to vary across geographical regions, an understanding of its population genetic structure is still incipient. In light of this, we evaluated the genetic diversity of Cx. quinquefasciatus and Cx. pipiens x Cx. quinquefasciatus hybrids collected from nine localities in Brazil and one site in Argentina. We used mitochondrial genes cox1 and nd4, along with the coxA and wsp genes of the maternally-inherited Wolbachia endosymbiont. The nd4 fragment was invariant between samples, whilst cox1 exhibited four haplotypes that separated two types of Cx. quinquefasciatus, one clustered in southern Brazil. Low sequence diversity was generally observed, being discussed. Both Brazilian and Argentinian mosquitoes were infected with a single Wolbachia strain. As reported in previous studies with these populations, cox1 and nd4 diversity is not congruent with the population structure revealed by nuclear markers or alar morphology. Future Cx. quinquefasciatus research should, if possible, evaluate mtDNA diversity in light of other markers.

MOLECULAR CHARACTERIZATION AND SEQUENCE PHYLOGENETIC ANALYSIS OF SURFACE ANTIGEN 3 (SAG3) GENE OF LOCAL INDIAN ISOLATES (CHENNAI AND IZATNAGAR) OF Toxoplasma gondii

Context and objective:The molecular characterization of local isolates of Toxoplasma gondii is considered significant so as to assess the homologous variations between the different loci of various strains of parasites.Design and setting:The present communication deals with the molecular cloning and sequence analysis of the 1158 bp entire open reading frame (ORF) of surface antigen 3 (SAG3) of two Indian T. gondii isolates (Chennai and Izatnagar) being maintained as cryostock at the IVRI.Method:The surface antigen 3 (SAG3) of two local Indian isolates were cloned and sequenced before being compared with the available published sequences.Results:The sequence comparison analysis revealed 99.9% homology with the standard published RH strain sequence of T. gondii. The strains were also compared with other established published sequences and found to be most related to the P-Br strain and CEP strain (both 99.3%), and least with PRU strain (98.4%). However, the two Indian isolates had 100% homology between them.Conclusion:Finally, it was concluded that the Indian isolates were closer to the RH strain than to the P-Br strain (Brazilian strain), the CEP strain and the PRU strains (USA), with respect to nucleotide homology. The two Indian isolates used in the present study are known to vary between themselves, as far as homologies related to other genes are concerned, but they were found to be 100% homologous as far as SAG3 locus is concerned. This could be attributed to the fact that this SAG3 might be a conserved locus and thereby, further detailed studies are thereby warranted to exploit the use of this particular molecule in diagnostics and immunoprophylactics. The findings are important from the point of view of molecular phylogeny.

Detection of enteroviral sequence in endomyocardial tissues from patients with cardiac diseases in Northern Brazil

In the present report we describe the results from a pilot study aimed at detecting enterovirus sequence in cardiac tissues, obtained through endomyocardial biopsies, from patients suffering from cardiac diseases in the Amazon region. Six samples that were collected from three patients were analysed by RT-PCR showing 3 positive and 3 negative results. These preliminary findings suggest the participation of enteroviruses in the etiology of cardiac diseases, mainly myocarditis, and warrant further and broader local studies on this subject.

A first case of protease codon 35 amino acid insertion in a HIV-1 subtype B sequence detected in the Bauru region, state of São Paulo, Brazil: case report

Amino acid insertions in the protease have rarely been described in HIVinfected patients. One of these insertions has recently been described in codon 35, although its impact on resistance remains unknown. This study presents a case of an HIV variant with an insertion in codon 35 of the protease, described for the first time in Bauru, State of Sao Paulo, Brazil, circulating in a 38-year-old caucasian male with asymptomatic HIV infection since 1997. The variant isolated showed a codon 35 insertion of two amino acids in the protease: a threonine and an aspartic acid, resulting in the amino acid sequence E35E_TD.

Ressonância nuclear magnética no diagnóstico de pseudoaneurisma de ventrículo esquerdo. Antigo problema, nova opção

Paciente feminina, 67 anos, internada por infarto agudo do miocárdio de parede lateral, com edema agudo de pulmão e evolução para choque cardiogênico nas primeiras horas. Ecocardiograma transesofágico e ressonância nuclear magnética confirmaram o diagnóstico de pseudoaneurisma de ventrículo esquerdo. A paciente foi submetida a tratamento cirúrgico com sucesso.

