Release of insulin from PLGA-alginate dressing stimulates regenerative healing of burn wounds in rats

Burn wound healing involves a complex set of overlapping processes in an environment conducive to ischemia, inflammation, and infection costing $7.5 billion/year in the US alone, in addition to the morbidity and mortality that occur when the burns are extensive. We previously showed that insulin, when topically applied to skin excision wounds, accelerates re-epithelialization, and stimulates angiogenesis. More recently, we developed an alginate sponge dressing (ASD) containing insulin encapsulated in PLGA microparticles that provides a sustained release of bioactive insulin for >20days in a moist and protective environment. We hypothesized that insulin-containing ASD accelerates burn healing and stimulates a more regenerative, less scarring, healing. Using a heat-induced burn injury in rats, we show that burns treated with dressings containing 0.04mg insulin/cm2, every three days for 9 days, have faster closure, faster rate of disintegration of dead tissue, and decreased oxidative stress.In addition, in insulin-treated wounds the pattern of neutrophil inflammatory response suggests faster clearing of the burn dead tissue. We also observe faster resolution of the pro-inflammatory macrophages. We also found that insulin stimulates collagen deposition and maturation with the fibers organized more like a basket weave (normal skin) than aligned and crosslinked (scar tissue). In summary , application of ASD-containing insulin-loaded PLGA particles on burns every three days stimulates faster and more regenerative healing. These results suggest insulin as a potential therapeutic agent in burn healing and, because of its long history of safe use in humans, insulin could become one of the treatments of choice when repair and regeneration are critical for proper tissue function.

Clivagem e caracterização de frutalina recombinante produzida em Escherichia coli

Dissertação de mestrado integrado em Engenharia Biomédica (área de especialização em Engenharia Clínica)

Low frequency of TERT promoter mutations in gastrointestinal stromal tumors (GISTs).

Somatic mutations in the promoter region of telomerase reverse transcriptase (TERT) gene, mainly at positions c. − 124 and c. − 146 bp, are frequent in several human cancers; yet its presence in gastrointestinal stromal tumor (GIST) has not been reported to date. Herein, we searched for the presence and clinicopathological association of TERT promoter mutations in genomic DNA from 130 bona fide GISTs. We found TERT promoter mutations in 3.8% (5/130) of GISTs. The c. − 124C4T mutation was the most common event, present in 2.3% (3/130), and the c. − 146C4T mutation in 1.5% (2/130) of GISTs. No significant association was observed between TERT promoter mutation and patient’s clinicopathological features. The present study establishes the low frequency (4%) of TERT promoter mutations in GISTs. Further studies are required to confirm our findings and to elucidate the hypothetical biological and clinical impact of TERT promoter mutation in GIST pathogenesis.

Minilaparoscopy and sentinel lymph node in uterine cancer

CRSLS MIS Case Reports from SLS.org.

The role of the mammalian DNA end-processing enzyme polynucleotide kinase 3'-phosphatase in spinocerebellar ataxia Type 3 pathogenesis

DNA strand-breaks (SBs) with non-ligatable ends are generated by ionizing radiation, oxidative stress, various chemotherapeutic agents, and also as base excision repair (BER) intermediates. Several neurological diseases have already been identified as being due to a deficiency in DNA end-processing activities. Two common dirty ends, 3'-P and 5'-OH, are processed by mammalian polynucleotide kinase 3'-phosphatase (PNKP), a bifunctional enzyme with 3'-phosphatase and 5'-kinase activities. We have made the unexpected observation that PNKP stably associates with Ataxin-3 (ATXN3), a polyglutamine repeat-containing protein mutated in spinocerebellar ataxia type 3 (SCA3), also known as Machado-Joseph Disease (MJD). This disease is one of the most common dominantly inherited ataxias worldwide; the defect in SCA3 is due to CAG repeat expansion (from the normal 14-41 to 55-82 repeats) in the ATXN3 coding region. However, how the expanded form gains its toxic function is still not clearly understood. Here we report that purified wild-type (WT) ATXN3 stimulates, and by contrast the mutant form specifically inhibits, PNKP's 3' phosphatase activity in vitro. ATXN3-deficient cells also show decreased PNKP activity. Furthermore, transgenic mice conditionally expressing the pathological form of human ATXN3 also showed decreased 3'-phosphatase activity of PNKP, mostly in the deep cerebellar nuclei, one of the most affected regions in MJD patients' brain. Finally, long amplicon quantitative PCR analysis of human MJD patients' brain samples showed a significant accumulation of DNA strand breaks. Our results thus indicate that the accumulation of DNA strand breaks due to functional deficiency of PNKP is etiologically linked to the pathogenesis of SCA3/MJD.