New Exercise-Dipyridamole Combined Test for Nuclear Cardiology in Insufficient Effort: Appropriate Diagnostic Sensitivity Keeping Exercise Prognosis

AbstractBackground:Myocardial perfusion scintigraphy (MPS) in patients not reaching 85% of the maximum predicted heart rate (MPHR) has reduced sensitivity.Objectives:In an attempt to maintain diagnostic sensitivity without losing functional exercise data, a new exercise and dipyridamole combined protocol (EDCP) was developed. Our aim was to evaluate the feasibility and safety of this protocol and to compare its diagnostic sensitivity against standard exercise and dipyridamole protocols.Methods:In patients not reaching a sufficient exercise (SE) test and with no contraindications, 0.56 mg/kg of dipyridamole were IV administered over 1 minute simultaneously with exercise, followed by 99mTc-MIBI injection.Results:Of 155 patients, 41 had MPS with EDCP, 47 had a SE test (≥ 85% MPHR) and 67 underwent the dipyridamole alone test (DIP). They all underwent coronary angiography within 3 months. The three stress methods for diagnosis of coronary lesions had their sensitivity compared. For stenosis ≥ 70%, EDCP yielded 97% sensitivity, SE 90% and DIP 95% (p = 0.43). For lesions ≥ 50%, the sensitivities were 94%, 88% and 95%, respectively (p = 0.35). Side effects of EDCP were present in only 12% of the patients, significantly less than with DIP (p < 0.001).Conclusions:The proposed combined protocol is a valid and safe method that yields adequate diagnostic sensitivity, keeping exercise prognostic information in patients unable to reach target heart rate, with fewer side effects than the DIP.

Estudos citológicos em Hemíoteros da família Coreidae

A more or less detailed study of the spermatogenesis in six species of Hemiptera belonging to the Coreid Family is made in the present paper. The species studied and their respective chromosome numbers were: 1) Diactor bilineatus (Fabr.) : spermatogonia with 20 + X, primary spermatocytes with 10 + X, X dividing equationaliv in the first division and passing undivided to one pole in the second. 2) Lcptoglossus gonagra (Fabr.) : spermatogonia with 20 + X, primary spermatocytes with 10 + X, X dividing equationally in the first division and passing undivided to one pole in the second. 3) Phthia picta (Drury) : spermatogonia with 20 + X, primary spermatocytes with 10 + X, X dividing equationally in the first division and passing undivided to one pole in the second. 4) Anisocelis foliacea Fabr. : spermatogonia with 26 + X fthe highest mumber hitherto known in the Family), primary .spermatocytes with 13 + X, X dividing equationally in the first division an passing undivided to one pole in the second. 5) Pachylis pharaonis (Herbtst) : spermatogonia with 16 + X, primary spermatocytes with 8 + X. Behaviour of the heteroehromosome not referred. 6) Pachylis laticornis (Fabr.) : spermatogonia with 14 + X, primary spermatocytes with 7 + X, X passing undivided to one pole in the first division and therefore secondary spermatocytes with 7 + X and 7 chromosomes. General results and conclusions a) Pairing modus of the chromosomes (Telosynapsis or Farasynapsis ?) - In several species of the Coreld bugs the history of the chromosomes from the diffuse stage till diakinesis cannot be follewed in detail due specially to the fact that lhe bivalents, as soon as they begin to be individually distinct they appear as irregular and extremely lax chromatic areas, which through an obscure process give rise to the diakinesis and then to the metaphase chomosomes. Fortunately I was able to analyse the genesis of the cross-shaped chromosomes, becoming thus convinced that even in the less favorable cases like that of Phthia, in which the crosses develop from four small condensation areas of the diffuse chromosomes, nothing in the process permit to interpret the final results as being due to a previous telosynaptic pairing. In the case of long bivalents formed by two parallel strands intimately united at both endsegments and more or less widely open in the middle (Leptoglossus, Pachylis), I could see that the lateral arms of the crosses originate from condensation centers created by a torsion or bending in the unpaired parts of the chromosomes In the relatively short bivalents the lateral branches of the cross are formed in the middle but in the long ones, whose median opening is sometimes considerable, two asymetrical branches or even two independent crosses may develop in the same pair. These observations put away the idea of an end-to-end pairing of the chromosomes, since if it had occured the lateral arms of the crosses would always be symetrical and median and never more than two. The direct observation of a side- toside pairing of the chromosomal threads at synizesis, is in foil agreement with the complete lack of evidence in favour of telosynapsis. b) Anaphasic bridges and interzonal connections - The chromosomes as they separate from each other in anaphase they remain connected by means of two lateral strands corresponding to the unpaired segmenas observed in the bivalents at the stages preceding metaphase. In the early anaphase the chromosomes again reproduce the form they had in late diafcinesis. The connecting threads which may be thick and intensely coloured are generally curved and sometimes unequal in lenght, one being much longer than the other and forming a loop outwardly. This fact points to a continuous flow of chromosomal substance independently from both chromosomes of the pair rather than to a mechanical stretching of a sticky substance. At the end of anaphase almost all the material which formed the bridges is reduced to two small cones from whose vertices a very fine and pale fibril takes its origin. The interzonal fibres, therefore, may be considered as the remnant of the anaphasic bridges. Abnormal behaviour of the anaphase chromosomes showed to be useful in aiding the interpretation of normal aspects. It has been suggested by Schrader (1944) "that the interzonal is nothing more than a sticky coating of the chromosome which is stretched like mucilage between the daughter chromosomes as they move further and further apart". The paired chromosomes being enclosed in a commom sheath, as they separate they give origin to a tube which becomes more and more stretched. Later the walls of the tube collapse forming in this manner an interzonal element. My observations, however, do not confirm Schrader's tubular theory of interzonal connections. In the aspects seen at anaphase of the primary spermatocytes and described in this paper as chromosomal bridges nothing suggests a tubular structure. There is no doubt that the chromosomes are here connected by two independent strands in the first division of the spermatocytes and by a single one in the second. The manner in which the chromosomes separate supports the idea of transverse divion, leaving little place for another interpretation. c) Ptafanoeomc and chromatoid bodies - The colourabtlity of the plasmosome in Diactor and Anisocelis showed to be highly variable. In the latter species, one may find in the same cyst nuclei provided with two intensely coloured bodies, the larger of which being the plasmosome, sided by those in which only the heterochromosome took the colour. In the former one the plasmosome strongly coloured seen in the primary metaphase may easily be taken for a supernumerary chromosome. At anaphase this body stays motionless in the equator of the cell while the chromosomes are moving toward the poles. There, when intensely coloured ,it may be confused with the heterochromosome of the secondary spermatocytes, which frequently occupies identical position in the corresponding phase, thus causing missinterpretation. In its place the plasmosome may divide into two equal parts or pass undivided to one cell in whose cytoplasm it breaks down giving rise to a few corpuscles of unequal sizes. In Pachylis pharaonis, as soon as the nuclear membrane breate down, the plasmosome migrates to a place in the periphery of the cell (primary spermatocyte), forming there a large chromatoid body. This body is never found in the cytoplasm prior to the dissolution of the nuclear membrane. It is certain that chromatoid bodies of different origin do exist. Here, however, we are dealing, undoubtedly, with true plasmosomes. d) Movement of the heterochromosome - The heterochromosome in the metaphase of the secondary spermatocytes may occupy the most different places. At the time the autosomes prient themselves in the equatorial plane it may be found some distance apart in this plane or in any other plane and even in the subpolar and polar regions. It remains in its place during anaphase. Therefore, it may appear at the same level with the components of one of the anaphase plates (synchronism), between both plates (succession) or between one plate and tbe pole (precession), what depends upon the moment the cell was fixed. This does not mean that the heterochromosome sometimes moves as quickly as the autosomes, sometimes more rapidly and sometimes less. It implies, on the contrary, that, being anywhere in the cell, the heterochromosome m he attained and passed by the autosomes. In spite of being almost motionless the heterochromosome finishes by being enclosed in one of the resulting nuclei. Consequently, it does move rapidly toward the group formed by the autosomes a little before anaphase is ended. This may be understood assuming that the heterochromosome, which do not divide, having almost inactive kinetochore cannot orient itself, giving from wherever it stays, only a weak response to the polar influences. When in the equator it probably do not perform any movement in virtue of receiving equal solicitation from both poles. When in any other plane, despite the greater influence of the nearer pole, the influence of the opposite pole would permit only so a slow movement that the autosomes would soon reach it and then leave it behind. It is only when the cell begins to divide that the heterochromosome, passing to one of the daughter cells scapes the influence of the other and thence goes quickly to join the autosomes, being enclosed with them in the nucleus formed there. The exceptions observed by BORING (1907) together with ; the facts described here must represent the normal behavior of the heterocromosome of the Hemiptera, the greater frequency of succession being the consequence of the more frequent localization of the heterochromosome in the equatorial plane or in its near and of the anaphase rapidity. Due to its position in metaphase the heterochromosome in early anaphase may be found in precession. In late anaphase, oh the contrary ,it appears almost always in succession. This is attributed to the fact of the heterochromosome being ordinairily localized outside the spindle area it leaves the way free to the anaphasic plate moving toward the pole. Moreover, the heterochromosome being a round element approximately of the size of the autosomes, which are equally round or a little longer in the direction of the movement, it can be passed by the autosomes even when it stands in the area of the spindle, specially if it is not too far from the equatorial plane. e) The kinetochore - This question has been fully discussed in another paper (PIZA 1943a). The facts treated here point to the conclusion that the chromosomes of the Coreidae, like those of Tityus bahiensis, are provided with a kinetochore at each end, as was already admitted by the present writer with regard to the heterochromosome of Protenor. Indeed, taking ipr granted the facts presented in this paper, other cannot be the interpretation. However, the reasons by which the chromosomes of the species studied here do not orient themselves at metaphase of the first division in the same way as the heterochromosome of Protenor, that is, with the major axis parallelly to the equatorial plane, are claiming for explanation. But, admiting that the proximity of the kinetochores at the ends of chromosomes which do not separate until the second division making them respond to the poles as if they were a single kinetochore ,the explanation follows. (See PIZA 1943a). The median opening of the diplonemas when they are going to the diffuse stage as well as the reappearance of the bivalents always united at the end-segments and open in the middle is in full agreement with the existence of two terminal kinetochores. The same can be said with regard to the bivalents which join their extremities to form a ring.