Finalidades do estudo de um instrumento musical: intervenção pedagógica com recurso a estratégias no ensino do violino para desenvolver a atenção seletiva

Relatório de estágio de mestrado em Ensino de Música

Maternal adjustment and maternal attitudes in adolescent and adult pregnant women

Study Objective: This study analyzes differences between adolescent and adult pregnant women and the contribution of maternal age to maternal adjustment and maternal attitudes during pregnancy. Design, Setting, and Participants: A sample of 398 Portuguese pregnant women (111 younger than 19 years) was recruited in a Portuguese Maternity Hospital and completed the Maternal Adjustment and Maternal Attitudes Questionnaire between the 24th and 36th weeks of gestation. Main Outcome Measures: Maternal Adjustment and Maternal Attitudes Questionnaire. Results: Adolescent pregnant women show lower maternal adjustment (poorer body image and worse marital relationship) and poorer maternal attitudes (more negative attitudes to sex) than adult pregnant women. When controlling for socio-demographics, age at pregnancy predicts poorer body image and more negative attitudes to sex, but not a worse marital relationship, more somatic symptoms or negative attitudes to pregnancy and the baby. A worse marital relationship was better predicted by living without the partner, and more somatic symptoms and negative attitudes to pregnancy and the baby was predicted by higher education. Conclusion: Adolescent pregnant women show lower maternal adjustment and poorer maternal attitudes than adult pregnant women according to socio-demographics and unfavorable developmental circumstances.

Estudo preliminar da variação dos níveis de cortisol em doentes da espécie Canis familiaris sujeitos a mastectomia por doença oncológica da mama

Dissertação de Mestrado Integrado em Medicina Veterinária

Ultrasound Blocks for the Anterior Abdominal Wall - Adult and Pediatric Surgery

Somatic post-surgical pain is invalidating and distressing to patients and carries the risk of important complications. The anterior abdominal wall is involved in most surgical procedures in general, gynecologic, obstetric, urological, vascular and pediatric surgery. Combined multimodal strategies involving nerve blocks, opiates, and non-steroidal anti-inflammatory drugs for systemic analgesia are necessary for optimal pain modulation. Anterior abdominal wall blocks, transverse abdominal plexus block, iliohypogastric and ilioinguinal nerveblock, genitofemoral nerve block and rectus sheath block have an important role as components of multimodal analgesia for somatic intraoperative and postoperative pain control. Ultrasound visualization has improved the efficacy and safety of abdominal blocks and implemented the application in the clinical setting. For this reason, they are a very important tool for all anesthesiologists who aim to treat effectively patients’ pain. This guide provides an evidence based comprehensive and necessary overview of anatomical, anesthesiological and technical information needed to safely perform these blocks.

Mortality and Embolic Potential of Cardiac Tumors

Background: Cardiac tumors are rare, mostly benign with high embolic potential. Objectives: To correlate the histological type of cardiac masses with their embolic potential, implantation site and long term follow up in patients undergoing surgery. Methods: Between January 1986 and December 2011, we retrospectively analyzed 185 consecutive patients who underwent excision of intracardiac mass (119 females, mean age 48±20 years). In 145 patients, the left atrium was the origin site. 72% were asymptomatic and prior embolization was often observed (19.8%). The diagnosis was established by echocardiography, magnetic resonance and histological examination. Results: Most tumors were located in the left side of the heart. Myxoma was the most common (72.6%), followed by fibromas (6.9%), thrombi (6.4%) and sarcomas (6.4%). Ranging from 0.6cm to 15cm (mean 4.6 ± 2.5cm) 37 (19.8%) patients had prior embolization, stroke 10.2%, coronary 4.8%, peripheral 4.3% 5.4% of hospital death, with a predominance of malignant tumors (40% p < 0.0001). The histological type was a predictor of mortality (rhabdomyomas and sarcomas p = 0.002) and embolic event (sarcoma, lipoma and fibroelastoma p = 0.006), but not recurrence. Tumor size, atrial fibrillation, cavity and valve impairment were not associated with the embolic event. During follow-up (mean 80±63 months), there were 2 deaths (1.1%) and two recurrences 1 and 11 years after the operation, to the same cavity. Conclusion: Most tumors were located in the left side of the heart. The histological type was predictor of death and preoperative embolic event, while the implantation site carries no relation with mortality or to embolic event.