Uma nova modalidade de sexo-determinação no grilo sul-americano eneoptera surinamensis

The male of Eneoptera surinamensis (Orthoptera-Eneopteridae) is provided with 9 chromosomes, that is, with 3 pairs of autosomes and 3 sex chromosomes. Spermatogonia. - The autosomes of the spermatogonia are of the same size and U-shaped. One of the sex chromosomes approximately equalling the autosomes in size is telocentric, while the other two are much larger and V-shaped. One of the latter is smaller than the other. The sex chromosomes as showed in Figs. 1 and 2 are designated by X, Yl and Y2, X being the larger V, Yl the smaller one and Y2 the rod-shaped. Primary spermatocytes. - Before the growth period of the spermatocytes all the three sex chromosomes are visible in a state of strong heteropycnosis. X is remarkable in this stage in having two long arms well separated by a wide commissural segment. (Figs. 4, 5 and 6). During the growth period Y2 disappears, while X and Yl remain in a condensed form until metaphase. These may be separated from one another or united in the most varied and irregular manner. (Fig. 7 to 12). In the latter case the segments in contact seem to be always different so that we cannot recognize any homology of parts in the sense os genetics. At diplotene Y2 reappears together with the autosomal tetrads. X and Yl may again be seen as separate or united elements. (Figs. 13 and 14). At later diakinesis and metaphase the three sex chromosomes are always independent from each other, Y2 being typically rod-shaped, X and Yl V-shaped, X being a little larger than Yl. (Fig. 15 to 18). At metaphase the three condensed tetrads go to the equatorial plane, while the sex chromosomes occupy any position at both sides of this plane. In almost all figures which could be perfectly analysed X appeared at one side of the autosomal plate an Yl together with Y2 far apart at the other side. (Figs. 16 and 18). Only a few exception have been found. (Figs. 17 and 19). At anaphase X goes in precession to one pole, Yl and Y2 to the other (Figs. 20 and 21). As it is suggested by the few figures in which a localization of the sex chromosomes different from the normal has been observed, the possibility of other types of segregation of these elements cannot be entirely precluded. But, if this does happen, the resulting gametes should be inviable or give inviable zygotes. Early in anaphase autosomes and sex chromosomes divide longitudinally, being maintained united only by the kinetochore. (Figs. 20 and 21). At metaphase the three sex chromosomes seem to show no special repulsion against each other, X being found in the proximity of Yl or Y2 indifferently. At anaphase, however, the evidences in hand point to a stronger repulsion between X on the one side and both Ys on the other, so that in spite of the mutual repulsion of the latter they finish by going to the same pole. Secondary spermatocytes. - At telophase of the primary spermatocytes all the chromosomes enter into distension without disappearing of view. A nuclear membrane is formed around the chromosomes. All the chromosomes excepting Y2 which has two arms, are four-branched. (Fig. 22). Soon the chromosomes enter again into contraction giving rise to the secondary metaphase plate. Secondary spermatocytes provided as expected with four and five chromosomes are abundantly found. (Figs. 23 and 24). In the former all chromosomes are X-shaped while in the latter there is one which is V-shaped. This is the rod- shaped Y2. In the anaphase of the spermatocytes with four chromosomes all the chromosomes are V-shaped, one of them (X) being much larger than the others. In those with five there is one rod-shaped chromosome (Y2). (Fig. 25), Spermatids. Two classes of spermatids are produced, one with X and other with Yl and Y2. All the autosomes as well as Y2 soon enter into solution, X remaining visible for long time in one class and Yl in the other. (Figs. 26 and 27). Since both are very alike at this stage, one cannot distinguish the two classes of spermatids. Somatic chromosomes in the famale. - In the follicular cells of the ovary 8 chromosomes were found, two of which are much larger than the rest. (Figs. 29 and 30). These are considered as being sex chromosomes. CONCLUSION: Eneoptera surinamensis has a new type of sex-determining mechanism, the male being X Yl Y2 and the female XX. The sex chromosomes segregate without entering into contact at metaphase or forming group. After a review of the other known cases of complex sex chromosome mechanism the author held that Eneoptera is the unique representative of a true determinate segregation of sex chromosomes. Y2 behaving as sex chromosome and as autosome is considered as representing an intermediary state of the evolution of the sex chromosomes.