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.

Provas adicionais da dicentricidade dos cromossômios dos Hemípteros

In order to test Piza's conclusions regarding the dicentricity of Hemipteran chromosomes, two species of bugs of the family Coreidae, namely, Anasa sp. and Leptoglossus stigma (Herbst), are studied in the present paper. a) Anasa sp. - The male of this species has 21 chromosomes, that is, 20 pairs of autosomes and a single sex chromosome. The latter divides equationally in the first division of the spermatocytes and passes undivided to one cell in the second division. In this it moves with its longer axis parallelly to the spindle axis and shows fibrillar connections with both poles. Special attention was paid to the behavior of the chromosomes in the anaphase of the spermatogonia. As it was previously stated (Piza 1946 and 1946a) with regard to other species, the chromosomes are here attached to the spindle by both ends and begin to move toward the poles strongly curved to them. No intercalary fibers could be detected although their existente may not be denied by theoretical reasons developed in another paper (Piza 1946). Mitoses in somatic tissues of the embryo were equally studied. Careful examination of anaphase chromosomes in a great number of cells showed that the chromosomes behave exactly as in the spermatogonia, being equally attached to the spindle by the extremities alone and moving with their ends looking to the pole. A weak median constriction sometimes replaced by a slightly clearer space was observed in prometaphase and even in metaphase chromosomes of the spermatogonia as well as the somatic cells, having already been referred to in the case of Diactor bilineatus. (Piza 1945). Hemipteran chromosomes being considered as iso-chromosomes originated by a longitudinal spliting of the monocentric chromosomes resulting from the second division of the spermatocytes, the median aspect just mentioned may be regarded as the point of union of the separated halves. (See origin of dicentricity in Piza 1946). b) Leptoglossus stigma - This species has spermatogonia provided with 20 pairs of autosomes and one sex chromosome whose behavior differs in nothing from what was stated in regard of the preceding species. In the primary spermatocytes nothing meriting special mention was observed. Orientation, connection with the poles and movements of the sex chromosome in the secondary spermatocytes confirm the views already developed.

Notas sobre cromossômios de alguns escorpiões brasileiros

Three species of Scorpions beloging to two different families were studied cytologically: a) Tityus mattogrossensis Borelli (Fam. Buthidae), - This species presents spermatogonia provided with 20 short chromosomes which orient at metaphase with their axis parallelly to the plane of the equator and move toward the poles without changing this position, from the stage pachytene to metaphase the bivalents become, as in Tityus bahiensis, progressivery shorter and thicker, without showing that chiasmata occured at any time. The paired chromosomes never open themselves, out to form loops as in orthodox meioses. As in Tityus bahiensis the bivalents are inserted In the spindle before reaching their maxim contraction. No diakinesis has been observed. The primary spermatocyte metaphases are provided, with 10 pairs of chromosones, two of which are larger and two smaller than the rest. The bivalents orient as in Tityus bahiensis with their length in the plane of the equator and separate parallelly. Spindle fibres are seen alongst their entire body. While, in Tityus bahiensis the ends of the chromosomes are pronouncedly turned to opposite poles at metaphase, nothing like this was observed in the present species. Only late in anaphase the chromosomes of Tityus mattogrossensis show a bending to the poles. The secondary spermatocytes present 10 short chromosomes, two being larger than, the others. Here, on the contrary, the chromosomes are strongly curved toward the poles since the beginning of anaphase. Some chromosomal anomalies have been noticed. Primary spermatocytes with 14 bivalents, some of which representing probably free fragments, were observed. Primary spermatocytes with 8 bivalents and one cross of 4 chromosomes were interpreted as resulting from breakages followed by translocations Primary spermatocytes with 9 bivalents, one of which being much longer than the longst of the normal plates, show that fusion by the extremities of two non homologous chromosomes on the onde side, and of their respective homologous in the same way on tre other, have occured. Orientation of bivalents with their body parallelly to the spindle axis and anaphasic bridges have been encountered. All in all points to the conclusion that the chromosomes of Tityus mattogrossesis, like those of Tityus bahiensia are provided with one kinetochore at each end. Ananteris balzani Thorell - (Fam. Buthidae). - This species which belongs to the same family as Tityus, is provided with 12 chromosomes (diploid). These studied in embryonic tissues, showed the same behavior as the somatic chromosomes of Tityus bahiensis. Bothrirus sp. (Bothriuridae). - Only spermatogonia were found in the testis, of the single male hitherto investigated. The chromosomes, in number of 36, are of different sizes but small and provided, as ordinarily, with a single kinetochore. They behave therefore in an orthodox manner in mitosis.

Meiose em Rhinocricus Padbergi Verhoeff, 1938

In the present paper the behaviour of the chromosomes in the spermatogenesis of the Myriapod Rhinocricus Padbergi Verhoeff, 1938 is studied. The primary spermatocytes are provided with 10 independent bivalents which separate normally giving rise to equivalent secondary spermatocytes. No indication of sex chromosomes has been found. Fusion of two bivalents or of four, two by two, has been observed, giving origin to secondary spermatocytes with 9 and 8 chromosomes respectively, in which fused chromosomes could be discovered. For analysing the facts the chomosomes of both, primary and secondary metaphases were separately counted from a total of 190 celis of four individuals and statistically treted. The X2-test gave insignificant results. Twenty chomosomes were counted in somatic tissues. The heterochròmatic parts of the leptotene threads were usually arranged in the periphery of the nucleus. In resting nuclei chromocenters can be observed in varyng number. Their chromosomal nature is revealed by the fact that when treated by KCÑ or KNOS they begin uncoiling.

Sobre o comportamento mitótico e meiótico de cromossomas policêntricos ou com ponto de inserção difuso

Material: Studies were made mainly with Ascaris megalocephála Cloq. univalens and bivalens, and also with Tityus bahiensis Perty. 1) Somatic pairing of heterochromatic regions. The heterochromatic ends of the somatic chromosomes in Ascaris show a very strong tendency for unspecifical somatic pairing which may occur between parts of different chromosomes (Figs. 1, 2, 3, 7, 10, 11, 12, 13, 14, 16, 18,), between the two ends of the same chromosome either directly (Figs. 4, 5, 7, 8, 11, 12, 13, 15, 16, 17, 18) or inversely (Fig. 8, in the arrow) and also within a same chromosomal arm (Fig. 6). 2) During the early first cleavage division the chomosomes are an isodiametric cylinder (Figs. 6, 9, 11, 13, 14). But in later metaphase the ends become club shaped (Figs. 1, 2, 3, 4, 5, 7, 10) which is interpreted as the beginning of migration of chromatic substance from the central euchromatic region towards the heterochromatic regions. This migration becomes more and accentuated in anaphase (Figs. 19, 22, 23) and in the vegetative cells where euchromatic region looses more and more staing power, especially in the intersititial zones between the individual small spherical chromosomes into which the euchromatic region desintegrates. The emigrated chromatin material is finally eliminated with the heterochromatic chromosome ends (Fig. 23 and 24). 3) It seems a general rule that during mitotic anaphase all chromosomes with diffuse or multiple spindle fiber attachement (Ascaris, Tityus, Luzula, Steatococcus, Homoptera and Heteroptera in general) move to the poles in the form of an U with precedence of the chromosomal ends. In Ascaris, the heterocromatic regions are pulled passively towards the poles and only the euchromatic central portion may be U-shaped (Fig. 19, 22, 25). While in the other species this U-shape is perfect since the beginning of anaphase, giving the impression that movement towards the poles begins at both ends of a chromosome simultaneously, this is not the case in Ascaris. There the euchromatic region is at first U-shaped, passing then to form a straight or zig-zag line and becoming again U-shaped during late anaphase. This is explained by the fact that the ends of the euchromatic regions have to pull the weight of the passive heterochromatic portions. 4) While it is generally accepted that, during first meio-tic division untill second anaphase, all attachement regions remain either undivided or at least united closely, this is not the case in chromosomes with diffused or multiple attachment. Here one clearly sees in all cases so far studied four parallel chromatids at first metaphase. In Luzula and Tityus (for Tityus all figs. 26 to 31) this division is allready quite clear in paraphase (pro-metaphase) and it cannot be said wether in other species the division in sister chromatids is allready present, but not visible at this stage. During first anaphase the sister chromatids of Titbits remain more or less in contact, while in Luzula and especially in Ascaris they are quite separated. Thus one can count in late anaphase or telophase of Ascaris megalocephala bivalens, nearly allways, four separate chromosomes near each pole, or a total of eight chromatids per division figure (Figs. 35, 36, 37, 38, 39, 40, 41).